The present invention provides methods and compositions for identifying soybean plants that are tolerant or have improved tolerance, or those that are susceptible to, iron deficient growth conditions. The methods use molecular markers to identify, select, and/or introgress genetic loci modulating phenotypic expression of an iron deficiency tolerance trait in soybean plant breeding. Methods are provided for screening germplasm entries for the performance and expression of this trait.

Description

25 parts

›CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a 35 U.S.C. § 371 National Phase application of International Patent Application No. PCT/US2012/052871, filed Aug. 29, 2012 and incorporated herein by reference in its entirety which claims the benefit of U.S. Provisional Patent Application No. 61/529,838, filed Aug. 31, 2011, which is incorporated herein by reference in its entirety.

›INCORPORATION OF SEQUENCE LISTING

A sequence listing is contained in the file named “46_21_57867.txt” which is 96,920 bytes (measured in MS-Windows) and was created on Aug. 29, 2012, and comprising 186 nucleotide sequences and is electronically filed herewith and is incorporated herein by reference.

›INCORPORATION OF APPENDIX

A listing of various soybean markers is provided herewith in an Appendix to the Specification as Tables 12, 14, 17, 20, 23, 26, and 29.

›BACKGROUND OF INVENTION

Soybean, Glycine max (L.) Merril, is a major economic crop worldwide and is a primary source of vegetable oil and protein (Sinclair and Backman, Compendium of Soybean Diseases, 3 rd Ed. APS Press, St. Paul, Minn., p. 106. (1989). Growing demand for low cholesterol and high fiber diets has increased soybean's importance as a health food.

Soybean varieties grown in the United States have a narrow genetic base. Six introductions, ‘Mandarin,’ ‘Manchu,’ ‘Mandarin’ (Ottawa), “Richland,’ ‘AK’ (Harrow), and ‘Mukden,’ contributed nearly 70% of the germplasm represented in 136 cultivar releases. To date, modern day cultivars can be traced back from these six soybean strains from China. In a study conducted by Cox et al., Crop Sci. 25:529-532 (1988), the soybean germplasm is comprised of 90% adapted materials, 9% un-adapted, and only 1% from exotic species. The genetic base of cultivated soybean could be widened through exotic species. In addition, exotic species may possess such key traits as disease, stress, and insect resistance.

The availability of a specific micronutrient, such as iron (Fe), is often related to soil characteristics. Soil pH has a major impact on the availability of Fe. Iron deficiency has been a common, serious, and yield limiting problem for soybean production in some parts of the United States.

Iron is one of the necessary micronutrients for soybean plant growth and development. Iron is needed for the development of chlorophyll. It is involved in energy transfer, plant respiration, and plant metabolism. It is a constituent of certain enzymes and proteins in plants. Iron is also necessary for soybean root nodule formation and has a role in N-fixation, thus, low levels of Fe can lead to reduction in N-fixation.

When Fe is limited, iron deficiency chlorosis (IDC) can be expressed in soybean plants. IDC in soybean is the result of a complex interaction among many factors including soil chemistry, environmental conditions, and soybean physiology and genetics. The most common IDC symptom is interveinal chlorosis in which leaf tissue of newly developed soybean leaves turn yellow, while the veins remain green. The leaves may develop necrotic spots that eventually coalesce and fall off the plant. Iron deficiency symptoms are similar to that of Manganese (Mn), therefore, only soil and tissue analysis can confirm the deficiency.

Severe yield reductions have been reported from IDC throughout the North-Central U.S with losses estimated to be around $120 million annually. Soybean IDC symptoms typically occur between the first and third trifoliate stage. Depending on the severity of the problem, symptoms might improve later in the season. Severe stress can stunt soybean plants causing more than 50% or more yield reduction and may even kill the plants.

Some calcareous soils with pH more than 7.4, heavy, poorly drained, and compacted soils may exhibit IDC symptoms, due to insufficient Fe uptake. However, soil pH is not a good indicator and does not correlate very well with IDC. Symptoms are highly variable between years and varieties and depend on other soil factors and weather conditions.

There is a direct relationship between IDC and high concentrations of calcium carbonate and soluble salts. Iron uptake is adversely impacted by high concentrations of phosphorous (P), manganese (Mn), and zinc (Zn). High levels of calcium (Ca) in the soil cause Fe molecules to bind tightly to the soil particles and become unavailable for uptake. It is important to measure the percentage of calcium carbonate and soluble salts in the soil as some combinations of percentage of free calcium carbonate and soluble salts can cause severe IDC. Sandy soils with low organic matter also may exhibit IDC symptoms.

Weather also plays a role in IDC symptoms. Cool soil temperature and wet weather, combined with soils that have marginal levels of available Fe can increase IDC symptoms.

Soybean producers have sought to develop plants tolerant to low iron growth conditions (thus not exhibiting IDC) as a cost-effective alternative or supplement to standard foliar, soil and/or seed treatments (e.g., Hintz et al. (1987) “Population development for the selection of high-yielding soybean cultivars with resistance to iron deficiency chlorosis,” Crop Sci. 28:369-370). Studies also suggest that cultivar selection is more reliable and universally applicable than foliar sprays or iron seed treatment methods, though environmental and cultivar selection methods can also be used effectively in combination. See also, Goos and Johnson (2000) “A Comparison of Three Methods for Reducing Iron-Deficiency Chlorosis in Soybean” Agronomy Journal 92:1135-1139; and Goos and Johnson “Seed Treatment, Seeding Rate, and Cultivar Effects on Iron Deficiency Chlorosis of Soybean” Journal of Plant Nutrition 24 (8) 1255-1268. U.S. Pat. No. 7,977,533 discloses genetic loci associated with iron deficiency tolerance in soybean.

Soybean cultivar improvement for IDC tolerance can be performed using classical breeding methods, or, more preferably, using marker assisted selection (MAS). Genetic markers for low iron growth condition tolerance/susceptibility have been identified (e.g., Lin et al. (2000) “Molecular characterization of iron deficiency chlorosis in soybean” Journal of Plant Nutrition 23:1929-1939). Recent work suggests that marker assisted selection is particularly beneficial when selecting plants because the strength of environmental effects on chlorosis expression impedes progress in improving tolerance. See also, Charlson et al., “Associating SSR Markers with Soybean Resistance to Iron Chlorosis,” Journal of Plant Nutrition, vol. 26, nos. 10 & 11; 2267-2276 (2003). Molecular Markers and Marker Assisted Selection. U.S. Pat. No. 7,977,533 also discloses genetic loci associated with iron deficiency tolerance in soybean.

There is a need in the art of plant breeding to identify additional markers linked to genomic regions associated with tolerance to low iron growth conditions (e.g., IDC tolerance) in soybean. There is in particular a need for numerous markers that are closely associated with low iron growth condition tolerance in soybean that permit introgression of such regions in the absence of extraneous linked DNA from the source germplasm containing the regions. Additionally, there is a need for rapid, cost-efficient method to assay the absence or presence of IDC tolerance loci in soybean.

›SUMMARY OF INVENTION · 1 of 3

In certain embodiments, the present invention provides for compositions and methods for identifying soybean plants or germplasm with tolerance to low iron growth conditions. Certain embodiments of the invention provide for polymorphic nucleic acids useful for identifying or producing soybean plants or germplasm with tolerance to low iron growth conditions. Certain embodiments of the invention also provide for methods for selecting, producing, and/or breeding soybean plants or germplasm with tolerance to low iron growth conditions. In certain embodiments, the present invention further relates to methods to determine the presence or absence of quantitative trait loci (QTL) conferring tolerance to low iron growth conditions to soybean plants, including but not limited to exotic germplasm, populations, lines, elite lines, cultivars and varieties. In certain embodiments, the invention relates to methods that provide for identification of molecular markers associated with low iron growth condition tolerance trait QTL. In certain embodiments, the present invention relates to the use of molecular markers to screen and select for low iron growth condition tolerance within soybean plants, including but not limited to exotic germplasm, populations, lines, elite lines, and varieties.

Methods of identifying a soybean plant that comprises a genotype associated with a low iron growth condition tolerant phenotype are provided. In certain embodiments, these methods of identifying a soybean plant that comprises a genotype associated with a low iron growth condition tolerant phenotype can comprise: i) detecting in the soybean plant an allele in at least one polymorphic nucleic acid marker locus associated with the low iron growth condition tolerant phenotype wherein the marker locus is in a: a) linkage group N genomic region flanked by loci Glyma03g34510 and Glyma03g42250, b) linkage group L genomic region flanked by loci Glyma19g32880 and Contig9146, c) linkage group O genomic region flanked by loci NS0116559 and Gm_W82_CR10.G236520, d) linkage group O genomic region flanked by loci NS0116559 and TA68568_3847, e) linkage group O genomic region flanked by loci Glyma10g37560 and Gm_W82_CR10.G236520, f) linkage group M genomic region flanked by loci Gm_W82_CR07.G4950 and Glyma070g33560, g) linkage group M genomic region flanked by loci Gm_W82_CR07.G4950 and Glyma07g12210, or h) linkage group M genomic region flanked by loci Glyma07g29640 and Glyma070g33560; and ii) denoting that said plant comprises a genotype associated with a low iron growth condition tolerant phenotype. In certain embodiments, these methods can further comprise the step of selecting the denoted plant from a population of plants. In certain embodiments of these methods, a denoted and/or selected plant exhibits a low iron growth condition tolerant phenotype. In certain embodiments of any of the aforementioned methods, a genotype associated with a low iron growth condition phenotype comprises at least one allele associated with a low iron growth condition tolerant phenotype of a polymorphic nucleic acid marker selected from the group consisting of SEQ ID NOs: 1-7, 20-30, and 47-51.

Also provided herein are methods for obtaining a soybean plant comprising in its genome at least one low iron growth condition tolerance locus. In certain embodiments, the methods for obtaining a soybean plant comprising in its genome at least one low iron growth condition tolerance locus can comprise genotyping a plurality of soybean plants with respect to at least one locus in a: a) linkage group N genomic region flanked by loci Glyma03g34510 and Glyma03g42250, b) linkage group L genomic region flanked by loci Glyma19g32880 and Contig9146, c) linkage group O genomic region flanked by loci NS0116559 and Gm_W82_CR10.G236520, d) linkage group O genomic region flanked by loci NS0116559 and TA68568_3847, e) linkage group O genomic region flanked by loci Glyma10g37560 and Gm_W82_CR10.G236520, f) linkage group M genomic region flanked by loci Gm_W82_CR07.G4950 and Glyma070g33560, g) linkage group M genomic region flanked by loci Gm_W82_CR07.G4950 and Glyma07g12210, or h) linkage group M genomic region flanked by loci Glyma07g29640 and Glyma070g33560; and ii) selecting a soybean plant comprising in its genome at least one low iron growth condition tolerance locus comprising a genotype associated with a low iron growth condition tolerant phenotype. In certain embodiments of these methods, the selected soybean plant exhibits tolerance to low iron growth conditions. In certain embodiments of the aforementioned methods, the methods can further comprise assaying the selected plant of step (ii) for a low iron growth condition tolerant phenotype. In certain embodiments of any of the aforementioned methods, the methods can further comprise a step wherein a low iron growth condition tolerance locus is genotyped for at least one allele of a polymorphic nucleic acid marker selected from the group consisting of SEQ ID NOs: 1-7, 20-30, and 47-51.

Also provided are methods for identifying a soybean plant comprising in its genome at least one introgressed low iron growth condition tolerance locus. In certain embodiments, methods for identifying a soybean plant comprising in its genome at least one introgressed low iron growth condition tolerance locus can comprise crossing a first soybean plant with a second soybean plant, the second plant comprising: i) a low iron growth condition tolerance locus in a: a) linkage group N genomic region flanked by loci Glyma03g34510 and Glyma03g42250, b) linkage group L genomic region flanked by loci Glyma19g32880 and Contig9146, c) linkage group O genomic region flanked by loci NS0116559 and Gm_W82_CR10.G236520, d) linkage group O genomic region flanked by loci NS0116559 and TA68568_847, e) linkage group O genomic region flanked by loci Glyma10g37560 and Gm_W82_CR10.G236520, f) linkage group M genomic region flanked by loci Gm_W82_CR07.G4950 and Glyma070g33560, g) linkage group M genomic region flanked by loci Gm_W82_CR07.G4950 and Glyma07g12210, or h) linkage group M genomic region flanked by loci Glyma07g29640 and Glyma070g33560; and ii) detecting the polymorphic nucleic acid in at least one soybean plant from the population of soybean plants, wherein the one soybean plant lacks the additional polymorphic locus, thereby identifying a soybean plant comprising in its genome at least one introgressed low iron growth condition tolerance locus. In certain embodiments, these methods can further comprise the step of selecting the one soybean plant, thereby obtaining a soybean plant comprising in its genome at least one introgressed low iron growth condition tolerance locus. In certain embodiments of any of the aforementioned methods, the identified or the selected plant is tolerant to low iron growth conditions. In certain embodiments of any of the aforementioned methods, the identified or selected plant is assayed for tolerance to low iron growth conditions. In certain embodiments of any of the aforementioned methods, the additional polymorphic locus is detected with a genotypic marker, a phenotypic marker, or both.

›SUMMARY OF INVENTION · 2 of 3

In certain embodiments of any of the aforementioned methods, the additional polymorphic locus is a linked polymorphic locus located on linkage group N, L, O, or M, but not within a: a) linkage group N genomic region flanked by loci Glyma03g34510 and Glyma03g42250, b) linkage group L genomic region flanked by loci Glyma19g32880 and Contig9146, c) linkage group O genomic region flanked by loci NS0116559 and Gm_W82_CR10.G236520, d) linkage group O genomic region flanked by loci NS0116559 and TA68568_3847, e) linkage group O genomic region flanked by loci Glyma10g37560 and Gm_W82_CR10.G236520, f) linkage group M genomic region flanked by loci Gm_W82_CR07.G4950 and Glyma070g33560, g) linkage group M genomic region flanked by loci Gm_W82_CR07.G4950 and Glyma07g12210, or h) linkage group M genomic region flanked by loci Glyma07g29640 and Glyma070g33560.

Also provided herein are soybean plants obtainable by any of the aforementioned methods. Soybean plants comprising genomic regions associated with a low iron growth condition tolerant phenotype wherein immediately adjacent genomic regions and/or one or more adjacent genomic regions characteristic of soybean germplasms that lack the genomic regions associated with a low iron growth condition phenotype and/or that are distinct from the germplasm from which the genomic region is derived are also provided.

In certain embodiments, a soybean plant comprising i) a low iron growth condition tolerance locus in a: a) linkage group N genomic region flanked by loci Glyma03g34510 and Glyma03g42250, b) linkage group L genomic region flanked by loci Glyma19g32880 and Contig9146, c) linkage group O genomic region flanked by loci NS0116559 and Gm_W82_CR10.G236520, d) linkage group O genomic region flanked by loci NS0116559 and TA68568_3847, e) linkage group O genomic region flanked by loci Glyma10g37560 and Gm_W82_CR10.G236520, f) linkage group M genomic region flanked by loci Gm_W82_CR07.G4950 and Glyma070g33560, g) linkage group M genomic region flanked by loci Gm_W82_CR07.G4950 and Glyma07g12210, or h) linkage group M genomic region flanked by loci Glyma07g29640 and Glyma070g33560; and ii) one or more polymorphic loci comprising alleles or combinations of alleles that are not found in a low iron growth condition tolerant soybean varieties harboring said tolerance locus, and that are linked to said locus, wherein said soybean plant is tolerant to low iron growth conditions is provided.

In any of the aforementioned embodiments, the soybean plant can comprise an allele of one or more polymorphic markers selected from the group consisting of SEQ ID NOs: 1-7, 20-30, and 47-51. In certain embodiments, the nucleic acid can further comprise a detectable moiety. In certain embodiments, the detectable moiety can be selected from the group consisting of a chromophore, a fluorophore, and a hapten.

Certain other methods of identifying a soybean plant that comprises a genotype associated with a low iron growth condition tolerant phenotype are provided. In certain embodiments, these methods comprise: i) detecting in said soybean plant an allele in at least one polymorphic nucleic acid marker locus associated with the low iron growth condition tolerant phenotype wherein the marker locus is in a: a) linkage group G genomic region flanked by loci FE898349 and Cf14688d, b) linkage group A1 genomic region flanked by loci Mt7 and BARCSOYSSR_05_0640, c) linkage group M genomic region flanked by loci BQ081048 and TA53357_3847, d) linkage group D2 genomic region flanked by loci BG726970 and Contig41076, e) linkage group L genomic region flanked by loci Glyma19g01630 and TC27203, or f) linkage group K genomic region flanked by loci Cf4289d and Cf974d; and ii) denoting that said plant comprises a genotype associated with a low iron growth condition tolerant phenotype. In certain embodiments, these methods can further comprise the step of selecting said denoted plant from a population of plants. In certain embodiments of these methods, a denoted and/or selected plant exhibits a low iron growth condition tolerant phenotype. In certain embodiments of any of the aforementioned methods, a genotype associated with a low iron growth condition tolerant phenotype comprises at least one allele associated with a low iron growth condition tolerant phenotype of a polymorphic nucleic acid marker selected from the group consisting of SEQ ID NOs: 52-62, 71-93, 94-110, 115-123, 124-138, and 139-174.

Also provided herein are methods for obtaining a soybean plant comprising in its genome at least one locus associated with a low iron growth condition tolerant phenotype. In certain embodiments, these methods can compromising the steps of: i) genotyping a plurality of soybean plants with respect to at least one low iron growth condition tolerance locus in a:

a) linkage group G genomic region flanked by loci FE898349 and Cf14688d, b) linkage group A1 genomic region flanked by loci Mt7 and BARCSOYSSR_05_0640, c) linkage group M genomic region flanked by loci BQ081048 and TA53357_3847, d) linkage group D2 genomic region flanked by loci BG726970 and Contig41076, e) linkage group L genomic region flanked by loci Glyma19g01630 and TC27203, or f) linkage group K genomic region flanked by loci Cf4289d and Cf974d; and ii) selecting a soybean plant comprising in its genome at least one low iron growth condition tolerance locus comprising a genotype associated with a low iron growth condition tolerant phenotype. In certain embodiments of these methods, the selected soybean plant exhibits tolerance to low iron growth conditions. In certain embodiments of the aforementioned methods, the methods can further comprise assaying said selected plant of step (ii) for a low iron growth condition tolerant phenotype. In certain embodiments of any of the aforementioned methods, the methods can further comprises a step wherein said low iron growth condition tolerance locus is genotyped for at least one polymorphic allele of any of the markers selected from the group consisting of SEQ ID NOs: 52-62, 71-93, 94-110, 115-123, 124-138, and 139-174.

›SUMMARY OF INVENTION · 3 of 3

Also provided are methods for identifying a soybean plant comprising in its genome at least one introgressed low iron growth condition tolerance locus. In certain embodiments, these methods can comprising crossing a first soybean plant with a second soybean plant, the second plant comprising: i) a low iron growth condition tolerance locus in a: a) linkage group G genomic region flanked by loci FE898349 and Cf14688d, b) linkage group A1 genomic region flanked by loci Mt7 and BARCSOYSSR_05_0640, c) linkage group M genomic region flanked by loci BQ081048 and TA53357_3847, d) linkage group D2 genomic region flanked by loci BG726970 and Contig41076, e) linkage group L genomic region flanked by loci Glyma19g01630 and TC27203, or f) linkage group K genomic region flanked by loci Cf4289d and Cf974d; and ii) at least one additional polymorphic locus located outside of said region, to obtain a population of soybean plants segregating for the low iron growth condition tolerance loci and said at least one additional polymorphic locus; and detecting said polymorphic nucleic acid in at least one soybean plant from said population of soybean plants, wherein said one soybean plant lacks said additional polymorphic locus, thereby identifying a soybean plant comprising in its genome at least one introgressed low iron growth condition tolererance locus. In certain embodiments, these methods can further comprise the step of selecting said one soybean plant, thereby obtaining a soybean plant comprising in its genome at least one introgressed low iron growth condition tolerance locus. In certain embodiments of any of the aforementioned methods, the identified or selected plant is tolerant to low iron growth conditions. In certain embodiments of any of the aforementioned methods, the identified or selected plant is assayed for tolerance to low iron growth conditions. In certain embodiments of any of the aforementioned inventions, the additional polymorphic locus is detected with a genotypic marker, a phenotypic marker, or both.

In certain embodiments of any of the aforementioned methods, the additional polymorphic locus is a linked polymorphic locus located on linkage group G, A1, M, D2, L, or K, but not within a: a) linkage group G genomic region flanked by loci FE898349 and Cf14688d, b) linkage group A1 genomic region flanked by loci Mt7 and BARCSOYSSR_05_0640, c) linkage group M genomic region flanked by loci BQ081048 and TA53357_3847, d) linkage group D2 genomic region flanked by loci BG726970 and Contig41076, e) linkage group L genomic region flanked by loci Glyma19g01630 and TC27203, or f) linkage group K genomic region flanked by loci Cf4289d and Cf974d.

Also further provided herein are soybean plants obtainable by any of the aforementioned methods. Soybean plants comprising genomic regions associated with a low iron growth condition tolerant phenotype wherein immediately adjacent genomic regions and/or one or more adjacent genomic regions characteristic of soybean germplasms that lack the genomic regions associated with a low iron growth condition phenotype and/or that are distinct from the germplasm from which the genomic region is derived are also provided.

In certain embodiments, a soybean plant comprises: i) a low iron growth condition tolerance locus in a: a) linkage group G genomic region flanked by loci FE898349 and Cf14688d, b) linkage group A1 genomic region flanked by loci Mt7 and BARCSOYSSR_05_0640, c) linkage group M genomic region flanked by loci BQ081048 and TA53357_3847, d) linkage group D2 genomic region flanked by loci BG726970 and Contig41076, e) linkage group L genomic region flanked by loci Glyma19g01630 and TC27203, or f) linkage group K genomic region flanked by loci Cf4289d and Cf974d; and ii) one or more polymorphic loci comprising alleles or combinations of alleles that are not found in a low iron growth condition tolerant soybean varieties harboring said tolerance locus, and that are linked to said locus, wherein said soybean plant is tolerant to low iron growth conditions is provided.

