KR20010005625A - A Method to Identify and Breed Corn with Increased Kernel Oil Concentration - Google Patents

Abstract

A method for breeding corn with increased kernel oil concentrations is described. The method involves the use of genetic markers associated with transgenic loci that control kernel oil concentration. These genetic markers are used to make selections regarding kernel oil concentrations in a breeding population. Also described are methods for selecting complementary oil matrix sources using genetic markers that are likely to produce better offspring. In addition, a transgenic locus that regulates corn kernel oil concentration is described.

Description

A Method to Identify and Breed Corn with Increased Kernel Oil Concentration

Field of Invention

The present invention belongs to the field of plant breeding and molecular biology. More specifically, the present invention relates to the identification of corn loci conferring increased kernel oil concentrations using genetic markers and to the breeding of corns with increased kernel oil concentrations, using the genetic markers as an aid in their identification.

〈Inventive Background〉

Corn is the main crop used as a food source for humans, animal feed and also as a source of carbohydrates, fats, proteins and fiber. Corn is mainly used as an energy source in animal feed or as a raw material for obtaining starch, protein feed fractions, fiber, flaking flour, flour and oil.

Most commercial corn is produced from hybrid seeds throughout the United States. Corn hybrid production requires the development of superior maize breeding species that produce agriculturally superior hybrids at mating. In developing maize allied breeders, plant growers choose many different traits that affect agricultural performance. Such traits include, but are not limited to, stem strength, lodging, disease resistance, grain moisture and yield. Agricultural traits tend to be quantitatively measured in continuous rather than distinct distributions. Quantitative traits are generally described as being regulated by several genes with equivalent and small effects. In addition, the observed phenotype is due in part to these genetic and environmental factors.

The heritability of a characteristic is broadly defined by the ratio of genetic variation to total phenotypic variation. Many agricultural traits show low heredity, ie the performance of offspring cannot be predicted from the performance of parent plants. Thus, traits with low heritability have fewer genetic variation elements compared to the observed mutations. A problem that affects plant growers is that the value of the genetic composition of the plant is difficult to determine from agricultural trait measurements. In an attempt to maximize the discriminative power of these measures, growers collect a variety of measurements from both genetically related individuals and various environments. This strategy is a beneficial source because it involves the use of a wide range of attempts to produce even minor benefits for plant improvement. This is associated with the fact that improved corn strains are selected simultaneously for multiple traits, leading to the development of superior maize breeding varieties for both wasteful and expensive labor.

The addition of new traits in corn breeding programs places additional burdens on plant growers. Depending on the genetic complexity of the new trait (ie single gene versus many genes), considerable time and effort is required to produce a good lineage with the new trait. One such trait is kernel oil concentration.

Corn with increased kernel oil oil concentrations was found in poultry (Han Y. et al., Poultry Sci. 66: 103-111 (1987)) and livestock (Nordstrom, J. W.). , JW) et al., J An. Sci 35 (2): 357-361 (1972)). Typical corn hybrid grains contain 4% oil. In 1896, in an effort to increase kernel oil concentrations. G. Open pollinated cv. By C.G. Hopkins. A long-term recurrent selection program was initiated in Burr's White. This reproducibly selected population known as Illinois High Oil (IHO) using modified material selection methods has been selected for increased oil concentrations for over 90 generations (Dudley, JW and RJ Lambert). (1992) Afaydica 37: 1-7]). As a result, the oil concentration increased by more than 20% in the population. Since the derived material is obtained substantially less than conventional variants, little germ plasm has been used (Alexander, DE (1988) In: Proc. 43rd Ann. Corn and Sorghum Res. Conf. Am. Seed Trade Assoc). , Washington, DC pp 97-105).

Using 38 open polluted cultivars and hybrids, Alexander launched a second reproduction selection program (Alexo hybridization) to increase kernel oil (Alexander, DE (1988) In: Corn). and Corn Improvement.GF Spraque and JW Dudley eds.American Society of Agronomy, Madison WI. Pp 869-880]. Based on the oil concentration of one ear, the same generation of oil farms were obtained for IHO in 28 generations using a selection method based on the oil concentration of one kernel in the next generation. Although the performance was not the same as for conventional hybrids, due to the larger genetic diversity initially available, the yield capability of the Alexo-derived material (high concentration oil breeding species X conventional breeding species) for IHO in a single hybrid hybrid was Improved. The development of agriculturally good corn germplasm with increased kernel oil oil concentrations using conventional plant breeding methods is also an obvious challenge.

