CA2280933A1 - A method to identify and breed corn with increased kernel oil concentration - Google Patents
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Abstract
A method for breeding with high oil corn germplasm is disclosed. The method involves the use of genetic markers associated with trait loci controlling kernel oil concentration. These genetic markers are used to select for kernel oil concentration in breeding populations. Also disclosed is a method for selecting complementary oil parent sources using genetic markers, which are likely to produce superior offspring. Also disclosed are the trait loci controlling corn kernel oil concentration.
Description
TITLE
A METHOD TO IDENTIFY AND BREED CORN WITH
INCREASED KERNEL OIL CONCENTRATION
FIELD OF INVENTION
The invention is in the fields of plant breeding and molecular biology.
More specifically, the invention relates to the identification of corn loci conferring increased kernel oil concentration using genetic markers and the use of genetic markers as an aid to the identification and breeding of corn with increased kernel oil concentration.
BACKGROUND OF INVENTION
Corn is a major crop used as a human food source, an animal feed, and as a source of carbohydrate, oil, protein, and fiber. It is principally used as an energy source in animal feeds, or as a raw material for the recovery of starch, protein feed fractions, fiber, flaking grits, flour, and oil.
Most commercial corn produced throughout the United States is produced from hybrid seed. The production of corn hybrids requires the development of elite corn inbreds that upon intermating produce agronomically superior hybrids.
During the development of corn inbreds, plant breeders select for a number of different traits affecting agronomic performance. These traits include but are not limited to stalk strength, lodging, disease resistance, grain moisture and grain yield. Agronomic traits tend to be quantitatively measured with continuous rather than discrete distributions. It is theorized that quantitative traits are controlled by several genes with small and generally equivalent effects. Further, the observed phenotype is due partially to this genetic component and an environmental component.
The heritability of a trait is defined in the broad sense as the ratio of the genetic variance to the total phenotypic variance. Many agronomic traits display low heritability; i.e., the performance of parent plants is a poor predictor of offspring performance. Thus, traits with low heritability have small genetic variance components in comparison with observed variation. The impact on the plant breeder is that in breeding populations, the value of a plant's genetic composition is difficult to determine from agronomic trait measurements. In an attempt to maximize their discriminative abilities, breeders collect multiple measurements both from individuals related by descent and from many environments. This strategy is resource intensive because it involves the use of extensive trialing to make even small gains in plant improvement. This, coupled with the fact that improved corn lines are selected for multiple traits simultaneously, makes the development of superior corn inbreds both a time-consuming and an expensive labor.
The addition of novel traits in a corn breeding program imposes an additional burden upon the plant breeder. Depending upon the genetic complexity of the novel trait (i.e., single gene versus many genes), a signif cant increase in time and effort is required to produce elite lines containing novel traits.
One such trait is kernel oil concentration.
Com with increased kernel oil concentration is important because it possesses improved feeding value for poultry (Han Y. et ai. ( 1987) Poultry Sci.
66:103-111 ) and livestock (Nordstrom, J. W. et al. ( 1972) J. An. Sci 3.5(2):357-361 ). Grain from conventional corn hybrids typically contains 4%
oil.
In an effort to increase the kernel oil concentration, a long-term recurrent selection program was initiated in the open-pollinated cv. Burr's White by C.G. Hopkins in 1896. This recurrently-selected population known as Illinois High Oil (IHO), has been selected for increased oil concentration for over ninety generations (Dudley, J.W. and R.J. Lambert. (1992) Maydica 37:1-7) using modified mass selection.
As a result, oil concentration was increased in the population over 20%. The germplasm was little used because derived materials had yields substantially lower than conventional varieties (Alexander, D.E. (1988) In: Proc. 43rd Ann.
Com and Sorghum Res. Conf. Am. Seed Trade Assoc., Washington, D.C.
pp 97-105).
Using thirty-eight open-pollinated cultivars and synthetics, Alexander initiated a second recurrent selection program (Alexho synthetic) to increase kernel oil (Alexander, D.E. (1988) In: Corn and Corn Improvement. G.F. Sprague and J.W. Dudley eds. American Society of Agronomy, Madison WI. Pp 869-880).
Equivalent oil levels to IHO were achieved in twenty-eight cycles using selection based upon the oil concentration of single ears and in later generations based upon the oil concentration of single kernels. Yield performance of Alexho-derived material in single cross hybrids (high oil inbred x conventional inbred) is improved over IHO, presumably due to the greater genetic variability initially available, although performance was not equivalent to conventional hybrids.
The development of agronomically elite corn germplasm also containing increased kernel oil concentration is clearly a challenge using conventional plant breeding methods.
Kernel oil concentration can be phenotypically measured using a variety of analytical methods. Oil concentration displays a non-discrete distribution, 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 oil is limited in most of these lines because it is impossible to discriminate between lines based upon their true genetic composition. This situation is further aggrevated when simultaneous selection for agronomic performance is practiced. It would therefore be advantageous to base selection upon the genotype of the plants in the population.
Genetic markers, especially nucleic acid markers, may be used to advantage as an indirect selection method for complex quantitative traits. Genetic markers identifying alleles conferring increased oil would therefore be an advantageous tool for plant breeding programs developing elite high oil corn germplasm.
There is limited published information on the identification of genetic markers predictive for increased oil yield. Kahler (Kahler, A.L. (1985) In:
Proc.
40th Ann. Corn and Sorghum Res. Conf. Am. Seed Trade Assoc., Washington D.C. pp. 66-89) measured isozyme allelic frequency changes following twenty-five cycles of selection in Alexho synthetic and found eight significant loci.
Most of these allele frequency changes were also significant for tests measuring random genetic drift, making it difficult to conclude that selection based upon these isozyme alleles would be useful. More recently Goldman et al. (Goldman, LL., et al. (1994) Crop Sci. 34:908-915) and Berke and Rocheford (Berke, T.G. and Rocheford, T.R. (1995) Crop Sci. 35:1542-1549) used RFLP markers to identify significant marker loci associated with oil concentration in the Illinois long-term selection populations. These studies identified twenty-five and thirty-one markers respectively, in populations derived from Burr's White, which were significantly associated with increased oil. Some of the regions identified by significant RFLP
marker loci may be in common between the two studies, however of the fifteen RFLP markers which were used in both studies, six were in disagreement for their effect on oil concentration. In these studies the populations used were derived from common ancestry (Burr's White); however, the populations were selected for different traits (oil and protein) over many generations. It is not surprising that many identified oil loci would be unique to each population analyzed. It is therefore desirable to identify those genetic markers which are uniquely predictive of getznplasm being used in the breeding program.
SUMMARY OF INVENTION
A method is disclosed for reliably and predictably breeding for corn with increased kernel oil concentration. The method comprises a) using one or more genetic markers to select a corn plant from a corn breeding population by marker-assisted selection, wherein the genetic markers are selected from the group consisting of s 1375, s 13 84, s 13 94, s 1416, s 1422, s 1432, s 1457, s 1480, s 1476, s 1478, s 1484, s 1500, s 1513, s 1529, s 1544, s 1545, s 1630, s 1633, s 1647, s 1750, s 1756, s 1757, s 1767, s 1772, s 1774, s 1780, s 1797, s 1813, s 1816, s 1817, s 1836, s1853, s1860, sI870, s1921, s1922, s1925, s1931, s1933, s1939, s1946, s1949, s2054, s2055, s2057, s2058, s2097, s2122, s2I25, s2150, s2156 and s2175; and b) crossing the selected corn plant with a second corn plant wherein the progeny of the cross displays increased kernel oil concentration. A preferred source of high oil corn germplasm is a member of an Alexho synthetic population or a progeny thereof.
Also disclosed is a method for identifying corn plants or corn lines for use as parents for creation of a breeding population, the method comprising a) genotyping corn plants or corn lines with one or more genetic markers wherein the genetic markers are selected from the group consisting of x1375, x1384, s1394, s I 416, s 1422, s 1432, s I 457, s 1480, s 1476, s 1478, s 1484, s 1500, s 15 I 3, s 1529, s 1544, s 1545, s 1630, s 1633, s 1647, s 1750, s 1756, s 1757, s 1767, s 1772, s 1774, x1780, s1797, x1813, x1816, x1817, x1836, x1853, x1860, s1870, x1921, x1922, x1925, s1931, x1933, x1939, s1946, x1949, s2054, s2055, x2057, x2058, x2097, x2122, x2125, x2150, x2156 and x2175; and b) identifying corn plants or corn lines which, based upon their genotype, are predicted to produce transgressive segregants for kernel oil concentration.
The present invention provides a method for the identification of and selection for genes controlling increased corn kernel oil concentration. These oil alleles were initially identified in materials composed of or derived from the Alexho synthetic breeding populations. Further, the method facilitates the use of this high oil material in breeding programs with the objective of developing new high oil corn germplasm.
Specifically, the method uses genetic markers to predict the oil breeding value of lines in a corn breeding program. By indirect selection of oil loci using these markers, those lines with the greatest genetic potential for increased kernel oil concentration are chosen.
According to the method, any type of genetic marker may be used to identify an association with kernel oil concentration. The method is only limited by the ability to measure polymorphism at a given marker locus. Those skilled in the art will recognize that the various genetic markers which may be used includes but is not limited to restriction fragment length polymorphisms (RFLPs), random amplified polymorphic DNAs (RAPDs), simple sequence repeats (SSRs), AFLPs, various single base pair detection methods, allozymes, and phenotypic markers.
SSR markers useful in the practice of the instant method include s1375, s1384, s 1394, s 1416, s 1422, s 1432, s 1457, s 1476, s 1478, s 1480, s 1484, s 1500, s 1513, s 1529, s 1544, s 1545, s I 630, s 1633, s 1647, s 1750, s 1756, s I 757, s 1767, s 1772, x1774, s1780, s1797, x1813, x1816, x1817, x1836, x1853, x1860, s1870, s1921, x1922, x1925, x1931, x1933, x1939, x1946, s1949, x2054, x2055, x2057, x2058, x2097, x2122, s2125, x2150, x2156 and x2175.
A further embodiment of the present invention are the trait loci controlling the expression of corn kernel oiI concentration. These loci are identified and defined (i.e., mapped) by the marker loci of the present invention.
An additional embodiment of the present invention are corn plants and high oil corn germplasm that are produced using the instant breeding method.
DETAILED DESCRIPTION OF THE INVENTION
Table 1 provides a brief description of the genetic markers that form a part of the instant invention. Each marker is defined by it's constituent nucleic acid primers (forward and reverse) that facilitate amplification of the specific marker locus of the corn genome. Also indicated is the required identifier for each sequence. The identifiers listed in Table 1 correspond to those listed in the Sequence Listing (infra) as required by 37 C.F.R. ~ 1.821 et seq.
Table 1 Genetic markers useful for defining the location of trait loci controllin corn kernel oil concentration Marker Sequence (S'-3') Primer Type SEQ ID NO.