In any of the aforementioned embodiments, the soybean plant can comprise an allele of one or more markers selected from the group consisting of SEQ ID NOs: 52-62, 71-93, 94-110, 115-123, 124-138, or 139-174. In certain embodiments, the nucleic acid can further comprise a detectable moiety. In certain embodiments, the detectable moiety can be selected from the group consisting of a chromophore, a fluorophore, and a hapten. Also provide herewith are isolated nucleic acid molecules selected from the group consisting SEQs ID NO: 1-186.

Further areas of applicability will become apparent from the description provided herein. It should be understood that the description and specific examples are intended for purposes of illustration only and are not intended to limit the scope of the present disclosure.

›DESCRIPTION OF INVENTION · 1 of 17

I. Definitions

Unless otherwise indicated herein, nucleic acid sequences are written left to right in 5′ to 3′ orientation. Numeric ranges recited within the specification are inclusive of the numbers defining the range and include each integer or any non-integer fraction within the defined range. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains.

As used herein, an “allele” refers to one of two or more alternative forms of a genomic sequence at a given locus on a chromosome. When all the alleles present at a given locus on a chromosome are the same, that plant is homozygous at that locus. If the alleles present at a given locus on a chromosome differ, that plant is heterozygous at that locus.

As used herein, the term “bulk” refers to a method of managing a segregating population during inbreeding that involves growing the population in a bulk plot, harvesting the self-pollinated seed of plants in bulk, and using a sample of the bulk to plant the next generation.

As used herein, the term “comprising” means “including but not limited to”.

As used herein, the term “denoting” when used in reference to a plant genotype refers to any method whereby a plant is indicated to have a certain genotype. Such indications of a certain genotype include, but are not limited to, any method where a plant is physically marked or tagged. Physical markings or tags that can be used include, but not limited to, a barcode, a radio-frequency identification (RFID), a label or the like. Indications of a certain genotype also include, but are not limited to, any entry into any type of written or electronic database whereby the plant's genotype is provided.

As used herein, the term “locus” refers to a position on a genomic sequence that is usually found by a point of reference; e.g., a short DNA sequence that is a gene, or part of a gene or intergenic region. A locus may refer to a nucleotide position at a reference point on a chromosome, such as a position from the end of the chromosome.

As used herein, “linkage group N” corresponds to the soybean linkage group N described in Choi, et al., Genetics. 2007 May; 176(1): 685-696. Linkage group N, as used herein, also corresponds to soybean chromosome 3 (as described on the World Wide Web at soybase.org/LG2Xsome.php).

As used herein, “linkage group M” corresponds to the soybean linkage group M described in Choi, et al., Genetics. 2007 May; 176(1): 685-696. Linkage group M, as used herein, also corresponds to soybean chromosome 7 (as described on the World Wide Web at soybase.org/LG2Xsome.php).

As used herein, “linkage group L” corresponds to the soybean linkage group L described in Choi, et al., Genetics. 2007 May; 176(1): 685-696. Linkage group L, as used herein, also corresponds to soybean chromosome 19 (as described on the World Wide Web at soybase.org/LG2Xsome.php).

As used herein, “linkage group O” corresponds to the soybean linkage group O described in Choi, et al., Genetics. 2007 May; 176(1): 685-696. Linkage group O, as used herein, also corresponds to soybean chromosome 10 (as described on the World Wide Web at soybase.org/LG2Xsome.php).

As used herein, “linkage group G” corresponds to the soybean linkage group G described in Choi, et al., Genetics. 2007 May; 176(1): 685-696. Linkage group G, as used herein, also corresponds to soybean chromosome 18 (as described on the World Wide Web at soybase.org/LG2Xsome.php).

As used herein, “linkage group A1” corresponds to the soybean linkage group A1 described in Choi, et al., Genetics. 2007 May; 176(1): 685-696. Linkage group A1, as used herein, also corresponds to soybean chromosome 5 (as described on the World Wide Web at soybase.org/LG2Xsome.php).

As used herein, “linkage group D2” corresponds to the soybean linkage group D2 described in Choi, et al., Genetics. 2007 May; 176(1): 685-696. Linkage group D2, as used herein, also corresponds to soybean chromosome 17 (as described on the World Wide Web at soybase.org/LG2Xsome.php).

As used herein, “linkage group K” corresponds to the soybean linkage group K described in Choi, et al., Genetics. 2007 May; 176(1): 685-696. Linkage group K, as used herein, also corresponds to soybean chromosome 9 (as described on the World Wide Web at soybase.org/LG2Xsome.php).

As used herein, “polymorphism” means the presence of one or more variations of a nucleic acid sequence at one or more loci in a population of at least two members. The variation can comprise, but is not limited to, one or more nucleotide base substitutions, the insertion of one or more nucleotides, a nucleotide sequence inversion, and/or the deletion of one or more nucleotides.

As used herein, “genotype” means the genetic component of the phenotype and it can be indirectly characterized using markers or directly characterized by nucleic acid sequencing.

As used herein, the term “introgressed”, when used in reference to a genetic locus, refers to a genetic locus that has been introduced into a new genetic background. Introgression of a genetic locus can thus be achieved through both plant breeding methods or by molecular genetic methods. Such molecular genetic methods include, but are not limited to, various plant transformation techniques and/or methods that provide for homologous recombination, non-homologous recombination, site-specific recombination, and/or genomic modifications that provide for locus substitution or locus conversion. In certain embodiments, introgression could thus be achieved by substitution of a locus not associated with tolerance to low iron growth conditions with a corresponding locus that is associated with low iron growth condition tolerance or by conversion of a locus from a non-tolerant genotype to a tolerant genotype.

›DESCRIPTION OF INVENTION · 2 of 17

As used herein, “linkage” refers to relative frequency at which types of gametes are produced in a cross. For example, if locus A has genes “A” or “a” and locus B has genes “B” or “b” and a cross between parent I with AABB and parent B with aabb will produce four possible gametes where the genes are segregated into AB, Ab, aB and ab. The null expectation is that there will be independent equal segregation into each of the four possible genotypes, i.e. with no linkage ¼ of the gametes will of each genotype. Segregation of gametes into a genotypes differing from ¼ are attributed to linkage.

As used herein, the termed “linked”, when used in the context of markers and/or genomic regions, means that the markers and/or genomic regions are located on the same linkage group or chromosome.

As used herein, “marker” means a detectable characteristic that can be used to discriminate between organisms. Examples of such characteristics include, but are not limited to, genetic markers, biochemical markers, fermentation yield, fermentation efficiency, energy yield, secondary compounds, metabolites, morphological characteristics, and agronomic characteristics.

As used herein, “marker assay” means a method for detecting a polymorphism at a particular locus using a particular method. Marker assays thus include, but are not limited to, measurement of at least one phenotype (such as seed color, flower color, or other visually detectable trait as well as any biochemical trait), restriction fragment length polymorphism (RFLP), single base extension, electrophoresis, sequence alignment, allelic specific oligonucleotide hybridization (ASO), random amplified polymorphic DNA (RAPD), microarray-based polymorphism detection technologies, and the like.

As used herein, “phenotype” means the detectable characteristics of a cell or organism which can be influenced by gene expression.

As used herein, a “nucleic acid molecule,” of naturally occurring origins or otherwise, may be an “isolated” nucleic acid molecule. An isolated nucleic acid molecule is one removed from its native cellular and chromosomal environment. The term “isolated” is not intended to encompass molecules present in their native state. If desired, an isolated nucleic acid may be substantially purified, meaning that it is the predominant species present in a preparation. A substantially purified molecule may be at least about 60% free, preferably at least about 75% free, more preferably at least about 90% free, and most preferably at least about 95% free from the other molecules (exclusive of solvent) present in the preparation.

As used herein, “quantitative trait locus (QTL)” means a locus that controls to some degree numerically representable traits that are usually continuously distributed.

As used herein, the term “soybean” means Glycine max and includes all plant varieties that can be bred with soybean, including wild soybean species. In certain embodiments, soybean plants from the species Glycine max and the subspecies Glycine max L. ssp. max or Glycine max ssp. formosana can be genotyped using the compositions and methods of the present invention. In an additional aspect, the soybean plant is from the species Glycine soja , otherwise known as wild soybean, can be genotyped using these compositions and methods. Alternatively, soybean germplasm derived from any of Glycine max, Glycine max L. ssp. max, Glycine max ssp. Formosana , and/or Glycine soja can be genotyped using compositions and methods provided herein.

As used herein, the term “single nucleotide polymorphism,” also referred to by the abbreviation “SNP,” means a polymorphism at a single site wherein the polymorphism constitutes any or all of a single base pair change, an insertion of one or more base pairs, and/or a deletion of one or more base pairs.

As used herein, the phrases “low iron,” “low-available iron,” “low soluble iron,” “low iron conditions,” “low iron growth conditions,” iron shortage” or “iron deficiency” or the like refer to conditions where iron availability is less than optimal for soybean growth, and can cause plant pathology, e.g., IDC, due to the lack of metabolically-available iron. It is recognized that under “iron deficient” conditions, the absolute concentration of atomic iron may be sufficient, but the form of the iron (e.g., its incorporation into various molecular structures) and other environmental factors may make the iron unavailable for plant use. For example, high carbonate levels, high pH, high salt content, herbicide applications, cool temperatures, saturated soils, or other environmental factors can decrease iron solubility, and reduce the solubilized forms of iron that the plant requires for uptake. One of skill in the art is familiar with assays to measure iron content of soil, as well as those concentrations of iron that are optimal or sub-optimal for plant growth.

As used herein, the terms “tolerance” or “improved tolerance” in reference to a soybean plant grown in low iron growth conditions is an indication that the soybean plant is less affected by the low-available iron conditions with respect to yield, survivability and/or other relevant agronomic measures, compared to a less tolerant, more “susceptible” plant. Tolerance is a relative term, indicating that a tolerant” plant survives and/or produces better yield of soybean in low-available iron growth conditions compared to a different (less tolerant) plant (e.g., a different soybean strain) grown in similar low-available iron conditions. That is, the low-available iron growth conditions cause a reduced decrease in soybean survival and/or yield in a tolerant soybean plant, as compared to a susceptible soybean plant. As used in the art, iron-deficiency “tolerance” is sometimes used interchangeably with iron-deficiency “resistance.”

One of skill will appreciate that soybean plant tolerance to low-available iron conditions varies widely, and can represent a spectrum of more-tolerant or less-tolerant phenotypes. However, by simple observation, one of skill can generally determine the relative tolerance or susceptibility of different plants, plant lines or plant families under low-available iron conditions, and furthermore, will also recognize the phenotypic gradations of “tolerant.”

›DESCRIPTION OF INVENTION · 3 of 17

In one example, a plant's tolerance can be approximately quantitated using a chlorosis scoring system. In such a system, a plant that is grown in a known iron-deficient area, or in low-available iron experimental conditions, and is assigned a tolerance rating of between 1 (highly susceptible; most or all plants dead; those that live are stunted and have little living tissue) to 9 (highly tolerant; yield and survivability not significantly affected; all plants normal green color). See also, Dahiya and Singh (1979) “Effect of salinity, alkalinity and iron sources on availability of iron,” Plant and Soil 51:13-18.

II. Description of the Invention: Overview

In accordance with the present invention, Applicants have discovered genomic regions, associated markers, and associated methods for identifying and associating genotypes that effect an iron deficient growth condition tolerance trait. For example, in one embodiment, a method of the invention comprises screening for alleles of polymorphic nucleic acid markers, including, but not limited to NS0202842 (SEQ ID NO:4), NS0206055 (SEQ ID NO: 22), NS0092960 (SEQ ID NO: 26), and NS0202730 (SEQ ID NO: 50) that are associated with low iron growth condition tolerance loci described herein.

The advent of molecular genetic markers has facilitated mapping and selection of agriculturally important traits in soybean. Markers tightly linked to tolerance genes are an asset in the rapid identification of tolerant soybean lines on the basis of genotype by the use of marker assisted selection (MAS). Introgressing tolerance genes into a desired cultivar is also facilitated by using suitable nucleic acid markers.

The use of markers to infer a phenotype of interest results in the economization of a breeding program by substituting costly, time-intensive phenotyping assays with genotyping assays. Further, breeding programs can be designed to explicitly drive the frequency of specific, favorable phenotypes by targeting particular genotypes (U.S. Pat. No. 6,399,855). Fidelity of these associations may be monitored continuously to ensure maintained predictive ability and, thus, informed breeding decisions (US Patent Application 2005/0015827). In this case, costly, time-intensive phenotyping assays required for determining if a plant or plants contains a genomic region associated with a low iron growth condition tolerant phenotype can be supplanted by genotypic assays that provide for identification of a plant or plants that contain the desired genomic region.

III. QTL Associated with Tolerance to Low Iron Growth Conditions

Provided herewith are certain QTL that have been identified as associated with a desirable phenotype of tolerance to growth in low iron conditions when present in certain allelic forms.

The several soybean QTL provided—that can be associated with a desirable low iron growth condition tolerant phenotype when present in certain allelic forms—are located on soybean chromosome 3 (soybean linkage group N), soybean chromosome 7 (soybean linkage group M), soybean chromosome 19 (soybean linkage group L), and soybean chromosome 10 (soybean linkage group O).

A series of public and other markers useful in practicing the methods of this invention are provided herewith in Tables 1-10. Nucleic acid sequences for certain non-public markers useful in the practice of the invention are provided herewith in the accompanying sequence listing, which is incorporated herein by reference in its entirety.

Tables 1, 4, 6, and 9 (corresponding to chromosomes 3, 19, 10, and 7, respectively) shows the relative positions of certain markers that have been disclosed in public databases and non-public (bolded) polymorphic nucleic acid markers, designated SEQ ID NOs, genetic positions (cM) on the chromosome, the allelic forms of certain polymorphic nucleic acid markers associated with a low iron growth condition tolerant phenotype, the allelic forms of those polymorphic nucleic acid markers not associated with the low iron growth condition tolerant phenotype, the polymorphic position within the sequence of the polymorphic nucleic acid marker, and a designation of whether a particular marker has been identified as within a genomic region associated with a low iron growth condition tolerant phenotype.

Tables 2, 5, 7, and 10 (corresponding to chromosomes 3, 19, 10, and 7, respectively) provides for each polymorphic nucleic acid marker/SEQ ID NO the linkage group corresponding to the chromosome and the relative physical map positions of the markers.

Tables 3, 8 (corresponding to chromosomes 3 and 10, respectively) provide for certain polymorphic nucleic acid markers, the type of marker, and primer and probe sequences useful in detecting such markers.

Table 12 of the Appendix to the Specification, which is incorporated herein by reference in its entirety, discloses the sources of certain of the markers contained in Tables 1-10.

Provided herein is a QTL on chromosome 3—that is flanked by loci Glyma03g34510 and Glyma03g42250—that is associated with a low iron growth condition tolerant phenotype. Sub-regions of chromosome 3 that flank a QTL associated with a low iron growth condition tolerant phenotype include, but are not limited to sub-regions defined by any of the following sets of loci:

a) loci Glyma03g34510 and Glyma03g41830;

b) loci Gm_W82_CR03.G378360 and Glyma03g42250;

c) loci Gm_W82_CR03.G378360 and Glyma03g41830;

d) loci Glyma03g34760 and Glyma03g42250;

e) loci Glyma03g34760 and Glyma03g41830;

f) loci Glyma03g34860 and Glyma03g42250; or

g) loci Glyma03g34860 and Glyma03g41830.

(Table 1).

These loci flank a genomic interval that spans telomere proximal nucleotide 41914204 to centromere proximal nucleotide 47499828 in the physical map of linkage group N provided in the Table 2. Polymorphic nucleic acid markers located in this genomic interval that are associated with a low iron growth condition tolerant phenotype include, but are not limited to: NS0202712 (SEQ ID NO: 1); NS0129403 (SEQ ID NO: 2); NS0115624 (SEQ ID NO: 3); NS0202842 (SEQ ID NO: 4); NS0122122 (SEQ ID NO: 5); NS0205984 (SEQ ID NO: 6); and NS0202698 (SEQ ID NO: 7).

›DESCRIPTION OF INVENTION · 4 of 17

Provided herein is a QTL on chromosome 19—that is flanked by loci Glyma19g32880 and Contig9146—that is associated with a low iron growth condition tolerant phenotype. Sub-regions of chromosome 19 that flank a QTL associated with a low iron growth condition tolerant phenotype include, but are not limited to sub-regions defined by any of the following sets of loci:

a) loci Glyma19g32880 and Gm_W82_CR19.G249630;

b) loci Glyma19g32880 and Glyma19g34750;

c) loci Glyma19g32880 and BARCSOYSSR_19_1192;

d) loci Glyma19g32880 and Contig13742;

e) loci Glyma19g32880 and 220298_3433_1666;

f) loci Glyma19g32880 and NGMAX005710149;

g) loci Glyma19g32880 and Glyma19g34740;

h) loci Glyma19g32880 and Gm_W82_CR19.G249620;

i) loci Glyma19g33540 and Contig9146;

j) loci Glyma19g33540 and Gm_W82_CR19.G249630;

k) loci Glyma19g33540 and Glyma19g34750;

l) loci Glyma19g33540 and BARCSOYSSR_19_1192;

m) loci Glyma19g33540 and Contig13742;

n) loci Glyma19g33540 and 220298_3433_1666;

o) loci Glyma19g33540 and NGMAX005710149;

p) loci Glyma19g33540 and Glyma19g34740; or

q) loci Glyma19g33540 and Gm_W82_CR19.G249620.

(Table 4).

These loci flank a genomic interval that spans telomere proximal nucleotide 40562661 to centromere proximal nucleotide 42351748 in the physical map of linkage group L provided in the Table 5. Polymorphic nucleic acid markers located in this genomic interval that are associated with a low iron growth condition tolerant phenotype can be detected with polymorphic nucleic acid markers that include, but are not limited to: NS0206298_(SEQ ID NO: 20); NS0205620 (SEQ ID NO: 21); NS0206055 (SEQ ID NO: 22); and NS0204985 (SEQ ID NO: 23).

Provided herein is a genomic interval on chromosome 10 spanning two QTL—that is flanked by loci NS0116559 and Gm_W82_CR10.G236520—that is associated with a low iron growth condition tolerant phenotype. Sub-regions of chromosome 10 that flank QTL associated with a low iron growth condition tolerant phenotype include, but are not limited to sub-regions defined by any of the following sets of loci:

a) loci NS0116559 and TC412490;

b) loci NS0116559 and Glyma10g41470;

c) loci NS0116559 and Glyma10g41460;

d) loci NS0116559 and BF631841;

e) loci NS0116559 and Glyma10g40520;

f) loci NS0116559 and Glyma10g38600;

g) loci Glyma10g33710 and Gm_W82_CR10.G236520;

h) loci Glyma10g33710 and TC412490;

i) loci Glyma10g33710 and Glyma10g41470;

j) loci Glyma10g33710 and Glyma10g41460;

k) loci Glyma10g33710 and BF631841;

l) loci Glyma10g33710 and Glyma10g40520;

m) loci Glyma10g33710 and Glyma10g38600;

n) loci TA42233_3847 and Gm_W82_CR10.G236520;

o) loci TA42233_3847 and TC412490;

p) loci TA42233_3847 and Glyma10g41470;

q) loci TA42233_3847 and Glyma10g41460;

r) loci TA42233_3847 and BF631841;

s) loci TA42233_3847 and Glyma10g40520;

t) loci TA42233_3847 and Glyma10g38600;

u) loci AF108084.1 and Gm_W82_CR10.G236520;

v) loci AF108084.1 and TC412490;

w) loci AF108084.1 and Glyma10g41470;

x) loci AF108084.1 and Glyma10g41460;

y) loci AF108084.1 and BF631841;

z) loci AF108084.1 and Glyma10g40520;

aa) loci AF108084.1 and Glyma10g38600;

bb) loci AW734581 and Gm_W82_CR10.G236520;

cc) loci AW734581 and TC412490;

dd) loci AW734581 and Glyma10g41470;

ee) loci AW734581 and Glyma10g41460;

ff) loci AW734581 and BF631841;

gg) loci AW734581 and Glyma10g40520;

hh) loci AW734581 and Glyma10g38600;

ii) loci Glyma10g34260 and Gm_W82_CR10.G236520;

jj) loci Glyma10g34260 and TC412490;

kk) loci Glyma10g34260 and Glyma10g41470;

ll) loci Glyma10g34260 and Glyma10g41460;

mm) loci Glyma10g34260 and BF631841;

nn) loci Glyma10g34260 and Glyma10g40520;

oo) loci Glyma10g34260 and Glyma10g38600;

pp) loci Glyma10g34280 and Gm_W82_CR10.G236520;

qq) loci Glyma10g34280 and TC412490;

rr) loci Glyma10g34280 and Glyma10g41470;

ss) loci Glyma10g34280 and Glyma10g41460;

tt) loci Glyma10g34280 and BF631841;

›DESCRIPTION OF INVENTION · 5 of 17

uu) loci Glyma10g34280 and Glyma10g40520;

vv) loci Glyma10g34280 and Glyma10g38600;

ww) loci Glyma10g34290 and Gm_W82_CR10.G236520;

xx) loci Glyma10g34290 and TC412490;

yy) loci Glyma10g34290 and Glyma10g41470;

zz) loci Glyma10g34290 and Glyma10g41460;

aaa) loci Glyma10g34290 and BF631841;

bbb) loci Glyma10g34290 and Glyma10g40520;

ccc) loci Glyma10g34290 and Glyma10g38600;

ddd) loci Glyma10g34460 and Gm_W82_CR10.G236520;

eee) loci Glyma10g34460 and TC412490;

fff) loci Glyma10g34460 and Glyma10g41470;

ggg) loci Glyma10g34460 and Glyma10g41460;

hhh) loci Glyma10g34460 and BF631841;

iii) loci Glyma10g34460 and Glyma10g40520;

jjj) loci Glyma10g34460 and Glyma10g38600;

kkk) loci Glyma10g34600 and Gm_W82_CR10.G236520;

lll) loci Glyma10g34600 and TC412490;

mmm) loci Glyma10g34600 and Glyma10g41470;

nnn) loci Glyma10g34600 and Glyma10g41460;

ooo) loci Glyma10g34600 and BF631841;

ppp) loci Glyma10g34600 and Glyma10g40520;

qqq) loci Glyma10g34600 and Glyma10g38600;

rrr) loci Glyma10g34630 and Gm_W82_CR10.G236520;

sss) loci Glyma10g34630 and TC412490;

ttt) loci Glyma10g34630 and Glyma10g41470;

uuu) loci Glyma10g34630 and Glyma10g41460;

vvv) loci Glyma10g34630 and BF631841;

www) loci Glyma10g34630 and Glyma10g40520;

xxx) loci Glyma10g34630 and Glyma10g38600;

yyy) loci Glyma10g34850 and Gm_W82_CR10.G236520;

zzz) loci Glyma10g34850 and TC412490;

aaaa) loci Glyma10g34850 and Glyma10g41470;

bbbb) loci Glyma10g34850 and Glyma10g41460;

cccc) loci Glyma10g34850 and BF631841;

dddd) loci Glyma10g34850 and Glyma10g40520; or

eeee) loci Glyma10g34850 and Glyma10g38600.

