MXPA99008723A - A method to identify and breed corn with increased kernel oil concentration - Google Patents

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 locicontrolling corn kernel oil concentration.

Description

METHOD TO IDENTIFY AND PRODUCE A MAIZE WITH A GREATER CONCENTRATION OF OIL IN THE GRAIN FIELD OF THE INVENTION The invention is found in the field of plant production and molecular biology. More specifically, the invention relates to the identification of maize sites that confer a greater concentration of oil in the grain using genetic markers and the use of genetic markers to help the identification and production of maize with a higher concentration of oil in the grain.

BACKGROUND OF THE INVENTION Corn is one of the main crops used as a source of human food, an animal feed, and as a source of carbohydrates, oil, protein and fiber. This is typically used as a source of energy in animal fodder, or as a raw material for the recovery of starch, protein feed fractions, fiber, dehusked kernels, 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 REF: 31003 elite corn spawns that, after interbreeding, produce agronomically superior hybrids. During the development of maize spawn, plant producers select a number of different traits that affect agronomic performance or performance. Those traits include, but are not limited to stem resistance, deposit, disease resistance, grain moisture and grain yield. Agronomic traits tend to be measured quantitatively by continuous rather than discrete distributions. There is a theory that "quantitative traits are controlled by several genes with small and generally equivalent effects.In addition, the phenotype observed is partially due to this genetic component and an environmental component.The ability to inherit a trait is defined in the broad sense As the relationship of genetic variance to total phenotypic variance, many agronomic traits have a low inheritance capacity, that is, the functioning or performance of the original plants is a poor estimator of the performance or performance of the progeny. , traits with low inheritance capacity have components of small genetic variance compared to the observed variation.The impact on the plant producer is that in breeding populations, the value of a genetic composition of the plant is difficult to determine from of measurements of agronomic traits, in an attempt to maximize their capacities dis In the case of criminology, producers collect multiple measurements of individuals related by their offspring as well as of many environments. This strategy requires intensive resources because it involves the use of extensive trials to obtain "even small gains in plant improvement." This coupled with the fact that improved maize lineages are selected from multiple trials simultaneously, makes the development of spawn The addition of novel traits in a corn production program includes an additional burden on the producer of plants, depending on the genetic complexity of the novel trait (ie, a single trait). gene vs. many genes), a significant increase in time and effort is required to produce elite lines that contain novel traits.One of these features is the concentration of oil in the grain.Maize with a higher concentration of oil in the grain is important because it has an improved feed value for poultry (Han Y. et al. (1987) Poul try Sci. 65: 103-111) and cattle (Nordstrom, J., et al. (1972) J. An. Sci. 35 (2): 357-361). The grain of conventional corn hybrids typically contains 4% oil. In an effort to increase the concentration of oil in the grain, a long-term recurrent selection program was initiated in the field of cv. Burr White Openly pollinated by White CG Hopkins in 1986. This recurrently selected population known as Illinois High Oil Content (IHO) has been selected to increase 'oil concentration for ninety generations (Dudley, JW and RJ Lambert. (1992) Maydica 37: 1-1) using the modified mass selection. As a result, oil concentration increased in the population by more than 20%. Germplasm was little used because the derived material had substantially lower yields than conventional varieties (Alexander, DE (1988) In: Proc. 43rd Ann. Corn and Sorghum Res. Conf. Am. Seed Trade Assoc. Washington, DC pp 97-105). Using thirty-eight open and synthetic pollinated crops, Alexander initiated a second recurrent selection program (Alexho Synthetic) to increase the grain yield (Alexander, DE (1988) In: Corn and Corn Improvement, GF Sprague and JW Dudley eds American Society of Agronomy, Madison Wl. Pp 869-880). Oil levels equivalent to those of the IHO were achieved in twenty-eight cycles using the selection based on the oil concentration of single spikes and in later generations based on the concentration of simple grains. The performance of Alexho derived material in simple cross hybrids (spawn with high oil content x conventional spawn) improved over IHO, presumably due to the greater genetic variability initially available, although the performance was not equivalent to that of conventional hybrids. The development of agronomically elite corn germplasm. which also contains higher concentration of oil in the grain is clearly a challenge using conventional plant production methods. The concentration of oil in the grain can be measured phenotypically using a variety of analytical methods. The concentration of oil has a non-discrete distribution, common for inherited traits quantitatively controlled by several sites. The oil measurements of the grain select those producing lineages with the highest phenotypic expression. Unfortunately, the genetic potential for a high oil content is limited in most of these lineages because it is impossible to discriminate between lineages based on their true genetic makeup. This situation is further aggravated when simultaneous selection is practiced for agronomic performance or performance. Therefore, it would be advantageous to base the selection on the phenotype of the plants in the population. Genetic markers, especially nucleic acid markers, can advantageously be used as an indirect selection method for complex quantitative traits. The genetic markers that identify the alleles that confer a higher concentration of oil would therefore be an advantageous tool for the production programs of plants that develop maize germplasm with a high content of oil, elite. There is limited published information on the identification of predictive genetic markers for the increase in oil yield. Kahler (Kahler, AL (1985) In: Proc. 40th Ann. Corn And Sorghum Res. Conf. Am. Seed Trade Assoc. Washington DC pp. 66-89) measured the changes in the allelic frequency of the isozyme after the twenty-five selection cycles in a synthetic Alexho and found eight significant sites. Most of these changes in allele frequency were also significant for the random genetic shift in the tests, making it difficult to conclude that selection based on these isozyme alleles would be useful. More recently Goldman et al. (Goldman, I. L., 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 sites associated with oil concentration in 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 an increase in oil. Some of the regions identified by the RFLP marker site may be common between the two studies; however, of the fifteen RFLP markers that were used in both studies, six disagreed for their effect on oil concentration. In those studies the populations used were derived from common ancestors (Banco de Burr); however, populations were selected for different traits (oil and protein) over many generations. It is not surprising that many identified oil sites are unique to each population analyzed. It is therefore desirable to identify those genetic markers that are uniquely predictive of the germplasm that is being used in the production program.