sI375 TTTATGGGTTGGGAGATACTTG forward 1 AGATGTGTGCGTTTZTGAGAG reverse 2 s1384 TTACGGCCTAGACATTTCGAC fo~~d 3 CACTTGCTTTCAGGTACCCA reverse 4 s1394 CTGCCCAGTCCGTAATGAA forward TAGATTTATTTTCTGAACGATTGG reverse 6 s 1416GATCTCTCTGAGGCTTGTCC forward 7 TGTAGTTGAGGATGCTCCC reverse g s 1422AGGCAAGGCTTTCTTCATAC forward 9 CGGACGACGACTGTGTTC reverse 10 s1432 ACATGAGAAACAAGATAGAACCAG forward 11 AAAATGTAAGAACTTGTTTGGGA reverse 12 s1457 ____ _ 13 CTGCTTATTGCTTTCGTCATA forward TGCTGCACTACTTGAACCTAG reverse 14 s 1476ACACAGAGATGACAA.AAGCAA forward 15 GCAGGCGTGCTATGAGAG _ reverse 16 s1478 .-._._ ___- 17 AGCGGTGAAACCCTTATG forward CTGTGGCTGGTTCCTCTC reverse 1 g s1480 GCTCTTGATAAAAAGGCAAGT forward 19 _._._ CTTGTTGTAATGGATGAGTGAG_ reverse 20 __ _ ___ ___ s 1484GCTCGTAGTAGGGGTTACG forward _ GACAGCCTCACCTCAAGA reverse 22 s ACAGATCTTGACACGTACATACC forward 23 GGACGTGTATCCTCAAATCAT reverse 24 s CAGCGAATACTGAATAACGC forward 25 TGTTGGATGAGCACTGAAC reverse 26 s TGTTCTCAACAACCACCG forward 27 CGTTTAGCGATATCATTTTCC reverse 28 _____ s GATCCTACCAAAATCTTATAGGC forward 29 ACAGCTAGCCAAGATCTGATT reverse 30 s CGATACTAATGGAAGCCCTAA forward 31 ATGGCCCATTAAGTTTATCAC reverse 32 s1630AAAGCGTAGTCGGAAAGC forward 33 ACCAATGATCTTTACGCAGAT reverse 34 si633TAATCAGAGCGTACATCAGGA forward 35 AGGGCATCAATCAAGAATG reverse 36 s1647GAGACTTI"TGAGGAGAAAGCA forward 37 GATCAAAAGAGCAAAAGGAGA reverse 38 si750AACTGATGAATACCTTCCCAG forward 39 TGATTAACTTCTCCCTTTGGT reverse 40 s1756TCGGCACAACATATGAGTTAC forward 41 CCCCCATAGAGAGAGATAGAG reverse 42 s1757AAGCACGGCCCAATAGAAT forward 43 AGGATGTCCCTAGCTTTATTG reverse 44 s TCATTGCCCAAAGTGTTG forward 45 CTCATCACCCCTCCAGAG reverse 46 s1772GATCCACGCCATTTAAAC forward 47 TGATACTCTGGTGCATGTTC reverse 48 s1774GATCGCTCCGATCTATCC forward 49 AGCGGCATCTATGTTCTATG reverse 50 s1780CCCAGTGCGAAGAGACTC forward 51 ACACCTGCTCTGCACCAC reverse 52 si797CTAACCCACGACGACCCT forward 53 GCATGAGTGCATGTGCAT reverse 54 si813CTGCCACATGCTTTTCTG forward 55 CTGTAAAGAAGCTGGTCTGGA reverse 56 si816TTCTCCTCATGGATGCGT forward 57 CTATTTGGAAGTATGGGCTTCA reverse 58 s 1817GAGGGCATCTATGTGCAAC . forward GCTCAGAAGTTGCGTTTATG . reverse 60 s 1836TTCCTTCACGTTTCTCTGTTAA _ 61 forward CACATAAACCTAATGGGGTACA reverse 62 s1853 CCCAAAGGCGATACCTATT forward 63 _-__-.__,. CCCACTTTCTCACTCTTTTCT - reverse 6 -__..._. .. ________ s 1860GAGGTGAGTACTATGCAAATGC forward _ _ CAGGCTTACCTAGCCTTCTC reverse 66 s 1870CTATGGATGGCTGCTTGC forward 67 _._____________._."GTCAGGCAGCAGAATGTG __ ___...___reverse6g s1921 AAACCGTCCAGCGACTAC forward GGAAGAACCAATCCCATATCT reverse s1922 AACATCCTGTCGGAAACAG forward 71 TCATCACGTCTCTCTTTCAAC reverse 72 s 1925TTGTGGCAGAATCTCAAATTA forward 73 CGACTGGTGACATGTGAAG reverse 74 s 1931AGTGAGGAAAGAATATGCTGG forward 75 __ TGGACTGAGAAACTGATTTGA reverse 76 s1933 CACAAATGTGAAGGTAAACACT forward 77 AATGGTACGGTTCAGGATG reverse 7g s1939 AGATGACGCACGGAACAC forward 79 _ AGCATCATGTAGCAGGAGG reverse g0 s 1946TTGCAGCACTGTCGTAGTC fo,-~,~.d g 1 GCGCGAGTGGAGTAGTAAG reverse g2 s 1949AAGATTATGCAGATGAGACACC forward g3 GTTCCATGCTTTCCTTGG reverse g4 s2054 GCCGATACCATGTAAGAGAAT forward g5 CTCTGGGCTCTGTGTTAGAGT reverse g6 s2055 CTGCTTTCTCTGTTCCAGC forward g7 _-. __AATCGCTTACTTGTAACCCAC reverse gg s2057 AAGAACGTACGTCCCATAAAG __ _ ____ g9 ' forward CAAGGTAAAGTGACAAAGCAG reverse 90 s2058 GTTCAGGATGAGGCGGAA forward 91 ___. GTGATCATCGCAG_GA_GA_CC_____ reverse 92 ____ s2097 _ __ 93 GGAGCCTGGAGTGAGAAC __ __ forward ' CATGCTCACCTAACGTGG reverse 94 s2122ATCTGAACACTTGAGCAACAA forward 95 ATAGACCGGACCCATCAC reverse 96 s2125CGAACAGCGGGTACACCT forward 97 GAGGTCAGCTTCCTCGATCT reverse __98 __ _ s2150GGAATCGTTCCTCCACAC forward 99 CTTCCTCGGTGTCAGACG reverse _100 _ __ __ s2156ATGGAAACATCAAAGTGGATT forward 101 TGCTACCCTGATGACCTGAT reverse 102 s2175ACCACTAGTCTCATATGAAGGG forward 103 GGTAGGTGGGTAGGGGTT reverse 104 For the purposes of this invention, we define the following terms:
Corn. Any variety, cultivar, or population of Zea mays L.
S Elite. This term characterizes a plant or variety possessing favorable traits, such as, but not limited to high yield, good grain quality, and disease resistance.
This enables its use in commercial production of seed or grain at a profit.
The term also characterizes parents giving rise to such plants or varieties.
High Oil Com Germplasm. This term characterizes corn plants which, when either self pollinated or used as either the male or the female parent in a variety of outcrossing combinations, produce kernels with increased oil when compared to kernels produced by non-high oil germplasm. Examples of high oil corn germplasm include but are not limited to open-pollinated varieties, hybrids, synthetics, inbred lines, races, and populations or corn plants derived from one of the aforementioned.
Variety or cultivar. These terms refer to a group of similar plants that by structural features and performance can be identified from other varieties or cultivars within the same species.
Line. This term refers to a group of individuals from a common ancestry;
a more narrowly defined group than a variety.
Synthetic. This term refers to a genetically heterogeneous collection of plants of known ancestry created by the intermating of any combination of inbreds, hybrids, varieties, populations, races or other synthetics.
Inbred. This term refers to a substantially homozygous individual, variety or line.
Recombinant Inbreds. A population of independently derived lines developed by repeated selfing each generation until complete homozygosity is approached. Each recombinant inbred is derived from a single F2 plant using a breeding method commonly referred to as single seed descent.
Breeding. The art and science of improving a species of plant or animal through controlled genetic manipulation.
Marker-Assisted Selection. The use of genetic markers to identify and select plants with superior phenotypic potential. Genetic markers) determined previously to be associated with a trait locus or trait loci are used to uncover the genotype at trait loci by virtue of linkage between the marker locus and the trait locus. Plants containing desired trait alleles are chosen based upon their genotypes at linked marker loci.
Alexho Synthetic. Recurrently selected, high oil corn germplasm developed by Denton Alexander at the University of Illinois. Alexho synthetic high oil corn germplasm is composed of multiple synthetic populations defined by their cycle of advancement in the recurrent selection breeding program.
Breeding Population. A genetically heterogeneous collection of plants created for the purpose of identifying one or more individuals with desired phenotypic characteristics.
Phenotype. The observed expression of one or more plant characteristics.
Phenotypic Value. A measure of the expected expression of an allele at a trait locus. The phenotypic value of an allele at a trait locus is dependent upon its expressive strength in comparison to alternative alleles. The phenotypic value of an individual, and hence its phenotypic potential, is based upon its total genotypic composition at all loci for a given trait.
Transgressive Segregants. Individuals whose phenotype exceeds the phenotypic variation predicted by the parents.
Genetic Marker. Any morphological, biochemical, or nucleic acid based phenotypic difference which reveals a DNA polymorphism. Examples of genetic markers includes but is not limited to RFLPs, RAPDs, allozymes, SSRs, and AFLPs.
Marker locus. The genetically defined location of DNA polymorphisms as revealed by a genetic marker.
Trait Locus. A genetically defined location for a collection of one or more genes (alleles) which contribute to an observed characteristic.
Genotype. The allelic composition of an individual at genetic loci under study.
Restriction Fragment Length Polymorphism (RFLP). A DNA-based genetic marker in which size differences in restriction endonuclease generated DNA fragments are observed via hybridization (Botstein, D. et al. 1980. Am. J.
Hum. Genet. 3Z: 314-331.
Random Amplified Polymorphic DNA (RAPD}. A DNA amplification-based genetic marker in which short, sequence arbitrary primers are used and the resulting amplification products are size separated and differences in amplification patterns observed (Williams J.G.K. et al. 1990. Nucleic Acids Res. 18:631-6535).
Simple Sequence Repeat (SSR). A DNA amplification-based genetic marker in which short stretches of tandemly repeated sequence motifs are amplified and the resulting amplification products are size separated and differences in length of the nucleotide repeat are observed (Tautz D. 1989.
Nucleic Acids Res. 112:4127-4138).
AFLP. A DNA amplification-based genetic marker in which restriction endonuclease generated DNA fragments are ligated to short DNA fragments which facilitate the amplification of the restricted DNA fragments (Vos, P. et al.
1995. Nucleic Acids Res. 23:4407-4414). The amplified fragments are size separated and differences in amplification patterns observed.
Allozymes. Enzyme variants which are electrophoretically separated and detected via staining for enzymatic 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 trait loci controlling kernel oil concentration through the use of genetic markers. In populations in which variation for both kernel oil concentration and genetic marker alleles exist, oil measurements and marker-based genotypes were generated for members of the populations. Using least squares methods, the locations of oil concentration loci were determined in relation to markers genetically linked to these trait Loci.
Indirect selection of preferred oil alleles may now be practiced using the information at one or more linked genetic markers. Selected corn plants comprise one or more alleles encoding a high oil phenotype.
It is recognized that several different populations and population types could be used to locate trait loci of interest. Some of the population types include but are not limited to recombinant inbreds, backcrosses, F2's or their self pollinated or intermated derivatives, and synthetics. Further, it is understood that an alternative to measuring phenotypic and genotypic variation within populations is the measurement of genotypes and phenotypes between populations. In this alternative the second population is a selected derivative of the first population, selection being either on the trait of interest (phenotypic selection) or on specif c marker alleles (genotypic selection). It is also recognized by those skilled in the art that alternative statistical approaches may be used to determine a linkage relationship between marker loci and trait loci.
EXAMPLES
The present invention is further defined in the following Examples. It should be understood that these Examples, while indicating preferred embodiments of the invention, are given by way of illustration only. From the above discussion and these Examples. one skilled in the art can ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.
LOCATION OF LOCI CONFERRING INCREASED
KERNEL OIL CONCENTRATION
Population development and trait measurement LH119wx and LH51, two inbred corn lines developed by Holden's Foundation Seed Co., Williamsburg, IA were independently intermated with individual plants from the synthetic population ASKC28wx (deposited at the American Type Culture Collection, Rockville, MD; Accession No. ATCC 75105) (waxy kernels are highly represented in ASKC28 and as such I have designated the ASKC28 as being waxy). The F1 plants were selfed and resulting F2 populations were grown. Individual F2 plants were selfed and derived kernels were advanced using single seed decent through six generations of selfing (S6) to produce recombinant inbred lines. Up to twenty kernels from the S6 generation were grown and selfed producing a family of S7 ears representing each recombinant inbred line. Oil values were determined for each ear within a family using near infrared transmitance (Williams, P.C. (1987) In: Near Infrared Technology in the Agricultural and Food Industries; P.C. Williams and C.
Norris, eds. American Association of Cereal Chemists).
Geno is determination Ten seeds from single ears representing each of one hundred ninety-four (LH1 l9wx x ASKC28wx) or two hundred and four (LH51 x ASKC28wx) recombinant inbred lines were germinated on moistened filter paper. Root segments were excised from germinated seeds, pooled for each ear and extracted using an automated DNA extraction machine. The instrument uses a modification of the Murray and Thompson CTAB procedure (Murray, M.G. and Thompson, W.F. (1980) Nucl. Acids Res. 8:4321-4325). DNA samples were quantified via fluorescence using YoPro-1T"" iodide (Molecular Probes, Inc., Eugene, OR) and diluted to 4 pg/ml.
SSR regions for each DNA sample were analyzed using the following protocol:
1. Ten pI of amplification cocktail (see Table 2) was added to ~ pl (20 ng) of extracted DNA;
2. The DNA fragment flanked by sequences complementary to the primers present in the amplification cocktail was amplified by PCR (U.S.
Patent No. 4,683,202 and U.S. Patent No. 4,683,195) using the following protocol:
1 ) ~5 cycles of 50 sec at 95°C, 50 sec at 54°C and 80 sec at 72°C and 2) 1 cycle of 300 sec at 72°C;
A METHOD TO IDENTIFY AND BREED CORN WITH
INCREASED KERNEL OIL CONCENTRATION
FIELD OF INVENTION
The invention is in the fields of plant breeding and molecular biology.
More specifically, the invention relates to the identification of corn loci conferring increased kernel oil concentration using genetic markers and the use of genetic markers as an aid to the identification and breeding of corn with increased kernel oil concentration.