(Table 6).

These loci flank a genomic interval that spans telomere proximal nucleotide 42075167 to centromere proximal nucleotide 49718518 in the physical map of linkage group O provided in the Table 7. Polymorphic nucleic acid markers located in this genomic interval that are associated with a low iron growth condition tolerant phenotype can be detected with polymorphic nucleic acid markers that include, but are not limited to: NS0120070 (SEQ ID NO: 24); NS0097952 (SEQ ID NO: 25); NS0092960 (SEQ ID NO: 26); NS0118907 (SEQ ID NO: 27); NS0204740 (SEQ ID NO: 28); NS0205036 (SEQ ID NO: 29); and NS0206252 (SEQ ID NO: 30).

Provided herein is a first QTL on chromosome 10—that is flanked by loci NS0116559 and TA68568_3847—that is associated with a low iron growth condition tolerant phenotype. Sub-regions of chromosome 10 that flank this first QTL associated with a low iron growth condition tolerant phenotype include, but are not limited to sub-regions defined by any of the following sets of loci:

a) loci NS0116559 and BG511181;

b) loci NS0116559 and Glyma10g37610;

c) loci NS0116559 and CV528982;

d) loci NS0116559 and TC130824;

e) loci NS0116559 and BW595896;

f) loci NS0116559 and Glyma10g37600;

g) loci NS0116559 and TC354083;

h) loci NS0116559 and Glyma10g37560;

i) loci Glyma10g33710 and TA68568_3847;

j) loci Glyma10g33710 and BG511181;

k) loci Glyma10g33710 and Glyma10g37610;

l) loci Glyma10g33710 and CV528982;

m) loci Glyma10g33710 and TC130824;

n) loci Glyma10g33710 and BW595896;

o) loci Glyma10g33710 and Glyma10g37600;

p) loci Glyma10g33710 and TC354083;

q) loci Glyma10g33710 and Glyma10g37560;

r) loci TA42233_3847 and TA68568_3847;

s) loci TA42233_3847 and BG511181;

t) loci TA42233_3847 and Glyma10g37610;

u) loci TA42233_3847 and CV528982;

v) loci TA42233_3847 and TC130824;

w) loci TA42233_3847 and BW595896;

x) loci TA42233_3847 and Glyma10g37600;

y) loci TA42233_3847 and TC354083;

z) loci TA42233_3847 and Glyma10g37560;

aa) loci AF108084.1 and TA68568_3847;

bb) loci AF108084.1 and BG511181;

cc) loci AF108084.1 and Glyma10g37610;

dd) loci AF108084.1 and CV528982;

ee) loci AF108084.1 and TC130824;

ff) loci AF108084.1 and BW595896;

gg) loci AF108084.1 and Glyma10g37600;

hh) loci AF108084.1 and TC354083;

ii) loci AF108084.1 and Glyma10g37560;

jj) loci AW734581 and TA68568_3847;

kk) loci AW734581 and BG511181;

ll) loci AW734581 and Glyma10g37610;

mm) loci AW734581 and CV528982;

nn) loci AW734581 and TC130824;

oo) loci AW734581 and BW595896;

pp) loci AW734581 and Glyma10g37600;

qq) loci AW734581 and TC354083;

rr) loci AW734581 and Glyma10g37560;

ss) loci Glyma10g34260 and TA68568_3847;

tt) loci Glyma10g34260 and BG511181;

uu) loci Glyma10g34260 and Glyma10g37610;

vv) loci Glyma10g34260 and CV528982;

ww) loci Glyma10g34260 and TC130824;

xx) loci Glyma10g34260 and BW595896;

yy) loci Glyma10g34260 and Glyma10g37600;

zz) loci Glyma10g34260 and TC354083;

aaa) loci Glyma10g34260 and Glyma10g37560;

bbb) loci Glyma10g34280 and TA68568_3847;

ccc) loci Glyma10g34280 and BG511181;

ddd) loci Glyma10g34280 and Glyma10g37610;

eee) loci Glyma10g34280 and CV528982;

fff) loci Glyma10g34280 and TC130824;

ggg) loci Glyma10g34280 and BW595896;

hhh) loci Glyma10g34280 and Glyma10g37600;

iii) loci Glyma10g34280 and TC354083;

jjj) loci Glyma10g34280 and Glyma10g37560;

kkk) loci Glyma10g34290 and TA68568_3847;

›DESCRIPTION OF INVENTION · 6 of 17

lll) loci Glyma10g34290 and BG511181;

mmm) loci Glyma10g34290 and Glyma10g37610;

nnn) loci Glyma10g34290 and CV528982;

ooo) loci Glyma10g34290 and TC130824;

ppp) loci Glyma10g34290 and BW595896;

qqq) loci Glyma10g34290 and Glyma10g37600;

rrr) loci Glyma10g34290 and TC354083;

sss) loci Glyma10g34290 and Glyma1437560;

ttt) loci Glyma10g34460 and TA68568_3847;

uuu) loci Glyma10g34460 and BG511181;

vvv) loci Glyma10g34460 and Glyma10g37610;

www) loci Glyma10g34460 and CV528982;

xxx) loci Glyma10g34460 and TC130824;

yyy) loci Glyma10g34460 and BW595896;

zzz) loci Glyma10g34460 and Glyma10g37600;

aaaa) loci Glyma10g34460 and TC354083;

bbbb) loci Glyma10g34460 and Glyma10g37560;

cccc) loci Glyma10g34600 and TA68568_3847;

dddd) loci Glyma10g34600 and BG511181;

eeee) loci Glyma10g34600 and Glyma10g37610;

ffff) loci Glyma10g34600 and CV528982;

gggg) loci Glyma10g34600 and TC130824;

hhhh) loci Glyma10g34600 and BW595896;

iiii) loci Glyma10g34600 and Glyma10g37600;

jjjj) loci Glyma10g34600 and TC354083;

kkkk) loci Glyma10g34600 and Glyma10g37560;

llll) loci Glyma10g34630 and TA68568_3847;

mmmm) loci Glyma10g34630 and BG511181;

nnnn) loci Glyma10g34630 and Glyma10g37610;

oooo) loci Glyma10g34630 and CV528982;

pppp) loci Glyma10g34630 and TC130824;

qqqq) loci Glyma10g34630 and BW595896;

rrrr) loci Glyma10g34630 and Glyma10g37600;

ssss) loci Glyma10g34630 and TC354083;

tttt) loci Glyma10g34630 and Glyma10g37560;

uuuu) loci Glyma10g34850 and TA68568_3847;

vvvv) loci Glyma10g34850 and BG511181;

wwww) loci Glyma10g34850 and Glyma10g37610;

xxxx) loci Glyma10g34850 and CV528982;

yyyy) loci Glyma10g34850 and TC130824;

zzzz) loci Glyma10g34850 and BW595896;

aaaaa) loci Glyma10g34850 and Glyma10g37600;

bbbbb) loci Glyma10g34850 and TC354083; or

ccccc) loci Glyma10g34850 and Glyma10g37560.

(Table 6).

These loci flank a genomic interval that spans telomere proximal nucleotide 42075167 to centromere proximal nucleotide 46157806 in the physical map of linkage group O provided in the Table 7. Polymorphic nucleic acid markers located in this genomic interval that are associated with a low iron growth condition tolerant phenotype can be detected with polymorphic nucleic acid markers that include, but are not limited to: NS0120070 (SEQ ID NO: 24); NS0097952 (SEQ ID NO: 25); NS0092960 (SEQ ID NO: 26); and NS0118907 (SEQ ID NO: 27).

Provided herein is a second QTL on chromosome 10—that is flanked by loci Glyma10g37560 and Gm_W82_CR10.G236520—that is associated with a low iron growth condition tolerant phenotype. Sub-regions of chromosome 10 that flank this second QTL associated with a low iron growth condition tolerant phenotype include, but are not limited to sub-regions defined by any of the following sets of loci:

a) loci Glyma10g37560 and TC412490;

b) loci Glyma10g37560 and Glyma10g41470;

c) loci Glyma10g37560 and Glyma10g41460;

d) loci Glyma10g37560 and BF631841;

e) loci Glyma10g37560 and Glyma10g40520;

f) loci Glyma10g37560 and Glyma10g38600;

g) loci TC354083 and Gm_W82_CR10.G236520

h) loci TC354083 and TC412490;

i) loci TC354083 and Glyma10g41470;

j) loci TC354083 and Glyma10g41460;

k) loci TC354083 and BF631841;

l) loci TC354083 and Glyma10g40520;

m) loci TC354083 and Glyma10g38600;

n) loci Glyma10g37600 and Gm_W82_CR10.G236520

o) loci Glyma10g37600 and TC412490;

p) loci Glyma10g37600 and Glyma10g41470;

q) loci Glyma10g37600 and Glyma10g41460;

r) loci Glyma10g37600 and BF631841;

s) loci Glyma10g37600 and Glyma10g40520;

t) loci Glyma10g37600 and Glyma10g38600;

u) loci BW595896 and Gm_W82_CR10.G236520

v) loci BW595896 and TC412490;

w) loci BW595896 and Glyma10g41470;

x) loci BW595896 and Glyma10g41460;

y) loci BW595896 and BF631841;

z) loci BW595896 and Glyma10g40520;

aa) loci BW595896 and Glyma10g38600;

bb) loci TC130824 and Gm_W82_CR10.G236520

cc) loci TC130824 and TC412490;

dd) loci TC130824 and Glyma10g41470;

ee) loci TC130824 and Glyma10g41460;

ff) loci TC130824 and BF631841;

gg) loci TC130824 and Glyma10g40520;

hh) loci TC130824 and Glyma10g38600;

ii) loci CV528982 and Gm_W82_CR10.G236520

jj) loci CV528982 and TC412490;

kk) loci CV528982 and Glyma10g41470;

ll) loci CV528982 and Glyma10g41460;

mm) loci CV528982 and BF631841;

nn) loci CV528982 and Glyma10g40520;

oo) loci CV528982 and Glyma10g38600;

pp) loci Glyma10g37610 and Gm_W82_CR10.G236520

qq) loci Glyma10g37610 and TC412490;

rr) loci Glyma10g37610 and Glyma10g41470;

ss) loci Glyma10g37610 and Glyma10g41460;

tt) loci Glyma10g37610 and BF631841;

uu) loci Glyma10g37610 and Glyma10g40520;

vv) loci Glyma10g37610 and Glyma10g38600;

ww) loci BG511181 and Gm_W82_CR10.G236520

xx) loci BG511181 and TC412490;

yy) loci BG511181 and Glyma10g41470;

zz) loci BG511181 and Glyma10g41460;

aaa) loci BG511181 and BF631841;

bbb) loci BG511181 and Glyma10g40520;

ccc) loci BG511181 and Glyma10g38600;

ddd) loci TA68568_3847 and Gm_W82_CR10.G236520

ccc) loci TA68568_3847 and TC412490;

ddd) loci TA68568_3847 and Glyma10g41470;

eee) loci TA68568_3847 and Glyma10g41460;

fff) loci TA68568_3847 and BF631841;

ggg) loci TA68568_3847 and Glyma10g40520; or

hhh) loci TA68568_3847 and Glyma10g38600.

(Table 6).

These loci flank a genomic interval that spans telomere proximal nucleotide 46112579 to centromere proximal nucleotide 49718518 in the physical map of linkage group O provided in the Table 7. Polymorphic nucleic acid markers located in this genomic interval that are associated with a low iron growth condition tolerant phenotype can be detected with polymorphic nucleic acid markers that include, but are not limited to: NS0204740 (SEQ ID NO: 28); NS0205036 (SEQ ID NO:29); and NS0206252 (SEQ ID NO: 30).

Provided herein is a genomic interval on chromosome 7 spanning two QTL—that is flanked by loci Gm_W82_CR07.G4950 and Glyma07g33560—that is associated with a low iron growth condition tolerant phenotype. Sub-regions of chromosome 7 that flank QTL associated with a low iron growth condition tolerant phenotype include, but are not limited to sub-regions defined by any of the following sets of loci:

a) loci Gm_W82_CR07.G4950 and any of loci: Glyma07g33090, Glyma07g33070, Glyma07g32330, or Glyma07g31610;

›DESCRIPTION OF INVENTION · 7 of 17

b) loci TC112538 and any of loci: Glyma07g33560, Glyma07g33090, Glyma07g33070, Glyma07g32330, or Glyma07g31610;

c) loci Glyma07g03790 and any of loci: Glyma07g33560, Glyma07g33090, Glyma07g33070, Glyma07g32330, or Glyma07g31610;

d) loci Gm_W82_CR07.08460 and any of loci: Glyma07g33560, Glyma07g33090, Glyma07g33070, Glyma07g32330, or Glyma07g31610;

e) loci Glyma07g03800 and any of loci: Glyma07g33560, Glyma07g33090, Glyma07g33070, Glyma07g32330, or Glyma07g31610; or

f) loci Glyma07g03810 and any of loci: Glyma07g33560, Glyma07g33090, Glyma07g33070, Glyma07g32330, or Glyma07g31610.

(Table 9).

These loci flank a genomic interval that spans telomere proximal nucleotide 1582889 to centromere proximal nucleotide 38480663 in the physical map of linkage group M provided in the Table 10. Polymorphic nucleic acid markers located in this genomic interval that are associated with a low iron growth condition tolerant phenotype can be detected with polymorphic nucleic acid markers that include, but are not limited to: NS0202654 (SEQ ID NO: 47), NS0206351 (SEQ ID NO: 48), NS0202966 (SEQ ID NO: 49), NS0202730 (SEQ ID NO: 50), and NS0206314 (SEQ ID NO: 51).

Provided herein is a first QTL on chromosome 7—that is flanked by loci Gm_W82_CR07.G4950 and Glyma07g12210—that is associated with a low iron growth condition tolerant phenotype. Sub-regions of chromosome 7 that flank this first QTL associated with a low iron growth condition tolerant phenotype include, but are not limited to sub-regions defined by any of the following sets of loci:

a) loci Gm_W82_CR07.G4950 and any of loci: Glyma07g11180, TA57919_3847, NS0119842, Glyma07g09970, Glyma07g09960, Glyma07g09900, Glyma07g09170, Glyma07g09160, Glyma07g09150, Glyma07g09110, Glyma07g08950, TC385708, Glyma07g07560, TC119399, Pvcon9217, Gm_W82_CR07.G30600, Glyma07g07380, Glyma07g06560, Glyma07g06510, Glyma07g06150, NS0102362, Glyma07g05820, and Glyma07g05420;

b) loci TC112538 and any of loci: Glyma07g12210, Glyma07g11180, TA57919_3847, NS0119842, Glyma07g09970, Glyma07g09960, Glyma07g09900, Glyma07g09170, Glyma07g09160, Glyma07g09150, Glyma07g09110, Glyma07g08950, TC385708, Glyma07g07560, TC119399, Pvcon9217, Gm_W82_CR07.G30600, Glyma07g07380, Glyma07g06560, Glyma07g06510, Glyma07g06150, NS0102362, Glyma07g05820, and Glyma07g05420;

c) loci Glyma07g03790 and any of loci: Glyma07g12210, Glyma07g11180, TA57919_3847, NS0119842, Glyma07g09970, Glyma07g09960, Glyma07g09900, Glyma07g09170, Glyma07g09160, Glyma07g09150, Glyma07g09110, Glyma07g08950, TC385708, Glyma07g07560, TC119399, Pvcon9217, Gm_W82_CR07.G30600, Glyma07g07380, Glyma07g06560, Glyma07g06510, Glyma07g06150, NS0102362, Glyma07g05820, and Glyma07g05420;

d) loci Gm_W82_CR07.G8460 and any of loci: Glyma07g12210, Glyma07g11180, TA57919_3847, NS0119842, Glyma07g09970, Glyma07g09960, Glyma07g09900, Glyma07g09170, Glyma07g09160, Glyma07g09150, Glyma07g09110, Glyma07g08950, TC385708, Glyma07g07560, TC119399, Pvcon9217, Gm_W82_CR07.G30600, Glyma07g07380, Glyma07g06560, Glyma07g06510, Glyma07g06150, NS0102362, Glyma07g05820, and Glyma07g05420;

e) Glyma07g03800 and any of loci: Glyma07g12210, Glyma07g11180, TA57919_3847, NS0119842, Glyma07g09970, Glyma07g09960, Glyma07g09900, Glyma07g09170, Glyma07g09160, Glyma07g09150, Glyma07g09110, Glyma07g08950, TC385708, Glyma07g07560, TC119399, Pvcon9217, Gm_W82_CR07.G30600, Glyma07g07380, Glyma07g06560, Glyma07g06510, Glyma07g06150, NS0102362, Glyma07g05820, and Glyma07g05420; and

f) Glyma07g03810 and any of loci: Glyma07g12210, Glyma07g11180, TA57919_3847, NS0119842, Glyma07g09970, Glyma07g09960, Glyma07g09900, Glyma07g09170, Glyma07g09160, Glyma07g09150, Glyma07g09110, Glyma07g08950, TC385708, Glyma07g07560, TC119399, Pvcon9217, Gm_W82_CR07.G30600, Glyma07g07380, Glyma07g06560, Glyma07g06510, Glyma07g06150, NS0102362, Glyma07g05820, and Glyma07g05420.

(Table 9).

These loci flank a genomic interval that spans telomere proximal nucleotide 1582889 to centromere proximal nucleotide 10530235 in the physical map of linkage group M provided in the Table 10. Polymorphic nucleic acid markers located in this genomic interval that are associated with a low iron growth condition tolerant phenotype can be detected with polymorphic nucleic acid markers that include, but are not limited to: NS00202654 (SEQ ID NO: 47); NS0206351 (SEQ ID NO: 48); and NS0202966 (SEQ ID NO: 49).

Provided herein is a second QTL on chromosome 7—that is flanked by loci Glyma07g29640 and Glyma07g33560—that is associated with a low iron growth condition tolerant phenotype. Sub-regions of chromosome 7 that flank this second QTL associated with a low iron growth condition tolerant phenotype include, but are not limited to sub-regions defined by any of the following sets of loci:

a) loci Glyma07g29640 and any of loci: Glyma07g33090, Glyma07g33070, Glyma07g32330, and Glyma07g31610;

b) loci Glyma07g29650 and any of loci: Glyma07g33560, Glyma07g33090, Glyma07g33070, Glyma07g32330, and Glyma07g31610;

›DESCRIPTION OF INVENTION · 8 of 17

c) loci Glyma07g29940 and any of loci: Glyma07g33560, Glyma07g33090, Glyma07g33070, Glyma07g32330, and Glyma07g31610;

d) loci Glyma07g31380 and any of loci: Glyma07g33560, Glyma07g33090, Glyma07g33070, Glyma07g32330, and Glyma07g31610; or

e) loci Glyma07g31390 and any of loci: Glyma07g33560, Glyma07g33090, Glyma07g33070, Glyma07g32330, and Glyma07g31610.

(Table 9).

These loci flank a genomic interval that spans telomere proximal nucleotide 34635418 to centromere proximal nucleotide 38480663 in the physical map of linkage group M provided in the Table 10. Polymorphic nucleic acid markers located in this genomic interval that are associated with a low iron growth condition tolerant phenotype can be detected with polymorphic nucleic acid markers that include, but are not limited to: NS0202730 (SEQ ID NO: 50) and NS0206314 (SEQ ID NO: 51).

Further provided herewith are certain other QTL that have also been identified as associated with a desirable phenotype of tolerance to growth in low iron conditions when present in certain allelic forms.

These several soybean QTL provided—that can be associated with a desirable low iron growth condition tolerant phenotype when present in certain allelic forms—are located on soybean chromosome 18 (soybean linkage group G), soybean chromosome 5 (soybean linkage group A1), soybean chromosome 7 (soybean linkage group M), soybean chromosome 17 (soybean linkage group D2), soybean chromosome 19 (soybean linkage group L), and soybean chromosome 9 (soybean linkage group K).