BRIEF DESCRIPTION OF THE INVENTION A method is described for reliably and predictably reproducing corn with a higher concentration of oil in the grain. The method comprises a) using one or more genetic markers selected from a corn plant of a maize breeding population by marker-assisted selection, wherein the genetic markers are selected from the group consisting of sl375, sl384, sl394, S1416, sl422 , sl432, sl457, S1480, sl476, sl478, sl484, sl500, sl513, sl529, sl544, sl545, sl630, sl633, sl647, sl750, sl756, sl757, sl767, sl772, sl774, sl780, sl797, sl813, sl816, sl817 , sl836, sl853, sl860, sl870, sl921, sl922, sl925, sl931, sl933, sl939, sl946, sl949, s2054, s2055, s2057, s2058, s2097, s2122, s2125, s2150, s2156 and s2175; and b) crossing the selected maize plant with a second maize plant, where the progeny of the cross has a higher concentration of oil in the grain. A preferred source of maize germplasm with a high oil content is a member of a synthetic population of Alexho or a progeny thereof. A method to identify maize plants or maize lineages to be used as parents for the creation of a breeding population is also described, the method comprising a) determining the genotype of maize plants or maize lineages with one or more genetic markers, where the genetic markers are selected from the group consisting of S1375, S1384, sl394, sl416, sl422, sl432, sl457, sl480, S1476, S1478, sl484, sl500, sl513, sl529, sl544, sl545, S1630, sl633, sl647, sl750, sl756, sl757, sl767, sl772, S1774, sl780, sl797, sl813, sl816, sl817, sl836, sl853, S1860, sl870, sl921, sl922, sl925, sl931, sl933, sl939, S1946, sl949, s2054, s2055, s2057, s2058, s2097, s2122, s2125, s2150. s2156 and s2175; and b) identify maize plants or maize lineages, which, based on their genotype, produce transgressive segregants for the concentration of oil in the grain. The present invention provides a method for the identification and selection of genes to control the increase of the oil concentration in the corn kernel. Those alleles of the oil were initially identified in materials composed of, or derived from, synthetic Alexho breeding populations. In addition, the method facilitates the use of material with a high oil content in breeding programs, with the aim of developing corn germplasm with a high content of novel oil. Specifically, the method uses genetic markers to predict the value in an oil lineages in a maize breeding program. By indirectly selecting oil sites using those markers, those lineages with the highest genetic potential were selected for a higher concentration of oil in the grain. According to the method, any type of genetic marker can be used to identify an association with the concentration of oil in the grain. The method is limited only by the ability to measure the polymorphism at the site of a given marker. Those skilled in the art will recognize that different genetic markers that may be used include, but are not limited to, restriction fragment length polymorphisms (RFLP), randomly amplified polymorphic DNA (RAPD), single sequence repeats (SSR), AFLP. , several detection methods of simple base pairs, allozymes, and phenotypic markers. The SSR markers useful in the practice of the present invention include S1375, sl384, sl394, sl416, sl422, sl432, sl457, sl476, s1478, s1480, sl484, sl500, sl513, sl529, sl544, sl545, s1630, sl633, sl647 , sl750, sl756, sl757, sl767, sl772, S1774, S1780, sl797, sl813, sl816, sl817, sl836, sl853, s1860, sl870, sl921, sl922, sl925, sl931, sl933, sl939, sl946, sl949, s2054, s2055 , s2057, s2058, s2097, s2122, s2125, s2150, s2156 and s2175. A further embodiment of the present invention are the sites of the traits that control the expression of the oil concentration in the grain. These sites are identified and defined (that is, mapped) by the sites > markers of the present invention. A further embodiment of the present invention are maize plants and corn germplasm with a high oil content, which are produced using the breeding method herein.