BACKGROUND OF INVENTION
Corn is a major crop used as a human food source, an animal feed, and as a source of carbohydrate, oil, protein, and fiber. It is principally used as an energy source in animal feeds, or as a raw material for the recovery of starch, protein feed fractions, fiber, flaking grits, flour, and oil.
Most commercial corn produced throughout the United States is produced from hybrid seed. The production of corn hybrids requires the development of elite corn inbreds that upon intermating produce agronomically superior hybrids.
During the development of corn inbreds, plant breeders select for a number of different traits affecting agronomic performance. These traits include but are not limited to stalk strength, lodging, disease resistance, grain moisture and grain yield. Agronomic traits tend to be quantitatively measured with continuous rather than discrete distributions. It is theorized that quantitative traits are controlled by several genes with small and generally equivalent effects. Further, the observed phenotype is due partially to this genetic component and an environmental component.
The heritability of a trait is defined in the broad sense as the ratio of the genetic variance to the total phenotypic variance. Many agronomic traits display low heritability; i.e., the performance of parent plants is a poor predictor of offspring performance. Thus, traits with low heritability have small genetic variance components in comparison with observed variation. The impact on the plant breeder is that in breeding populations, the value of a plant's genetic composition is difficult to determine from agronomic trait measurements. In an attempt to maximize their discriminative abilities, breeders collect multiple measurements both from individuals related by descent and from many environments. This strategy is resource intensive because it involves the use of extensive trialing to make even small gains in plant improvement. This, coupled with the fact that improved corn lines are selected for multiple traits simultaneously, makes the development of superior corn inbreds both a time-consuming and an expensive labor.
The addition of novel traits in a corn breeding program imposes an additional burden upon the plant breeder. Depending upon the genetic complexity of the novel trait (i.e., single gene versus many genes), a signif cant increase in time and effort is required to produce elite lines containing novel traits.
One such trait is kernel oil concentration.
Com with increased kernel oil concentration is important because it possesses improved feeding value for poultry (Han Y. et ai. ( 1987) Poultry Sci.
66:103-111 ) and livestock (Nordstrom, J. W. et al. ( 1972) J. An. Sci 3.5(2):357-361 ). Grain from conventional corn hybrids typically contains 4%
oil.
In an effort to increase the kernel oil concentration, a long-term recurrent selection program was initiated in the open-pollinated cv. Burr's White by C.G. Hopkins in 1896. This recurrently-selected population known as Illinois High Oil (IHO), has been selected for increased oil concentration for over ninety generations (Dudley, J.W. and R.J. Lambert. (1992) Maydica 37:1-7) using modified mass selection.
As a result, oil concentration was increased in the population over 20%. The germplasm was little used because derived materials had yields substantially lower than conventional varieties (Alexander, D.E. (1988) In: Proc. 43rd Ann.
Com and Sorghum Res. Conf. Am. Seed Trade Assoc., Washington, D.C.
pp 97-105).
Using thirty-eight open-pollinated cultivars and synthetics, Alexander initiated a second recurrent selection program (Alexho synthetic) to increase kernel oil (Alexander, D.E. (1988) In: Corn and Corn Improvement. G.F. Sprague and J.W. Dudley eds. American Society of Agronomy, Madison WI. Pp 869-880).
Equivalent oil levels to IHO were achieved in twenty-eight cycles using selection based upon the oil concentration of single ears and in later generations based upon the oil concentration of single kernels. Yield performance of Alexho-derived material in single cross hybrids (high oil inbred x conventional inbred) is improved over IHO, presumably due to the greater genetic variability initially available, although performance was not equivalent to conventional hybrids.
The development of agronomically elite corn germplasm also containing increased kernel oil concentration is clearly a challenge using conventional plant breeding methods.
Kernel oil concentration can be phenotypically measured using a variety of analytical methods. Oil concentration displays a non-discrete distribution, 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 oil is limited in most of these lines because it is impossible to discriminate between lines based upon their true genetic composition. This situation is further aggrevated when simultaneous selection for agronomic performance is practiced. It would therefore be advantageous to base selection upon the genotype of the plants in the population.
Genetic markers, especially nucleic acid markers, may be used to advantage as an indirect selection method for complex quantitative traits. Genetic markers identifying alleles conferring increased oil would therefore be an advantageous tool for plant breeding programs developing elite high oil corn germplasm.
There is limited published information on the identification of genetic markers predictive for increased oil yield. Kahler (Kahler, A.L. (1985) In:
Proc.
40th Ann. Corn and Sorghum Res. Conf. Am. Seed Trade Assoc., Washington D.C. pp. 66-89) measured isozyme allelic frequency changes following twenty-five cycles of selection in Alexho synthetic and found eight significant loci.
Most of these allele frequency changes were also significant for tests measuring random genetic drift, making it difficult to conclude that selection based upon these isozyme alleles would be useful. More recently Goldman et al. (Goldman, LL., et al. (1994) Crop Sci. 34:908-915) and Berke and Rocheford (Berke, T.G. and Rocheford, T.R. (1995) Crop Sci. 35:1542-1549) used RFLP markers to identify significant marker loci associated with oil concentration in the Illinois long-term selection populations. These studies identified twenty-five and thirty-one markers respectively, in populations derived from Burr's White, which were significantly associated with increased oil. Some of the regions identified by significant RFLP
marker loci may be in common between the two studies, however of the fifteen RFLP markers which were used in both studies, six were in disagreement for their effect on oil concentration. In these studies the populations used were derived from common ancestry (Burr's White); however, the populations were selected for different traits (oil and protein) over many generations. It is not surprising that many identified oil loci would be unique to each population analyzed. It is therefore desirable to identify those genetic markers which are uniquely predictive of getznplasm being used in the breeding program.
SUMMARY OF INVENTION
A method is disclosed for reliably and predictably breeding for corn with increased kernel oil concentration. The method comprises a) using one or more genetic markers to select a corn plant from a corn breeding population by marker-assisted selection, wherein the genetic markers are selected from the group consisting of s 1375, s 13 84, s 13 94, s 1416, s 1422, s 1432, s 1457, s 1480, s 1476, s 1478, s 1484, s 1500, s 1513, s 1529, s 1544, s 1545, s 1630, s 1633, s 1647, s 1750, s 1756, s 1757, s 1767, s 1772, s 1774, s 1780, s 1797, s 1813, s 1816, s 1817, s 1836, s1853, s1860, sI870, s1921, s1922, s1925, s1931, s1933, s1939, s1946, s1949, s2054, s2055, s2057, s2058, s2097, s2122, s2I25, s2150, s2156 and s2175; and b) crossing the selected corn plant with a second corn plant wherein the progeny of the cross displays increased kernel oil concentration. A preferred source of high oil corn germplasm is a member of an Alexho synthetic population or a progeny thereof.
Also disclosed is a method for identifying corn plants or corn lines for use as parents for creation of a breeding population, the method comprising a) genotyping corn plants or corn lines with one or more genetic markers wherein the genetic markers are selected from the group consisting of x1375, x1384, s1394, s I 416, s 1422, s 1432, s I 457, s 1480, s 1476, s 1478, s 1484, s 1500, s 15 I 3, s 1529, s 1544, s 1545, s 1630, s 1633, s 1647, s 1750, s 1756, s 1757, s 1767, s 1772, s 1774, x1780, s1797, x1813, x1816, x1817, x1836, x1853, x1860, s1870, x1921, x1922, x1925, s1931, x1933, x1939, s1946, x1949, s2054, s2055, x2057, x2058, x2097, x2122, x2125, x2150, x2156 and x2175; and b) identifying corn plants or corn lines which, based upon their genotype, are predicted to produce transgressive segregants for kernel oil concentration.
The present invention provides a method for the identification of and selection for genes controlling increased corn kernel oil concentration. These oil alleles were initially identified in materials composed of or derived from the Alexho synthetic breeding populations. Further, the method facilitates the use of this high oil material in breeding programs with the objective of developing new high oil corn germplasm.
Specifically, the method uses genetic markers to predict the oil breeding value of lines in a corn breeding program. By indirect selection of oil loci using these markers, those lines with the greatest genetic potential for increased kernel oil concentration are chosen.
According to the method, any type of genetic marker may be used to identify an association with kernel oil concentration. The method is only limited by the ability to measure polymorphism at a given marker locus. Those skilled in the art will recognize that the various genetic markers which may be used includes but is not limited to restriction fragment length polymorphisms (RFLPs), random amplified polymorphic DNAs (RAPDs), simple sequence repeats (SSRs), AFLPs, various single base pair detection methods, allozymes, and phenotypic markers.
SSR markers useful in the practice of the instant method include s1375, s1384, s 1394, s 1416, s 1422, s 1432, s 1457, s 1476, s 1478, s 1480, s 1484, s 1500, s 1513, s 1529, s 1544, s 1545, s I 630, s 1633, s 1647, s 1750, s 1756, s I 757, s 1767, s 1772, x1774, s1780, s1797, x1813, x1816, x1817, x1836, x1853, x1860, s1870, s1921, x1922, x1925, x1931, x1933, x1939, x1946, s1949, x2054, x2055, x2057, x2058, x2097, x2122, s2125, x2150, x2156 and x2175.
A further embodiment of the present invention are the trait loci controlling the expression of corn kernel oiI concentration. These loci are identified and defined (i.e., mapped) by the marker loci of the present invention.
An additional embodiment of the present invention are corn plants and high oil corn germplasm that are produced using the instant breeding method.
DETAILED DESCRIPTION OF THE INVENTION
Table 1 provides a brief description of the genetic markers that form a part of the instant invention. Each marker is defined by it's constituent nucleic acid primers (forward and reverse) that facilitate amplification of the specific marker locus of the corn genome. Also indicated is the required identifier for each sequence. The identifiers listed in Table 1 correspond to those listed in the Sequence Listing (infra) as required by 37 C.F.R. ~ 1.821 et seq.
Table 1 Genetic markers useful for defining the location of trait loci controllin corn kernel oil concentration Marker Sequence (S'-3') Primer Type SEQ ID NO.