Various assays can be used to determine kernel oil concentrations phenotypicly. Oil concentrations represent a non-individual distribution and are common for quantitatively inherited traits controlled by several loci. Kernel oil measurements select those breeding lines with the highest phenotypic expression. Unfortunately, the genetic potential for high concentrations of oil is limited in most of these lines because it is impossible to distinguish between lines based on their true genetic composition. This condition is exacerbated when simultaneous screening is performed for this agricultural performance. Therefore, it would be advantageous to base the selection on genotypes of plants in the population. It may be advantageous to use genetic markers, in particular nucleic acid markers, as direct selection methods for complex quantitative traits. Therefore, genetic markers to find alleles that confer increased oil would be an advantageous tool for plant breeding programs that develop good high concentration oil corn germplasm.

Limited information is available on the identification of prognostic genetic markers for increased oil yield. Kahul, a. Kahler, A. L., Proc. 40th Ann. Corn and Sorghum Res. Conf. Am. Seed Trade Assoc .. Washington D.C. 66-89 (1985) measured the change in the number of isozyme alleles after 25 generations of the selection in the Alexo hybridization method and found eight significant loci. Changes in most of these allele numbers are also significant in tests that measure random gene transfer, making it difficult to conclude that selection methods based on these isozyme alleles would be useful. More recently, Goldman, child. Goldman, I. L., Crop Sci. 34: 908-915 (1994), and Berke, T. G. and Rocheford, T. R. Crop Sci. 35: 1542-1549 (1995) used RFLP markers to identify significant marker loci associated with oil concentration in the Illinois long-term selection population. These studies identified 25 and 31 markers, respectively, in the population derived from Burj White, which were highly associated with increasing oil. Some regions identified by important RFLP marker loci may be common between the two studies, but six of the 15 RFLP markers used in both studies did not match their effects on oil concentration. In this study, the population used was derived from the common line (Burze White), but the population was selected for different traits (oils and proteins) over numerous generations. It is not surprising that the large number of oil loci identified are specific for each analysis population. Therefore, it is desirable to find genetic markers that are specific precursors of the germ plasm used in breeding programs.

<Summary of invention>

The present invention discloses a reliable and predictable breeding method for corn with increased kernel oil concentration. This method

a) s1375, s1384, s1394, s14l6, s1422, s1432. to select corn plants from the maize breeding population by marker-assisted selection; s1457, s1480, s1476, s1478, s1484, s1500, s1513, s1529, s1544, s1545, s1630, s1633, s1647, s1750, s1756, s1757, s1767, s1772, s1774, 13, 18, 16, 18 s1853, s1860, s1870, s1921, s1922, sl925, s1931, s1933, s1939, s1946, s1949, s2O54, s2O55, s2O57, s2O58, s2O97, s2122, s2125, s2156 and s2125 Using markers,

b) crossing the selected corn plant with a second corn plant

Included, characterized in that the crossed offspring exhibit an increased kernel oil concentration. Alexo hybrid populations or their offspring are preferred sources of high concentrations of oil corn germplasm.

Disclosed is a method for finding a corn plant or corn lineage for use as a parent for breeding populations.

a) s1375, s1384, s1394, s14l6, s1422, s1432. s1457, s1480, s1476, s1478, s1484, s1500, s1513, s1529, s1544, s1545, s1630, s1633, s1647, s1750, s1756, s1757, s1767, s1772, s1774, 13, 18, 16, 18 s1853, s1860, s1870, s1921, s1922, sl925, s1931, s1933, s1939, s1946, s1949, s2O54, s2O55, s2O57, s2O58, s2O97, s2122, s2125, s2156 and s2125 Determining the genotype of a corn plant or maize lineage with a marker,

b) identifying corn plants or corn lines that are expected to produce a transgressive segregant for kernel oil concentration based on their genotypes

It describes a method of identifying a corn plant or corn line for use as a parent for the production of breeding population, including.

The present invention provides a method for identifying and identifying genes that control increased kernel oil concentrations of corn. These oil alleles were initially found in individuals composed or derived from the Alex hybrid breeding group. It is also a method of facilitating the use of high concentration oil materials in breeding programs for the purpose of developing new high concentration corn germplasm.