DETAILED DESCRIPTION OF THE INVENTION Table 1 provides a brief description of the genetic markers that form part of the present invention. Each marker is defined by its constituent nucleic acid primers (forward and backward) that facilitate amplification of the specific marker site in the maize genome. The identifier required for each sequence is also indicated. 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 Useful genetic markers to define the location of the sites of the traits that control the oil concentration in the corn kernel Marker Sequence (5 '- 3') Type of Primer SEQ ID NO. sl375 TTTATGGGTTGGGAGATACTTG forward? AGATGTGTGCGTTTTTGAGAG backward "2 sl384 TTACGGCCTAGACATTTCGAC forward 3 CACTTGCTTTCAGGTACCCA backward 4 S1394 CTGCCCAGTCCGTAATGAA forward 5 TAGATTTATTTTCTGAACGATTGG backward 6 S1416 GATCTCTCTGAGGCTTGTCC forward TGTAGTTGAGGATGCTCCC backward si422 AGGCAAGGCTTTCTTCATAC forward CGGACGACGACTGTGTTC backward 10 S1432 ACATGAGAAACAAGATAGAACCAG forward 11 AAAATGTAAGAACTTGTTTGGGA backward 12 sl457 CTGCTTATTGCTTTCGTCATA forward 13 TGCTGCACTACTTGAACCTAG backward 14 s1476 ACACAGAGATGACAAAAGCAA forward 15 GCAGGCGTGCTATGAGAG backward 16 S1478 AGCGGTGAAACCCTTATG forward 17 CTGTGGCTGGTTCCTCTC backwards 18 S1480 GCTCTTGATAAAAAGGCAAGT forward 19 CTTGTTGTAATGGATGAGTGAG backwards 20 sl484 GCTCGTAGTAGGGGTTACG forward 21 GACAGCCTCACCTCAAGA backwards 22 S1500 ACAGATCTTGACACGTACATACC forward 23 GGACGTGTATCCTCAAATCAT backward 24 • yes 513 CAGCGAMACTGAATAACGC forward 25 TGTTGGATGAGCACTGAAC backward 26 s1529 TGTTCTCAACAACCACCG forward 27 CGTTTAGCGATATCATTTTCC backward 28 sl544 GATCCTACCAAAATCTTATAGGC forward 29 ACAGCTAGCCAAGATCTGATT backward 30 si545 CGATACTAATGGAAGCCCTAA forward 31 ATGGCCCATTAAGTTTATCAC backward 32 si63O AAAGCGTAGTCGGAAAGC forward 33 ACCAATGATCTTTACGCAGAT back 34 forward si633 TAATCAGAGCGTACATCAGGA 35 AGGGCATCAATCAAGAATG back 36 forward s1647 GAGACTTTTGAGGAGAAAGCA 37 GATCAAAAGAGCAAAAGGAGA back 38 forward s175O AACTGATGAATACCTTCCCAG 39 TGATTAACTTCTCCCTTTGGT back 40 forward s1756 TCGGCACAACATATGAGTTAC 41 CCCCCATAGAGAGAGATAGAG back 42 S1757 AAGCACGGCCCAATAGAAT forward 43 AGGATGTCCCTAGCTTTATTG back 44 S1767 TCATTGCCCAAAGTGTTG forward 45 CTCATCACCCCTCCAGAG backward 46 sl772 GATCCACGCCATTTAAAC forward 47 TGATACTCTGGTGCATGTTC backward 48 S1774 GATCGCTCCGATCTATCC forward 49 AGCGGCATCTATGTTCTATG backwards 50 S1780 CCCAGTGCGAAGAGACTC forward 51 ACACCTGCTCTGCACCAC rearward 52 s1797 CTAACCCACGACGACCCT forward 53 GCATGAGTGCATGTGCAT rearward 54 S1813 CTGCCACATGCTTTTCTG forward 55 CTGTAAAGAAGCTGGTCTGGA backward 56 sl816 TTCTCCTCATGGATGCGT forward 57 CTATTTGGAAGTATGGGCTTCA backward 58 S1817 GAGGGCATCTATGTGCAAC forward 59 GCTCAGAAGTTGCGTTTATG backward 60 sl836 TTCCTTCACGTTTCTCTGTTAA forward 61 CACATAAACCTAATGGGGTACA rearward 62 si853 CCCAAAGGCGATACCTATT forward 63 CCCACTTTCTCACTCTTTTCT backward 64 s1860 GAGGTGAGTACTATGCAAATGC forward 65 CAGGCTTACCTAGCCTTCTC backward 66 si870 CTATGGATGGCTGCTTGC forward 67 GTCAGGCAGCAGAATGTG rearward 68 s1921 AAACCGTCCAGCGACTAC forward 69 GGAAGAACCAATCCCATATCT backward 70 s1922 AACATCCTGTCGGAAACAG forward 71 TCATCACGTCTCTCTTTCAAC back 72 -s1925 TTGTGGCAGA? TCTCA TTA forward CGACTGGTGACATGTGAAG back 73 74 75 si931 AGTGAGGAAAGAATATGCTGG forward TGGACTGAGAAACTGATTTGA back 76 forward si933 CACAAATGTGAAGGTAAACACT 77 AATGGTACGGTTCAGGATG back 78 forward si939 AGATGACGCACGGAACAC 79 AGCATCATGTAGCAGGAGG back 80 forward s1946 TTGCAGCACTGTCGTAGTC GCGCGAGTGGAGTAGTAAG back 82 sl949 AAGATTATGGCAGATGAGACACC forward 83 GTTCCATGCTTTCCTTGG backward 84 s2054 GCCGATACCATGTAAGAGAAT forward 85 CTCTGGGCTCTGTGTTAGAGT backward 86 s2055 CTGCTTTCTCTGTTCCAGC forward 87 AATCGCTTACTTGTAACCCAC backward 88 s2057 AAGAACGTACGTCCCATAAAG forward 89 CAAGGTAAAGTGACAAAGCAG backward 90 S2058 GTTCAGGATGAGGCGGAA forward 91 GTGATCATCGCAGGAGACC rearward 92 s2097 GGAGCCTGGAGTGAGAAC forward 93 CATGCTCACCTAACGTGG rearward 94 s2122 ATCTGAACACTTGAGCAACAA forward 95 ATAGACCGGACCCATCAC rearward 96 s2125 CGAACAGCGGGTACACCT forward 97 GAGGTCAGCTTCCTCGATCT backward 98 s2150 GGAATCGTTCCTCCACAC forward 99 CTTCCTCGGTGTCAGACG backward 100 s2156 ATGGAAACATCAAAGTGGATT forward 101 TGCTACCCTGATGACCTGAT backward 102 s2175 ACCACTAGTCTCATATGAAGGG forward 103 GGTAGGTGGGTAGGGGTT backward 104 For the purposes of this invention, we define the following terms: Corn. Any variety, cultivation or population of Zea mays L. Elite. This term characterizes a plant or variety that has favorable traits, such as, but not limited to, high yield, good quality. grain and disease resistance. This allows its use in the commercial production of seeds or grains in a lucrative manner. The term also characterizes the parents that give rise to such plants or varieties. Corn Germplasm with a High Content of Oil. This term characterizes corn plants, which, when they are self-pollinated or used as the male or female parent in a variety of cross combinations, produce grains with higher oil content; when compared to the. grains produced by germplasm without a high oil content. Examples of corn germplasm with a high oil content include, but are not limited to, pollinated varieties of. open way, engendered lineages, races and populations or maize plants derived from one of the aforementioned.

Variety or crop. These terms refer to a group of similar plants that by structural and functional characteristics can be identified from other varieties or crops within the same species. Lineage. This term refers to a group of individuals of a common ancestor; a group more closely than a variety. 'Synthetic. This term refers to a genetically heterogeneous condition of plants of known ancestors, created by the intertwinement of any combination of spawns, hybrids, varieties, populations, races or other synthetics. Spawn. This term refers to an individual, variety or lineages substantially homozygous. Recombinant spawn. A population of independently derived lineages, developed by themselves repeatedly each generation until it approaches a homozygous state. Each recombinant spawn is derived from a single F2 plant using a harvest method commonly known as a single-seed descendant. Reproduction. The art and science of improving a plant or animal species through controlled genetic manipulation. Selection Helped by a Marker. The use of genetic markers to identify and select plants with superior phenotypic potential. The genetic markers determined previously associated with a site of a trait or trait sites were used to discover the genotype at the site of the trait by virtue of the link between the marker site and the site of the trait. The plants that contain the alleles of the desired trait are chosen based on their genotypes at the linked marker sites. Synthetic Alexho. Corn germplasm with a high content of oil, selected recurrently, developed by Alexander Denton at the University of Illinois. Alexho's maize germplasm with a high content of synthetic oil is composed of multiple synthetic populations defined by its cycle of advancement in the recurrent selection breeding program. Reproductive Population. A genetically heterogeneous collection of plants created for the purpose of / identifying one or more individuals with the desired phenotypic characteristics. Phenotype The observed expression of .one or more characteristics of the plant. Phenotypic value. A measurement of the expected expression of an allele from the site of the trait. The phenotypic value of an allele at the site of the trait depends on its expression strength compared to alternative alleles. The phenotypic value of an individual, which sequence its phenotypic potential, is based on its total genotypic composition at all sites for a given trait. Transgressive Segregants. Individuals whose phenotype exceeds the phenotypic variation predicted by the parents. Genetic marker Any morphological, biochemical or phenotypic difference based on the nucleic acid that reveals a DNA polymorphism. Examples of genetic markers include but are not limited to RFLPs, RAPDs, allozymes, SSRs, and AFLPs. Bookmark Site The genetically defined place of DNA polymorphisms revealed by a genetic marker. Trait Site. A genetically defined site for a collection of one or more genes (alleles) that contribute to an observed characteristic. Genotype. The allelic composition of an individual in the genetic sites under study. Restriction Fragment Length Polymorphism (RFLP). A genetic marker based on the DNA in which size differences are observed in the. DNA fragments generated by restriction endonucleases via hybridization (Botstein, D. et al., 1980. Am., J. Hum. Genet, 32: 314-331). Random Amplified Polymorphic DNA (RAPD).