sI375 TTTATGGGTTGGGAGATACTTG forward 1 AGATGTGTGCGTTTZTGAGAG reverse 2 s1384 TTACGGCCTAGACATTTCGAC fo~~d 3 CACTTGCTTTCAGGTACCCA reverse 4 s1394 CTGCCCAGTCCGTAATGAA forward TAGATTTATTTTCTGAACGATTGG reverse 6 s 1416GATCTCTCTGAGGCTTGTCC forward 7 TGTAGTTGAGGATGCTCCC reverse g s 1422AGGCAAGGCTTTCTTCATAC forward 9 CGGACGACGACTGTGTTC reverse 10 s1432 ACATGAGAAACAAGATAGAACCAG forward 11 AAAATGTAAGAACTTGTTTGGGA reverse 12 s1457 ____ _ 13 CTGCTTATTGCTTTCGTCATA forward TGCTGCACTACTTGAACCTAG reverse 14 s 1476ACACAGAGATGACAA.AAGCAA forward 15 GCAGGCGTGCTATGAGAG _ reverse 16 s1478 .-._._ ___- 17 AGCGGTGAAACCCTTATG forward CTGTGGCTGGTTCCTCTC reverse 1 g s1480 GCTCTTGATAAAAAGGCAAGT forward 19 _._._ CTTGTTGTAATGGATGAGTGAG_ reverse 20 __ _ ___ ___ s 1484GCTCGTAGTAGGGGTTACG forward _ GACAGCCTCACCTCAAGA reverse 22 s ACAGATCTTGACACGTACATACC forward 23 GGACGTGTATCCTCAAATCAT reverse 24 s CAGCGAATACTGAATAACGC forward 25 TGTTGGATGAGCACTGAAC reverse 26 s TGTTCTCAACAACCACCG forward 27 CGTTTAGCGATATCATTTTCC reverse 28 _____ s GATCCTACCAAAATCTTATAGGC forward 29 ACAGCTAGCCAAGATCTGATT reverse 30 s CGATACTAATGGAAGCCCTAA forward 31 ATGGCCCATTAAGTTTATCAC reverse 32 s1630AAAGCGTAGTCGGAAAGC forward 33 ACCAATGATCTTTACGCAGAT reverse 34 si633TAATCAGAGCGTACATCAGGA forward 35 AGGGCATCAATCAAGAATG reverse 36 s1647GAGACTTI"TGAGGAGAAAGCA forward 37 GATCAAAAGAGCAAAAGGAGA reverse 38 si750AACTGATGAATACCTTCCCAG forward 39 TGATTAACTTCTCCCTTTGGT reverse 40 s1756TCGGCACAACATATGAGTTAC forward 41 CCCCCATAGAGAGAGATAGAG reverse 42 s1757AAGCACGGCCCAATAGAAT forward 43 AGGATGTCCCTAGCTTTATTG reverse 44 s TCATTGCCCAAAGTGTTG forward 45 CTCATCACCCCTCCAGAG reverse 46 s1772GATCCACGCCATTTAAAC forward 47 TGATACTCTGGTGCATGTTC reverse 48 s1774GATCGCTCCGATCTATCC forward 49 AGCGGCATCTATGTTCTATG reverse 50 s1780CCCAGTGCGAAGAGACTC forward 51 ACACCTGCTCTGCACCAC reverse 52 si797CTAACCCACGACGACCCT forward 53 GCATGAGTGCATGTGCAT reverse 54 si813CTGCCACATGCTTTTCTG forward 55 CTGTAAAGAAGCTGGTCTGGA reverse 56 si816TTCTCCTCATGGATGCGT forward 57 CTATTTGGAAGTATGGGCTTCA reverse 58 s 1817GAGGGCATCTATGTGCAAC . forward GCTCAGAAGTTGCGTTTATG . reverse 60 s 1836TTCCTTCACGTTTCTCTGTTAA _ 61 forward CACATAAACCTAATGGGGTACA reverse 62 s1853 CCCAAAGGCGATACCTATT forward 63 _-__-.__,. CCCACTTTCTCACTCTTTTCT - reverse 6 -__..._. .. ________ s 1860GAGGTGAGTACTATGCAAATGC forward _ _ CAGGCTTACCTAGCCTTCTC reverse 66 s 1870CTATGGATGGCTGCTTGC forward 67 _._____________._."GTCAGGCAGCAGAATGTG __ ___...___reverse6g s1921 AAACCGTCCAGCGACTAC forward GGAAGAACCAATCCCATATCT reverse s1922 AACATCCTGTCGGAAACAG forward 71 TCATCACGTCTCTCTTTCAAC reverse 72 s 1925TTGTGGCAGAATCTCAAATTA forward 73 CGACTGGTGACATGTGAAG reverse 74 s 1931AGTGAGGAAAGAATATGCTGG forward 75 __ TGGACTGAGAAACTGATTTGA reverse 76 s1933 CACAAATGTGAAGGTAAACACT forward 77 AATGGTACGGTTCAGGATG reverse 7g s1939 AGATGACGCACGGAACAC forward 79 _ AGCATCATGTAGCAGGAGG reverse g0 s 1946TTGCAGCACTGTCGTAGTC fo,-~,~.d g 1 GCGCGAGTGGAGTAGTAAG reverse g2 s 1949AAGATTATGCAGATGAGACACC forward g3 GTTCCATGCTTTCCTTGG reverse g4 s2054 GCCGATACCATGTAAGAGAAT forward g5 CTCTGGGCTCTGTGTTAGAGT reverse g6 s2055 CTGCTTTCTCTGTTCCAGC forward g7 _-. __AATCGCTTACTTGTAACCCAC reverse gg s2057 AAGAACGTACGTCCCATAAAG __ _ ____ g9 ' forward CAAGGTAAAGTGACAAAGCAG reverse 90 s2058 GTTCAGGATGAGGCGGAA forward 91 ___. GTGATCATCGCAG_GA_GA_CC_____ reverse 92 ____ s2097 _ __ 93 GGAGCCTGGAGTGAGAAC __ __ forward ' CATGCTCACCTAACGTGG reverse 94 s2122ATCTGAACACTTGAGCAACAA forward 95 ATAGACCGGACCCATCAC reverse 96 s2125CGAACAGCGGGTACACCT forward 97 GAGGTCAGCTTCCTCGATCT reverse __98 __ _ s2150GGAATCGTTCCTCCACAC forward 99 CTTCCTCGGTGTCAGACG reverse _100 _ __ __ s2156ATGGAAACATCAAAGTGGATT forward 101 TGCTACCCTGATGACCTGAT reverse 102 s2175ACCACTAGTCTCATATGAAGGG forward 103 GGTAGGTGGGTAGGGGTT reverse 104 For the purposes of this invention, we define the following terms:
Corn. Any variety, cultivar, or population of Zea mays L.
S Elite. This term characterizes a plant or variety possessing favorable traits, such as, but not limited to high yield, good grain quality, and disease resistance.
This enables its use in commercial production of seed or grain at a profit.
The term also characterizes parents giving rise to such plants or varieties.
High Oil Com Germplasm. This term characterizes corn plants which, when either self pollinated or used as either the male or the female parent in a variety of outcrossing combinations, produce kernels with increased oil when compared to kernels produced by non-high oil germplasm. Examples of high oil corn germplasm include but are not limited to open-pollinated varieties, hybrids, synthetics, inbred lines, races, and populations or corn plants derived from one of the aforementioned.
Variety or cultivar. These terms refer to a group of similar plants that by structural features and performance can be identified from other varieties or cultivars within the same species.
Line. This term refers to a group of individuals from a common ancestry;
a more narrowly defined group than a variety.
Synthetic. This term refers to a genetically heterogeneous collection of plants of known ancestry created by the intermating of any combination of inbreds, hybrids, varieties, populations, races or other synthetics.
Inbred. This term refers to a substantially homozygous individual, variety or line.
Recombinant Inbreds. A population of independently derived lines developed by repeated selfing each generation until complete homozygosity is approached. Each recombinant inbred is derived from a single F2 plant using a breeding method commonly referred to as single seed descent.
Breeding. The art and science of improving a species of plant or animal through controlled genetic manipulation.
Marker-Assisted Selection. The use of genetic markers to identify and select plants with superior phenotypic potential. Genetic markers) determined previously to be associated with a trait locus or trait loci are used to uncover the genotype at trait loci by virtue of linkage between the marker locus and the trait locus. Plants containing desired trait alleles are chosen based upon their genotypes at linked marker loci.
Alexho Synthetic. Recurrently selected, high oil corn germplasm developed by Denton Alexander at the University of Illinois. Alexho synthetic high oil corn germplasm is composed of multiple synthetic populations defined by their cycle of advancement in the recurrent selection breeding program.
Breeding Population. A genetically heterogeneous collection of plants created for the purpose of identifying one or more individuals with desired phenotypic characteristics.
Phenotype. The observed expression of one or more plant characteristics.
Phenotypic Value. A measure of the expected expression of an allele at a trait locus. The phenotypic value of an allele at a trait locus is dependent upon its expressive strength in comparison to alternative alleles. The phenotypic value of an individual, and hence its phenotypic potential, is based upon its total genotypic composition at all loci for a given trait.
Transgressive Segregants. Individuals whose phenotype exceeds the phenotypic variation predicted by the parents.
Genetic Marker. Any morphological, biochemical, or nucleic acid based phenotypic difference which reveals a DNA polymorphism. Examples of genetic markers includes but is not limited to RFLPs, RAPDs, allozymes, SSRs, and AFLPs.
Marker locus. The genetically defined location of DNA polymorphisms as revealed by a genetic marker.
Trait Locus. A genetically defined location for a collection of one or more genes (alleles) which contribute to an observed characteristic.
Genotype. The allelic composition of an individual at genetic loci under study.
Restriction Fragment Length Polymorphism (RFLP). A DNA-based genetic marker in which size differences in restriction endonuclease generated DNA fragments are observed via hybridization (Botstein, D. et al. 1980. Am. J.
Hum. Genet. 3Z: 314-331.
Random Amplified Polymorphic DNA (RAPD}. A DNA amplification-based genetic marker in which short, sequence arbitrary primers are used and the resulting amplification products are size separated and differences in amplification patterns observed (Williams J.G.K. et al. 1990. Nucleic Acids Res. 18:631-6535).
Simple Sequence Repeat (SSR). A DNA amplification-based genetic marker in which short stretches of tandemly repeated sequence motifs are amplified and the resulting amplification products are size separated and differences in length of the nucleotide repeat are observed (Tautz D. 1989.
Nucleic Acids Res. 112:4127-4138).
AFLP. A DNA amplification-based genetic marker in which restriction endonuclease generated DNA fragments are ligated to short DNA fragments which facilitate the amplification of the restricted DNA fragments (Vos, P. et al.
1995. Nucleic Acids Res. 23:4407-4414). The amplified fragments are size separated and differences in amplification patterns observed.
Allozymes. Enzyme variants which are electrophoretically separated and detected via staining for enzymatic 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 trait loci controlling kernel oil concentration through the use of genetic markers. In populations in which variation for both kernel oil concentration and genetic marker alleles exist, oil measurements and marker-based genotypes were generated for members of the populations. Using least squares methods, the locations of oil concentration loci were determined in relation to markers genetically linked to these trait Loci.
Indirect selection of preferred oil alleles may now be practiced using the information at one or more linked genetic markers. Selected corn plants comprise one or more alleles encoding a high oil phenotype.
It is recognized that several different populations and population types could be used to locate trait loci of interest. Some of the population types include but are not limited to recombinant inbreds, backcrosses, F2's or their self pollinated or intermated derivatives, and synthetics. Further, it is understood that an alternative to measuring phenotypic and genotypic variation within populations is the measurement of genotypes and phenotypes between populations. In this alternative the second population is a selected derivative of the first population, selection being either on the trait of interest (phenotypic selection) or on specif c marker alleles (genotypic selection). It is also recognized by those skilled in the art that alternative statistical approaches may be used to determine a linkage relationship between marker loci and trait loci.
EXAMPLES
The present invention is further defined in the following Examples. It should be understood that these Examples, while indicating preferred embodiments of the invention, are given by way of illustration only. From the above discussion and these Examples. one skilled in the art can ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.
LOCATION OF LOCI CONFERRING INCREASED
KERNEL OIL CONCENTRATION
Population development and trait measurement LH119wx and LH51, two inbred corn lines developed by Holden's Foundation Seed Co., Williamsburg, IA were independently intermated with individual plants from the synthetic population ASKC28wx (deposited at the American Type Culture Collection, Rockville, MD; Accession No. ATCC 75105) (waxy kernels are highly represented in ASKC28 and as such I have designated the ASKC28 as being waxy). The F1 plants were selfed and resulting F2 populations were grown. Individual F2 plants were selfed and derived kernels were advanced using single seed decent through six generations of selfing (S6) to produce recombinant inbred lines. Up to twenty kernels from the S6 generation were grown and selfed producing a family of S7 ears representing each recombinant inbred line. Oil values were determined for each ear within a family using near infrared transmitance (Williams, P.C. (1987) In: Near Infrared Technology in the Agricultural and Food Industries; P.C. Williams and C.
Norris, eds. American Association of Cereal Chemists).
Geno is determination Ten seeds from single ears representing each of one hundred ninety-four (LH1 l9wx x ASKC28wx) or two hundred and four (LH51 x ASKC28wx) recombinant inbred lines were germinated on moistened filter paper. Root segments were excised from germinated seeds, pooled for each ear and extracted using an automated DNA extraction machine. The instrument uses a modification of the Murray and Thompson CTAB procedure (Murray, M.G. and Thompson, W.F. (1980) Nucl. Acids Res. 8:4321-4325). DNA samples were quantified via fluorescence using YoPro-1T"" iodide (Molecular Probes, Inc., Eugene, OR) and diluted to 4 pg/ml.
SSR regions for each DNA sample were analyzed using the following protocol:
1. Ten pI of amplification cocktail (see Table 2) was added to ~ pl (20 ng) of extracted DNA;
2. The DNA fragment flanked by sequences complementary to the primers present in the amplification cocktail was amplified by PCR (U.S.
Patent No. 4,683,202 and U.S. Patent No. 4,683,195) using the following protocol:
1 ) ~5 cycles of 50 sec at 95°C, 50 sec at 54°C and 80 sec at 72°C and 2) 1 cycle of 300 sec at 72°C;
3. Approximately 8 pl of each sample was loaded onto agarose gels composed of 2% Metaphor (FMC Corp., Rockland, ME), 1X TBE, and 0.5 ~g/ml S ethidium bromide, and electrophoresed for 2 h at 6.1 V/cm in horizontal electrophoresis units to which IX TBE buffer and 0.5 ~Cg/ml ethidium bromide was added; and 4. DNA bands were visualized by LJV fluorescence.
Table 2 Amplification Cocktail Reagent Stock Concentration Final Concentration Buffer* 1 OX 1.SX
dNTPs 2 mM 0.3 mM
Forward Primer 40 ~M 0.45 ~M
Reverse Primer 40 ~M 0.45 ~M
AmpliTaq PolymeraseTM 5 U/~1 0.05 U/p.l * 1 OX Buffer is a pH 9.0 solution composed of 800 mM Tris-OH, 200 mM
~4)2SD4~ ~d 25 mM MgCl2.
Localization of oil loci One hundred thirty three polymorphic SSR marker loci were used to genotype the recombinant inbreds from the LH 1 l9wx x ASKC28wx cross and one hundred and three polymorphic SSR marker loci were used to genotype the LH51 x ASKC28wx-derived population. In addition, twenty publicly available polymorphic SSR loci with previously established chromosome locations and covering all ten maize chromosomes (available from Research Genetics, Huntsville, AL) were also mapped in both populations.