In particular, the method can use genetic markers to predict phylogenetic oil breeding values in the maize seedling program. Indirect selection of oil loci using such markers selects lines with the greatest genetic potential for increased kernel oil concentrations.

In accordance with the present invention, any form of genetic marker may be used to confirm association with kernel oil concentration. The present invention is only limited by the ability to measure polymorphism at a given marker locus. Those skilled in the art will appreciate that various genetic markers that can be used include, but are not limited to, restriction enzyme fragment polymorphisms (RFLPs), random amplified polymorphic DNA (RAPD), single sequence repeat (SSR), AFLP, various single base pair detection methods, allo It will be appreciated that it includes both azyme and phenotype markers. SSR markers useful in the practice of the present invention are s1375, s1384, s1394, s14l6, s1422, s1432. s1457, s1480, s1476, s1478, s1484, s1500, s1513, s1529, s1544, s1545, s1630, s1633, s1647, s1750, s1756, s1757, s1767, s1772, s1774, 13, 18, 16, 18 s1853, s1860, s1870, s1921, s1922, sl925, s1931, s1933, s1939, s1946, s1949, s2O54, s2O55, s2O57, s2O58, s2O97, s2122, s2125, s2156,

A further embodiment of the invention is a transgenic locus that regulates the expression of corn kernel oil concentration. These loci are found and defined (mapped) by the marker locus of the present invention.

Additional embodiments of the invention are corn plants and high concentration oil corn germplasm produced using this breeding method.

<Detailed Description of the Invention>

Table 1 provides a brief description of the genetic markers that form part of the present invention. Each marker is defined by its constitutive nucleic acid primers (forward and reverse) that facilitate the amplification of specific marker loci of the corn genome. In addition, the identifier required for each sequence is shown. The identifiers listed in Table 1 are found in 37 C.F.R.§1. As required by 821 et seq, they correspond to those recorded in the sequence listing below.

Gene markers useful for determining the locus of the genetic locus that controls corn kernel oil concentration

For the purposes of the present invention we define the following terms.

Maize: any variety, cultivars or Zea mays L. population

Elite: The term is characterized by plants or varieties having advantageous traits such as, but not limited to, good grain quality and disease resistance. It can be used to produce commercially advantageous seeds or grains. The term also refers to the parent from which this plant or variety is derived.

Concentrated oil corn germplasm: This term is used as one of the male or female parents in various cross-breeding combinations, or increased kernel of oil when compared to kernel oil produced by a high concentration of oil germplasm at self-pollination. It is characterized by corn plants that produce oil. Examples of high concentration oil corn germplasm are, but are not limited to, open polluted varieties, hybrids, hybrids, breeding lines, species, and corn plants or populations derived from one of the above.

Variety or cultivar: This term refers to a similar plant group that can be identified as another variety or cultivar within the same class by structural features and performance.

Lineage: This term refers to a population from a common lineage, and refers to a group defined more narrowly than a variant.

Hybrid species: The term refers to a group of genetically heterologous plants of known lineage produced by crossing any breed, hybrid, variety, colony, species or other hybrid species in any combination.

Homogeneous species: This term refers to almost homogeneous individuals, varieties or strains.

Recombinant Homogeneous Species: Independently derived lineages developed by repeatedly self-pollinating each generation until full homozygosity is achieved. Each recombinant allogeneic species is derived from a single F2 plant using a breeding method commonly referred to as single seed offspring.

Breeding: Technology and science that improves plant or animal species through controlled genetic engineering.

Marker-Assisted Selection: Use of genetic markers to find and select plants with good phenotypic potential. Genotypes are identified at the transgenic locus by binding between the marker locus and the transgenic locus using predetermined genetic markers to be associated with the transgenic locus or the transgenic loci. Plants containing preferred trait alleles are selected based on their genotype in the associated marker gene sin.

Alexo hybrids: Reproduced high concentration oil corn germplasm developed by Denton Alexander of the University of Illinois. Alexo mating High concentration oil maize germplasm consists of multiple mating groups defined by their previous generation in reproducible selective breeding programs.

Breeding group: A group of genetically heterologous plants for finding one or more individuals with desirable phenotypic characteristics.

Phenotype: Observed expression of one or more plant traits.