A series of public and other markers useful in practicing the methods of this invention are provided herewith in Tables 14-31. Nucleic acid sequences for certain non-public markers useful in the practice of the invention are provided herewith in the accompanying sequence listing, which is incorporated herein by reference in its entirety.

Tables 15, 18, 21, 24, 27, and 30 (corresponding to chromosomes 18, 5, 7, 17, 19, and 9, respectively) shows the relative positions of certain markers that have been disclosed in public databases and non-public (bolded) polymorphic nucleic acid markers, designated SEQ ID NOs, genetic positions (cM) on the chromosome, the allelic forms of certain polymorphic nucleic acid markers associated with a low iron growth condition tolerant phenotype, the allelic forms of those polymorphic nucleic acid markers not associated with the low iron growth condition tolerant phenotype, and the polymorphic position within the sequence of the polymorphic nucleic acid marker. The bolded markers have been identified as within a genomic region associated with a low iron growth condition tolerant phenotype.

Tables 16, 19, 22, 25, 28 and 31 (corresponding to chromosomes 18, 5, 7, 17, 19, and 9, respectively) provides for each polymorphic nucleic acid marker/SEQ ID NO the linkage group corresponding to the chromosome and the relative physical map positions of the markers.

Tables 32, 33, and 34 (corresponding to chromosomes 18, 7 and 9, respectively) provide for certain polymorphic nucleic acid markers, the type of marker, and primer and probe sequences useful in detecting such markers.

Tables 14, 17, 20, 23, 26, and 29 of the Appendix to the Specification, which is incorporated herein by reference in its entirety, discloses the sources of certain of the markers contained in Tables 15, 16, 18, 19, 21, 22, 24, 25, 27, 28, 30, 31, and 32-34.

Provided herein is a QTL on chromosome 18—that is flanked by loci FE898349 and Cf14688d—that is associated with a low iron growth condition tolerant phenotype. Sub-regions of chromosome 18 that flank a QTL associated with a low iron growth condition tolerant phenotype include, but are not limited to sub-regions defined by any of the following sets of loci:

a) loci FE898349 and 108285_2293_0177;

b) loci FE898349 and Cf17723d;

c) loci TC126194 and 108285_2293_0177;

d) loci TC126194 and Cf14688d;

e) loci TC126194 and Cf17723d;

f) loci Glyma18g50900 and 108285_2293_0177;

g) loci Glyma18g50900 and Cf14688d; or

h) loci Glyma18g50900 and Cf17723d.

(Table 15).

These loci flank a genomic interval that spans telomere proximal nucleotide 59900436 to centromere proximal nucleotide 62095680 in the physical map of linkage group G provided in the Table 16. Polymorphic nucleic acid markers located in this genomic interval that are associated with a low iron growth condition tolerant phenotype include, but are not limited to: NGMAX008183850 (SEQ ID NO: 52); NGMAX008184740 (SEQ ID NO: 53); NS0204945 (SEQ ID NO: 54); NGMAX008190659 (SEQ ID NO: 55); NGMAX008190985 (SEQ ID NO: 56); NS0205638 (SEQ ID NO: 57); NGMAX008191317 (SEQ ID NO: 58); NS0092671 (SEQ ID NO: 59); NS0118292 (SEQ ID NO: 60); NGMAX008194551 (SEQ ID NO: 61); and NGMAX008194952 (SEQ ID NO: 62).

›DESCRIPTION OF INVENTION · 9 of 17

Provided herein is a QTL on chromosome 5—that is flanked by loci Mt7 and BARCSOYSSR_05_0640—that is associated with a low iron growth condition tolerant phenotype. Sub-regions of chromosome 5 that flank a QTL associated with a low iron growth condition tolerant phenotype include, but are not limited to sub-regions defined by any of the following sets of loci:

a) loci Mt7 and BARCSOYSSR_05_0638;

b) loci Mt7 and BARCSOYSSR_05_0639;

c) loci WmFPC_Contig7872 and BARCSOYSSR_05_0638;

d) loci WmFPC_Contig7872 and BARCSOYSSR_05_0639; or

e) loci WmFPC_Contig7872 and BARCSOYSSR_05_0640.

(Table 18).

These loci flank a genomic interval that spans telomere proximal nucleotide 1030287 to centromere proximal nucleotide 26176471 in the physical map of linkage group A1 provided in Table 19. Polymorphic nucleic acid markers located in this genomic interval that are associated with a low iron growth condition tolerant phenotype can be detected with polymorphic nucleic acid markers that include, but are not limited to: NGMAX006398059 (SEQ ID NO: 71); NGMAX006398075 (SEQ ID NO: 72); NGMAX006398171 (SEQ ID NO: 73); NGMAX006398585 (SEQ ID NO: 74); NGMAX006398641 (SEQ ID NO: 75); NGMAX006398662 (SEQ ID NO: 76); NGMAX006398682 (SEQ ID NO: 77); NS0203176 (SEQ ID NO: 78); NGMAX006398725 (SEQ ID NO: 79); NGMAX006398726 (SEQ ID NO: 80); NGMAX006398800 (SEQ ID NO: 81); NGMAX006398923 (SEQ ID NO: 82); NGMAX006399042 (SEQ ID NO: 83); NGMAX006399063 (SEQ ID NO: 84); NGMAX006399066 (SEQ ID NO: 85); NGMAX006399168 (SEQ ID NO: 86); NGMAX006399482 (SEQ ID NO: 87); NGMAX006400296 (SEQ ID NO: 88); NGMAX006401034 (SEQ ID NO: 89); NGMAX006401429 (SEQ ID NO: 90); NGMAX006401765 (SEQ ID NO: 91); NGMAX008341472 (SEQ ID NO: 92); and NGMAX006401896 (SEQ ID NO: 93).

In certain embodiments of the invention, it is useful to detect in, or determine, whether a soybean plant has an allelic state that is associated with a low iron growth condition tolerant phenotype (Table 18). Certain non-limiting exemplary examples include identifying a soybean plant that has any one or more of: the CC allelic state of the polymorphic nucleic acid of NGMAX006398059_(SEQ ID NO: 71); the GG allelic state of the polymorphic nucleic acid of NGMAX006398075 (SEQ ID NO: 72); the AA allelic state of the polymorphic nucleic acid of NGMAX006398171 (SEQ ID NO: 73); the GG allelic state of the polymorphic nucleic acid of NGMAX006398585 (SEQ ID NO: 74); the CC allelic state of the polymorphic nucleic acid of NGMAX006398641 (SEQ ID NO: 75); the GG allelic state of the polymorphic nucleic acid of NGMAX006398662 (SEQ ID NO: 76); the CC allelic state of the polymorphic nucleic acid of NGMAX006398682 (SEQ ID NO: 77); the TT allelic state of the polymorphic nucleic acid of NS0203176 (SEQ ID NO: 78); the CC allelic state of the polymorphic nucleic acid of NGMAX006398725 (SEQ ID NO: 79); the GG allelic state of the polymorphic nucleic acid of NGMAX006398726 (SEQ ID NO: 80); the AA allelic state of the polymorphic nucleic acid of NGMAX006398800 (SEQ ID NO: 81); the CC allelic state of the polymorphic nucleic acid of NGMAX006398923 (SEQ ID NO: 82); the CC allelic state of the polymorphic nucleic acid of NGMAX006399042 (SEQ ID NO: 83); the AA allelic state of the polymorphic nucleic acid of NGMAX006399063 (SEQ ID NO: 84); the GG allelic state of the polymorphic nucleic acid of NGMAX006399066 (SEQ ID NO: 85); the CC allelic state of the polymorphic nucleic acid of NGMAX006399168 (SEQ ID NO: 86); the CC allelic state of the polymorphic nucleic acid of NGMAX006399482 (SEQ ID NO: 87); the CC allelic state of the polymorphic nucleic acid of NGMAX006400296 (SEQ ID NO: 88); the GG allelic state of the polymorphic nucleic acid of NGMAX006401034 (SEQ ID NO: 89); the GG allelic state of the polymorphic nucleic acid of NGMAX006401429 (SEQ ID NO: 90); the GG allelic state of the polymorphic nucleic acid of NGMAX006401765 (SEQ ID NO: 91); the CC allelic state of the polymorphic nucleic acid of NGMAX008341472 (SEQ ID NO: 92); and the GG allelic state of the polymorphic nucleic acid of NGMAX006401896 (SEQ ID NO: 93). One of skill in the art will also recognize that it can be useful to identify at a genetic locus a polymorphic nucleic acid marker that is not associated with a low iron growth condition tolerant phenotype (Table 18) in a plant or germplasm, such as when introgressing a QTL associated with a low iron growth tolerant phenotype into a genetic background not associated with such a phenotype.

Provided herein is a QTL on chromosome 7—that is flanked by loci BQ081048 and TA53357_3847—that is associated with a low iron growth condition tolerant phenotype. Sub-regions of chromosome 7 that flank QTL associated with a low iron growth condition tolerant phenotype include, but are not limited to sub-regions defined by any of the following sets of loci:

a) loci BQ081048 and NGMAX006626085;

b) loci BQ081048 and TC381430;

c) loci BQ081048 and BI785468;

d) loci Cf9876d and NGMAX006626085;

e) loci Cf9876d and TC381430;

f) loci Cf9876d and BI785468;

g) loci Cf9876d and TA53357_3847;

h) loci BARCSOYSSR_07_0184 and NGMAX006626085;

i) loci BARCSOYSSR_07_0184 and TC381430;

j) loci BARCSOYSSR_07_0184 and BI785468;

k) loci BARCSOYSSR_07_0184 and TA53357_3847;

l) loci NGMAX006621532 and NGMAX006626085;

m) loci NGMAX006621532 and TC381430;

n) loci NGMAX006621532 and BI785468;

o) loci NGMAX006621532 and TA53357_3847;

p) loci NGMAX006621610 and NGMAX006626085;

q) loci NGMAX006621610 and TC381430;

r) loci NGMAX006621610 and BI785468;

s) loci NGMAX006621610 and TA53357_3847;

t) loci NGMAX006621674 and NGMAX006626085;

u) loci NGMAX006621674 and TC381430;

v) loci NGMAX006621674 and BI785468; or

w) loci NGMAX006621674 and NGMAX006626085.

(Table 21).

These loci flank a genomic interval that spans telomere proximal nucleotide 3391037 to centromere proximal nucleotide 5460346 in the physical map of linkage group M provided in the Table 22. Polymorphic nucleic acid markers located in this genomic interval that are associated with a low iron growth condition tolerant phenotype can be detected with polymorphic nucleic acid markers that include, but are not limited to: NGMAX006621699 (SEQ ID NO: 97); NGMAX006621706 (SEQ ID NO: 98); NGMAX006621720 (SEQ ID NO: 99); NGMAX006621726 (SEQ ID NO: 100); NGMAX006621767 (SEQ ID NO: 101); NGMAX006621777 (SEQ ID NO: 102); NGMAX006622006 (SEQ ID NO: 103); NGMAX006622614 (SEQ ID NO: 104); NGMAX006622952 (SEQ ID NO: 105); NGMAX006623558 (SEQ ID NO: 106); NGMAX006623995 (SEQ ID NO: 107); NGMAX006624271 (SEQ ID NO: 108); and NGMAX006625883 (SEQ ID NO: 109).

›DESCRIPTION OF INVENTION · 10 of 17

Provided herein is a QTL on chromosome 17—that is flanked by loci BG726970 and Contig41076—that is associated with a low iron growth condition tolerant phenotype. Sub-regions of chromosome 17 that flank this first QTL associated with a low iron growth condition tolerant phenotype include, but are not limited to sub-regions defined by any of the following sets of loci:

a) loci BG726970 and BARCSOYSSR_17_0274;

b) loci BG726970 and 086788_3391_0381;

c) loci BARCSOYSSR_17_0169 and BARCSOYSSR_17_0274;

d) loci BARCSOYSSR_17_0169 and 086788_3391_0381; or

e) loci BARCSOYSSR_17_0169 and Contig41076.

(Table 24).

These loci flank a genomic interval that spans telomere proximal nucleotide 3146028 to centromere proximal nucleotide 8774159 in the physical map of linkage group D2 provided in the Table 25. Polymorphic nucleic acid markers located in this genomic interval that are associated with a low iron growth condition tolerant phenotype can be detected with polymorphic nucleic acid markers that include, but are not limited to: NGMAX007790014 (SEQ ID NO: 115); NGMAX007790381 (SEQ ID NO: 116); NGMAX007790411 (SEQ ID NO: 117); NGMAX007790440 (SEQ ID NO: 118); NS0263544 (SEQ ID NO: 119); NGMAX007790913 (SEQ ID NO: 120); NGMAX007790961 (SEQ ID NO: 121); NGMAX007790982 (SEQ ID NO: 122); and NGMAX007793044 (SEQ ID NO: 123).

Provided herein is a QTL on chromosome 19—that is flanked by loci Glyma19g01630 and TC27203—that is associated with a low iron growth condition tolerant phenotype. Sub-regions of chromosome 19 that flank QTL associated with a low iron growth condition tolerant phenotype include, but are not limited to sub-regions defined by any of the following sets of loci:

a) loci Glyma19g01630 and NGMAX008200913;

b) loci Glyma19g01630 and 354114_3517_2150;

c) loci Glyma19g01630 and TC409049;

d) loci Glyma19g01630 and Contig30217;

e) loci AI37921 and NGMAX008200913;

f) loci AI37921 and 354114_3517_2150;

g) loci AI37921 and TC409049;

h) loci AI37921 and Contig30217;

i) loci AI37921 and TC27203;

j) loci BARCSOYSSR_19_0083 and NGMAX008200913;

k) loci BARCSOYSSR_19_0083 and 354114_3517_2150;

l) loci BARCSOYSSR_19_0083 and TC409049;

m) loci BARCSOYSSR_19_0083 and Contig30217; or

n) loci BARCSOYSSR_19_0083 and TC27203.

(Table 27).

These loci flank a genomic interval that spans telomere proximal nucleotide 1239090 to centromere proximal nucleotide 1818955 in the physical map of linkage group L provided in the Table 28. Polymorphic nucleic acid markers located in this genomic interval that are associated with a low iron growth condition tolerant phenotype can be detected with polymorphic nucleic acid markers that include, but are not limited to: NGMAX008199658 (SEQ ID NO: 124), NGMAX008199750 (SEQ ID NO: 125), NGMAX008199864 (SEQ ID NO: 126), NGMAX008199973 (SEQ ID NO: 127), NGMAX008200015 (SEQ ID NO: 128); NGMAX008200022 (SEQ ID NO: 129); NGMAX008200044 (SEQ ID NO: 130); NGMAX008200052 (SEQ ID NO: 131); NGMAX008200275 (SEQ ID NO: 132); NGMAX008200335 (SEQ ID NO: 133); NGMAX008200568 (SEQ ID NO: 134); NGMAX008200652 (SEQ ID NO: 135); NGMAX008200656 (SEQ ID NO: 136); and NGMAX008200785 (SEQ ID NO: 137).

›DESCRIPTION OF INVENTION · 11 of 17

Provided herein is a QTL on chromosome 9—that is flanked by loci Cf4289d and Cf974d—that is associated with a low iron growth condition tolerant phenotype. Sub-regions of chromosome 9 that flank this first QTL associated with a low iron growth condition tolerant phenotype include, but are not limited to sub-regions defined by any of the following sets of loci:

a) loci Cf4289d and Cf5524d;

b) loci Cf4289d and Glyma09g34600;

c) loci Cf4289d and FE711739;

d) loci Cf9351d and Cf5524d;

e) loci Cf9351d and Glyma09g34600;

f) loci Cf9351d and FE711739;

g) loci Cf9351d and Cf974d;

h) loci BARCSOYSSR_09_1350 and Cf5524d;

i) loci BARCSOYSSR_09_1350 and Glyma09g34600;

j) loci BARCSOYSSR_09_1350 and FE711739; or

k) loci BARCSOYSSR_09_1350 and Cf974d.

(Table 30).

These loci flank a genomic interval that spans telomere proximal nucleotide 40236306 to centromere proximal nucleotide 40284995 in the physical map of linkage group K provided in the Table 31. Polymorphic nucleic acid markers located in this genomic interval that are associated with a low iron growth condition tolerant phenotype can be detected with polymorphic nucleic acid markers that include, but are not limited to: NGMAX006925911 (SEQ ID NO: 139), NGMAX006925967 (SEQ ID NO: 140), NGMAX006926063 (SEQ ID NO: 141), NGMAX006926228 (SEQ ID NO: 142), NGMAX006926426 (SEQ ID NO: 143); NGMAX006926451 (SEQ ID NO: 144); NGMAX006926709 (SEQ ID NO: 145); NGMAX006926784 (SEQ ID NO: 146); NGMAX006926934 (SEQ ID NO: 147); NGMAX006926989 (SEQ ID NO: 148); NGMAX006927044 (SEQ ID NO: 149); NGMAX006927083 (SEQ ID NO: 150); NGMAX006927293 (SEQ ID NO: 151); NGMAX006927454 (SEQ ID NO: 152); NGMAX006927578 (SEQ ID NO: 153); NS0094370 (SEQ ID NO: 154); NS0202727 (SEQ ID NO: 155); NGMAX006927737 (SEQ ID NO: 156); NGMAX006927783 (SEQ ID NO: 157); NGMAX006927836 (SEQ ID NO: 158); NGMAX006928046 (SEQ ID NO: 159); NS0123372 (SEQ ID NO: 160); NGMAX006928148 (SEQ ID NO: 161); NGMAX006928238 (SEQ ID NO: 162); NGMAX006928269 (SEQ ID NO: 163); NGMAX006928358 (SEQ ID NO: 164); NGMAX006928507 (SEQ ID NO: 165); NGMAX006928537 (SEQ ID NO: 166); NGMAX006928614 (SEQ ID NO: 167); NGMAX006928650 (SEQ ID NO: 168); NGMAX006928755 (SEQ ID NO: 169); NS0202939 (SEQ ID NO: 170); NS0119073 (SEQ ID NO: 171); NGMAX006928836 (SEQ ID NO: 172); NS0202984 (SEQ ID NO: 173); and NGMAX006928848 (SEQ ID NO: 174).

In certain embodiments of the invention, it is useful to detect in, or determine whether, a soybean plant has an allelic state that is associated with a low iron growth condition tolerant phenotype (Table 30). Certain non-limiting exemplary examples include identifying a soybean plant that has any one or more of: the GG allelic state of the polymorphic nucleic acid of NGMAX006926228 (SEQ ID NO: 142), the TT allelic state of the polymorphic nucleic acid of NGMAX006926426 (SEQ ID NO: 143); the CC allelic state of the polymorphic nucleic acid of NGMAX006926451 (SEQ ID NO: 144); the TT allelic state of the polymorphic nucleic acid of NGMAX006926709 (SEQ ID NO: 145); the GG allelic state of the polymorphic nucleic acid of NGMAX006926784 (SEQ ID NO: 146); the CC allelic state of the polymorphic nucleic acid of NGMAX006926934 (SEQ ID NO: 147); the TT allelic state of the polymorphic nucleic acid of NGMAX006926989 (SEQ ID NO: 148); the TT allelic state of the polymorphic nucleic acid of NGMAX006927044 (SEQ ID NO: 149); the GG allelic state of the polymorphic nucleic acid of NGMAX006927083 (SEQ ID NO: 150); the AA allelic state of the polymorphic nucleic acid of NGMAX006927393 (SEQ ID NO: 151); the GG allelic state of the polymorphic nucleic acid of NGMAX006927454 (SEQ ID NO: 152); the AA allelic state of the polymorphic nucleic acid of NGMAX006927578 (SEQ ID NO: 153); the CC allelic state of the polymorphic nucleic acid of NS0094370 (SEQ ID NO: 154); the CC allelic state of the polymorphic nucleic acid of NS0202727 (SEQ ID NO: 155); the CC allelic state of the polymorphic nucleic acid of NGMAX006927737 (SEQ ID NO: 156); the TT allelic state of the polymorphic nucleic acid of NGMAX006927783 (SEQ ID NO: 157); the GG allelic state of the polymorphic nucleic acid of NGMAX006927836 (SEQ ID NO: 158); the CC allelic state of the polymorphic nucleic acid of NGMAX006928046 (SEQ ID NO: 159); the TT allelic state of the polymorphic nucleic acid of NS0123372 (SEQ ID NO: 160); the CC allelic state of the polymorphic nucleic acid of NGMAX006928148 (SEQ ID NO: 161); the TT allelic state of the polymorphic nucleic acid of NGMAX006928238 (SEQ ID NO: 162); the CC allelic state of the polymorphic nucleic acid of NGMAX006928269 (SEQ ID NO: 163); the CC allelic state of the polymorphic nucleic acid of NGMAX006928358 (SEQ ID NO: 164); the CC allelic state of the polymorphic nucleic acid of NGMAX006928507 (SEQ ID NO: 165); the GG allelic state of the polymorphic nucleic acid of NGMAX006928537 (SEQ ID NO: 166); the TT allelic state of the polymorphic nucleic acid of NGMAX006928614 (SEQ ID NO: 167); the AA allelic state of the polymorphic nucleic acid of NGMAX006928650 (SEQ ID NO: 168); the CC allelic state of the polymorphic nucleic acid of NGMAX006928755 (SEQ ID NO: 169); the GG allelic state of the polymorphic nucleic acid of NS0202939 (SEQ ID NO: 170); the GG allelic state of the polymorphic nucleic acid of NS0119073 (SEQ ID NO: 171); the CC allelic state of the polymorphic nucleic acid of NGMAX006928836 (SEQ ID NO: 172); the GG allelic state of the polymorphic nucleic acid of NS0202984 (SEQ ID NO: 173); and the GG allelic state of the polymorphic nucleic acid of NGMAX006928848 (SEQ ID NO: 174). One of skill in the art will also recognize that it can be useful to identify at a genetic locus a polymorphic nucleic acid marker that is not associated with a low iron growth condition tolerant phenotype (Table 30) in a plant or germplasm, such as when introgressing a QTL associated with a low iron growth tolerant phenotype into a genetic background not associated with such a phenotype.