A genetic marker based on DNA amplification in which primers of arbitrary, short sequence are used and the products resulting from the amplification are separated by size and the differences in the amplification patterns are observed (Williams, JGK et al., 1990. Nucleic Acids Res. 18: 6531-6535). Simple Sequence Repetition (SSR). A genetic marker based on DNA amplification in which short extensions of repeated sequence motifs are amplified in cascade and the products resulting from the amplification are separated by size and the differences in length of the repeated nucleotide are observed (Tautz D. 1989. Nucleic Acids Res. 122: 4127-4138). AFLP. A genetic marker based on the amplification of DNA in which the DNA fragments generated by the restriction endonuclease are linked to short DNA fragments which facilitate the amplification of restricted DNA fragments (Vos, P. et al., 1995. Nucleic Acids Res. 23: 4407-4414). The amplified fragments are separated by size and the differences in the amplification patterns are observed. - Alozymes. Enzyme variants which are electrophoretically separated and detected via staining for enzymatic activity (Stuber, C.W. and M.M. Goodman, 1983. USDA Agrie. Res. Results, Southern Ser., No. 16). The present invention relates to the discovery of sites of traits that control the concentration of oil in the grain through the use of genetic markers. In populations in which there is variation in the concentration of oil in the grain and the alleles of the genetic marker, oil measurements and genotypes based on the marker were generated for the members of the populations. Using the least squares method, the locations of the oil concentration sites were determined in relation to the markers genetically linked to those sites of the trait. Indirect selection of the preferred oil alleles can now be practiced using the information in one or more linked genetic markers. The selected maize plants comprise one or more alleles that code for a phenotype of high oil content. It is recognized that several different populations and types of populations could be used to locate sites of the trait of interest. Some of the population types include, but are not limited to, recombinant, retroactive, F2 or their pollinated or interaparelated and synthetic derivatives. In addition, you should understand that an alternative to measure genotypic and phenotypic variation within populations is the measurement of genotypes and phenotypes among populations. In this alternative, the second population is a derivative selected from the first population, the selection is either on the trait of interest (phenotypic selection) or supernatants specific markers (genotypic selection). It will also be recognized by those skilled in the art that alternative statistical methods may be used to determine a linkage relationship between the marker sites and the trait sites or traits.

EXAMPLES The present invention is better defined in the following Examples. It should be understood that these Examples, while indicating the preferred embodiments of the invention, are given by way of illustration only. From the foregoing discussion and those Examples, one skilled in the art can determine 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 this to various uses and terms.

EXAMPLE 1 LOCATION OF SITES CONFERRING A GREATER CONCENTRATION OF GRAIN OIL Development of the population and the trait Two maize lineages spawned LH119wx and LH51, developed by the Holden's Foundation Seed Co., Williambsburg, IA, were independently paired with individual plants of the synthetic population ASKC28wx (deposited in the American Type Culture Collection, Rockville, MD; Accession No. ATCC 75105) (waxy seeds are highly represented in ASKC28 and therefore, we have designated ASKC28 as waxy). The Fl plants were individualized, and the resulting F2 populations were grown. The individual F2 plants were individualized and the derived grains were advanced using a single seed descended through six generations of individualization (S6), to produce recombinant sired lineages. Until they grew, twenty grains of the S6 generation were individualized producing a family of S7 spikes representing each recombinant sired lineage. Oil values were determined for each ear within a family using near infrared transmittance (Williams, PC (1987) In: Near Infrared Technology in Food and Agricultural Industries, PC Williams and C. Norris, eds. Association of Cereal Chemists).

Genotypic Determination The single-cob seeds representing each of the one hundred ninety-four (LH119wx x ASKC28wx) or two hundred four (LH51 x ASKC28wx) recombinant sired lineages were germinated on wet filter paper. Root fragments of the germinated seeds were excised, gathered on the cob and extracted using an automated DNA extraction machine. The instrument uses a modification of the CTAB procedure of Murray and Thompson (Murray, M. G. and Thompson, W. F. (1980) Nucí.Aids Res. 8: 4321-4325). DNA samples were quantified via fluorescence using YoPro-lMR iodine (Molecular Probes, Inc., Eugene, OR) and diluted to 4 μg / ml. The SSR regions of each sample of DNA using the following protocol: 1. 10 μl of amplification cocktail (see Table 2) was added to 5 μl (20 ng) of extracted DNA. 2. The DNA fragment flanked by the 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) 45 es of 50 sec. At 95 ° C, 50 sec. At 54 ° C and 80 sec. At 72 ° C and 2) 1 e of 300 sec. At 72 ° C: 3. Approximately 8 μl of each sample was loaded onto agarose gels composed of 2% Metaphor (FMC, Corp., Rockland, ME), IX TBE, and 0.5 μg / ml ethidium bromide, and electrophoresed for 2 h at 6.1 V / cm in horizontal electrophoresis units, to which buffer IX was added. TBE and 0.5 μg / ml ethidium bromide; 4. The DNA bands were visualized by UV fluorescence.