Phenotypic Value: A measure of the expected expression of an allele at a transgenic locus. The phenotype of an allele as compared to another allele at the transgenic locus depends on its expression capacity. The phenotype value of an individual, and its phenotypic potential, is based on its total genotype composition at all loci for a given trait.

Transgressive segregant: An entity whose phenotype exceeds the phenotypic variation expected by both parents.

Genetic Marker: Any morphological, biochemical or nucleic acid based on differences in phenotypes representing DNA polymorphism. Examples of genetic markers include, but are not limited to, RFLP, RAPD, allozyme, SSR, and AFLP.

Marker locus: A genetically defined position of DNA polymorphism as revealed by genetic markers.

Transgenic locus: A genetically defined position for one or more gene (allele) groups that contribute to an observed characteristic.

Genotype: Individual allele composition at the locus under study.

Restriction Fragment Length Polymorphism (RFLP): DNA-based genetic markers in which size differences of DNA produced by restriction endonucleases are observed through hybridization (Botstein, D. et al. 1980). Am. J. Hum. Genet. 32: 314-331].

Random Amplified Polymorphic DNA (RAPD): DNA amplification based genetic markers using random primers of short sequence, resulting amplification products are size-separated, and differences in amplification patterns observed (Williams JGK et al. 1990). Nucleic 4cids Res. 18: 6531-6535]).

Simple Sequence Repeat (SSR): Genetic markers based on DNA amplification where short stretches of tandem repeated sequence motifs are amplified, resulting amplification products are separated by size and length differences of nucleotide repeats are observed (Tautz D. 1989. Nucleic Acids Res. 112: 4127-4138].

AFLP: Gene markers based on DNA amplification, wherein the DNA fragments generated by restriction nucleotides are bound to short DNA fragments that facilitate amplification of restricted DNA fragments (Vos, P. et al. 1995. Nucleic Acids Res. 23: 4407-4414]. Amplified fragments are separated by size and differences in amplification patterns are observed.

Allozyme: Enzyme variants isolated by electrophoresis and detected through staining for enzyme activity (Stuber, C.W. and M.M. Goodman. 1983. USDA Agric. Res. Results, Southern Ser., No. 16).

The present invention relates to the discovery of transgenic loci that control kernel oil concentration using genetic markers. In populations with variants for both kernel oil concentrations and gene marker alleles, oil measurements and marker-based genotypes were generated for population members. The minimum squares method was used to determine the location of the oil concentration locus for markers genetically associated with this transgenic locus. Indirect selection of preferred oil alleles can be performed using this information in one or more linked gene markers. The corn plants selected include one or more alleles that encode a high concentration oil phenotype.

It is appreciated that several different populations and population types can be used to find important trait loci. Some population forms are, but are not limited to, recombinant homologous breeding species, backcrossing species, F2 or their self pollination or intercrossing induced species and hybrids. It is also understood that other methods of measuring phenotype and genotype variation in a population are measurements of genotype and phenotype between populations. In this alternative, the second population is a selected derived species of the first population and the selection can be either an important trait (phenotype selection) or a specific marker allele (genotype selection). In addition, one skilled in the art knows that another statistical approach can be used to determine the binding relationship between the marker locus and the transgenic locus.

In addition, the present invention is defined in the following examples. It is to be understood that these examples, which represent preferred embodiments of the present invention, are for illustration only. Those skilled in the art from the foregoing discussion and these examples are claiming important features of the present invention without departing from the spirit and scope of the invention, and various modifications and variations are made to the present invention to enable its use in a variety of uses and conditions.

<Example 1>

Find loci conferring increased kernel oil concentration

Cluster Development and Characterization

LH119wx and LH51, two types of allogeneic maize strains developed by Holden Foundation Sid, Williamsburg, Iowa, USA, were independently bred with plant objects from breeding group ASKC28wx (American Type Culture Collection (Rockville, MD); Identification number ATCC75105) (The wax kernel is usually referred to as ASKC28 and I myself state that ASKC28 is waxy as well). The F2 group produced by self-crossing F1 plants was grown. The kernel, derived from the self-breeding of each F2 plant, was developed using six offspring, with one offspring, through six generations. Up to 20 kernels were grown and selfcrossed from six generations to produce seventh generation ear species representing each recombinant native lineage. Near-infrared light was used to determine the oil values for each group of ears within a certain class (Williams, PC (I 987) In: Near Infrared Technology in the Agricultural and Food Industries; PC Williams and C. Norris, eds). American Association of Cereal Chemists]).