›DESCRIPTION OF INVENTION · 12 of 17

Additional genetic markers can be used either in conjunction with the polymorphic nucleic acid markers provided in any of the Tables herein or independently of such markers. Publicly available marker databases from which useful markers can be obtained include, but are not limited to, the soybase.org website on the internet (World Wide Web) that is administered by the United States Agricultural Research Service, the United States Department of Agriculture, and Iowa State University. Additional soybean markers that can be used and that have been described in the literature include, but are not limited to, Hyten et al., BMC Genomics. 11:38, 2010; Choi et al., Genetics. 176(1):685-96, 2007; Yoon et al., Theor Appl Genet. 2007 March; 114(5):885-99; and Hyten et al. Crop Sci. 2010 50: 960-968.

Given the provision herein of genomic regions, QTL, and polymorphic markers identified herein as well as an assortment of soybean germplasms and their decedents from which tolerance to low iron growth conditions has been observed, additional markers located either within or near this genomic region that are associated with these phenotypes can be obtained by merely typing the new markers in the various germplasms provided herewith. The genomic regions, QTL, and polymorphic markers identified herein can also be mapped relative to markers provided in any publicly available or other soybean physical or genetic map to place this genetic locus on that map.

IV. Identification of Plants Exhibiting Tolerance to Low Iron Growth Conditions

To observe the presence or absence of low iron growth condition tolerant phenotypes, soybean plants comprising genotypes of interest can be exposed to low iron or iron deficient growth conditions in seedling stages, early to mid-vegetative growth stages, or in early reproductive stages. Experienced plant breeders can recognize tolerant soybean plants in the field, and can select the tolerant individuals or populations for breeding purposes or for propagation. In this context, the plant breeder recognizes “tolerant” and “susceptible” soybean plants in fortuitous naturally-occurring filed observations.

Breeders will appreciate, however, that plant tolerance is a phenotypic spectrum consisting of extremes in tolerance, susceptibility, and a continuum of intermediate phenotypes. Tolerance also varies due to environmental effects. Evaluation of phenotypes using reproducible assays and tolerance scoring methods are of value to scientists who seek to identify genetic loci that impart tolerance, conduct marker assisted selection to create tolerant soybean populations, and for introgression techniques to breed a tolerance trait into an elite soybean line, for example.

In contrast to fortuitous field observations that classify plants as either “tolerant” or “susceptible,” various methods are known in the art for determining (and quantitating) the tolerance of a soybean plant to iron-deficient growth conditions. These techniques can be applied to different fields at different times, or to experimental greenhouse or laboratory settings, and provide approximate tolerance scores that can be used to characterize the tolerance of a given strain or line regardless of growth conditions or location. See, for example, Diers et al. (1992) “Possible identification of quantitative trait loci affecting iron efficiency in soybean,” J. Plant Nutr. 15:217-2136; Dahiya and M. Singh (1979) “Effect of salinity, alkalinity and iron sources on availability of iron,” Plant and Soil 51:13-18; and Gonzalez-Vallejo et al. (2000) “Iron Deficiency Decreases the Fe(III)-Chelate Reducing Activity of Leaf Protoplasts” Plant Physiol. 122 (2): 337-344.

The degree of IDC in a particular plant or stand of plants can be quantitated by using a system to score the severity of the disease in each plant. A plant strain or variety or a number of plant strains or varieties are planted and grown in a single stand in soil that is known to produce chlorotic plants as a result of iron deficiency (“field screens,” i.e., in filed that have previously demonstrated IDC), or alternatively, in controlled nursery conditions. When the assay is conducted in controlled nursery conditions, defined soils can be used, where the concentration of iron (e.g., available iron) has been previously measured. The plants can be scored at maturity, or at any time before maturity. The scoring system rates each plant on a scale of one (most susceptible—most severe disease) to nine (most tolerant—no disease), as shown in Table 11.

It will be appreciated that any such scale is relative, and furthermore, there may be variability between practitioners as to how the individual plants and the entire stand as a whole are scored. Optionally, the degree of chlorosis can be measured using a chlorophyll meter, e.g., a Minolta SPAD-502 Chlorophyll Meter, where readings off a single plant or a stand of plants can be made. Optionally, multiple readings can be obtained and averaged.

The IDC scoring of soybean stands can occur at any time. For example, plots can be scored in the early season, typically mid-July (depending on geographic latitude), so that the results can be used in making crossing decisions. Alternatively, soybean plots can be scored in the late season, which generally yields more precise data.

In general, while there is a certain amount of subjectivity to assigning severity measurements for disease symptoms, assignment to a given scale as noted above is well within the ordinary skill of a practitioner in the field. Measurements can also be averaged across multiple scores to reduce variation in field measurements.

Although protocols using field nurseries known to produce chlorotic plants can be used in assessing tolerance, it is typical for tolerance ratings to be based on actual field observations of fortuitous natural disease incidence, with the information corresponding to disease incidence for a cultivar being averaged over many locations and, typically, several season of crop plantings. Optionally, field stands or nursery/greenhouse plantings can be co-cultivated with IDC susceptibility “reference checks.” A reference check is a planting of soybean strains or varieties with known susceptibilities to low iron growth conditions, for example, highly tolerant strains and highly susceptible strains. This parallel planting can aid the breeder to compare the plant pathology in the experimental stands with the plant pathology in the reference stands.

›DESCRIPTION OF INVENTION · 13 of 17

When plants are studied in a fortuitous natural field setting, if there is no chlorosis present, the rating system in Table 11 cannot be used, because the existence of iron-deficient soil cannot be ascertained. However, if some number of plants demonstrate IDC symptoms, the growth conditions in that filed can be assumed to be iron-deficient, and the entire stand can be scored. These scores can be accumulated over multiple locations and years to show disease tolerance for given varieties or cultivars. Relative tolerance measurements between different strains in the same field at the same time can easily be made using the described or equivalent scoring system. Furthermore, the tolerance rating can be updated and refined each year based on the previous year's observation in the field.

V. Introgression of a Genomic Region Associated with a Low Iron Growth Condition Tolerance Phenotype

Provided herewith are unique soybean germplasms comprising one or more introgressed genomic regions, QTL, or polymorphic nucleic acid markers associated with a low iron growth condition tolerant phenotype and methods of obtaining the same. Marker-assisted introgression involves the transfer of a chromosomal region, defined by one or more markers, from one germplasm to a second germplasm. Offspring of a cross that contain the introgressed genomic region can be identified by the combination of markers characteristic of the desired introgressed genomic locus from a first germplasm (e.g., a low iron growth condition tolerant germplasm) and both linked and unlinked markers characteristic of the desired genetic background of a second germplasm (e.g., a low iron growth condition susceptible germplasm). In addition to the polymorphic nucleic acid markers provided herewith that identify alleles of certain QTL associated with a low iron growth condition tolerant phenotype, flanking markers that fall on both the telomere proximal end and the centromere proximal end of the genomic intervals comprising the QTL are also provided in Tables 1-10 and 14-31. Such flanking markers are useful in a variety of breeding efforts that include, but are not limited to, introgression of genomic regions associated with a low iron growth condition tolerant phenotype into a genetic background comprising markers associated with germplasm that ordinarily contains a genotype associated with a susceptible phenotype. Numerous markers that are linked and either immediately adjacent or adjacent to a low iron growth condition tolerant QTL in soybean that permit introgression of low iron growth condition tolerant QTL in the absence of extraneous linked DNA from the source germplasm containing the QTL are provided herewith. In certain embodiments, the linked and immediately adjacent markers are within about 105 kilobases (kB), 80 kB, 60 kB, 50 kB, 40 kB, 30 kB, 20 kB, 10 kB, 5 kB, 1 kB, 0.5 kB, 0.2 kB, or 0.1 kB of the introgressed genomic region. In certain embodiments, the linked and adjacent markers are within 1,000 kB, 600 kB, 500 kB, 400 kB, 300 kB, 200 kB, 150 kB of the introgressed genomic region. In certain embodiments, genomic regions comprising some or all of one or more of a low iron growth condition tolerant QTL described herein can be introgressed into the genomes of susceptible varieties by using markers that include, but are not limited to, adjacent markers and/or immediately adjacent markers provided in Tables 1-10 and 14-31. Those skilled in the art will appreciate that when seeking to introgress a smaller genomic region comprising a low iron growth condition tolerant QTL locus described herein, that any of the telomere proximal or centromere proximal markers that are immediately adjacent to a larger genomic region comprising a low iron growth condition tolerant QTL locus can also be used to introgress that smaller genomic region.

Provided herein are methods of introgressing any of the genomic regions comprising a low iron growth condition tolerance QTL locus of Tables 1-10 and 14-31 into soybean germplasm that lacks such a locus. In certain embodiments, the soybean germplasm that lacks such a genomic region comprising a low iron growth condition tolerance QTL locus of Tables 1-10 and 14-31 is susceptible or has less than optimal levels of tolerance to low iron growth conditions. In certain embodiments, the methods of introgression provided herein can yield soybean plants comprising introgressed genomic regions comprising one or more low iron growth condition tolerance QTL loci of Tables 1-10 and 14-31, where the immediately adjacent genomic DNA and/or some or all of the adjacent genomic DNA between the introgressed genomic region and the telomere or centromere will comprise allelic forms of the markers of Tables 1-10 and 14-31 that are characteristic of the germplasm into which the genomic region is introgressed and distinct from the germplasm from which the genomic region is derived. In certain embodiments, the soybean germplasm into which the genomic region is introgressed is germplasm that lacks such a low iron growth condition tolerance locus. In certain embodiments, the soybean germplasm into which the genomic region is introgressed is germplasm that lacks such a low iron growth condition tolerance locus and is either susceptible to low iron growth conditions or has less than optimal tolerance to low iron growth conditions.

Also provided herein are soybean plants produced by the aforementioned methods of introgression. In certain embodiments, such soybean plants will comprising introgressed genomic regions comprising a low iron growth condition tolerance QTL locus of Tables 1-10 and 14-31, where the immediately adjacent genomic DNA and/or some or all of the adjacent genomic DNA between the introgressed genomic region and the telomere or centromere will comprise allelic forms of the markers of Tables 1-10 that are characteristic of the germ plasm into which the genomic region is introgressed and distinct from the germplasm from which the genomic region is derived.

›DESCRIPTION OF INVENTION · 14 of 17

Soybean plants or germplasm comprising an introgressed genomic region that is associated with a low iron growth condition tolerant phenotype, wherein at least 10%, 25%, 50%, 75%, 90%, or 99% of the remaining genomic sequences carry markers characteristic of soybean plants or germplasm that are otherwise or ordinarily comprise a genomic region associated with susceptibility to low iron growth conditions, are thus provided. Furthermore soybean plants comprising an introgressed region where closely linked regions adjacent and/or immediately adjacent to the genomic regions, QTL, and markers provided herewith that comprise genomic sequences carrying markers characteristic of soybean plants or germplasm that are otherwise or ordinarily comprise a genomic region associated with the susceptibility to low iron growth conditions are also provided.

VI. Soybean Donor Plants Comprising Genomic Region Associated with Low Iron Growth Condition Phenotypes

Low iron growth condition tolerance QTL allele or alleles can be introduced from any plant that contains that allele (donor) to any recipient soybean plant. In one aspect, the recipient soybean plant can contain additional low iron growth condition tolerance loci. In another aspect, the recipient soybean plant can contain a transgene. In another aspect, while maintaining the introduced QTL, the genetic contribution of the plant providing the low iron growth condition tolerance QTL can be reduced by back-crossing or other suitable approaches. In one aspect, the nuclear genetic material derived from the donor material in the soybean plant can be less than or about 50%, less than or about 25%, less than or about 13%, less than or about 5%, 3%, 2% or 1%, but that genetic material contains the low iron growth condition tolerance locus or loci of interest

Plants containing one or more of the low iron growth condition tolerance loci described herein can be donor plants. In certain embodiments, a donor plant can be a susceptible line. In certain embodiments, a donor plant can also be a recipient soybean plant. A non-limiting and exemplary list of soybean varieties that are believed to comprise genomic regions associated with a low iron growth condition tolerance phenotype include, but are not limited to AG00501, AG00901, AG0131, AG0202, AG0231, AG0331, AG0401, AG801, AG0808, AG1031, AG1102, AG1230, AG2131, DKB22-52, AG3039, and AG3830 (Branded names of Asgrow (designated “AG”) and DELAKB soybean varieties from Monsanto CO. 800 N. Lindbergh Blvd., St. Louis, Mo., USA.)

In a preferred embodiment, the donor soybean plant is AG801 and derivatives thereof, and is used as the resistant parent in a bi-parental mapping population to select for genomic regions associated with a low iron growth condition tolerance phenotype.

Also provided herewith are additional soybean plants that comprise a genomic region associated with a low iron growth condition tolerance phenotype that are identified by use of the markers provided in Tables 1-10 and 14-31 and/or methods provided herein. Any of the soybean plants identified above or other soybean plants that are otherwise identified using the markers or methods provided herein can be used in methods that include, but are not limited to, methods of obtaining soybean plants with an introgressed low iron growth condition tolerance locus, obtaining a soybean plant that exhibits a low iron growth condition tolerance phenotype, or obtaining a soybean plant comprising in its genome a genetic region associated with a low iron growth condition tolerance phenotype.

In certain embodiments, the soybean plants provided herein or used in the methods provided herein can comprise a transgene that confers tolerance to glyphosate. Transgenes that can confer tolerance to glyphosate include, but are not limited to, transgenes that encode glyphosate tolerant Class I EPSPS (5-enolpyruvylshikimate-3-phosphate synthases) enzymes or glyphosate tolerant Class II EPSPS (5-enolpyruvylshikimate-3-phosphate synthases) enzymes. Useful glyphosate tolerant EPSPS enzymes provided herein are disclosed in U.S. Pat. No. 6,803,501, RE39,247, U.S. Pat. Nos. 6,225,114, 5,188,642, and 4,971,908. In certain embodiments, the glyphosate tolerant soybean plants can comprise a transgene encoding a glyphosate oxidoreductase or other enzyme which degrades glyphosate. Glyphosate oxidoreductase enzymes had been described in U.S. Pat. No. 5,776,760 and U.S. Reissue Pat. RE38,825. In certain embodiments the soybean plant can comprise a transgene encoding a glyphosate N-acetyltransferase gene that confers tolerance to glyphosate. In certain embodiments, the soybean plant can comprise a glyphosate n-acetyltransferase encoding transgene such as those described in U.S. Pat. No. 7,666,644. In still other embodiments, soybean plants comprising combinations of transgenes that confer glyphosate tolerance are provided. Soybean plants comprising both a glyphosate resistant EPSPS and a glyphosate N-acetyltransferase are also provided herewith. In certain embodiments, it is contemplated that the soybean plants used herein can comprise one or more specific genomic insertion(s) of a glyphosate tolerant transgene including, but not limited to, as those found in: i) MON89788 soybean (deposited under ATCC accession number PTA-6708 and described in US Patent Application Publication Number 20100099859), ii) GTS 40-3-2 soybean (Padgette et al., Crop Sci. 35: 1451-1461, 1995), iii) event 3560.4.3.5 soybean (seed deposited under ATCC accession number PTA-8287 and described in US Patent Publication 20090036308), or any combination of i (MON89788 soybean), ii (GTS 40-3-2 soybean), and iii (event 3560.4.3.5 soybean).

A low iron growth condition tolerance associated QTL of the present invention may also be introduced into an soybean line comprising one or more transgenes that confer tolerance to herbicides including, but not limited to, glufosinate, dicamba, chlorsulfuron, and the like, increased yield, insect control, fungal disease resistance, virus resistance, nematode resistance, bacterial disease resistance, mycoplasma disease resistance, modified oils production, high oil production, high protein production, germination and seedling growth control, enhanced animal and human nutrition, low raffinose, environmental stress resistant, increased digestibility, industrial enzymes, pharmaceutical proteins, peptides and small molecules, improved processing traits, improved flavor, nitrogen fixation, hybrid seed production, reduced allergenicity, biopolymers, and biofuels among others. These agronomic traits can be provided by the methods of plant biotechnology as transgenes in soybean.

›DESCRIPTION OF INVENTION · 15 of 17

In certain embodiments, it is contemplated that genotypic assays that provide for non-destructive identification of the plant or plants can be performed either in seed, the emergence stage, the “VC” stage (i.e. cotyledons unfolded), the V1 stage (appearance of first node and unifoliate leaves), the V2 stage (appearance of the first trifoliate leaf), and thereafter. In certain embodiments, non-destructive genotypic assays are performed in seed using apparati and associated methods as described in U.S. Pat. Nos. 6,959,617; 7,134,351; 7,454,989; 7,502,113; 7,591,101; 7,611,842; and 7,685,768, which are incorporated herein by reference in their entireties. In certain embodiments, non-destructive genotypic assays are performed in seed using apparati and associated methods as described in US Patent Application Publications 20100086963, 20090215060, and 20090025288, which are incorporated herein by reference in their entireties. Published U.S. Patent Applications US 2006/0042527, US 2006/0046244, US 2006/0046264, US 2006/0048247, US 2006/0048248, US 2007/0204366, and US 2007/0207485, which are incorporated herein by reference in their entirety, also disclose apparatus and systems for the automated sampling of seeds as well as methods of sampling, testing and bulking seeds. Thus, in a certain embodiments, any of the methods provided herein can comprise screening for markers in individual seeds of a population wherein only seed with at least one genotype of interest is advanced.

VII. Molecular Assisted Breeding Techniques

Genetic markers that can be used in the practice of the instant invention include, but are not limited to, are Restriction Fragment Length Polymorphisms (RFLP), Amplified Fragment Length Polymorphisms (AFLP), Simple Sequence Repeats (SSR), Single Nucleotide Polymorphisms (SNP), Insertion/Deletion Polymorphisms (Indels), Variable Number Tandem Repeats (VNTR), and Random Amplified Polymorphic DNA (RAPD), and others known to those skilled in the art. Marker discovery and development in crops provides the initial framework for applications to marker-assisted breeding activities (US Patent Applications 2005/0204780, 2005/0216545, 2005/0218305, and 2006/00504538). The resulting “genetic map” is the representation of the relative position of characterized loci (DNA markers or any other locus for which alleles can be identified) along the chromosomes. The measure of distance on this map is relative to the frequency of crossover events between sister chromatids at meiosis.

As a set, polymorphic markers serve as a useful tool for fingerprinting plants to inform the degree of identity of lines or varieties (U.S. Pat. No. 6,207,367). These markers can form abasis for determining associations with phenotype and can be used to drive genetic gain. The implementation of marker-assisted selection is dependent on the ability to detect underlying genetic differences between individuals.

Certain genetic markers for use in the present invention include “dominant” or “codominant” markers. “Codominant markers” reveal the presence of two or more alleles (two per diploid individual). “Dominant markers” reveal the presence of only a single allele. The presence of the dominant marker phenotype (e.g., a band of DNA) is an indication that one allele is present in either the homozygous or heterozygous condition. The absence of the dominant marker phenotype (e.g., absence of a DNA band) is merely evidence that “some other” undefined allele is present. In the case of populations where individuals are predominantly homozygous and loci are predominantly dimorphic, dominant and codominant markers can be equally valuable. As populations become more heterozygous and multiallelic, codominant markers often become more informative of the genotype than dominant markers.

In another embodiment, markers that include. but are not limited, to single sequence repeat markers (SSR), AFLP markers, RFLP markers, RAPD markers, phenotypic markers, isozyme markers, single nucleotide polymorphisms (SNPs), insertions or deletions (Indels), single feature polymorphisms (SFPs, for example, as described in Borevitz et al. 2003 Gen. Res. 13:513-523), microarray transcription profiles, DNA-derived sequences, and RNA-derived sequences that are genetically linked to or correlated with low iron growth condition tolerance loci, regions flanking low iron growth condition tolerance loci, regions linked to low iron growth condition tolerance loci, and/or regions that are unlinked to low iron growth condition tolerance loci can be used in certain embodiments of the instant invention.

In one embodiment, nucleic acid-based analyses for determining the presence or absence of the genetic polymorphism (i.e. for genotyping) can be used for the selection of seeds in a breeding population. A wide variety of genetic markers for the analysis of genetic polymorphisms are available and known to those of skill in the art. The analysis may be used to select for genes, portions of genes, QTL, alleles, or genomic regions (Genotypes) that comprise or are linked to a genetic marker that is linked to or correlated with low iron growth condition tolerance loci, regions flanking low iron growth condition tolerance loci, regions linked to low iron growth condition tolerance loci, and/or regions that are unlinked to low iron growth condition tolerance loci can be used in certain embodiments of the instant invention.

Herein, nucleic acid analysis methods include, but are not limited to, PCR-based detection methods (for example, TaqMan assays), microarray methods, mass spectrometry-based methods and/or nucleic acid sequencing methods. In one embodiment, the detection of polymorphic sites in a sample of DNA, RNA, or cDNA may be facilitated through the use of nucleic acid amplification methods. Such methods specifically increase the concentration of polynucleotides that span the polymorphic site, or include that site and sequences located either distal or proximal to it. Such amplified molecules can be readily detected by gel electrophoresis, fluorescence detection methods, or other means.

›DESCRIPTION OF INVENTION · 16 of 17

A method of achieving such amplification employs the polymerase chain reaction (PCR) (Mullis et al. 1986 Cold Spring Harbor Symp. Quant. Biol. 51:263-273; European Patent 50,424; European Patent 84,796; European Patent 258,017; European Patent 237,362; European Patent 201,184; U.S. Pat. Nos. 4,683,202; 4,582,788; and 4,683,194), using primer pairs that are capable of hybridizing to the proximal sequences that define a polymorphism in its double-stranded form.