Table 2 Reagent Amplification Cocktail Concentration Final Pattern Concentration Shock Absorber * 10X 1.5X DNTP 2 mM 0.3 mM Primer Forward 40 μM 0.45 μM Primer To 'Back 40 μM 0.45 μM Polymerase AmpliTaq .1MR 5 U / μl 0.05 U / μl * Shock Absorber 10X is a pH 9.0 solution of 800 mM Tris-OH, 200 mM (NH4) 2 SO, and 25 mM MgCl2.

Location of sites for oil One hundred thirty-three polymorphic SSR markers were used to determine the phenotype of the recombinant spawn for the LH119wx x ASKC28wx junction and one hundred three polymorphic SSR marker sites were used to determine the genotype of the population derived from LH51 x ASKC28 x. In addition, maps were also drawn from twenty publicly available polymorphic SSR sites with the locations of the chromosomes established above and covering the ten corn chromosomes (available from Research Genetics, Huntsville, AL) in both populations. The genetic linkage and the distance between the marker sites were determined independently of 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 corn chromosomes. Each link group was assigned to a chromosome based on the link to the public SSR markers. Twenty-three and ten markers in the populations LH119wx x ASKC28wx and LH51 x ASKC28 x, respectively, were not assigned to the positions of the chromosomes because the genetic link could not be clearly established. The analysis of variance was used to identify the marker sites in the link with the sites of the trait or traits that confer a higher concentration of oil. The concentration of the oil was used as a dependent variable and the ANOVAS were calculated, separated with the SAS Proc GLM (SAS inst., Cary, NC) using each marker site as a single independent variable (Edwards., MD, et al. 1987) Genetics 11 6: 113-125). Therefore, for each ANOVA test, the average oil values of the marker allele classes were compared. The marker sites were declared significant if p < 0.05. We examined link or connection data for significant marker sites to determine both the number of feature sites present, as its probable location. Significant marker sites on the same linkage group detected the same site of the trait or alternatively different sites of the trait or traits. By careful examination of the phenotypic variation explained by each marker site along the chromosome, a determination was made of the number of trait sites on a linkage group. It was stated that significant marker sites, on the same linkage group not interrupted by non-significant marker sites, detect the same site of the trait on the chromosome. If significant marker sites on the same chromosome were interrupted by non-significant marker sites, then it would be declared that each significant region contains a trait site that results in multiple trait sites on the same chromosome.

To confirm the number of trait sites, the marker data assigned to linkage groups and oil data were also analyzed with Mapmaker / QTL 1.0 (Lincoln, SE et al., (1990) Whitehead Inst. Biomed. Res. , Cambridge, MA). The results with the Mapmaker agreed with the initial analysis of the number of trait sites on each 'chromosome. Eleven and twelve sites that control oil concentration in the grain were located in recombinant sired populations LH119wx x "ASKC28wx and LH51 x ASKC28wx, respectively. Each oil site is defined by one or more sites of the linked marker. In cases where the same marker sites were used in both populations, alignment of linkage groups is possible. It was found that in most cases both populations located the same sites for the oil. Considering the common marker sites, a total of seventeen sites were found that control the concentration of oil in the grain. To each site for the. Oil was assigned an arbitrary alphabetical designation (Table 3).

Table 3 Markers sites genetically linked to, and that predict the location by means of the sites of the trait that confer greater oil concentration in the grain Site for the Oil Chromosome Sites Markers _. 1 S1922 B 1 sl478, S1853, sl949 C 1 S1860, sl925, sl931, s2150 D 2 s2175 E 3 S1394 F 4 S1476, S1772, S1816, S2122, S1836 G 4 S1939, S1946 H 4 sl870 I 5 sl529 J 5 s2054, sl647, sl500, sl545, S1774, s2097 K 6 S1457, s20"55, S1757, S2125, S1780, S1375, S1797, S1416, S1432, S1921 7 S1630, S1422, S2156 M 8 sl817, s2057 Table 3 (Continued) Site for the Chromosome Oil Sites Bookmarks Ñ 9 sl544, sl633, sl384, S1813, S1767, s2058, S1933, S1513, sl484 OR 10 S1756 P 10 yes480 (positive allele of the oil in LH51) Q NA * sl750 * NA - location of the unknown chromosome In cases where comparisons could be made, sites for the oil that were identified in a population were identified in the same place as the second population In two exceptions, the site for the oil was found in one population, but not in the second In the first case, the allele with a positive effect on the oil was found in LH51, and of this node , it would not be expected to identify the same site in the population LH119 x x 'ASKC28 x. In the second case, it was found that: different marker alleles derived from ASKC28wx were segregating in the populations. Therefore, the effect of oil on each population of a different allele ASKC28 x at the site of the trait was measured. The allele for the more abundant ASKC28 x oil secreted in the LH119 xx ASKC28wx had a positive effect on the oil on the allele derived from alternative LH119 x, whereas in the LH51 x ASKC28wx population, the ASKC28 x abundant allele did not have a positive effect on the oil. With the exception of the site for oil bound to marker S1480, all alleles with positive effects on oil concentration were derived from ASKC28wx. i EXAMPLE 2 SELECTION OF REPRODUCTIVE LINES HELPED BY THE MARKER USING GENETIC MARKERS FOR GREATER CONCENTRATION OF GRAIN OIL The genetic marker sites related to the trait sites for the oil are highly predictive of the oil concentration and as such can be used as an indirect measurement of the oil in the grain in a selection program aided by a marker. Consequently, the genotypic information of linked marker sites would facilitate the selection of reproductive lineages with a higher concentration of oil. Direct measurements of oil can not differentiate between different compositions of the genotypic trait site with equivalent phenotypic effects. This is especially problematic in the first generation that segregates the breeding populations, where only a limited fixation of the sites for the oil has occurred. As an example, an objective of a corn breeding program could be the creation of new elite sired lineages, which contain alleles of traits that confer a higher concentration of oil in the grain. Those alleles of the trait would be introduced by 'the inter-pairing of germplasm with a high oil content with one or more elite corn spawn. The resulting hybrid could be self-pollinated to produce an F2 population for the purpose of initiating a conventional pedigree breeding program (Allard R. W. (1960) Principles of Plant Breeding, John Wiley &; Sons, Inc. New York, pp 115-128). To identify those F2 individuals with the desired genotypes, tissues of the plant of each F2 individual would be collected in the population and the genotype would be determined with the SSR marker sites listed in Table 1. Those F2 individuals with the highest frequency of SSR marker alleles derived from a source with a high oil content would also be selected and chosen based on their agronomic convenience. Continuing reproduction and segregation, those sites for oil in a heterozygous state, could be set for the allele of high oil content or low oil content. Therefore, it is likely that the genotype determination and material selection of later generations could be practiced to further segregate breeding lineages based on their marker allele and consequently the composition of the allele for the oil. Depending on the size and serendipity of the population, the spawning offspring of the pedigree production program may not demonstrate sufficient agronomic competitiveness or sufficient expression of oil in the grain because an inadequate number of alleles was recovered for the oil. These new monsters could therefore be used as parent material and initiate new breeding projects. The SSR markers could be used again for a greater oil selection as described. It is obvious to those skilled in the art that many variants to the selection methodology can be devised. The selection could be based on the allelic composition of one or more marker sites that identify the sites for the oil trait present in a population. Additional selection would be made by examining and selecting genotypes of individual plants, families or their progeny. Predictive models could be developed using genotypic information, which could generate several selection indices. These models would allow to weight the effect predicted by the marker sites. This is because the predictive value of an individual marker site depends on its genetic distance from the corresponding trait site as well as the ability to express the trait site. Selection strategies that combine selection based on the phenotype and based on the genotype can also be contemplated. The marker sites presented here are predictive for sites for oil in synthetic populations of Alexho. Because the ASKV28wx represent the 28th reproduction cycle for the oil of a genetically close population, any reproduction cycles are composed of the same sites for the oil. It is expected that the cycles will simply differ in their allelic frequency at the identified oil sites. Therefore, in breeding populations derived from the first Alexho cycles, the marker sites described in this invention will be useful for the identification of the sites for the oil and the prediction of the oil concentration.