Genetic linkage and distance between marker loci was determined independently for each population using MAPMAKER 3.0 (Lincoln S.E., et al.
( 1993) Whitehead Inst. Biomed. Res., Cambridge, MA). This resulted in the establishment of ten linkage groups for each population corresponding to the ten chromosomes of maize. Each linkage group was assigned to a chromosome based upon linkage to the public SSR markers. Twenty-three and ten markers in the LHI l9wx x ASKC28wx and LH51 x ASKC28wx populations, respectively, were not assigned chromosome positions because genetic linkage could not be clearly established.
Analysis of variance was used to identify marker loci in linkage with trait loci conferring increased oil concentration. Oil concentration was used as a dependent variable and separate ANOVAs were calculated with SAS Proc GLM
(SAS Inst., Cary, NC) using each marker locus as a single independent variable (Edwards, M.D., et aI. ( 1987) Genetics 116:113-I25). Therefore, for each ANOVA test the mean oil values of marker allele classes were compared. Marker loci were declared signif cant if p < 0.05.
Linkage data for significant marker loci was examined to determine both the number of trait Ioci present and their probable location. Significant marker loci on the same linkage group are either detecting the same trait locus or alternatively different trait loci. By careful examination of the phenotypic variation explained by each marker locus along the chromosome, a determination of the number trait loci on a linkage group was made. Significant marker loci, on the same linkage group and uninterrupted by non-significant marker loci, were declared to be detecting the same trait locus on the chromosome. If significant marker loci on the same chromosome were interrupted by non-significant marker loci then each significant region was declared to contain a trait locus resulting in multiple trait loci on the same chromosome.
To confirm the number of trait loci, marker data assigned to linkage groups and oil data were also analyzed with Mapmaker/QTL 1.0 (Lincoln, S.E. et al. (1990) Whitehead Inst. Biomed. Res., Cambridge, MA). Results with Mapmaker/QTL were in agreement with the initial analysis for the number trait loci on each chromosome.
Eleven and twelve loci controlling kernel oil concentration were located in the LH1 l9wx x ASKC28wx and LH51 x ASKC28wx recombinant inbred populations, respectively. Each oil locus is defined by one or more linked marker loci.
In instances where the same marker loci were used in both populations, alignment of linkage groups is possible. It was found that in most instances both populations localized the same oil loci. By considering common marker loci, a total of seventeen loci controlling kernel oil concentration were found. Each oil locus was assigned an arbitrary letter designation (Table 3}.
Table 3 Marker loci genetically linked to and predictive of the location of trait loci conferring increased kernel oil concentration Oil locus Chromosome Marker loci A I s1922 B I s 1478, s I 853, s I 949 C I S I 860, s 1925, s 1931, s2150 D 2 s2175 E 3 s1394 F 4 s 1476, s 1772, s 1816, s2122, s 1836 G 4 s1939,s1946 H 4 s1870 I 5 s1529 J ~ s2054, s 1647, s 1500, s 1 X45, s 1774, s2097 K 6 s1457, s2055, s1757, s2125, s1780, s1375, s 1797, s 1416, s 1432, s 1921 L 7 s1630, s1422, s2156 M 8 s1817,s2057 N 9 s1544, s1633, s1384, s1813, s1767, s2058, s1933,s1513,s1484 O 10 s1756 P 10 s1480 (positive oil allele in LH51) Q N.A.* s1750 *N.A. - chromosome location not known In instances where comparisons could be made, oil loci which were identified in one population were identified at the same location in the second population. In two exceptions, an oil locus was found in one population, but not in the second population. In the first case, the allele with a positive oil effect was found in LH51 and thus it would be unexpected to identify the same locus in the LH 119wx x ASKC28wx population. In the second case, it was found that different ASKC28wx-derived marker alleles were segregating in the populations;
therefore, each population was measuring the oil effect of a different ASKC28wx allele at the trait locus. The most abundant ASKC28wx oil allele segregating in LH 119wx x ASKC28wx had a positive oil effect versus the alternative LH 119-derived allele, whereas in the LH51 x ASKC28wx population, the abundant ASKC28wx allele had no positive oil effect. With the exception of the oil locus linked to marker s1480, all alleles with positive effects on oil concentration were derived from ASKC28wx.
MARKER-ASSISTED SELECTION OF BREEDING LINES USING GENETIC
MARKERS FOR INCREASE KERNEL OIL CONCENTRATION
Genetic marker loci in linkage with oil trait loci are highly predictive of oil concentration and as such may be used as an indirect measurement of kernel oil in a marker-assisted selection program. Accordingly, genotypic information from linked marker loci would facilitate the selection of breeding lines with increased oil concentration. Direct oii measurements cannot differentiate between various genotypic trait locus compositions with equivalent phenotypic effects. This is especially problematic in early generation segregating breeding populations where only limited fixation of oil loci has occurred.
By way of example, an objective of a corn breeding program could be the creation of new elite inbred lines which contain trait alleles conferring increased kernel oil concentration. These trait alleles would be introduced by the intermating of high oil germplasm with one or more elite corn inbreds. The resultant hybrid could be self pollinated to produce an F2 population for the purposes of initiating a conventional pedigree breeding program (Allard, R.W.
( 1960) Principles of Plant Breeding. John Wiley & Sons, Inc. New York.
Pp 115-128}.
In order to identify those F2 individuals with the desired genotypes, plant tissue would be collected from each F2 individual in the population and genotyped with the SSR marker loci listed in Table 1. Those F2 individuals with the highest frequency of SSR marker alleles derived from the high oil source would be selected and further culled based upon their agronomic fitness. With continued inbreeding and segregation, those oil loci in a heterozygous state could become fixed for either the high oil or low oil allele. It is therefore likely that genotyping and selection of later generation materials would be practiced in order to further segregate breeding Iines based upon their marker allele and hence oil allele composition.
Depending upon population size and serendipity, the resulting inbreds from the pedigree breeding program may not demonstrate su~cient agronomic competitiveness or sufficient kernel oil expression because an inadequate number of oil alleles was recovered. These new inbreds could therefore be used as parental material and new breeding projects initiated. The SSR markers could again be used for further selection of oil as described.
It is obvious to those skilled in the art that many variants to selection methodology may by envisioned. Selection would be based upon the allelic composition of one or more marker loci which identify trait oil loci present in a population. Further selection would be performed by examination and selection of genotypes from individual plants, families, or their progeny. Various predictive models could be developed using genotypic information, which could generate various selection indices. These models would permit weighting the effect predicted by marker loci. This is because the predictive value of an individual marker locus is dependent upon its genetic distance from the corresponding trait locus as well as the expressivity of the trait locus. Selection strategies which combine phenotype-based and genotype-based selection may also be envisioned.
The marker loci presented here are predictive of oil loci in Alexho synthetic populations. Because ASKC28wx represents the ?8th oil breeding cycle of a genetically closed population, earlier breeding cycles are composed of the same oil Loci. It is expected that cycles differ simply in their allelic frequency at the identified oil loci. Therefore, in breeding populations derived from earlier Alexho cycles, the marker loci described in this invention will be useful in identification of oil loci and in prediction of oil concentration.
IDENTIFICATION OF CORN PLANTS FOR USE AS PARENTS
FOR THE PRODUCTION OF TRANSGRESSIVE SEGREGANTS FOR
KERNEL OIL CONCENTRATION
It is important to identify corn plants and lines which, when used as parents, have the greatest probability of producing offspring with superior performance. Transgressive segregant offspring of such parents would result from the crossing of parents with complementary sets of alleles conferring the high-oil phenotype. Using the information provided herein, marker alleles which predict desired trait performance (i.e., high oil) at a given marker locus are known.
By genotyping lines at those marker loci, the value of those lines as parents is revealed. For example, if one wanted to create an individual containing superior alleles at 5 separate oil loci (A-E), one could identify and cross a parent composed of desired alleles for locus A, B, and C with a parent composed of desired alleles at B, D, and E. These parents are complementary because they permit the recovery of progeny containing desired alleles at all 5 loci. Ideally, parents would be chosen which when combined ensure maximum complementation of loci, so that a high frequency of desired recombinants are recovered.
SEQUENCE LISTING
(1) GENERAL INFORMATION:
(i) APPLICANT:
(A) ADDRESSEE: E. I. DU PONT DE NEMOURS AND COMPANY
(B) STREET: 1007 MARKET STREET
(C) CITY: WILMINGTON
(D) STATE: DELAWARE
(E) COUNTRY: USA
(F) ZIP: 19898 (G) TELEPHONE: 302-992-9926 (H) TELEFAX: 302-773-0169 (I) TELEX: 6717325 (ii) TITLE OF INVENTION: A METHOD TO IDENTIFY AND BREED
CORN WITH INCREASED KERNEL OIL
CONCENTRATION
{iii) NUMBER OF SEQUENCES: 104 (iv} COMPUTER READABLE FORM:
(A) MEDIUM TYPE DISKETTE, 3.50 INCH
(B) COMPUTER: IBM PC COMPATIBLE
(C) OPERATING SYSTEM: MICROSOFT WINDOWS 95 (D) SOFTWARE: MICROSOFT WORD VERSION 7.OA
(v) CURRENT APPLICATION DATA:
(A) APPLICATION NUMBER:
(B} FILING DATE:
(C) CLASSIFICATION:
(vi) PRIOR APPLICATION DATA:
(A) APPLICATION NUMBER: 60/041,515 (B) FILING DATE: MARCH 24, 1997 (C) CLASSIFICATION:
(vii) ATTORNEY/AGENT INFORMATION:
(A) NAME: MAJARIAN, WILLIAM R.