Methods for typing DNA based on mass spectrometry can also be used. Such methods are disclosed in U.S. Pat. Nos. 6,613,509 and 6,503,710, and references found therein.

Polymorphisms in DNA sequences can be detected or typed by a variety of effective methods well known in the art including, but not limited to, those disclosed in U.S. Pat. Nos. 5,468,613, 5,217,863; 5,210,015; 5,876,930; 6,030,787; 6,004,744; 6,013,431; 5,595,890; 5,762,876; 5,945,283; 5,468,613; 6,090,558; 5,800,944; 5,616,464; 7,312,039; 7,238,476; 7,297,485; 7,282,355; 7,270,981 and 7,250,252 all of which are incorporated herein by reference in their entireties. However, the compositions and methods of the present invention can be used in conjunction with any polymorphism typing method to type polymorphisms in genomic DNA samples. These genomic DNA samples used include but are not limited to genomic DNA isolated directly from a plant, cloned genomic DNA, or amplified genomic DNA.

For instance, polymorphisms in DNA sequences can be detected by hybridization to allele-specific oligonucleotide (ASO) probes as disclosed in U.S. Pat. Nos. 5,468,613 and 5,217,863. U.S. Pat. No. 5,468,613 discloses allele specific oligonucleotide hybridizations where single or multiple nucleotide variations in nucleic acid sequence can be detected in nucleic acids by a process in which the sequence containing the nucleotide variation is amplified, spotted on a membrane and treated with a labeled sequence-specific oligonucleotide probe.

Target nucleic acid sequence can also be detected by probe ligation methods as disclosed in U.S. Pat. No. 5,800,944 where sequence of interest is amplified and hybridized to probes followed by ligation to detect a labeled part of the probe.

Microarrays can also be used for polymorphism detection, wherein oligonucleotide probe sets are assembled in an overlapping fashion to represent a single sequence such that a difference in the target sequence at one point would result in partial probe hybridization (Borevitz et al., Genome Res. 13:513-523 (2003); Cui et al., Bioinformatics 21:3852-3858 (2005). On any one microarray, it is expected there will be a plurality of target sequences, which may represent genes and/or noncoding regions wherein each target sequence is represented by a series of overlapping oligonucleotides, rather than by a single probe. This platform provides for high throughput screening a plurality of polymorphisms. A single-feature polymorphism (SFP) is a polymorphism detected by a single probe in an oligonucleotide array, wherein a feature is a probe in the array. Typing of target sequences by microarray-based methods is disclosed in U.S. Pat. Nos. 6,799,122; 6,913,879; and 6,996,476.

Target nucleic acid sequence can also be detected by probe linking methods as disclosed in U.S. Pat. No. 5,616,464, employing at least one pair of probes having sequences homologous to adjacent portions of the target nucleic acid sequence and having side chains which non-covalently bind to form a stem upon base pairing of the probes to the target nucleic acid sequence. At least one of the side chains has a photoactivatable group which can form a covalent cross-link with the other side chain member of the stem.

Other methods for detecting SNPs and Indels include single base extension (SBE) methods. Examples of SBE methods include, but are not limited, to those disclosed in U.S. Pat. Nos. 6,004,744; 6,013,431; 5,595,890; 5,762,876; and 5,945,283. SBE methods are based on extension of a nucleotide primer that is adjacent to a polymorphism to incorporate a detectable nucleotide residue upon extension of the primer. In certain embodiments, the SBE method uses three synthetic oligonucleotides. Two of the oligonucleotides serve as PCR primers and are complementary to sequence of the locus of genomic DNA which flanks a region containing the polymorphism to be assayed. Following amplification of the region of the genome containing the polymorphism, the PCR product is mixed with the third oligonucleotide (called an extension primer) which is designed to hybridize to the amplified DNA adjacent to the polymorphism in the presence of DNA polymerase and two differentially labeled dideoxynucleosidetriphosphates. If the polymorphism is present on the template, one of the labeled dideoxynucleosidetriphosphates can be added to the primer in a single base chain extension. The allele present is then inferred by determining which of the two differential labels was added to the extension primer. Homozygous samples will result in only one of the two labeled bases being incorporated and thus only one of the two labels will be detected. Heterozygous samples have both alleles present, and will thus direct incorporation of both labels (into different molecules of the extension primer) and thus both labels will be detected.

In another method for detecting polymorphisms, SNPs and Indels can be detected by methods disclosed in U.S. Pat. Nos. 5,210,015; 5,876,930; and 6,030,787 in which an oligonucleotide probe having a 5′ fluorescent reporter dye and a 3′ quencher dye covalently linked to the 5′ and 3′ ends of the probe. When the probe is intact, the proximity of the reporter dye to the quencher dye results in the suppression of the reporter dye fluorescence, e.g. by Forster-type energy transfer. During PCR forward and reverse primers hybridize to a specific sequence of the target DNA flanking a polymorphism while the hybridization probe hybridizes to polymorphism-containing sequence within the amplified PCR product. In the subsequent PCR cycle DNA polymerase with 5′→3′ exonuclease activity cleaves the probe and separates the reporter dye from the quencher dye resulting in increased fluorescence of the reporter.

›DESCRIPTION OF INVENTION · 17 of 17

In another embodiment, the locus or loci of interest can be directly sequenced using nucleic acid sequencing technologies. Methods for nucleic acid sequencing are known in the art and include technologies provided by 454 Life Sciences (Branford, Conn.), Agencourt Bioscience (Beverly, Mass.), Applied Biosystems (Foster City, Calif.), LI-COR Biosciences (Lincoln, Nebr.), NimbleGen Systems (Madison, Wis.), Illumina (San Diego, Calif.), and VisiGen Biotechnologies (Houston, Tex.). Such nucleic acid sequencing technologies comprise formats such as parallel bead arrays, sequencing by ligation, capillary electrophoresis, electronic microchips, “biochips,” microarrays, parallel microchips, and single-molecule arrays, as reviewed by R.F. Service Science 2006 311:1544-1546.

The markers to be used in the methods of the present invention should preferably be diagnostic of origin in order for inferences to be made about subsequent populations. Experience to date suggests that SNP markers may be ideal for mapping because the likelihood that a particular SNP allele is derived from independent origins in the extant populations of a particular species is very low. As such, SNP markers appear to be useful for tracking and assisting introgression of QTLs, particularly in the case of Genotypes.

APPENDIX TO THE SPECIFICATION

›Tables in the description — 21

TABLE 1 — Chromosome 3 - QTL on chromosome 3 associated with a low iron growth condition tolerant phenotype.

			Allelic	Allelic		Identified
			Form(s)	Form(s) Not-		Within
		cM Map	Associated	Associated		Region
	SEQ	Position on	With Low Fe	with Low Fe		Associated
	ID	Chromosome	Tolerance	Tolerance	Polymorphic	With Low Fe
Marker or Locus Name	NO	Three (3)	Phenotype 1	Phenotype 1	Position	Tolerance
Glyma03g34510	—	—	—	—	—	—
Gm_W82_CR03.G378360	—	—	—	—	—	—
Glyma03g34760	—	—	—	—	—	—
Glyma03g34860	—	—	—	—	—	—
NS0202712
1	122.3	AA	TT	218	***
Glyma03g35130	—	—	—	—	—	***
Glyma03g35280	—	—	—	—	—	***
NS0129403
2	130.7	GG	AA	83	***
NS0115624
3	131	TT	AA	582	***
Glyma03g36800	—	—	—	—	—	***
Glyma03g38030	—	—	—	—	—	***
NS0202842
4	141	CC	TT	145	***
NS0122122
5	141.3	CC	TT	271	***
NS0205984
6	141.9	GG	AA	244	***
Pvcon3607	—	—	—	—	—	***
Glyma03g39610	—	—	—	—	—	***
FE710890	—	—	—	—	—	***
NS0202698
7	149.8	GG	AA	421	***
Glyma03g41830	—	—	—	—	—	—
Glyma03g42250	—	—	—	—	—	—

TABLE 2 — Chromosome 3 - Physical positions of certain genetic markers on soybean chromosome 3 in proximity to QTL associated with a low iron growth condition tolerant phenotype.

	SEQ			Middle
Marker or Locus Name	ID NO	LG	Chromosome	Position	Start	Stop
Glyma03g34510	—	N	3	41915727	41914204	41917250
Gm_W82_CR03.G378360	—	N	3	41915733	41914204	41917263
Glyma03g34760	—	N	3	42086656	42085228	42088084
Glyma03g34860	—	N	3	42164646	42160939	42168354
NS0202712	1	N	3	42243449	42243249	42243649
Glyma03g35130	—	N	3	42445457	42444310	42446604
Glyma03g35280	—	N	3	42553329	42551024	42555635
NS0129403	2	N	3	43515346	43515925	43514768
NS0115624	3	N	3	43561673	43562124	43561223
Glyma03g36800	—	N	3	43651286	43650506	43652066
Glyma03g38030	—	N	3	44472415	44469992	44474838
NS0202842	4	N	3	45075121	45075320	45074923
NS0122122	5	N	3	45114092	45113696	45114489
NS0205984	6	N	3	45207137	45207317	45206958
Pvcon3607	—	N	3	45618970	45618216	45619724
Glyma03g39610	—	N	3	45621485	45618205	45624765
FE710890	—	N	3	45622786	45620962	45624610
NS0202698	7	N	3	46401175	46400931	46401420
Glyma03g41830	—	N	3	47201754	47200244	47203264
Glyma03g42250	—	N	3	47498375	47496922	47499828

TABLE 3 — Chromosome 3-Marker type and nucleic acid primers and probes useful in the detection of certain alleles of polymorphic nucleic acid markers associated with an iron deficiency growth condition tolerant phenotype. SEQ

Marker or	ID	Marker
Locus Name	NO	Type	Sequence
NS0129403	2	TAQMAN	—
Forward	8	—	GCGGATAGCCGGATAGCAT
Primer
Reverse	9	—	GCATACAACTTAGTTTTATATACATAT
Primer			ATAGTGTATATATATCA
Vic Probe	10	—	CATGTAACAGAGGTCA
Fam Probe	11	—	ATGTAGCAGAGGTCA
NS0115624	3	TAQMAN	—
Forward	12	—	GAGTCAAAGTAGGAATCTTTTCGCATA
Primer
Reverse	13	—	TTACTTTGTCCTCAGGGATGAACA
Primer
Vic Probe	14	—	TTTGCAAGTAGCGAGC
Fam Probe	15	—	TTGCATGTAGCGAGC
NS0122122	5	TAQMAN	—
Forward	16	—	GGTTCCGTCTGACAGTATTTTTGG
Primer
Reverse	17	—	ATGGAGAAAAGAACAGAGGTGGTT
Primer
Vic Probe	18	—	CCCAACCACCATTAT
Fam Probe	19	—	AACACCCAACTACC

TABLE 4 — Chromosome 19 - QTL on chromosome 19 associated with a low iron growth condition tolerant phenotype.

			Allelic	Allelic		Identified
		cM Map	Form(s)	Form(s) Not-		Within
		Position on	Associated	Associated		Region
	SEQ	chromosome	With Low Fe	with Low Fe		Associated
	ID	nineteen	Tolerance	Tolerance	Polymorphic	With Low Fe
Marker or Locus Name	NO	(19)	Phenotype 1	Phenotype 1	Position	Tolerance
Glyma19g32880	—	—	—	—	—	—
Glyma19g33540	—	—	—	—	—	—
NS0206298	20	107.5	AA	CC	1044	***
NS0205620	21	110.9	GG	CC	275	***
NS0206055	22	111.9	AA	GG	530	***
Glyma19g34480	—	—	—	—	—	***
NS0204985	23	114.1	CC	TT	619	***
Gm_W82_CR19.G249620	—	—	—	—	—	—
Glyma19g34740	—	—	—	—	—	—
NGMAX005710149	—	—	—	—	—	—
220298_3433_1666	—	—	—	—	—	—
Contig13742	—	—	—	—	—	—
BARCSOYSSR_19_1192	—	—	—	—	—	—
Glyma19g34750	—	—	—	—	—	—
Gm_W82_CR19.G249630	—	—	—	—	—	—
Contig9146	—	—	—	—	—	—

TABLE 5 — Chromosome 19 - Physical positions of certain genetic markers on soybean chromosome 19 in proximity to QTL associated with a low iron growth condition tolerant phenotype.

	SEQ			Middle
Marker or Locus Name	ID NO	LG	Chromosome	Position	Start	Stop
Glyma19g32880	—	L	19	40563828	40562661	40564996
Glyma19g33540	—	L	19	41122500	41119227	41125773
NS0206298	20	L	19	41609816	41609250	41610382
NS0205620	21	L	19	42012550	42012728	42012372
NS0206055	22	L	19	42018389	42018856	42017922
Glyma19g34480	—	L	19	42073448	42072643	42074253
NS0204985	23	L	19	42345212	42344620	42345805
Gm_W82_CR19.G249620	—	L	19	42345294	42344177	42346412
Glyma19g34740	—	L	19	42345351	42344291	42346412
NGMAX005710149	—	L	19	42345386	42345236	42345537
220298_3433_1666	—	L	19	42345865	42345741	42345989
Contig13742	—	L	19	42346076	42345967	42346185
BARCSOYSSR_19_1192	—	L	19	42347717	42347702	42347733
Glyma19g34750	—	L	19	42351644	42350923	42352365
Gm_W82_CR19.G249630	—	L	19	42351644	42350923	42352365
Contig9146	—	L	19	42351663	42351579	42351748

TABLE 6 — Chromosome 10 - QTL on chromosome 10 associated with a low iron growth condition tolerant phenotype.

			Allelic	Allelic		Identified
			Form(s)	Form(s) Not-		Within
		cM Map	Associated	Associated		Region
	SEQ	Position on	With Low Fe	with Low Fe		Associated
	ID	chromosome	Tolerance	Tolerance	Polymorphic	With Low Fe
Marker or Locus Name	NO	ten (10)	Phenotype 1	Phenotype 1	Position	Tolerance
NS0116559	—	—	—	—	—	—
Glyma10g33710	—	—	—	—	—	—
TA42233_3847	—	—	—	—	—	—
AF108084.1	—	—	—	—	—	—
AW734581	—	—	—	—	—	—
Glyma10g34260	—	—	—	—	—	—
Glyma10g34280	—	—	—	—	—	—
Glyma10g34290	—	—	—	—	—	—
Glyma10g34460	—	—	—	—	—	—
Glyma10g34600	—	—	—	—	—	—
Glyma10g34630	—	—	—	—	—	—
Glyma10g34850	—	—	—	—	—	—
NS0120070
24	148.6	TT	CC	468	***
NS0097952
25	148.6	GG	AA	420	***
NS0092960
26	149.8	TT	AA	89	***
Glyma10g36370	—	—	—	—	—	***
NS0118907
27	154.2	AA	CC	450	***
Glyma10g37560	—	—	—	—	—	—
TC354083	—	—	—	—	—	—
Glyma10g37600	—	—	—	—	—	—
BW595896	—	—	—	—	—	—
TC130824	—	—	—	—	—	—
CV528982	—	—	—	—	—	—
Glyma10g37610	—	—	—	—	—	—
BG511181	—	—	—	—	—	—
TA68568_3847	—	—	—	—	—	—
NS0204740
28	162.1	AA	CC	424	***
NS0205036
29	163.5	AA	TT	33	***
Glyma10g37910	—	—	—	—	—	***
Glyma10g37920	—	—	—	—	—	***
Gm_W82_CR10.G221330	—	—	—	—	—	***
NS0206252
30	165.8	CC	TT	100	***
Glyma10g38600	—	—	—	—	—	—
Glyma10g40520	—	—	—	—	—	—
BF631841	—	—	—	—	—	—
Glyma10g41460	—	—	—	—	—	—
Glyma10g41470	—	—	—	—	—	—
TC412490	—	—	—	—	—	—
Gm_W82_CR10.G236520	—	—	—	—	—	—

TABLE 7 — Chromosome 10 - Physical positions of certain genetic markers on soybean chromosome 10 in proximity to QTL associated with a low iron growth condition tolerant phenotype.

	SEQ			Middle
Marker or Locus Name	ID NO	LG	Chromosome	Position	Start	Stop
NS0116559	—	O	10	42075575	42075167	42075983
Glyma10g33710	—	O	10	42593023	42590873	42595174
TA42233_3847	—	O	10	42593038	42590903	42595173
AF108084.1	—	O	10	42593043	42590914	42595173
AW734581	—	O	10	42593522	42591892	42595153
Glyma10g34260	—	O	10	43021758	43021192	43022324
Glyma10g34280	—	O	10	43030489	43029802	43031177
Glyma10g34290	—	O	10	43035456	43034563	43036349
Glyma10g34460	—	O	10	43204121	43201964	43206279
Glyma10g34600	—	O	10	43366015	43361916	43370114
Glyma10g34630	—	O	10	43409208	43408273	43410144
Glyma10g34850	—	O	10	43607308	43606350	43608266
NS0120070
24	O	10	44567643	44567905	44567382
NS0097952
25	O	10	44571243	44571555	44570932
NS0092960
26	O	10	44742555	44742322	44742789
Glyma10g36370	—	O	10	45149778	45147589	45151968
NS0118907
27	O	10	45409273	45409660	45408887
Glyma10g37560	—	O	10	46113736	46112579	46114893
TC354083	—	O	10	46143267	46141283	46145251
Glyma10g37600	—	O	10	46143977	46141275	46146680
BW595896	—	O	10	46144779	46144294	46145265
TC130824	—	O	10	46145332	46144251	46146413
CV528982	—	O	10	46154071	46153277	46154865
Glyma10g37610	—	O	10	46156146	46154486	46157806
BG511181	—	O	10	46156618	46156462	46156775
TA68568_3847	—	O	10	46157451	46157096	46157806
NS0204740
28	O	10	46254426	46254082	46254770
NS0205036
29	O	10	46276912	46276995	46276829
Glyma10g37910	—	O	10	46405425	46403762	46407089
Glyma10g37920	—	O	10	46425433	46424402	46426465
Gm_W82_CR10.G221330	—	O	10	46735180	46732478	46737883
NS0206252
30	O	10	46749499	46749827	46749172
Glyma10g38600	—	O	10	46983790	46982907	46984673
Glyma10g40520	—	O	10	48498769	48497366	48500173
BF631841	—	O	10	48536884	48536690	48537079
Glyma10g41460	—	O	10	49116840	49114600	49119081
Glyma10g41470	—	O	10	49123712	49121296	49126128
TC412490	—	O	10	49124228	49122322	49126134
Gm_W82_CR10.G236520	—	O	10	49715508	49712499	49718518

TABLE 8 — Chromosome 10-Marker type and nucleic acid primer and probes useful in the detection of certain alleles of polymorphic nucleic acid markers associated with an iron deficiency growth condition tolerant phenotype. SEQ

Marker or	ID	Marker
Locus Name	NO	Type	Sequence
NS0120070	24	TAQMAN	—
Forward	31	—	AATATCCACGGTATGCTGTTTGAAT
Primer
Reverse	32	—	CTCGTTCTAATGGAAAAGCGAATC
Primer
Vic Probe	33	—	CTTGATAGCTGCCATTT
Fam Probe	34	—	AGCTGTCATTTCTT
NS0097952	25	TAQMAN	—
Forward	35	—	CGAAGCATTACACTATTTTCTGTCAAA
Primer
Reverse	36	—	AAAAAATCACATGATACGAGAAAAGA
Primer			TCT
Vic Probe	37	—	CAATTGAACCATTTCG
Fam Probe	38	—	TTGAACCGTTTCGAGC
NS0092960	26	TAQMAN	—
Forward	39	—	CCATTCTGAAGATGAGGCATTG
Primer
Reverse	40	—	ACAAGAGTTTGTAGGAAAACATGATGTT
Primer
Vic Probe	41	—	TCCATTGGGTACTCC
Fam Probe	42	—	CATTGGGAACTCC
NS0118907	27	TAQMAN	—
Forward	43	—	CCTTTCAAAACCTTTAAGGCATGTA
Primer
Reverse	44	—	GTTCCTAGCCAACAATGAGTTTCTC
Primer
Vic Probe	45	—	AGCTCCAACATATGAT
Fam Probe	46	—	CTCCAACCTATGATTG

TABLE 9 — Chromosome 7 - QTL on chromosome 7 associated with a low iron growth condition tolerant phenotype.