EXAMPLE 3 IDENTIFICATION OF CORN PLANTS TO BE USED AS PARENTS FOR THE PRODUCTION OF TRANSGRESSIVE SEGREGANTES FOR THE CONCENTRATION OF OIL IN GRAIN It is important to identify maize plants and lineages that, when used as parents, have the highest probability of producing high performance descent.

The segregating transgressive descent of such parents would result from the crossing of parents with complementary sets of alleles conferring a phenotype of high oil content. Using the information provided herein, marker alleles are known to illustrate the performance or function of the desired trait (i.e., high concentration, oil) at a given marker site. By determining the genotypes of the lineages in those marker sites, the value of those lineages as parents is revealed. For example, if it is desired to create an individual containing higher alleles at 5 separate oil sites (AE), a parent or origin composed of the desired alleles A, B and C could be identified with a parent or origin composed of The desired alleles in B, D and E. These parents are complementary because they allow the recovery of a progeny that contains the desired alleles in the 5 sites. Ideally, the parents would be chosen from which, when combined, ensure the maximum complementation of the sites, so that a high frequency of the desired recombinants is recovered.

LIST OF SEQUENCES (1) GENERAL INFORMATION: (A) RECIPIENT: E. l. 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: 302-992-4926"( H) TELEFAX: 302-773-0164 (I) TELEX: 6717325 (ii) TITLE OF THE INVENTION: A METHOD FOR IDENTIFYING AND PRODUCING MAIZE WITH A MAJOR CONCENTRATION OF OIL IN THE GRAIN (iii) SEQUENCE NUMBER: 104 (iv) COMPUTER LEGIBLE FORM: (A) TYPE OF MEDIA: FLEXIBLE DISK, 3.5 INCHES (B) COMPUTER: IBM COMPATIBLE PC (C) OPERATING SYSTEM: MICROSOT WINDOWS 95 (D) PROGRAM: MICROSOFT WORD VERSION 7.0A (v) DATA OF THE CURRENT APPLICATION: (A) APPLICATION NUMBER: (B) DATE OF PRESENTATION: (C) CLASSIFICATION: (vi) DATA FROM THE PREVIOUS APPLICATION: (A) APPLICATION NUMBER: 60 / 041,515 (B) SUBMISSION DATE: MARCH 24, 1997 (C) CLASSIFICATION: (vii) INFORMATION OF THE APPORTER / AGENT (A) NAME: MAJARÍAN, WILLIAM R. (B) REGISTRATION NUMBER: P-41,173 (C) REFERENCE NUMBER / DOSSIER: BB-1076 (2) INFORMATION FOR SEQ ID NO: l: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: l: TTTATGGGTT GGGAGATACT TG 22 (2) INFORMATION FOR SEQ ID NO: 2: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: l: AGATGTGTGC GTTTTTGAGA G 21 (2) INFORMATION FOR SEQ ID NO: 3: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 3: TTACGGCCTA GACATTTCGA C 21 (2) INFORMATION FOR SEQ ID NO: 4: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 4: CACTTGCTTT CAGGTACCCA 20 (2) INFORMATION FOR SEQ ID NO: 5: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 5: CTGCCCAGTC CGTAATGAA 19 (2) INFORMATION FOR SEQ ID NO: 6: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 24 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 6: TAGATTTATT TTCTGAACGA TTGG 24 (2) INFORMATION FOR SEQ ID NO: 7: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 7: GATCTCTCTG AGGCTTGTCC 20 (2) INFORMATION FOR SEQ ID NO: 8: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 8: TGTAGTTGAG GATGCTCCC 19 (2) INFORMATION FOR SEQ ID NO: 9: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 9: AGGCAAGGCT TTCTTCATAC 20 (2) INFORMATION FOR SEQ ID NO: 10: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 10: CGGACGACGA CTGTGTTC 18 (2) INFORMATION FOR SEQ ID NO: 11: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 24 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 11: ACATGAGAAA CAAGATAGAA CCAG 24 (2) INFORMATION FOR SEQ ID NO: 12: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 12: AAAATGTAAG AACTTGTTTG GGA 23 (2) INFORMATION FOR SEQ ID NO: 13: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 13: CTGCTTATTG CTTTCGTCAT A 21 (2) INFORMATION FOR SEQ ID NO: 14:. (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 14: TGCTGCACTA CTTGAACCTA G 21 (2) INFORMATION FOR SEQ ID NO: 15: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 15: ACACAGAGAT GACAAAAGCA A 21 (2) INFORMATION FOR SEQ ID NO: 16: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple. (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 16: GCAGGCGTGC TATGAGAG 18 (2) INFORMATION FOR SEQ ID NO: 17: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 17: AGCGGTGAAA CCCTTATG 18 (2) INFORMATION FOR SEQ ID NO: 18: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 18: CTGTGGCTGG TTCCTCTC 18 (2) INFORMATION FOR SEQ ID NO: 19: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 19: GCTCTTGATA AAAAGGCAAG T 21 (2) INFORMATION FOR SEQ ID NO: 20: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 20: CTTGTTGTAA TGGATGAGTG AG 22 (2) INFORMATION FOR SEQ ID NO: 21: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 21: GCTCGTAGTA GGGGTTACG 19 (2) INFORMATION FOR SEQ ID NO: 22: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 22: GACAGCCTCA CCTCAAGA 18 (2) INFORMATION FOR SEQ ID NO: 23: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 23: ACAGATCTTG ACACGTACAT ACC 23 (2) INFORMATION FOR SEQ ID NO: 24: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 24: GGACGTGTAT CCTCAAATCA T 21 (2) INFORMATION FOR SEQ ID NO: 25: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid * (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 25: CAGCGAATAC TGAATAACGC 20 (2) INFORMATION FOR SEQ ID NO: 26: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 26: TGTTGGATGA GCACTGAAC 19 (2) INFORMATION FOR SEQ ID NO: 27: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base 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AAGATCTGAT T 21 (2) INFORMATION FOR SEQ ID NO: 31: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 31: CGATACTAAT GGAAGCCCTA A 21 (2) INFORMATION FOR SEQ ID NO: 32: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 32: ATGGCCCATT AAGTTTATCA C 21 (2) INFORMATION FOR SEQ ID NO: 33: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 33: AAAGCGTAGT CGGAAAGC 18 (2) INFORMATION FOR SEQ ID NO: 34: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 34: ACCAATGATC TTTACGCAGA T 21 (2) INFORMATION FOR SEQ ID NO: 35: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 35: TAATCAGAGC GTACATCAGG A 21 (2) INFORMATION FOR SEQ ID NO: 36: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 36: AGGGCATCAA TCAAGAATG 19 (2) INFORMATION FOR SEQ ID NO: 37: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 37: GAGACTTTTG AGGAGAAAGC A 21 (2) INFORMATION FOR SEQ ID NO: 38: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 38: GATCAAAAGA GCAAAAGGAG A 21 (2) INFORMATION FOR SEQ ID NO: 39: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 39: AACTGATGAA TACCTTCCCA G 21 (2) INFORMATION FOR SEQ ID NO: 40: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 40: TGATTAACTT CTCCCTTTGG T 21 (2) INFORMATION FOR SEQ ID NO: 41: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 41: TCGGCACAAC ATATGAGTTA C 21 (2) INFORMATION FOR SEQ ID NO: 42: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 42: CCCCCATAGA GAGAGATAGA G 21 (2) INFORMATION FOR SEQ ID NO: 43: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 43: AAGCACGGCC CAATAGAAT 19 (2) INFORMATION FOR SEQ ID NO: 44: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 44: AGGATGTCCC TAGCTTTATT G 21 (2) INFORMATION