(B) REGISTRATION NUMBER: P-41,173 (C) REFERENCE/DOCKET NUMBER: BB-1076 (2) INFORMATION FOR SEQ ID NO:1:
(i) SEQUENCE CHARACTERISTICS:
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SEQ ID
WO 98/42870 PCT/(1598/05550 (2) INFORMATION
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ID NO:
S:
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SEQ ID
CTGCCCAGTCCGTAATGAA
1g (2) INFORMATION
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TAGATTTATTTTCTGAACGA TTGG
(2) INFORMATION
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SEQ ID
GATCTCTCTGAGGCTTGTCC
(2) INFORMATION
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SEQ ID
(2) INFORMATION FOR SEQ ID N0:9:
( i ) SEQUENCE C!-iARACTERISTICS
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SEQ ID
(2) INFORMATION
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ID N0:12:
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SEQ ID
(2) INFORMATION
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(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION:
SEQ ID N0:13:
CTGCTTATTG CTTTCGTCAT
A
(2) INFORMATION
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(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:14:
SEQ ID
TGCTGCACTA CTTGAACCTA
G
(2) INFORMATION
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SEQ ID
ACACAGAGAT GACAAAAGCA A
(2) INFORMATION
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(A) LENGTH: 18 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:16:
SEQ ID
GCAGGCGTGC TATGAGAG
(2) INFORMATION FOR SEQ ID N0:17:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID NO: I7:
(2) INFORMATION FOR SEQ ID NO:18:
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(2) INFORMATION FOR SEQ ID N0:19:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:19:
(2) INFORMATION FOR SEQ ID N0:20:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:20:
(2) _TNFORMATION FOR SEQ ID N0:21:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:21:
GCTCGTAGTA GGGGTTACG
(2) INFORMATION FOR SEQ ID N0:22:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:22:
GACAGCCTCA CCTCAAGA
(2) INFORMATION FOR SEQ ID N0:23:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS:. single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:23:
(2) INFORMATION FOR SEQ ID N0:24:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:24:
(2) INFORMATION FOR SEQ ID N0:25:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear iii) MOLECULE TYPE: other nucleicacid (xi) SEQUENCE DESCRIPTION: N0:25:
SEQ ID
(2) INFORMATION
FOR
SEQ
ID N0:26:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleicacid (xi) SEQUENCE DESCRIPTION: N0:26:
SEQ ID
(2) INFORMATION
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ID N0:27:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (8) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleicacid (xi) SEQUENCE DESCRIPTION: N0:27:
(2) INFORMATION
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SEQ
ID N0:28:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleicacid (xi) SEQUENCE DESCRIPTION: N0:28:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:29:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:29:
SEQ ID
GATCCTACCA
AAATCTTATA
GGC
(2) INFORMATION
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ID N0:30:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:30:
SEQ ID
ACAGCTAGCCAAGATCTGAT T
(2) INFO RMATION FOR SEQ ID N0:31:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:31:
SEQ ID
CGATACTAATGGAAGCCCTA A
(2) INFORMATION
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ID N0:32:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION:
SEQ ID N0:32:
ATGGCCCATTAAGTTTATCA C
(2) INFORMATION FOR SEQ ID N0:33:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:33:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:34:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi} SEQUENCE DESCRIPTION: N0:39:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:35:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:35:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:36:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:36:
SEQ ID
(2) INFORMATION
FOR
SEQ
ID N0:37:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION:
SEQ ID N0:37:
GAGACTTTTG
AGGAGAAAGC
A
(2) INFORMATION
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SEQ
ID N0:38:
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(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:38:
SEQ ID
GATCAAAAGA
GCAAAAGGAG
A
(2) INFORMATION FOR SEQ ID N0:39:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:39:
SEQ ID
AACTGATGAATACCTTCCCA G
(2) INFORMATION
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SEQ
ID N0:40:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: . other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:40:
SEQ ID
TGATTAACTTCTCCCTTTGG T
(2) INFORMATION
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ID N0:41:
(i) SEQUENCE CHARACTERISTICS:
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SEQ ID
(2) INFORMATION
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ID N0:42:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:42:
SEQ ID
(2) INFORMATION
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ID N0:43:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:43:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:49:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:49:
SEQ ID
(2) INFORMATION
FOR
SEQ
ID N0:45:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid tC) STRANDEDNESS: single (D) TOPOLOGY: linear tii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:45:
SEQ ID
TCATTGCCCA ~
AAGTGTTG
(2) INFORMATION
FOR
SEQ
ID N0:96:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:96:
SEQ ID
CTCATCACCCCTCCAGAG
(2) INFORMATION FOR SEQ ID N0:97:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs tB) TYPE: nucleic acid (C) STRANDEDNESS: single tD) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:47:
SEQ ID
GATCCACGCCATTTAAAC
(2) INFORMATION
FOR
SEQ
ID N0:48:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) .SEQUENCE DESCRIPTION: N0:98:
SEQ ID
TGATACTCTGGTGCATGTTC
(2) INFORMATION FOR SEQ ID N0:99:
(i) SEQUENCE CHARACTERISTICS:
(A} LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: iinear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:99:
SEQ ID
GATCGCTCCGATCTATCC lg (2) INFORMATION
FOR SEQ
ID N0:50:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear fii} MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:50:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:51:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:51:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:52:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:52:
SEQ ID
(2) INFORMATION FOR SEQ ID N0:53:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:53:
CTAACCCACG ACGACCCT
(2) INFORMATION FOR SEQ ID N0:59:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:54:
(2) INFORMATION FOR SEQ ID N0:55:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:55:
(2) INFORMATION FOR SEQ ID N0:56:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRT_PTION: SEQ ID N0:56:
(2) INFORMATION FOR SEQ ID N0:57:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:57:
SEQ ID
(2) INFORMATION
FOR
SEQ
ID N0:58:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:58:
SEQ ID
(2) INFORMATION
FOR
SEQ
ID N0:59:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:59:
SEQ ID
(2) INFORMATION FOR SEQ ID N0:60:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:60:
SEQ ID
(2) INFORMATION FOR SEQ ID N0:61:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:61:
TTCCTTCACG TTTCTCTGTT AA
(2) INFORMATION FOR SEQ ID N0:62:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:62:
(2) INFORMATION FOR SEQ ID N0:63:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:63:
CCCAAAGGCG ATACCTATT
(2) INFORMATION FOR SEQ ID N0:64:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:69:
(2) INFORMATION FOR SEQ ICJ N0:65:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:65:
(2) INFORMATION FOR SEQ ID N0:66:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:66:
(2) INFORMATION FOR SEQ ID N0:67:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1B base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:67:
CTATGGATGG CTGCTTGC lg (2) INFORMATION FOR SEQ ID N0:68:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:68:
GTCAGGCAGC AGAATGTG lg (2) INFORMATION FOR SEQ ID N0:69:
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AAACCGTCCA GCGACTAC
(2) INFORMATION FOR SEQ ID N0:70:
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GGAAGAACCA ATCCCATATC T
(2) INFORMATION FOR SEQ ID N0:71:
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AACATCCTGT CGGAAACAG
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TCATCACGTC TCTCTTTCAA C
(2) INFORMATION FOR SEQ ID N0:73:
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(2) INFORMATION FOR SEQ ID N0:76:
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(2) INFORMATION
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SEQ ID
CACAAATGTG
AAGGTAAACA
CT
(2) INFORMATION
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SEQ ID
AATGGTACGGTTCAGGATG
(2) INFO RMATION FOR SEQ ID N0:79:
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SEQ ID
AGATGACGCACGGAACAC
(2) INFORMATION
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(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:80:
SEQ ID
AGCATCATGTAGCAGGAGG
(2) INFORMATION FOR SEQ ID N0:81:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:81:
SEQ ID
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SEQ ID
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SEQ ID
(2) INFORMATION
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ID N0:84:
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(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:84:
SEQ ID
(2) INFORMATION FOR SEQ ID N0:85:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:85:
CTCTGGGCTC TGTGTTAGAG T
(2) INFORMATION FOR SEQ ID N0:86:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:86:
(2) INFORMATION FOR SEQ ID N0:87:
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(A) LENGTH: 19 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:87:
CTGCTTTCTC TGTTCCAGC 1g (2) INFORMATION FOR SEQ ID N0:88:
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(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:88:
(2) INFORMATION FOR SEQ ID N0:89:
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(A) LENGTH: 21 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:89:
SEQ ID
(2) INFORMATION
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(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:90:
SEQ ID
(2) INFORMATION
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ID N0:91:
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(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:91:
SEQ ID
(2) INFORMATION
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ID N0:92:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:92:
SEQ ID
(2) INFORMATION
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ID N0:93:
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SEQ ID
GGAGCCTGGA
GTGAGAAC
(2) INFORMATION
FOR
SEQ
ID N0:94:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C} STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:94:
SEQ ID
CATGCTCACCTAACGTGG
(2) INFORMATION
FOR
SEQ
ID N0:95:
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(A) LENGTH: 21 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:95:
SEQ ID
ATCTGAACACTTGAGCAACA A
(2) INFORMATION
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ID N0:96:
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(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:96:
SEQ ID
ATAGACCGGACCCATCAC
(2) INFORMATION FOR SEQ ID N0:97:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:97:
SEQ ID
(2) INFORMATION
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ID N0:98:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:98:
SEQ ID
(2) INFORMATION
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(A) LENGTH: 18 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:99:
SEQ ID
(2) INFORMATION
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(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: NO:100:
SEQ ID
(2) INFORMATION
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ID NO:101:
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(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:101:
ATGGAAACAT
CAAAGTGGAT
T
(2) INFORMATION
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ID N0:102:
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(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID NO:102:
TGCTACCCTGATGACCTGAT
(2} INFORMATION
FOR SEQ
ID N0:103:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:103:
ACCACTAGTCTCATATGAAG GG
(2) INFORMATION
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ID N0:109:
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(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:104:
GGTAGGTGGGTAGGGGTT
Table 2 Amplification Cocktail Reagent Stock Concentration Final Concentration Buffer* 1 OX 1.SX
dNTPs 2 mM 0.3 mM
Forward Primer 40 ~M 0.45 ~M
Reverse Primer 40 ~M 0.45 ~M
AmpliTaq PolymeraseTM 5 U/~1 0.05 U/p.l * 1 OX Buffer is a pH 9.0 solution composed of 800 mM Tris-OH, 200 mM
~4)2SD4~ ~d 25 mM MgCl2.
Localization of oil loci One hundred thirty three polymorphic SSR marker loci were used to genotype the recombinant inbreds from the LH 1 l9wx x ASKC28wx cross and one hundred and three polymorphic SSR marker loci were used to genotype the LH51 x ASKC28wx-derived population. In addition, twenty publicly available polymorphic SSR loci with previously established chromosome locations and covering all ten maize chromosomes (available from Research Genetics, Huntsville, AL) were also mapped in both populations.
Genetic linkage and distance between marker loci was determined independently for each population using MAPMAKER 3.0 (Lincoln S.E., et al.
( 1993) Whitehead Inst. Biomed. Res., Cambridge, MA). This resulted in the establishment of ten linkage groups for each population corresponding to the ten chromosomes of maize. Each linkage group was assigned to a chromosome based upon linkage to the public SSR markers. Twenty-three and ten markers in the LHI l9wx x ASKC28wx and LH51 x ASKC28wx populations, respectively, were not assigned chromosome positions because genetic linkage could not be clearly established.
Analysis of variance was used to identify marker loci in linkage with trait loci conferring increased oil concentration. Oil concentration was used as a dependent variable and separate ANOVAs were calculated with SAS Proc GLM
(SAS Inst., Cary, NC) using each marker locus as a single independent variable (Edwards, M.D., et aI. ( 1987) Genetics 116:113-I25). Therefore, for each ANOVA test the mean oil values of marker allele classes were compared. Marker loci were declared signif cant if p < 0.05.
Linkage data for significant marker loci was examined to determine both the number of trait Ioci present and their probable location. Significant marker loci on the same linkage group are either detecting the same trait locus or alternatively different trait loci. By careful examination of the phenotypic variation explained by each marker locus along the chromosome, a determination of the number trait loci on a linkage group was made. Significant marker loci, on the same linkage group and uninterrupted by non-significant marker loci, were declared to be detecting the same trait locus on the chromosome. If significant marker loci on the same chromosome were interrupted by non-significant marker loci then each significant region was declared to contain a trait locus resulting in multiple trait loci on the same chromosome.
To confirm the number of trait loci, marker data assigned to linkage groups and oil data were also analyzed with Mapmaker/QTL 1.0 (Lincoln, S.E. et al. (1990) Whitehead Inst. Biomed. Res., Cambridge, MA). Results with Mapmaker/QTL were in agreement with the initial analysis for the number trait loci on each chromosome.
Eleven and twelve loci controlling kernel oil concentration were located in the LH1 l9wx x ASKC28wx and LH51 x ASKC28wx recombinant inbred populations, respectively. Each oil locus is defined by one or more linked marker loci.
In instances where the same marker loci were used in both populations, alignment of linkage groups is possible. It was found that in most instances both populations localized the same oil loci. By considering common marker loci, a total of seventeen loci controlling kernel oil concentration were found. Each oil locus was assigned an arbitrary letter designation (Table 3}.
Table 3 Marker loci genetically linked to and predictive of the location of trait loci conferring increased kernel oil concentration Oil locus Chromosome Marker loci A I s1922 B I s 1478, s I 853, s I 949 C I S I 860, s 1925, s 1931, s2150 D 2 s2175 E 3 s1394 F 4 s 1476, s 1772, s 1816, s2122, s 1836 G 4 s1939,s1946 H 4 s1870 I 5 s1529 J ~ s2054, s 1647, s 1500, s 1 X45, s 1774, s2097 K 6 s1457, s2055, s1757, s2125, s1780, s1375, s 1797, s 1416, s 1432, s 1921 L 7 s1630, s1422, s2156 M 8 s1817,s2057 N 9 s1544, s1633, s1384, s1813, s1767, s2058, s1933,s1513,s1484 O 10 s1756 P 10 s1480 (positive oil allele in LH51) Q N.A.* s1750 *N.A. - chromosome location not known In instances where comparisons could be made, oil loci which were identified in one population were identified at the same location in the second population. In two exceptions, an oil locus was found in one population, but not in the second population. In the first case, the allele with a positive oil effect was found in LH51 and thus it would be unexpected to identify the same locus in the LH 119wx x ASKC28wx population. In the second case, it was found that different ASKC28wx-derived marker alleles were segregating in the populations;
therefore, each population was measuring the oil effect of a different ASKC28wx allele at the trait locus. The most abundant ASKC28wx oil allele segregating in LH 119wx x ASKC28wx had a positive oil effect versus the alternative LH 119-derived allele, whereas in the LH51 x ASKC28wx population, the abundant ASKC28wx allele had no positive oil effect. With the exception of the oil locus linked to marker s1480, all alleles with positive effects on oil concentration were derived from ASKC28wx.
MARKER-ASSISTED SELECTION OF BREEDING LINES USING GENETIC
MARKERS FOR INCREASE KERNEL OIL CONCENTRATION
Genetic marker loci in linkage with oil trait loci are highly predictive of oil concentration and as such may be used as an indirect measurement of kernel oil in a marker-assisted selection program. Accordingly, genotypic information from linked marker loci would facilitate the selection of breeding lines with increased oil concentration. Direct oii measurements cannot differentiate between various genotypic trait locus compositions with equivalent phenotypic effects. This is especially problematic in early generation segregating breeding populations where only limited fixation of oil loci has occurred.
By way of example, an objective of a corn breeding program could be the creation of new elite inbred lines which contain trait alleles conferring increased kernel oil concentration. These trait alleles would be introduced by the intermating of high oil germplasm with one or more elite corn inbreds. The resultant hybrid could be self pollinated to produce an F2 population for the purposes of initiating a conventional pedigree breeding program (Allard, R.W.
( 1960) Principles of Plant Breeding. John Wiley & Sons, Inc. New York.
Pp 115-128}.