Determination of genotype

Ten seeds were germinated on a damp filter paper from one ear representing each of 194 (LH119wx x ASKC28wx) or 204 (LH51 x ASKC28wx) recombinant homologous breeding lineages. Root portions were cut from the germinated seeds, collected for each ear and extracted using an automated DNA extractor. The instrument is Murray, M. Murray, M.G. and Thomson, W. The CTAB method of Thampson, W. F. was modified (NucL Acids Res. 8: 4321-4325 (1980)). DNA samples were quantified via fluorescence using YoPro-I ™ iodide (Molecular Probes, Inc., Eugene. OR) and diluted to 4 μg / ml.

The SSR region for each DNA sample was analyzed by the following protocol.

1. Add 10 [mu] l of amplification cocktail (see Table 2) to 5 [mu] l (20 ng) of extracted DNA;

2. Amplifying a DNA fragment (US Pat. No. 4,683,202 and US Pat. No. 4,68,195) using the following protocol using a protocol complementary to a primer present in the amplification cocktail;

1) Repetition of 45 seconds at 95 ° C., 50 seconds at 54 ° C. and 80 seconds at 72 ° C.

2) 300 seconds once at 72 ℃

3. About 8 μl of each sample was added drop wise to an agarose gel consisting of 2% Metaphor (FMC, Rockland, Mine), 1 × TBE and 0.5 ug / ml of Ethium bromide, and 1 × TBE buffer and Electrophoresis at 6.1 V / cm for 2 hours in a horizontal electrophorator filled with 0.5 ug / ml of ethidium bromide; And

4. Visualizing by irradiating the DNA band with UV fluorescence.

Amplified cocktail reagent Stock concentration Final concentration Buffer * 10 X 10 X dNTPs 2 mM 0.3 mM Forward primer 40 μM 0.45 μM Reverse primer 40 μM 0.45 μM AmpliTaq polymerase 5 U / μl 0.05 U / μl * 10X buffer is a solution of pH 9.0 consisting of 800 mM Tris-OH, 200 mM (NH ₄ ) ₂ SO ₂ and 25 mM MgCl ₂

Oil locus

133 polymorphic SSR marker genes were used to determine the genotype of recombinant allogeneic species from LH119wxx ASKC28wx crosses, and 103 polymorphic SSR marker genes were used to genotype the LH51 x ASKC28wx-derived group. It also maps both populations to 22 publicly available polymorphic SSR loci with already established chromosome locations and covering all 10 maize chromosomes (from Research Genetics, Huntsville, Alabama, USA). It was.

Genetic associations and distances between marker loci are independently for each group using MAPMAKER 3.0 (Lincoln S, E., et al. (1993) Whitehead Inst. Biomad. Res., Cambridge. MA). Decided. As a result, 10 associated groups were established for each group corresponding to 10 corn chromosomes. Each linkage group corresponded to chromosomes based on binding to known SSR markers. Each of the 23 and 10 markers in the LH119wx x ASKC28wx and LH51 x ASKC28wx populations did not correspond to chromosomal location because no genetic association was clearly established.

Mutation analysis was used to find marker loci associated with transgenic loci that confer increased oil concentration. Oil concentrations were used as dependent variables, and each marker locus was used as an independent variable, using SAS Proc GLM (Edwards, MD, et al. (1987) Genetics 116: 113-125 ]) To calculate a separate ANOVA. Therefore, the mean oil values of the marker allele types were compared for each Anova test. Marker locus was significant when p <0.05.

Associated data for significant marker loci were examined to determine both the number of transgenic loci and their expected positions. Marker loci that are significant for the same linkage group may find the same or different transgenic loci. Phenotypic variation described by each marker locus along the chromosome was carefully examined to determine the number of transgenic loci for the binding group. Significant marker loci uninterrupted by the marker locus, which is also not significant for the same binding group, have been found to find the same transgenic locus on the chromosome.

To confirm the number of transgenic loci, marker data and oil data for the binding group were also analyzed by Mapmaker / QTL 1.0 (Lincoln, SE et al. (1990) Whitehead Inst. Biomed. Res., Cambridge, MA]). The results of mapmaker / QTL were consistent with the initial analysis of the number of transgenic loci for each chromosome.