			Allelic	Allelic		Identified
			Form(s)	Form(s) Not-		Within
		cM Map	Associated	Associated		Region
		Position on	With Low Fe	with Low Fe		Associated
	SEQ	chromosome	Tolerance	Tolerance	Polymorphic	With Low Fe
Marker or Locus Name	ID NO	seven (7)	Phenotype 1	Phenotype 1	Position	Tolerance
Gm_W82_CR07.G4950	—	—	—	—	—	—
TC112538	—	—	—	—	—	—
Glyma07g03790	—	—	—	—	—	—
Gm_W82_CR07.G8460	—	—	—	—	—	—
Glyma07g03800	—	—	—	—	—	—
Glyma07g03810	—	—	—	—	—	—
NS0202654
47	27.6	TT	GG	117	***
NS0206351
48	27.7	CC	GG	131	***
Glyma07g04470	—	—	—	—	—	***
Glyma07g04840	—	—	—	—	—	***
NS0202966
49	36.2	TT	GG	180	***
Glyma07g05420	—	—	—	—	—	—
Glyma07g05820	—	—	—	—	—	—
NS0102362	—	—	—	—	—	—
Glyma07g06150	—	—	—	—	—	—
Glyma07g06510	—	—	—	—	—	—
Glyma07g06560	—	—	—	—	—	—
Glyma07g07380	—	—	—	—	—	—
Gm_W82_CR07.G30600	—	—	—	—	—	—
Pvcon9217	—	—	—	—	—	—
TC119399	—	—	—	—	—	—
Glyma07g07560	—	—	—	—	—	—
TC385708	—	—	—	—	—	—
Glyma07g08950	—	—	—	—	—	—
Glyma07g09110	—	—	—	—	—	—
Glyma07g09150	—	—	—	—	—	—
Glyma07g09160	—	—	—	—	—	—
Glyma07g09170	—	—	—	—	—	—
Glyma07g09900	—	—	—	—	—	—
Glyma07g09960	—	—	—	—	—	—
Glyma07g09970	—	—	—	—	—	—
NS0119842	—	—	—	—	—	—
TA57919_3847	—	—	—	—	—	—
Glyma07g11180	—	—	—	—	—	—
Glyma07g12210	—	—	—	—	—	—
Glyma07g29640	—	—	—	—	—	—
Glyma07g29650	—	—	—	—	—	—
Glyma07g29940	—	—	—	—	—	—
Glyma07g31380	—	—	—	—	—	—
Glyma07g31390	—	—	—	—	—	—
NS0202730
50	129.3	AA	GG	212	***
NS0206314
51	129.5	CC	TT	44	***
Glyma07g31610	—	—	—	—	—	—
Glyma07g32330	—	—	—	—	—	—
Glyma07g33070	—	—	—	—	—	—
Glyma07g33090	—	—	—	—	—	—
Glyma07g33560	—	—	—	—	—	—

TABLE 10 — Chromosome 7 - Physical positions of certain genetic markers on soybean chromosome 7 in proximity to QTL associated with a low iron growth condition tolerant phenotype. SEQ

Marker or Locus Name	ID NO	LG	Chromsome	Middle Position	Start	Stop
Gm_W82_CR07.G4950	—	M	7	1583569	1582889	1584249
TC112538	—	M	7	2300269	2295259	2305279
Glyma07g03790	—	M	7	2612066	2611352	2612781
Gm_W82_CR07.G8460	—	M	7	2612066	2611352	2612781
Glyma07g03800	—	M	7	2617193	2616471	2617916
Glyma07g03810	—	M	7	2625857	2624956	2626758
NS0202654
47	M	7	2799169	2799349	2798990
NS0206351
48	M	7	2884070	2883889	2884252
Glyma07g04470	—	M	7	3258830	3257169	3260492
Glyma07g04840	—	M	7	3566409	3565090	3567728
NS0202966
49	M	7	3874645	3874138	3875152
Glyma07g05420	—	M	7	4097016	4095962	4098070
Glyma07g05820	—	M	7	4509739	4508711	4510768
NS0102362	—	M	7	4840904	4840559	4841250
Glyma07g06150	—	M	7	4868984	4867327	4870641
Glyma07g06510	—	M	7	5248237	5245895	5250579
Glyma07g06560	—	M	7	5283366	5281545	5285187
Glyma07g07380	—	M	7	6062119	6060096	6064143
Gm_W82_CR07.G30600	—	M	7	6062119	6060096	6064143
Pvcon9217	—	M	7	6063220	6062920	6063521
TC119399	—	M	7	6063744	6063342	6064147
Glyma07g07560	—	M	7	6267199	6266247	6268152
TC385708	—	M	7	6660065	6659800	6660331
Glyma07g08950	—	M	7	7481983	7480599	7483367
Glyma07g09110	—	M	7	7584736	7583460	7586013
Glyma07g09150	—	M	7	7623295	7617936	7628654
Glyma07g09160	—	M	7	7634956	7632638	7637275
Glyma07g09170	—	M	7	7650900	7646391	7655409
Glyma07g09900	—	M	7	8354620	8353183	8356057
Glyma07g09960	—	M	7	8379199	8377095	8381304
Glyma07g09970	—	M	7	8384620	8383415	8385825
NS0119842	—	M	7	8964940	8965292	8964589
TA57919_3847	—	M	7	9404069	9399907	9408231
Glyma07g11180	—	M	7	9406330	9399842	9412818
Glyma07g12210	—	M	7	10528875	10527516	10530235
Glyma07g29640	—	M	7	34637242	34635418	34639067
Glyma07g29650	—	M	7	34646347	34640449	34652245
Glyma07g29940	—	M	7	35026011	35025068	35026954
Glyma07g31380	—	M	7	36403003	36400692	36405314
Glyma07g31390	—	M	7	36408650	36407590	36409711
NS0202730
50	M	7	36497284	36497794	36496774
NS0206314
51	M	7	36529671	36529465	36529877
Glyma07g31610	—	M	7	36591219	36590751	36591687
Glyma07g32330	—	M	7	37262246	37261063	37263430
Glyma07g33070	—	M	7	37970359	37969519	37971200
Glyma07g33090	—	M	7	37977896	37976947	37978845
Glyma07g33560	—	M	7	38479141	38477620	38480663

TABLE 32 — Chromosome 18-Marker type and nucleic acid primers and probes useful in the detection of certain alleles of polymorphic nucleic acid markers associated with an iron deficiency growth condition tolerant phenotype.

Marker or	SEQ
Locus	ID	Marker
Name	NO	Type	Sequence
NS0092671	59	TAQMAN	—
Primer 1	63	—	TCTCCATTGCTAATAATTGCTTGAGT
Primer 2	64	—	AACGAAAAGGGCTCAAAAACTAGTT
Probe 1	65	—	ATTTGTTCTTTTTCTTTTC
Probe 2	66	—	ATTTGTTTTTTTTCTTTTCC
N50118292	60	TAQMAN	—
Primer 1	67	—	GCGTCAATATCCTTTTCTAAAAGAAA
			ACT
Primer 2	68	—	GAAACGAGACGGATCGGGTT
Probe 1	69	—	TCTTAGTATGAAAACTTC
Probe 2	70	—	CTTAGTCTGAAAACTT

TABLE 33 — Chromosome 7-Marker type and nucleic acid primer and probes useful in the detection of certain alleles of polymorphic nucleic acid markers associated with an iron deficiency growth condition tolerant phenotype. SEQ

Marker or	ID	Marker
Locus Name	NO	Type	Sequence
NS0120070	108	TAQMAN	—
Primer 1	111	—	ATTACGAATACCGTTGTTCAGTGACT
Primer 2	112	—	ATCTTAGGTATATGAATTAAAACGAA
			AAATTAAGTTGA
Probe 1	113	—	TCATCCCTCATAGTCTCAT
Probe 2	114	—	CATCCCTCATTGTCTCAT

TABLE 34 — Chromosome 9-Marker type and nucleic acid primer and probes useful in the detection of certain alleles of polymorphic nucleic acid markers associated with an iron deficiency growth condition tolerant phenotype. SEQ

Marker or	ID	Marker
Locus Name	NO	Type	Sequence
NS0094370		TAQMAN	—
Primer 1	175	—	CCGTATGCTGCATTTGTGTATTC
Primer 2	176	—	TGCATAACCGTCCAATGTATTTTG
Probe 1	177	—	TAGTGCAGCAGGAAA
Probe 2	178	—	TGCAGTAGGAAATC
NS0123372		TAQMAN	—
Primer 1	179	—	CCCGATCAGCTGCTTTTTAGG
Primer 2	180	—	CGCCCTTCTCCTGGACAAC
Probe 1	181	—	CTGATCTAGTTGGAATAG
Probe 2	182	—	ACTGATCTAGTAGGAATAG
NS0119073		TAQMAN	—
Primer 1	183	—	GAATGGAAGCTGGTATTGATA
			TTTGATA
Primer 2	184	—	TTATCTCCCCTCTCACTTTCA
			CTACA
Probe 1	185	—	TGACTACGACATGCAC
Probe 2	186	—	TGACTATGACATGCACG

TABLE 11 — Plant or Plant Stand

Score	Symptoms
1	Most plants are completely dead. The plants that are still alive are
	approximately 10% of normal height, and have very little living tissue.
2	Most leaves are almost dead, most stems are still green. Plants are severely
	stunted (10-20% of normal height).
3	Most plants are yellow and necrosis is seen on most leaves. Most plants are
	approximately 20-40% of normal height.
4	Most plants are yellow, and necrosis is seen on the edges of less than half
	the leaves. Most plants are approximately 50% of normal height.
5	Most plants are light green to yellow, and no necrosis is seen on the leaves.
	Most plants are stunted (50-75% of normal height).
6	More than half the plants show moderate chlorosis, but no necrosis is seen
	on the leaves.
7	Less than half of the plants showing moderate chlorosis (light green leaves).
8	A few plants are showing very light chlorosis on one or two leaves.
9	All plants are normal green color.

TABLE 12 — SEQ ID

Marker or Locus Name	NO	Source
Chromosome 3 (LG N)
Glyma03g34510	—	Glyma1
Gm_W82_CR03.G378360	—	Gm_W82_CR03
Glyma03g34760	—	Glyma1
Glyma03g34860	—	Glyma1
NS0202712	1	—
Glyma03g35130	—	Glyma1
Glyma03g35280	—	Glyma1
NS0129403	2	—
NS0115624	3	—
Glyma03g36800	—	Glyma1
Glyma03g38030	—	Glyma1
NS0202842	4	—
NS0122122	5	—
NS0205984	6	—
Pvcon3607	—	Phaseolus_vulgaris
Glyma03g39610	—	Glyma1
FE710890	—	Phaseolus_vulgaris
NS0202698	7	—
Glyma03g41830	—	Glyma1
Glyma03g42250	—	Glyma1
Chromosome 19 (LG L)
Glyma19g32880	—	Glyma1
Glyma19g33540	—	Glyma1
NS0206298	20	—
NS0205620	21	—
NS0206055	22	—
Glyma19g34480	—	Glyma1
NS0204985	23	—
Gm_W82_CR19.G249620	—	Gm_W82_CR19
Glyma19g34740	—	Glyma1
NGMAX005710149	—	—
220298_3433_1666	—	cajanus_cajan
Contig13742	—	cajanus_cajan
BARCSOYSSR_19_1192	—	Wm82_potential_SSR
Glyma19g34750	—	Glyma1
Gm_W82_CR19.G249630	—	Gm_W82_CR19
Contig9146	—	cajanus_cajan
Chromosome 14 (LG O)
NS0116559	—	—
Glyma10g33710	—	Glyma1
TA42233_3847	—	Glycine_max_release_2
AF108084.1	—	GenBank
AW734581	—	Glycine_max_release_2
Glyma10g34260	—	Glyma1
Glyma10g34280	—	Glyma1
Glyma10g34290	—	Glyma1
Glyma10g34460	—	Glyma1
Glyma10g34600	—	Glyma1
Glyma10g34630	—	Glyma1
Glyma10g34850	—	Glyma1
NS0120070	24	—
NS0097952	25	—
NS0092960	26	—
Glyma10g36370	—	Glyma1
NS0118907	27	—
Glyma10g37560	—	Glyma1
TC354083	—	GMGI.042210
Glyma10g37600	—	Glyma1
BW595896	—	LJGI.070108
TC130824	—	MTGI.071708
CV528982	—	Phaseolus_vulgaris
Glyma10g37610	—	Glyma1
BG511181	—	GMGI.042210
TA68568_3847	—	Glycine_max_release_2
NS0204740	28	—
NS0205036	29	—
Glyma10g37910	—	Glyma1
Glyma10g37920	—	Glyma1
Gm_W82_CR10.G221330	—	Gm_W82_CR10
NS0206252	30	—
Glyma10g38600	—	Glyma1
Glyma10g40520	—	Glyma1
BF631841	—	MTGI.071708
Glyma10g41460	—	Glyma1
Glyma10g41470	—	Glyma1
TC412490	—	GMGI.042210
Gm_W82_CR10.G236520	—	Gm_W82_CR10
Chromosome 7 (LG M)
Gm_W82_CR07.G4950	—	Gm_W82_CR07
TC112538	—	MTGI.071708
Glyma07g03790	—	Glyma1
Gm_W82_CR07.G8460	—	Gm_W82_CR07
Glyma07g03800	—	Glyma1
Glyma07g03810	—	Glyma1
NS0202654	47	—
NS0206351	48	—
Glyma07g04470	—	Glyma1
Glyma07g04840	—	Glyma1
NS0202966	49	—
Glyma07g05420	—	Glyma1
Glyma07g05820	—	Glyma1
NS0102362	—	—
Glyma07g06150	—	Glyma1
Glyma07g06510	—	Glyma1
Glyma07g06560	—	Glyma1
Glyma07g07380	—	Glyma1
Gm_W82_CR07.G30600	—	Gm_W82_CR07
Pvcon9217	—	Phaseolus_vulgaris
TC119399	—	MTGI.071708
Glyma07g07560	—	Glyma1
TC385708	—	GMGI.042210
Glyma07g08950	—	Glyma1
Glyma07g09110	—	Glyma1
Glyma07g09150	—	Glyma1
Glyma07g09160	—	Glyma1
Glyma07g09170	—	Glyma1
Glyma07g09900	—	Glyma1
Glyma07g09960	—	Glyma1
Glyma07g09970	—	Glyma1
NS0119842	—	—
TA57919_3847	—	Glycine_max_release_2
Glyma07g11180	—	Glyma1
Glyma07g12210	—	Glyma1
Glyma07g29640	—	Glyma1
Glyma07g29650	—	Glyma1
Glyma07g29940	—	Glyma1
Glyma07g31380	—	Glyma1
Glyma07g31390	—	Glyma1
NS0202730	50	—
NS0206314	51	—
Glyma07g31610	—	Glyma1
Glyma07g32330	—	Glyma1
Glyma07g33070	—	Glyma1
Glyma07g33090	—	Glyma1
Glyma07g33560	—	Glyma1

TABLE 14 — Chromosome 18 (LG G) SEQ

	ID NO:		Start Base	End Base
Locus/DisplayName (1)	(2)	Source (3)	(4)	(5)
Gm_W82_CR18.G397200	—	Gm_W82_CR18	59900386	59906556
NGMAX008183850	52	—	59904411	59904712
Contig19662	—	cajanus_cajan	59905905	59906118
BARCSOYSSR_18_1832	—	Wm82_potential_SSR	60056533	60056556
NGMAX008184740	53	—	60061647	60061948
Pvcon7024	—	Phaseolus_vulgaris	60065999	60067897
TA4278_3885	—	Phaseolus_vulgaris_release_2	60245743	60247776
NS0204945	54	—	60247182	60246514
TC378862	—	GMGI.042210	60247386	60247845
Gm_W82_CR18.G400990	—	Gm_W82_CR18	61248610	61249504
NGMAX008190659	55	—	61262829	61263130
304729_3194_1618	—	cajanus_cajan	61262926	61263150
TC373110	—	GMGI.042210	61315062	61315579
NGMAX008190985	56	—	61322677	61322978
BM108253	—	Glycine_max_release_2	61329398	61329834
BARCSOYSSR_18_1891	—	Wm82_potential_SSR	61385362	61385381
NS0205638	57	—	61388179	61388758
Cf2356d	—	Chafa1_1clean	61388553	61388793
TC373562	—	GMGI.042210	61390489	61391050
NGMAX008191317	58	—	61392011	61392312
Cf18802d	—	Chafa1_1clean	61392112	61392386
NP7265835	—	MTGI.071708	61430944	61432625
NS0092671	59	—	61432518	61432769
Glyma18g53050	—	Glyma1	61434592	61439004
NS0118292	60	—
NGMAX008194551	61	—	61994075	61994376
NGMAX008194952	62	—	62082932	62083233
108285_2293_0177	—	cajanus_cajan	62090438	62090654

TABLE 17 — Chromosome 5 (LG A1) SEQ ID NO:

Locus/DisplayName (1)	(2)	Source (3)	Start Base (4)	End Base (5)
WmFPC_Contig7872	—	Wm82	8611032	8760634
NGMAX006398059	71	—	8726001	8726302
TA74988_3847	—	Glycine_max_release_2	8726509	8728196
TA11305_34305	—	Lotus_japonicus_release_1	8726743	8728281
Glyma05g08860	—	Glyma1	8731333	8733792
Gm_W82_CR05.G36010	—	Gm_W82_CR05	8731333	8733792
NGMAX006398075	72	—	8734373	8734674
Glyma05g08870	—	Glyma1	8749735	8751567
TC412701	—	GMGI.042210	8789875	8790358
CV534998	—	Phaseolus_vulgaris	8790063	8790596
BARCSOYSSR_05_0384	—	Wm82_potential_SSR	8791244	8791303
NGMAX006398171	73	—	8792560	8792861
Contig45244	—	cajanus_cajan	8793029	8793205
Contig16193	—	cajanus_cajan	8793151	8793206
Contig48186	—	cajanus_cajan	8972135	8973187
AW278035	—	Glycine_max_release_2	8972910	8973235
NGMAX006398585	74	—	8975992	8976293
442533_3845_3611	—	cajanus_cajan	8977523	8977782
TC414557	—	GMGI.042210	8977562	8977758
Cf7706d	—	Chafa1_1clean	8999539	8999973
Glyma05g09220	—	Glyma1	8995921	9005129
NGMAX006398641	75	—	9000470	9000771
TC416067	—	GMGI.042210	9000945	9001313
Glyma05g09240	—	Glyma1	9010402	9011072
Gm_W82_CR05.G38570	—	Gm_W82_CR05	9010402	9011072
169777_3462_2392	—	cajanus_cajan	9010970	9011057
NGMAX006398662	76	—	9012813	9013114
Cf3250d	—	Chafa1_1clean	9012500	9015254
Glyma05g09250	—	Glyma1	9014291	9015254
Gm_W82_CR05.G38580	—	Gm_W82_CR05	9014291	9015254
BP044357	—	Lotus_japonicus_release_1	9026484	9028268
AV764725	—	Lotus_japonicus_release_1	9026611	9028149
CB540591	—	Phaseolus_vulgaris	9026676	9028170
NGMAX006398682	77	—	9029664	9029965
Contig40310	—	cajanus_cajan	9032015	9032233
Contig32764	—	cajanus_cajan	9032145	9032358
Glyma05g09320	—	Glyma1	9048193	9049361
Gm_W82_CR05.G38650	—	Gm_W82_CR05	9048193	9049361
NGMAX006398725	78	—	9049111	9049412
NGMAX006398726	79	—	9049326	9049627
Contig42115	—	cajanus_cajan	9049922	9050265
Gm_W82_CR05.G38690	—	Gm_W82_CR05	9091842	9095724
NGMAX006398800	80	—	9096884	9097185
Cf13029d	—	Chafa1_1clean	9097111	9097317
BARC-044481-08709	—	marker_map4	9097066	9097473
BM568328	—	Glycine_soja_release_2	9231822	9232150
NGMAX006398923	82	—	9314174	9314475
Glyma05g09450	—	Glyma1	9314231	9314482
gi_251741293	—	cajanus_cajan	9957719	9957766
NGMAX006399042	83	—	9964942	9965243
Glyma05g10000	—	Glyma1	9966305	9967626
Cf16969d	—	Chafa1_1clean	10085119	10088679
NGMAX006399063	84	—	10105515	10105816
BARCSOYSSR_05_0410	—	Wm82_potential_SSR	10110781	10110830
Gm_W82_CR05.G41210	—	Gm_W82_CR05	10127470	10128074
NGMAX006399066	85	—	10128574	10128875
BW597367	—	LJGI.070108	10132125	10132681
BARCSOYSSR_05_0420	—	Wm82_potential_SSR	10747843	10747892
NGMAX006399168	86	—	10753693	10753994
138369_1624_0039	—	cajanus_cajan	10754650	10754859
033583_0211_2302	—	cajanus_cajan	12671045	12671171
BARCSOYSSR_05_0445	—	Wm82_potential_SSR	12706029	12706048
NGMAX006399482	87	—	12716333	12716634
Glyma05g12510	—	Glyma1	12744996	12745160
Gm_W82_CR05.G105990	—	Gm_W82_CR05	17286974	17287535
BARCSOYSSR_05_0506	—	Wm82_potential_SSR	17292930	17292983
NGMAX006400296	88	—	17294665	17294966
Glyma05g15680	—	Glyma1	17305698	17305960
Contig15035	—	cajanus_cajan	21200771	21201057
NGMAX006401034	89	—	21201143	21201444
199534_3796_0358	—	cajanus_cajan	21201471	21201711
152322_2470_3960	—	cajanus_cajan	22915341	22915534
BARCSOYSSR_05_0564	—	Wm82_potential_SSR	22945692	22945743
NGMAX006401429	90	—	23025046	23025347
TC368157	—	GMGI.042210	23027973	23029620
Satt454	—	marker_map4	25287587	25287839
BARCSOYSSR_05_0609	—	Wm82_potential_SSR	25287731	25287781
NGMAX006401765	91	—	25295318	25295619
381902_3512_4031	—	cajanus_cajan	25296404	25296662
Glyma05g21200	—	Glyma1	25627653	25634422
Gm_W82_CR05.G152350	—	Gm_W82_CR05	25627653	25634422
NGMAX008341472	92	—	25645423	25645724
BARCSOYSSR_05_0622	—	Wm82_potential_SSR	25648451	25648492
Cf5490d	—	Chafa1_1clean	26166473	26166936
318062_1525_0472	—	cajanus_cajan	26170236	26170357
NGMAX006401896	93	—	26170363	26170664
BARCSOYSSR_05_0638	—	Wm82_potential_SSR	26175536	26175569
BARCSOYSSR_05_0639	—	Wm82_potential_SSR	26175838	26175889
BARCSOYSSR_05_0640	—	Wm82_potential_SSR	26176422	26176471