In order to identify those F2 individuals with the desired genotypes, plant tissue would be collected from each F2 individual in the population and genotyped with the SSR marker loci listed in Table 1. Those F2 individuals with the highest frequency of SSR marker alleles derived from the high oil source would be selected and further culled based upon their agronomic fitness. With continued inbreeding and segregation, those oil loci in a heterozygous state could become fixed for either the high oil or low oil allele. It is therefore likely that genotyping and selection of later generation materials would be practiced in order to further segregate breeding Iines based upon their marker allele and hence oil allele composition.
Depending upon population size and serendipity, the resulting inbreds from the pedigree breeding program may not demonstrate su~cient agronomic competitiveness or sufficient kernel oil expression because an inadequate number of oil alleles was recovered. These new inbreds could therefore be used as parental material and new breeding projects initiated. The SSR markers could again be used for further selection of oil as described.
It is obvious to those skilled in the art that many variants to selection methodology may by envisioned. Selection would be based upon the allelic composition of one or more marker loci which identify trait oil loci present in a population. Further selection would be performed by examination and selection of genotypes from individual plants, families, or their progeny. Various predictive models could be developed using genotypic information, which could generate various selection indices. These models would permit weighting the effect predicted by marker loci. This is because the predictive value of an individual marker locus is dependent upon its genetic distance from the corresponding trait locus as well as the expressivity of the trait locus. Selection strategies which combine phenotype-based and genotype-based selection may also be envisioned.
The marker loci presented here are predictive of oil loci in Alexho synthetic populations. Because ASKC28wx represents the ?8th oil breeding cycle of a genetically closed population, earlier breeding cycles are composed of the same oil Loci. It is expected that cycles differ simply in their allelic frequency at the identified oil loci. Therefore, in breeding populations derived from earlier Alexho cycles, the marker loci described in this invention will be useful in identification of oil loci and in prediction of oil concentration.
IDENTIFICATION OF CORN PLANTS FOR USE AS PARENTS
FOR THE PRODUCTION OF TRANSGRESSIVE SEGREGANTS FOR
KERNEL OIL CONCENTRATION
It is important to identify corn plants and lines which, when used as parents, have the greatest probability of producing offspring with superior performance. Transgressive segregant offspring of such parents would result from the crossing of parents with complementary sets of alleles conferring the high-oil phenotype. Using the information provided herein, marker alleles which predict desired trait performance (i.e., high oil) at a given marker locus are known.
By genotyping lines at those marker loci, the value of those lines as parents is revealed. For example, if one wanted to create an individual containing superior alleles at 5 separate oil loci (A-E), one could identify and cross a parent composed of desired alleles for locus A, B, and C with a parent composed of desired alleles at B, D, and E. These parents are complementary because they permit the recovery of progeny containing desired alleles at all 5 loci. Ideally, parents would be chosen which when combined ensure maximum complementation of loci, so that a high frequency of desired recombinants are recovered.
SEQUENCE LISTING
(1) GENERAL INFORMATION:
(i) APPLICANT:
(A) ADDRESSEE: E. I. DU PONT DE NEMOURS AND COMPANY
(B) STREET: 1007 MARKET STREET
(C) CITY: WILMINGTON
(D) STATE: DELAWARE
(E) COUNTRY: USA
(F) ZIP: 19898 (G) TELEPHONE: 302-992-9926 (H) TELEFAX: 302-773-0169 (I) TELEX: 6717325 (ii) TITLE OF INVENTION: A METHOD TO IDENTIFY AND BREED
CORN WITH INCREASED KERNEL OIL
CONCENTRATION
{iii) NUMBER OF SEQUENCES: 104 (iv} COMPUTER READABLE FORM:
(A) MEDIUM TYPE DISKETTE, 3.50 INCH
(B) COMPUTER: IBM PC COMPATIBLE
(C) OPERATING SYSTEM: MICROSOFT WINDOWS 95 (D) SOFTWARE: MICROSOFT WORD VERSION 7.OA
(v) CURRENT APPLICATION DATA:
(A) APPLICATION NUMBER:
(B} FILING DATE:
(C) CLASSIFICATION:
(vi) PRIOR APPLICATION DATA:
(A) APPLICATION NUMBER: 60/041,515 (B) FILING DATE: MARCH 24, 1997 (C) CLASSIFICATION:
(vii) ATTORNEY/AGENT INFORMATION:
(A) NAME: MAJARIAN, WILLIAM R.
(B) REGISTRATION NUMBER: P-41,173 (C) REFERENCE/DOCKET NUMBER: BB-1076 (2) INFORMATION FOR SEQ ID NO:1:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:1:
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SEQ ID
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SEQ ID
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(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:4:
SEQ ID
WO 98/42870 PCT/(1598/05550 (2) INFORMATION
FOR
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ID NO:
S:
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SEQ ID
CTGCCCAGTCCGTAATGAA
1g (2) INFORMATION
FOR
SEQ
ID N0:6:
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(A) LENGTH: 24 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleicacid (xi) SEQUENCE DESCRIPTION: N0:6:
SEQ ID
TAGATTTATTTTCTGAACGA TTGG
(2) INFORMATION
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(A} LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleicacid (xi) SEQUENCE DESCRIPTION: N0:7:
SEQ ID
GATCTCTCTGAGGCTTGTCC
(2) INFORMATION
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SEQ ID
(2) INFORMATION FOR SEQ ID N0:9:
( i ) SEQUENCE C!-iARACTERISTICS
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:9:
SEQ ID
(2) INFORMATION
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ID N0:10:
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(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other. acid nucleic (xi) SEQUENCE DESCRIPTION: NO:10:
SEQ ID
(2) INFORMATION
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ID N0:11:
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(A) LENGTH: 24 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: NO:11:
SEQ ID
(2) INFORMATION
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ID N0:12:
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(A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:12:
SEQ ID
(2) INFORMATION
FOR SEQ ID N0:13:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION:
SEQ ID N0:13:
CTGCTTATTG CTTTCGTCAT
A
(2) INFORMATION
FOR SEQ ID N0:14:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:14:
SEQ ID
TGCTGCACTA CTTGAACCTA
G
(2) INFORMATION
FOR SEQ ID N0:15:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:15:
SEQ ID
ACACAGAGAT GACAAAAGCA A
(2) INFORMATION
FOR SEQ ID N0:16:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:16:
SEQ ID
GCAGGCGTGC TATGAGAG
(2) INFORMATION FOR SEQ ID N0:17:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID NO: I7:
(2) INFORMATION FOR SEQ ID NO:18:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:18:
(2) INFORMATION FOR SEQ ID N0:19:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:19:
(2) INFORMATION FOR SEQ ID N0:20:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:20:
(2) _TNFORMATION FOR SEQ ID N0:21:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:21:
GCTCGTAGTA GGGGTTACG
(2) INFORMATION FOR SEQ ID N0:22:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:22:
GACAGCCTCA CCTCAAGA
(2) INFORMATION FOR SEQ ID N0:23:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS:. single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:23:
(2) INFORMATION FOR SEQ ID N0:24:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:24:
(2) INFORMATION FOR SEQ ID N0:25:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear iii) MOLECULE TYPE: other nucleicacid (xi) SEQUENCE DESCRIPTION: N0:25:
SEQ ID
(2) INFORMATION
FOR
SEQ
ID N0:26:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleicacid (xi) SEQUENCE DESCRIPTION: N0:26:
SEQ ID
(2) INFORMATION
FOR
SEQ
ID N0:27:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (8) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleicacid (xi) SEQUENCE DESCRIPTION: N0:27:
(2) INFORMATION
FOR
SEQ
ID N0:28:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleicacid (xi) SEQUENCE DESCRIPTION: N0:28:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:29:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:29:
SEQ ID
GATCCTACCA
AAATCTTATA
GGC
(2) INFORMATION
FOR SEQ
ID N0:30:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:30:
SEQ ID
ACAGCTAGCCAAGATCTGAT T
(2) INFO RMATION FOR SEQ ID N0:31:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:31:
SEQ ID
CGATACTAATGGAAGCCCTA A
(2) INFORMATION
FOR SEQ
ID N0:32:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION:
SEQ ID N0:32:
ATGGCCCATTAAGTTTATCA C
(2) INFORMATION FOR SEQ ID N0:33:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:33:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:34:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi} SEQUENCE DESCRIPTION: N0:39:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:35:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:35:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:36:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:36:
SEQ ID
(2) INFORMATION
FOR
SEQ
ID N0:37:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION:
SEQ ID N0:37:
GAGACTTTTG
AGGAGAAAGC
A
(2) INFORMATION
FOR
SEQ
ID N0:38:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:38:
SEQ ID
GATCAAAAGA
GCAAAAGGAG
A
(2) INFORMATION FOR SEQ ID N0:39:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:39:
SEQ ID
AACTGATGAATACCTTCCCA G
(2) INFORMATION
FOR
SEQ
ID N0:40:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: . other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:40:
SEQ ID
TGATTAACTTCTCCCTTTGG T
(2) INFORMATION
FOR SEQ
ID N0:41:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:41:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:42:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:42:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:43:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:43:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:49:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:49:
SEQ ID
(2) INFORMATION
FOR
SEQ
ID N0:45:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid tC) STRANDEDNESS: single (D) TOPOLOGY: linear tii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:45:
SEQ ID
TCATTGCCCA ~
AAGTGTTG
(2) INFORMATION
FOR
SEQ
ID N0:96:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:96:
SEQ ID
CTCATCACCCCTCCAGAG
(2) INFORMATION FOR SEQ ID N0:97:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs tB) TYPE: nucleic acid (C) STRANDEDNESS: single tD) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:47:
SEQ ID
GATCCACGCCATTTAAAC
(2) INFORMATION
FOR
SEQ
ID N0:48:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) .SEQUENCE DESCRIPTION: N0:98:
SEQ ID
TGATACTCTGGTGCATGTTC
(2) INFORMATION FOR SEQ ID N0:99:
(i) SEQUENCE CHARACTERISTICS:
(A} LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: iinear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:99:
SEQ ID
GATCGCTCCGATCTATCC lg (2) INFORMATION
FOR SEQ
ID N0:50:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear fii} MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:50:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:51:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:51:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:52:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:52:
SEQ ID
(2) INFORMATION FOR SEQ ID N0:53:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:53:
CTAACCCACG ACGACCCT
(2) INFORMATION FOR SEQ ID N0:59:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:54:
(2) INFORMATION FOR SEQ ID N0:55:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:55:
(2) INFORMATION FOR SEQ ID N0:56:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRT_PTION: SEQ ID N0:56:
(2) INFORMATION FOR SEQ ID N0:57:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:57:
SEQ ID
(2) INFORMATION
FOR
SEQ
ID N0:58:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:58:
SEQ ID
(2) INFORMATION
FOR
SEQ
ID N0:59:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:59:
SEQ ID
(2) INFORMATION FOR SEQ ID N0:60:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:60:
SEQ ID
(2) INFORMATION FOR SEQ ID N0:61:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:61:
TTCCTTCACG TTTCTCTGTT AA
(2) INFORMATION FOR SEQ ID N0:62:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:62:
(2) INFORMATION FOR SEQ ID N0:63:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:63:
CCCAAAGGCG ATACCTATT
(2) INFORMATION FOR SEQ ID N0:64:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:69:
(2) INFORMATION FOR SEQ ICJ N0:65:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:65:
(2) INFORMATION FOR SEQ ID N0:66:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:66:
(2) INFORMATION FOR SEQ ID N0:67:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1B base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:67:
CTATGGATGG CTGCTTGC lg (2) INFORMATION FOR SEQ ID N0:68:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:68:
GTCAGGCAGC AGAATGTG lg (2) INFORMATION FOR SEQ ID N0:69:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:69:
AAACCGTCCA GCGACTAC
(2) INFORMATION FOR SEQ ID N0:70:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii} MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:70:
GGAAGAACCA ATCCCATATC T
(2) INFORMATION FOR SEQ ID N0:71:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:71:
AACATCCTGT CGGAAACAG
(2) INFORMATION FOR SEQ ID N0:72:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii} MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:72:
TCATCACGTC TCTCTTTCAA C
(2) INFORMATION FOR SEQ ID N0:73:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:73:
(2) INFORMATION FOR SEQ ID N0:74:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:74:
(2) INFORMATION FOR SEQ ID N0:75:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:75:
(2) INFORMATION FOR SEQ ID N0:76:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:76:
(2) INFORMATION
FOR SEQ
ID N0:77:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:77;
SEQ ID
CACAAATGTG
AAGGTAAACA
CT
(2) INFORMATION
FOR SEQ
ID N0:78:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:78:
SEQ ID
AATGGTACGGTTCAGGATG
(2) INFO RMATION FOR SEQ ID N0:79:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:79:
SEQ ID
AGATGACGCACGGAACAC
(2) INFORMATION
FOR SEQ
ID N0:80:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:80:
SEQ ID
AGCATCATGTAGCAGGAGG
(2) INFORMATION FOR SEQ ID N0:81:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:81:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:82:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:82:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:83:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:83:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:84:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:84:
SEQ ID
(2) INFORMATION FOR SEQ ID N0:85:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:85:
CTCTGGGCTC TGTGTTAGAG T
(2) INFORMATION FOR SEQ ID N0:86:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:86:
(2) INFORMATION FOR SEQ ID N0:87:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:87:
CTGCTTTCTC TGTTCCAGC 1g (2) INFORMATION FOR SEQ ID N0:88:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:88:
(2) INFORMATION FOR SEQ ID N0:89:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:89:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:90:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:90:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:91:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:91:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:92:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:92:
SEQ ID
(2) INFORMATION
FOR
SEQ
ID N0:93:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:93:
SEQ ID
GGAGCCTGGA
GTGAGAAC
(2) INFORMATION
FOR
SEQ
ID N0:94:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C} STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:94:
SEQ ID
CATGCTCACCTAACGTGG
(2) INFORMATION
FOR
SEQ
ID N0:95:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:95:
SEQ ID
ATCTGAACACTTGAGCAACA A
(2) INFORMATION
FOR
SEQ
ID N0:96:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:96:
SEQ ID
ATAGACCGGACCCATCAC
(2) INFORMATION FOR SEQ ID N0:97:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:97:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:98:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:98:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:99:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (H) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: N0:99:
SEQ ID
(2) INFORMATION
FOR SEQ
ID N0:100:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other acid nucleic (xi) SEQUENCE DESCRIPTION: NO:100:
SEQ ID
(2) INFORMATION
FOR SEQ
ID NO:101:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:101:
ATGGAAACAT
CAAAGTGGAT
T
(2) INFORMATION
FOR SEQ
ID N0:102:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID NO:102:
TGCTACCCTGATGACCTGAT
(2} INFORMATION
FOR SEQ
ID N0:103:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:103:
ACCACTAGTCTCATATGAAG GG
(2) INFORMATION
FOR SEQ
ID N0:109:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (xi) SEQUENCE DESCRIPTION: SEQ ID N0:104:
GGTAGGTGGGTAGGGGTT
Claims (20)
1. A method of breeding for corn with increased kernel oil concentration comprising:
a) using one or more genetic markers to select a corn plant from a corn breeding population by marker-assisted selection, wherein the genetic markers are selected from the group consisting of x1375, s1384, s1394, s1416, s1422, s1432, s1457, s1480, s1476, s1478, s1484, s1500, s1513, s1529, s1544, s1545, s1630, s1633, s1647, s1750, s1756, s1757, s1767, s1772, s1774, s1780, s1797, s1813, s1816, s1817, s1836, s1853, s1860, s1870, s1921, s1922, s1925, s1931, s1933, s1939, s1946, s1949, s2054, s2055, s2057, s2058, s2097, s2122, s2125, s2150, s2156 and s2175; and b) crossing the selected corn plant with a second corn plant wherein the progeny corn plants of the cross display increased kernel oil concentration.