FOR SEQ ID NO: 45: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 45: TCATTGCCCA AAGTGTTG 18 (2) INFORMATION FOR SEQ ID NO: 46: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TI O: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 46: CTCATCACCC CTCCAGAG 18 (2) INFORMATION FOR SEQ ID NO: 47: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid '(C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 47: GATCCACGCC ATTTAAAC 18 (2) INFORMATION FOR SEQ ID NO: 8: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 48: TGATACTCTG GTGCATGTTC 20 (2) INFORMATION FOR SEQ ID NO: 9: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 49: GATCGCTCCG ATCTATCC 18 (2) INFORMATION FOR SEQ ID NO: 50: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 50: AGCGGCATCT ATGTTCTATG 20 (2) INFORMATION FOR SEQ ID NO: 51: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 51: CCCAGTGCGA AGAGACTC 18 (2) INFORMATION FOR SEQ ID NO: 52: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 52: ACACCTGCTC TGCACCAC 18 (2) INFORMATION FOR SEQ ID NO: 53: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid "(C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 53: CTAACCCACG ACGACCCT 18 (2) INFORMATION FOR SEQ ID NO: 54: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 54: GCATGAGTGC ATGTGCAT 18 (2) INFORMATION FOR SEQ ID NO: 55: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 55: CTGCCACATG CTTTTCTG 18 (2) INFORMATION FOR SEQ ID NO: 56: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 56: CTGTAAAGAA GCTGGTCTGG A 21 (2) INFORMATION FOR SEQ ID NO: 57: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 57: TTCTCCTCAT GGATGCGT 18 (2) INFORMATION FOR SEQ ID NO: 58: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 58: CTATTTGGAA GTATGGGCTT CA 22 (2) INFORMATION FOR SEQ ID NO: 59: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 59: GAGGGCATCT ATGTGCAAC 19 (2) INFORMATION FOR SEQ ID NO: 60: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 60: GCTCAGAAGT TGCGTTTATG 20 (2) INFORMATION FOR SEQ ID NO: 61: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 61: TTCCTTCACG TTTCTCTGTT AA 22 (2) INFORMATION FOR SEQ ID NO: 62: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 62: CACATAAACC TAATGGGGTA CA 22 (2) INFORMATION FOR SEQ ID NO: 63: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 63: CCCAAAGGCG ATACCTATT 19 (2) INFORMATION FOR SEQ ID NO: 64: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 64: CCCACTTTCT CACTCTTTTC T 21 (2) INFORMATION FOR SEQ ID NO: 65: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 65: GAGGTGAGTA CTATGCAAAT GC 22 (2) INFORMATION FOR SEQ ID NO: 66: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 66: CAGGCTTACC TAGCCTTCTC 20 (2) INFORMATION FOR SEQ ID NO: 67: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 67: CTATGGATGG CTGCTTGC 18 (2) INFORMATION FOR SEQ ID NO: 68: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 68: GTCAGGCAGC AGAATGTG 18 (2) INFORMATION FOR SEQ ID NO: 69: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 69: AAACCGTCCA GCGACTAC 18 (2) INFORMATION FOR SEQ ID NO: 70: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 70: GGAAGAACCA ATCCCATATC T 21 (2) INFORMATION FOR SEQ ID NO: 71: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 71: AACATCCTGT CGGAAACAG 19 (2) INFORMATION FOR SEQ ID NO: 72: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 72: TCATCACGTC TCTCTTTCAA C 21 (2) INFORMATION FOR SEQ ID NO: 73: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid '(C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 73: TTGTGGCAGA ATCTCAAATT A 21 (2) INFORMATION FOR SEQ ID NO: 74: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 74: CGACTGGTGA CATGTGAAG 19 (2) INFORMATION FOR SEQ ID NO: 75: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 75: AGTGAGGAAA GAATATGCTG G 21 (2) INFORMATION FOR SEQ ID NO: 76: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 76: TGGACTGAGA AACTGATTTG A 21 (2) INFORMATION FOR SEQ ID NO: 77: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid "(C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 77: CACAAATGTG AAGGTAAACA CT 22 (2) INFORMATION FOR SEQ ID NO: 78: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 78: AATGGTACGG TTCAGGATG 19 (2) INFORMATION FOR SEQ ID NO: 79: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 79: AGATGACGCA CGGAACAC 18 (2) INFORMATION FOR SEQ ID NO: 80: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 80: AGCATCATGT AGCAGGAGG 19 (2) INFORMATION FOR SEQ ID NO: 8: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 81: TTGCAGCACT GTCGTAGTC 19 (2) INFORMATION FOR SEQ ID NO: 82: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 82: GCGCGAGTGG AGTAGTAAG 19 (2) INFORMATION FOR SEQ ID NO: 83: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 83: AAGATTATGC AGATGAGACA CC 22 (2) INFORMATION FOR SEQ ID NO: 84: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 84: GTTCCATGCT TTCCTTGG 18 (2) INFORMATION FOR SEQ ID NO: 85: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 85: CTCTGGGCTC TGTGTTAGAG T 21 (2) INFORMATION FOR SEQ ID NO: 86: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 86: CTCTGGGCTC TGTGTTAGAG T 21 (2) INFORMATION FOR SEQ ID NO: 87: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 87: CTGCTTTCTC TGTTCCAGC 19 (2) INFORMATION FOR SEQ ID NO: 88: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 88: AATCGCTTAC TTGTAACCCA C 21 (2) INFORMATION FOR SEQ ID NO: 89: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 89: AAGAACGTAC GTCCCATAAA G 21 (2) INFORMATION FOR SEQ ID NO: 90: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 90: CAAGGTAAAG TGACAAAGCA G 21 (2) INFORMATION FOR SEQ ID NO: 91: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 91: GTTCAGGATG AGGCGGAA 18 (2) INFORMATION FOR SEQ ID NO: 92: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 92: GTGATCATCG CAGGAGACC 19 (2) INFORMATION FOR SEQ ID NO: 93: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 93: GGAGCCTGGA GTGAGAAC 18 (2) INFORMATION FOR SEQ ID NO: 94: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 94: CATGCTCACC TAACGTGG 18 (2) INFORMATION FOR SEQ ID NO: 95: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 95: ATCTGAACAC TTGAGCAACA A 21 (2) INFORMATION FOR SEQ ID NO: 96: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 96: ATAGACCGGA CCCATCAC 18 (2) INFORMATION FOR SEQ ID NO: 97: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 97: CGAACAGCGG GTACACCT 18 (2) INFORMATION FOR SEQ ID NO: 98: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 98: GAGGTCAGCT TCCTCGATCT 20 (2) INFORMATION FOR SEQ ID NO: 99: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 99: GGAATCGTCC CTCCACAC 18 (2) INFORMATION FOR SEQ ID NO: 100: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 100: CTTCCTCGGT GTCAGACG 18 (2) INFORMATION FOR SEQ ID NO: 101: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 101: ATGGAAACAT CAAAGTGGAT T 21 (2) INFORMATION FOR SEQ ID NO: 102: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 102: TGCTACCCTG ATGACCTGAT 20 (2) INFORMATION FOR SEQ ID NO: 103: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 103: ACCACTAGTC TCATATGAAG GG 22 (2) INFORMATION FOR SEQ ID NO: 104: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 104: GGTAGGTGGG TAGGGGTT 18 It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (20)