TABLE 20 — Chromosome 7 (LG M) SEQ

	ID NO:		Start Base
Locus/DisplayName (1)	(2)	Source (3)	(4)	End Base (5)
NGMAX006621532	94	—	3393031	3393332
Glyma07g04610	—	Glyma1	3402388	3404721
Gm_W82_CR07.G10270	—	Gm_W82_CR07	3402388	3404721
TC403215	—	GMGI.042210	3412126	3412400
Glyma07g04620	—	Glyma1	3412764	3413309
NGMAX006621610	95	—	3416548	3416849
BARCSOYSSR_07_0185	—	Wm82_potential_SSR	3417142	3417201
NGMAX006621674	96	—	3427338	3427639
BARCSOYSSR_07_0186	—	Wm82_potential_SSR	3429880	3429909
Contig7081	—	cajanus_cajan	3430111	3430277
ss181360748	—	Wm82xPI468916	3433852	3433973
NGMAX006621699	97	—	3440093	3440394
AW720220	—	LJGI.070108	3441581	3444258
BM525303	—	Glycine_soja_release_2	3441548	3444605
112768_3355_0899	—	cajanus_cajan	3444787	3445772
213668_2891_0626	—	cajanus_cajan	3445730	3446583
NGMAX006621706	98	—	3446258	3446559
Contig9107	—	cajanus_cajan	3444989	3447842
TA5004_3885	—	Phaseolus_vulgaris_release_2	3446499	3448009
Glyma07g04800	—	Glyma1	3540566	3542258
Gm_W82_CR07.G10660	—	Gm_W82_CR07	3540566	3542258
NGMAX006621720	99	—	3541479	3541780
TC396277	—	GMGI.042210	3542083	3542437
TA49177_3847	—	Glycine_max_release_2	3541831	3543402
ss181360746	—	Wm82xPI468916	3543470	3543591
BARCSOYSSR_07_0190	—	Wm82_potential_SSR	3545904	3545967
NGMAX006621726	100	—	3547188	3547489
BF597028	—	Glycine_soja_release_2	3551690	3552575
BG043289	—	Glycine_max_release_2	3663176	3665198
TA2947_3886	—	Phaseolus_coccineus_release_2	3663077	3665376
AI443806	—	Glycine_max_release_2	3664939	3665318
Contig38368	—	cajanus_cajan	3664966	3665337
NGMAX006621767	101	—	3679813	3680114
NGMAX006621777	102	—	3682741	3683042
BARC-041667-08063	—	marker_map4	3683493	3683694
393760_2699_3151	—	cajanus_cajan	3809484	3809711
FK446225	—	GMGI.042210	3812085	3812372
WmFPC_Contig66	—	Wm82	3706750	3925221
NGMAX006622006	103	—	3816276	3816577
Contig45160	—	cajanus_cajan	3816479	3817114
TA40883_3847	—	Glycine_max_release_2	3816485	3817194
TC375163	—	GMGI.042210	3979825	3980713
Contig13143	—	cajanus_cajan	3980387	3980595
BARCSOYSSR_07_0207	—	Wm82_potential_SSR	3981130	3981165
NGMAX006622614	104	—	3981054	3981355
BARCSOYSSR_07_0208	—	Wm82_potential_SSR	3988651	3988670
Glyma07g05350	—	Glyma1	3988721	3990122
BI969369	—	Glycine_max_release_2	4122488	4124632
TC367324	—	GMGI.042210	4124193	4124669
NGMAX006622952	105	—	4126322	4126623
CD395608	—	Glycine_max_release_2	4126664	4126991
TC387188	—	GMGI.042210	4849393	4849841
NGMAX006623558	106	—	4850240	4850541
TA12767_34305	—	Lotus_japonicus_release_1	4854998	4857901
TA47785_3847	—	Glycine_max_release_2	5034853	5036117
asmbl_4174	—	Vigna_unguiculata	5034852	5036125
NGMAX006623995	107	—	5035388	5035689
259614_3432_2478	—	cajanus_cajan	5035714	5035850
Glyma07g06360	—	Glyma1	5084793	5084902
BARCSOYSSR_07_0271	—	Wm82_potential_SSR	5089392	5089423
NGMAX006624271	108	—	5092982	5093283
Contig23159	—	cajanus_cajan	5094525	5094763
Pvcon10792	—	Phaseolus_vulgaris	5418127	5418733
315592_2632_1694	—	cajanus_cajan	5418473	5418717
NGMAX006625883	109	—	5418776	5419077
Contig24687	—	cajanus_cajan	5419388	5419599
Cf18742d	—	Chafa1_1clean	5455985	5459507
NGMAX006626085	110	—	5457696	5457997
TC381430	—	GMGI.042210	5457600	5459580
BI785468	—	Glycine_max_release_2	5457683	5459525
TA53357_3847	—	Glycine_max_release_2	5457605	5460346
FE709341	—	Phaseolus_vulgaris	5458490	5459666

TABLE 23 — Chromosome 17 (LG D2) SEQ

	ID NO:		Start Base	End Base
Locus/DisplayName (1)	(2)	Source (3)	(4)	(5)
NGMAX007790014	115	—	3155696	3155997
BF425454	—	GMGI.042210	3155653	3156074
BF425454	—	Glycine_max_release_2	3155653	3156123
Cf14786d	—	Chafa1_1clean	3323205	3323429
BG511404	—	Glycine_max_release_2	3323134	3323620
105562_1346_1630	—	cajanus_cajan	3323270	3323498
Contig39552	—	cajanus_cajan	3323309	3323550
NGMAX007790381	116	—	3323744	3324045
Cf17659d	—	Chafa1_1clean	3320738	3328482
Contig37026	—	cajanus_cajan	3326324	3326599
Contig3555	—	cajanus_cajan	3326889	3327061
CA908533	—	Phaseolus_coccineus_release_2	3326779	3327187
Glyma17g04950	—	Glyma1	3326774	3328781
Gm_W82_CR17.G16430	—	Gm_W82_CR17	3326774	3328781
NGMAX007790411	117	—	3329853	3330154
Glyma17g04960	—	Glyma1	3330115	3332472
Gm_W82_CR17.G16440	—	Gm_W82_CR17	3329999	3332810
Contig24276	—	cajanus_cajan	3331849	3332089
125135_2424_0556	—	cajanus_cajan	3360193	3360344
NGMAX007790440	118	—	3364343	3364644
Cf16374d	—	Chafa1_1clean	3365597	3366588
BG838973	—	Glycine_max_release_2	3365233	3367059
BG507731	—	GMGI.042210	3611762	3612075
BARCSOYSSR_17_0195	—	Wm82_potential_SSR	3633036	3633099
NGMAX007790913	120	—	3633675	3633976
BW655748	—	GMGI.042210	3635257	3636145
Cf18958d	—	Chafa1_1clean	3636566	3636904
BARCSOYSSR_17_0196	—	Wm82_potential_SSR	3644289	3644308
NGMAX007790961	121	—	3647410	3647711
BG405534	—	Glycine_max_release_2	3654628	3655142
Cf13064d	—	Chafa1_1clean	3655848	3655983
Contig31396	—	cajanus_cajan	3655888	3656111
NGMAX007790982	122	—	3656282	3656583
Glyma17g05300	—	Glyma1	3657812	3658198
BARCSOYSSR_17_0197	—	Wm82_potential_SSR	3659872	3659903
085726_2424_0132	—	cajanus_cajan	4982475	4985016
NGMAX007793044	123	—	4984466	4984767
BARCSOYSSR_17_0274	—	Wm82_potential_SSR	4996872	4996919
086788_3391_0381	—	cajanus_cajan	4998989	4999064

TABLE 26 — Chromosome 19 (LG L) SEQ

	ID NO:		Start Base	End Base
Locus/DisplayName (1)	(2)	Source (3)	(4)	(5)
NGMAX008199658	124	—	1253726	1254027
Cf21314d	—	Chafa1_1clean	1254120	1255988
Gm_W82_CR19.G10090	—	Gm_W82_CR19	1271256	1272512
NGMAX008199750	125	—	1272430	1272731
BARCSOYSSR_19_0086	—	Wm82_potential_SSR	1272601	1272664
WmFPC_Contig3730	—	Wm82	1163579	1429575
NGMAX008199864	126	—	1296447	1296748
BARCSOYSSR_19_0088	—	Wm82_potential_SSR	1297586	1297645
BARCSOYSSR_19_0089	—	Wm82_potential_SSR	1308871	1308956
NGMAX008199973	127	—	1309280	1309581
Glyma19g01720	—	Glyma1	1309198	1310096
Glyma19g01750	—	Glyma1	1333537	1333700
NGMAX008200015	128	—	1333772	1334073
BARCSOYSSR_19_0092	—	Wm82_potential_SSR	1334292	1334333
NGMAX008200022	129	—	1337736	1338037
Gm_W82_CR19.G10370	—	Gm_W82_CR19	1343268	1344198
BI967966	—	Glycine_max_release_2	1387373	1388013
NGMAX008200044	130	—	1392143	1392444
Contig37264	—	cajanus_cajan	1396218	1396421
Glyma19g01820	—	Glyma1	1396249	1396497
NGMAX008200052	131	—	1414026	1414327
BI321279	—	GMGI.042210	1416432	1416851
BG511671	—	Glycine_max_release_2	1489429	1489735
NGMAX008200275	132	—	1490445	1490746
Glyma19g01870	—	Glyma1	1492838	1495764
Gm_W82_CR19.G15280	—	Gm_W82_CR19	1492838	1495764
Cf9954d	—	Chafa1_1clean	1501094	1501508
Glyma19g01880	—	Glyma1	1499643	1504582
Gm_W82_CR19.G15290	—	Gm_W82_CR19	1499643	1504595
NGMAX008200335	133	—	1502499	1502800
ss181360588	—	Wm82xPI468916	1502646	1502767
BARCSOYSSR_19_0099	—	Wm82_potential_SSR	1509013	1509038
BARCSOYSSR_19_0100	—	Wm82_potential_SSR	1526039	1526136
Contig11069	—	cajanus_cajan	1592437	1592670
Contig3143	—	cajanus_cajan	1592554	1592758
268933_0047_1009	—	cajanus_cajan	1592619	1592881
NGMAX008200568	134	—	1592869	1593170
BARCSOYSSR_19_0105	—	Wm82_potential_SSR	1593154	1593203
Contig1379	—	cajanus_cajan	1593217	1593635
Contig48254	—	cajanus_cajan	1593271	1593819
285107_3537_3801	—	cajanus_cajan	1593451	1593714
BARCSOYSSR_19_0106	—	Wm82_potential_SSR	1596261	1596300
BARCSOYSSR_19_0107	—	Wm82_potential_SSR	1599519	1599558
071040_1554_2187	—	cajanus_cajan	1610939	1610980
NGMAX008200652	135	—	1615160	1615461
BARCSOYSSR_19_0108	—	Wm82_potential_SSR	1615763	1615782
NGMAX008200656	136	—	1617406	1617707
BARCSOYSSR_19_0109	—	Wm82_potential_SSR	1630429	1630452
Satt446	—	marker_map4	1634051	1634348
Contig11069	—	cajanus_cajan	1651156	1651389
Contig3143	—	cajanus_cajan	1651273	1651470
NGMAX008200785	137	—	1651276	1651577
268933_0047_1009	—	cajanus_cajan	1651338	1651604
Contig1379	—	cajanus_cajan	1651725	1652180
Cf17202d	—	Chafa1_1clean	1809590	1809811
TC413656	—	GMGI.042210	1810134	1810736
NGMAX008200913	138	—	1813813	1814114
354114_3517_2150	—	cajanus_cajan	1815145	1815383
TC409049	—	GMGI.042210	1817061	1817307
BI970322	—	Glycine_max_release_2	7063345	7064079
Contig8569	—	cajanus_cajan	44569360	44571272
CB540932	—	Phaseolus_vulgaris	44569015	44571911

TABLE 29 — Chromosome 9 (LG K) SEQ

	ID NO:		Start	End Base
Locus/DisplayName (1)	(2)	Source (3)	Base (4)	(5)
NGMAX006925911	139	—	40243718	40244019
Contig41696_primers	—	cajanus_cajan	40257234	40257429
Contig41696	—	cajanus_cajan	40256966	40258176
BM108278	—	Glycine_max_release_2	40257091	40258183
Contig31037	—	cajanus_cajan	40258700	40259448
asmbl_6000	—	Vigna_unguiculata	40258719	40259446
BE823853	—	Glycine_max_release_2	40258861	40259409
NGMAX006925967	140	—	40259790	40260091
NGMAX006926063	141	—	40272402	40272703
BARCSOYSSR_09_1351	—	Wm82_potential_SSR	40274616	40274639
Cf13555d	—	Chafa1_1clean	40276310	40276615
NGMAX006926228	142	—	40308770	40309071
TC361317	—	GMGI.042210	40312128	40312691
TA56123_3847	—	Glycine_max_release_2	40312116	40312852
218500_2738_3544	—	cajanus_cajan	40312576	40312893
NGMAX006926426	143	—	40374697	40374998
BM307669	—	Glycine_max_release_2	40376529	40376938
351740_2368_1082	—	cajanus_cajan	40376719	40376915
Cf15642d	—	Chafa1_1clean	40376761	40376927
BARCSOYSSR_09_1359	—	Wm82_potential_SSR	40378752	40378773
BU080803	—	Glycine_max_release_2	40378097	40380883
NGMAX006926451	144	—	40381147	40381448
117697_1117_1653	—	cajanus_cajan	40385920	40386138
Contig915	—	cajanus_cajan	40387459	40387654
CA921370	—	MTGI.071708	40448774	40449433
Glyma09g34010	—	Glyma1	40448819	40451396
NGMAX006926709	145	—	40450053	40450354
BI320313	—	GMGI.042210	40466800	40466933
NGMAX006926784	146	—	40492973	40493274
Pvcon5634	—	Phaseolus_vulgaris	40493627	40494016
BP054761	—	Lotus_japonicus_release_1	40493628	40494077
BM892455	—	GMGI.042210	40538148	40539515
NGMAX006926934	147	—	40538904	40539205
BM892455	—	Glycine_max_release_2	40538148	40540469
BU545056	—	Glycine_max_release_2	40547146	40548442
NGMAX006926989	148	—	40548839	40549140
TA75773_3847	—	Glycine_max_release_2	40565516	40566166
TC356190	—	GMGI.042210	40565516	40566166
NGMAX006927044	149	—	40567379	40567680
Contig25820	—	cajanus_cajan	40571216	40571379
Contig4437	—	cajanus_cajan	40571319	40571422
008498_0634_0728	—	cajanus_cajan	40571401	40571432
NGMAX006927083	150	—	40573731	40574032
BARCSOYSSR_09_1372	—	Wm82_potential_SSR	40575734	40575771
368088_3993_0858	—	cajanus_cajan	40577562	40577807
Glyma09g34170	—	Glyma1	40606183	40615020
Cf16214d	—	Chafa1_1clean	40606988	40614651
NGMAX006927293	151	—	40611728	40612029
BU494285	—	Lotus_japonicus_release_1	40610299	40613740
BU494285	—	LJGI.070108	40610302	40613740
Glyma09g34190	—	Glyma1	40633379	40638472
NGMAX006927454	152	—	40636773	40637074
TC413502	—	GMGI.042210	40637328	40637485
TA74376_3847	—	Glycine_max_release_2	40660591	40660742
TC410021	—	GMGI.042210	40660591	40660742
NGMAX006927578	153	—	40660778	40661079
TC362522	—	GMGI.042210	40660852	40662410
TA49669_3847	—	Glycine_max_release_2	40660954	40662419
NS0094370	154	—
357928_2669_2557	—	cajanus_cajan	40663765	40664577
TA3851_3886	—	Phaseolus_coccineus_release_2	40663773	40664903
TC404180	—	GMGI.042210	40663478	40665282
asmbl_6018	—	Vigna_unguiculata	40663844	40665198
AI495278	—	Glycine_max_release_2	40664775	40665263
NS0202727	155	—	40667193	40666794
441702_2628_0479	—	cajanus_cajan	40667116	40667212
Contig27394	—	cajanus_cajan	40667116	40667212
BI424714	—	Glycine_max_release_2	40667593	40668779
BI424714	—	GMGI.042210	40667677	40668779
BI893945	—	Glycine_max_release_2	40677514	40677927
ss181358282	—	Wm82xPI468916	40679987	40680108
NGMAX006927737	156	—	40681901	40682202
TC396512	—	GMGI.042210	40682388	40685072
Glyma09g34250	—	Glyma1	40682388	40685488
TC368672	—	GMGI.042210	40682923	40685164
Gm_W82_CR09.G172960	—	Gm_W82_CR09	40682388	40685746
BARCSOYSSR_09_1385	—	Wm82_potential_SSR	40692420	40692461
NGMAX006927783	157	—	40693382	40693683
Glyma09g34260	—	Glyma1	40693196	40695274
Gm_W82_CR09.G172970	—	Gm_W82_CR09	40693196	40695274
BARCSOYSSR_09_1386	—	Wm82_potential_SSR	40695711	40695732
310291_2136_0449	—	cajanus_cajan	40699187	40699373
Contig29803	—	cajanus_cajan	40699810	40700422
NGMAX006927836	158	—	40700573	40700874
221522_3792_2185	—	cajanus_cajan	40701635	40701833
113453_3824_1247	—	cajanus_cajan	40701634	40701848
FE697104	—	Phaseolus_vulgaris	40701670	40702507
Glyma09g34320	—	Glyma1	40735795	40736472
BARCSOYSSR_09_1388	—	Wm82_potential_SSR	40737394	40737419
NGMAX006928046	159	—	40738085	40738386
asmbl_6023	—	Vigna_unguiculata	40738466	40741661
TA12621_34305	—	Lotus_japonicus_release_1	40757487	40758228
TC35672	—	LJGI.070108	40757487	40758228
NS0123372	160	—	40757850	40758506
339112_3138_3869	—	cajanus_cajan	40758893	40759121
381244_3433_2042	—	cajanus_cajan	40759267	40759490
074045_1887_1079	—	cajanus_cajan	40765413	40765510
200842_1421_3709	—	cajanus_cajan	40765605	40765747
NGMAX006928148	161	—	40771660	40771961
TC389778	—	GMGI.042210	40772519	40773261
TA44560_3847	—	Glycine_max_release_2	40772519	40773559
FE709357	—	Phaseolus_vulgaris	40775661	40776891
Contig34874	—	cajanus_cajan	40776637	40776916
NGMAX006928238	162	—	40780407	40780708
Contig39370	—	cajanus_cajan	40781564	40783018
asmbl_6024	—	Vigna_unguiculata	40786818	40787111
Contig6058	—	cajanus_cajan	40787205	40787407
NGMAX006928269	163	—	40790581	40790882
TA44261_3847	—	Glycine_max_release_2	40791854	40792498
BE801808	—	Glycine_max_release_2	40791923	40792444
Cf4870d	—	Chafa1_1clean	40806197	40806526
FK576691	—	GMGI.042210	40806485	40806679
NGMAX006928358	164	—	40808949	40809250
Glyma09g34430	—	Glyma1	40809022	40811799
Gm_W82_CR09.G173740	—	Gm_W82_CR09	40809022	40811799
BARC-059665-15981	—	marker_map4	40827693	40828086
NGMAX006928507	165		40834823	40835124
Gm_W82_CR09.G173770	—	Gm_W82_CR09	40839879	40843063
Glyma09g34460	—	Glyma1	40840286	40843063
NGMAX006928537	166	—	40841840	40842141
TA70211_3847	—	Glycine_max_release_2	40841283	40843007
TC349093	—	GMGI.042210	40841283	40843007
FE702368	—	Phaseolus_vulgaris	40884481	40886140
AM162189	—	Pisum_sativum_release_2	40885122	40885709
NGMAX006928614	167	—	40887274	40887575
BM139756	—	Glycine_max_release_2	40887540	40887691
DW246746	—	Glycine_max_release_2	40887692	40887860
271518_0250_0536	—	cajanus_cajan	40898791	40898892
139280_1004_2213	—	cajanus_cajan	40898807	40898893
NGMAX006928650	168	—	40901958	40902259
315453_0615_0207	—	cajanus_cajan	40909988	40910230
TC391225	—	GMGI.042210	40909479	40910839
Glyma09g34540	—	Glyma1	40915819	40917570
135326_1839_1598	—	cajanus_cajan	40919347	40919571
NGMAX006928755	169	—	40919514	40919815
CD418184	—	Glycine_max_release_2	40919571	40920074
NS0202939	170	—
Glyma09g34560	—	Glyma1	40925986	40926726
Pvcon4656	—	Phaseolus_vulgaris	40926434	40929444
NS0119073	171	—	40927939	40928620
BM309243	—	GMGI.042210	40928556	40928983
230306_1435_0813	—	cajanus_cajan	40928906	40929123
Cf15892d	—	Chafa1_1clean	40932401	40932559
Glyma09g34580	—	Glyma1	40932359	40933733
Gm_W82_CR09.G174470	—	Gm_W82_CR09	40932359	40933733
NGMAX006928836	172	—	40933766	40934067
Glyma09g34590	—	Glyma1	40933583	40935779
AW781873	—	Glycine_max_release_2	40934635	40935110
AI900149	—	Glycine_max_release_2	40934700	40935424
TC368843	—	GMGI.042210	40934635	40935779
TA55686_3847	—	Glycine_max_release_2	40934761	40935778
NS0202984	173	—	40935137	40935465
CD394340	—	Glycine_max_release_2	40935262	40935757
BARCSOYSSR_09_1395	—	Wm82_potential_SSR	40938697	40938724
BARCSOYSSR_09_1396	—	Wm82_potential_SSR	40941652	40941677
NGMAX006928848	174	—	40944266	40944567
Cf5524d	—	Chafa1_1clean	40948145	40948510
Glyma09g34600	—	Glyma1	40947262	40950520
FE711739	—	Phaseolus_vulgaris	40948145	40949768

Claims as granted

4 claims

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6 codes

IPC · International Patent Classification

Section A — Human necessities

A01H6/54
A01H1/04
A01H5/10

Section C — Chemistry; metallurgy

C12Q1/68
C07K14/415
C12Q1/6895

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