Eleven and twelve loci regulating kernel oil concentration were located in the LH119wx × ASKC28wx and LH51 × ASKC28wx recombinant allogeneic populations, respectively. Each oil locus is defined by one or more marker loci.

If the same marker locus is used in both populations, binding of the binding groups is possible. In most cases both groups were found to exhibit the same oil locus. Considering the common marker loci, a total of 17 loci that regulate kernel oil concentration were found. Each oil locus was assigned to any letter designation (Table 3)

`` Marker locus genetically binding to and prognosticing the locus of the locus that confers increased kernel oil concentration ''

If comparable, oil loci identified in one population were identified at the same location in the second population. With two exceptions, oil loci were found in one population but not in the second. In the first case, alleles with a positive oil effect were found in LH51, so it would be surprising to identify the same locus in the LH1199wx x ASKC28wx population. In the second case, different ASKC28wx-derived marker alleles were isolated from the population, and thus the oil effect of the ASKC28wx-allele at the transgenic locus was measured in each population. The most abundant ASKC28wx oil allele isolated from LH119wxx ASKC28wx had a positive oil effect on other LHI 19 induced alleles, whereas the LH51 x ASKC28wx population, abundant ASKC28wx alleles, did not have a positive oil effect. Except for the oil locus bound to marker sl480, all alleles with a positive effect on oil concentration were derived from ASKC28wx.

<Example 2>

Marker-assisted selection of breeding lines using genetic markers for increased kernel oil concentration

Genetic markers associated with oil transgenic loci are a high precursor to oil concentration and thus could be used as an indirect measure of kernel oil in a marker-assisted selection program. Thus, genotyping information from associated marker loci will be readily used to select breeding lines with increased oil concentrations. Direct oil measurement cannot distinguish between various genotype transgenic loci compositions and their phenotypic effects. This approach is problematic in early generations of breeding groups with limited oil loci.

For example, the purpose of the maize breeding program is to create a new good breeding line with alleles of traits that give increased kernel oil concentrations. These trait alleles will be introduced by crossing the high concentration oil germplasm with one or more good allogeneic maize. The resulting hybrids can be self-pollinated to generate F2 groups for the purpose of initiating conventional lineage breeding programs (Allard, RW (1960) Principles of Plant Breeding. John Wiley & Sons, Inc. New York. 128]).

To identify individuals among these F2s with the desired genotype, plant tissues were collected from each F2 individual in the population and genotyped with the SSR marker loci listed in Table 1. These F2 individuals with the highest number of SSR marker alleles derived from high concentration oil sources were selected and selected based on agricultural red crosses. With continued homocrossing and screening, the oil locus in a crossbreeding state can be immobilized with high or low oil alleles. Therefore, in order to further isolate the breeding lineage based on the marker allele and thus the oil allele composition, genotyping and separation of the next generation material should be performed.

Because the number of oil alleles is not properly recovered depending on population size and contingency, lineage breeding programs may not be able to demonstrate sufficient agricultural competition or sufficient kernel oil expression. Thus, these new breeding species can be used as parent material to initiate new breeding projects. SSR markers can be reused for oil sorting as described.

It is already apparent to those skilled in the art that many variations of the additional screening method can be planned. Selection will be based on the allele composition of one or more marker genes that locate the transgenic oil locus present in the population. Further selection will be made by screening the genotypes of plant individuals, species or their offspring. Genotype information can be used to develop a variety of predictive models, which can develop a variety of selectable markers. These models allow to measure the effects predicted by the marker locus. The reason is that the predictive value of each marker gene depends on the expression of the transgenic locus as well as the genetic distance from the corresponding transgenic locus. A combination of phenotypic and genotypic screening methods can also be performed.

The marker locus presented herein is a precursor of the oil locus in the Alexo crosses. ASKC28wx represents 28 generations of oil crossing in a genetically closed population, with the previous crossing cycle consisting of the same oil locus. It is expected that these generations will simply differ in their allele number at the identified oil loci. Thus, marker loci disclosed herein in crossbreeding derived from early Alleso generations will be useful for finding oil loci and predicting oil concentrations.