a) using one or more genetic markers to select a corn plant from a corn breeding population by marker-assisted selection, wherein the genetic markers are selected from the group consisting of x1375, s1384, s1394, s1416, s1422, s1432, s1457, s1480, s1476, s1478, s1484, s1500, s1513, s1529, s1544, s1545, s1630, s1633, s1647, s1750, s1756, s1757, s1767, s1772, s1774, s1780, s1797, s1813, s1816, s1817, s1836, s1853, s1860, s1870, s1921, s1922, s1925, s1931, s1933, s1939, s1946, s1949, s2054, s2055, s2057, s2058, s2097, s2122, s2125, s2150, s2156 and s2175; and b) crossing the selected corn plant with a second corn plant wherein the progeny corn plants of the cross display increased kernel oil concentration.
2. The method of claim 1 wherein the selected corn plant is member of an Alexho synthetic population or a progeny thereof.
3. A method for identifying corn plants or corn lines for use as parents for creation of a breeding population, the method comprising:
a) genotyping corn plants or corn lines with one or more genetic markers wherein the genetic markers are selected from the group consisting of s1375, s1384, s1394, s1416, s1422, s1432, s1457, s1480, s1476, s1478, s1484, s1500, s1513, s1529, s1544, s1545, s1630, s1633, s1647, s1750, s1756, s1757, s1767, s1772, s1774, s1780, s1797, s1813, s1816, s1817, s1836, s1853, s1860, s1870, s1921, s1922; s1925, s1931, s1933, s1939, s1946, s1949, s2054, s2055, s2057, s2058, s2097, s2122, s2125, s2150, s2156 and s2175; and b) identifying corn plants or corn lines which, based upon their genotype, are predicted to produce transgressive segregants for kernel oil concentration.
a) genotyping corn plants or corn lines with one or more genetic markers wherein the genetic markers are selected from the group consisting of s1375, s1384, s1394, s1416, s1422, s1432, s1457, s1480, s1476, s1478, s1484, s1500, s1513, s1529, s1544, s1545, s1630, s1633, s1647, s1750, s1756, s1757, s1767, s1772, s1774, s1780, s1797, s1813, s1816, s1817, s1836, s1853, s1860, s1870, s1921, s1922; s1925, s1931, s1933, s1939, s1946, s1949, s2054, s2055, s2057, s2058, s2097, s2122, s2125, s2150, s2156 and s2175; and b) identifying corn plants or corn lines which, based upon their genotype, are predicted to produce transgressive segregants for kernel oil concentration.
4. A trait locus controlling kernel oil concentration, the locus mapped by a genetic marker selected from the group consisting of s2054, s1647, s1500, s1545, s1774 and s2097.
5. A trait locus controlling kernel oil concentration, the locus mapped by a genetic marker selected from the group consisting of s1817 and s2057.
6. A trait locus controlling kernel oil concentration, the locus mapped by a genetic marker selected from the group consisting of s1860, s1931, s2150 and s1925.
7. A trait locus controlling kernel oil concentration, the locus mapped by a genetic marker selected from the group consisting of s1457, s2055, s1757, s2125, s1780, s1375, s1797, s1416, s1432 and s1921.
8. A trait locus controlling kernel oil concentration, the locus mapped by a genetic marker selected from the group consisting of s1544, s1633, s1384, s1813, s1767, s2058, s1933, s1513 and s1484.
9. A trait locus controlling kernel oil concentration, the locus mapped by a genetic marker selected from the group consisting of s1476, s1772, s1816, s2122 and s1836.
10. A trait locus controlling kernel oil concentration, the locus mapped by a genetic marker selected from the group consisting of s1939 and s1946.
11. A trait locus controlling kernel oil concentration, the locus mapped by a genetic marker selected from the group consisting of s1478, s1853 and s1949.
12. A trait locus controlling kernel oil concentration, the locus mapped by a genetic marker selected from the group consisting of s1630, s1422 and s2156.
13. A trait locus controlling kernel oil concentration, the locus mapped by the genetic marker s1756.
14. A trait locus controlling kernel oil concentration, the locus mapped by the genetic marker s1922.
15. A trait locus controlling kernel oil concentration, the locus mapped by the genetic marker s1529.
16. A trait locus controlling kernel oil concentration, the locus mapped by the genetic marker s1394.
17. A trait locus controlling kernel oil concentration, the locus mapped by the genetic marker s1750.
18. A trait locus controlling kernel oil concentration, the locus mapped by the genetic marker s1870.
19. A trait locus controlling kernel oil concentration, the locus mapped by the genetic marker s2175.
20. Corn plants that display increased kernel oil concentration produced by the method of Claim 1.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US4151597P | 1997-03-24 | 1997-03-24 | |
| US60/041,515 | 1997-03-24 | ||
| PCT/US1998/005550 WO1998042870A1 (en) | 1997-03-24 | 1998-03-19 | A method to identify and breed corn with increased kernel oil concentration |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2280933A1 true CA2280933A1 (en) | 1998-10-01 |
Family
ID=21916926
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002280933A Abandoned CA2280933A1 (en) | 1997-03-24 | 1998-03-19 | A method to identify and breed corn with increased kernel oil concentration |
Country Status (13)
| Country | Link |
|---|---|
| EP (1) | EP0972079A1 (en) |
| JP (1) | JP2001517951A (en) |
| KR (1) | KR20010005625A (en) |
| AR (1) | AR012152A1 (en) |
| AU (1) | AU734755B2 (en) |
| BR (1) | BR9815450A (en) |
| CA (1) | CA2280933A1 (en) |
| HU (1) | HUP0001745A3 (en) |
| IL (1) | IL131908A0 (en) |
| NZ (1) | NZ337906A (en) |
| PL (1) | PL335910A1 (en) |
| WO (1) | WO1998042870A1 (en) |
| ZA (1) | ZA982250B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6806399B1 (en) | 2000-04-19 | 2004-10-19 | Carmel-Haifa University Economic Corporation Ltd. | Pollen-mediated method for transformation of maize, tomato or melon |
| EP1392103B1 (en) * | 2001-02-06 | 2009-07-22 | Dow AgroSciences LLC | Methods of marker-assisted selection of high oil maize plants |
| AU2003304464A1 (en) * | 2002-03-15 | 2005-03-29 | Monsanto Technology Llc | Nucleic acid molecules associated with oil in plants |
| BRPI0708477B8 (en) | 2006-03-01 | 2022-12-06 | Pioneer Hi Bred Int | METHOD FOR IDENTIFYING A FIRST CORN PLANT OR A FIRST CORN GERMOPLASM COMPRISING IN ITS GENOME A DIAGLYCEROL O-ACYLTRANSFERASE TYPE 1 (DGAT) GENE, ISOLATED POLYNUCLEOTIDE, METHOD FOR PRODUCING A PLANT OR PLANT PART HAVING A SELECTED PHENOTYPE, METHOD TO IDENTIFY THE PRESENCE OF A MARKER LOCU |
| AU2007220007B2 (en) * | 2006-03-01 | 2013-02-21 | E.I. Du Pont De Nemours And Company | Compositions related to the quantitative trait locus 6 (QTL6) in maize and methods of use |
| EP1947198A1 (en) * | 2007-01-18 | 2008-07-23 | Syngeta Participations AG | Maize plants characterised by quantitative trait loci (QTL) |
| JP2008220269A (en) * | 2007-03-13 | 2008-09-25 | Japan Grassland Farming Forage Seed Association | Primer set for detecting dna marker linked to gene locus relating to fat-content in corn seed and its use |
| EP2242850B1 (en) * | 2008-01-18 | 2020-03-04 | Syngenta Participations AG | Maize plants characterised by quantitative trait loci (qtl) |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5492547B1 (en) * | 1993-09-14 | 1998-06-30 | Dekalb Genetics Corp | Process for predicting the phenotypic trait of yield in maize |
| US5476524A (en) * | 1993-12-09 | 1995-12-19 | Zeneca Limited | Oil producing sunflowers and production thereof |
-
1998
- 1998-03-17 ZA ZA9802250A patent/ZA982250B/en unknown
- 1998-03-19 PL PL98335910A patent/PL335910A1/en unknown
- 1998-03-19 EP EP98911903A patent/EP0972079A1/en not_active Withdrawn
- 1998-03-19 AU AU65751/98A patent/AU734755B2/en not_active Ceased
- 1998-03-19 CA CA002280933A patent/CA2280933A1/en not_active Abandoned
- 1998-03-19 KR KR1019997008691A patent/KR20010005625A/en not_active Application Discontinuation
- 1998-03-19 NZ NZ337906A patent/NZ337906A/en unknown
- 1998-03-19 JP JP54448798A patent/JP2001517951A/en active Pending
- 1998-03-19 HU HU0001745A patent/HUP0001745A3/en unknown
- 1998-03-19 WO PCT/US1998/005550 patent/WO1998042870A1/en not_active Application Discontinuation
- 1998-03-19 BR BR9815450-8A patent/BR9815450A/en not_active IP Right Cessation
- 1998-03-19 IL IL13190898A patent/IL131908A0/en unknown
- 1998-03-24 AR ARP980101333A patent/AR012152A1/en not_active Application Discontinuation
Also Published As
| Publication number | Publication date |
|---|---|
| IL131908A0 (en) | 2001-03-19 |
| AU734755B2 (en) | 2001-06-21 |
| NZ337906A (en) | 2001-08-31 |
| JP2001517951A (en) | 2001-10-09 |
| AU6575198A (en) | 1998-10-20 |
| HUP0001745A2 (en) | 2000-09-28 |
| EP0972079A1 (en) | 2000-01-19 |
| AR012152A1 (en) | 2000-09-27 |
| ZA982250B (en) | 1999-09-17 |
| BR9815450A (en) | 2001-10-23 |
| HUP0001745A3 (en) | 2002-04-29 |
| WO1998042870A1 (en) | 1998-10-01 |
| PL335910A1 (en) | 2000-05-22 |
| KR20010005625A (en) | 2001-01-15 |
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