<Example 3>

Identification of corn plants for use as parents for generating overexpression isolates for increased kernel oil concentration

It is important to identify corn plants and lines that, when used as parents, are most likely to produce offspring with good performance. Overexpression isolate progeny of such parents will be the result of parent crossing with a complementary allele set that confers a high concentration oil phenotype. Using the information provided herein, marker alleles are known that predict desirable transduction performance (ie, high concentration oils) at certain marker loci. By determining the genotypes of the lines at such marker loci, the value of those lines as parents is revealed. For example, if one wants to make an individual that contains a good allele at each of the five oil loci (AEs), a parent consisting of the preferred alleles for loci A, B and C can be found to find alleles B, D and It can be crossed with a parent consisting of the preferred allele in E. These parents are complementary because they enable the recovery of offspring containing the desired allele at all five loci. Ideally, the parents were chosen to ensure maximum complementarity at mating so that a large number of desired recombinants were recovered.

Claims (20)

a) s1375, s1384, s1394, s14l6, s1422, s1432. s1457, s1480, s1476, s1478, s1484, s1500, s1513, s1529, s1544, s1545, s1630, s1633, s1647, s1750, s1756, s1757, s1767, s1772, s1774, 13, 18, 16, 18 s1853, s1860, s1870, s1921, s1922, sl925, s1931, s1933, s1939, s1946, s1949, s2O54, s2O55, s2O57, s2O58, s2O97, s2122, s2125, s2156 and s2125 Selecting corn plants from the maize breeding population by marker-assisted selection using a marker, b) crossing the selected corn plant with a second corn plant Wherein the crossed offspring corn plants exhibit increased kernel oil concentration. The method of claim 1, wherein the selected corn plants are members of an Alexho breeding population or progeny thereof. a) s1375, s1384, s1394, s14l6, s1422, s1432. s1457, s1480, s1476, s1478, s1484, s1500, s1513, s1529, s1544, s1545, s1630, s1633, s1647, s1750, s1756, s1757, s1767, s1772, s1774, 13, 18, 16, 18 s1853, s1860, s1870, s1921, s1922, sl925, s1931, s1933, s1939, s1946, s1949, s2O54, s2O55, s2O57, s2O58, s2O97, s2122, s2125, s2156 and s2125 Determining the genotype of a corn plant or maize lineage with a marker, b) identifying corn plants or corn lines that are expected to produce a transgressive segregant for kernel oil concentration based on their genotypes Containing, Corn plant or corn line for use as a parent for the production of breeding populations identification method. A transgenic locus regulating kernel oil concentration, mapped by a genetic marker selected from the group consisting of s2O54, s1647, s1500, s1545, s1774 and s2O97. A transgenic locus regulating kernel oil concentration, mapped by a genetic marker selected from the group consisting of s1817 and s2O57. A transgenic locus regulating kernel oil concentration, mapped by a genetic marker selected from the group consisting of s1860, s1931, s2l5O and sl925. A transgenic locus that regulates kernel oil concentration, mapped by a genetic marker selected from the group consisting of sl457, s2O55, s1757, s2l25, s1780, s1375, s1797, s1416, s1432 and s1921. A locus regulating kernel oil concentration concentration, mapped by a genetic marker selected from the group consisting of s1544, s1633, s1384, s1813, s1767, s2O58, s1933, s1513 and s1484. A transgenic locus regulating kernel oil concentration, mapped by a genetic marker selected from the group consisting of s1476, s1772, s1816, s2l22 and s1836. A transgenic locus regulating kernel oil concentration, mapped by a genetic marker selected from the group consisting of s1939 and s1946. A transgenic locus regulating kernel oil concentration, mapped by a genetic marker selected from the group consisting of s1478, s1853 and s1949. A transgenic locus regulating kernel oil concentration, mapped by a genetic marker selected from the group consisting of sl630, sl422 and s2l56. A transgenic locus regulating kernel oil concentration, mapped by gene marker s1756. Transgenic locus regulating kernel oil concentration, mapped by gene marker s1922. Transgenic locus regulating kernel oil concentration, mapped by gene marker s1529. Transgenic locus regulating kernel oil concentration, mapped by gene marker s1394. Transgenic locus regulating kernel oil concentration, mapped by gene marker s1750. A transgenic locus regulating kernel oil concentration, mapped by gene marker s1870. Transgenic locus regulating kernel oil concentration, mapped by gene marker s2l75. Corn plants produced by the method of claim 1 exhibiting increased kernel oil concentration.