CLAIMS Having described the invention as above, the content of the following claims is claimed as property:

1. A method for producing corn, with a higher concentration of oil in the grain, characterized in that it comprises: a) using one or more genetic markers selected from a maize plant of a corn-producing population by selection assisted by the marker, where the Genetic markers are selected from the group consisting of sl375, sl384, sl394, sl416, sl422, sl432, sl457, s1480, s1476, sl478, sl484, sl500, sl513, sl529, sl544, s1545, s1630, sl633, sl647, sl750, sl756 , sl757, sl767, sl772, sl774, sl780, sl797, sl813, sl816, sl817, sl836, s1853, s1860, sl870, sl921, sl922, sl925, sl931, sl933, sl939, sl946, sl949, s2054, s2055, s2057, s2058 , s2097, s2122, s2125, s2150, s2156 and s2175; and b) crossing the selected maize plant with a second plant where the progeny of the maize plants of the crossing have a higher concentration of oil in the grain.

The method according to claim 1, characterized in that the selected maize plant is a member of an annexed synthetic population or progeny thereof.

3. A method to identify maize plants or maize lineages to be used as parents for the creation of a producing population, the method is characterized because it comprises: a) determining the genotype of 'maize plants or maize lineages with one or more genetic markers, where the genetic markers are selected from the group consisting of sl375, sl384, sl394, sl416, "sl422, sl432, sl457, sl480, s1476, sl478, sl484, sl500, sl513, sl529, sl544, s1545, s1630, sl633, sl647, sl750, sl756, sl757, sl767, s1772, s1774, sl780, sl797, sl813, sl816, sl817, sl836, s1853, sl860, sl870, sl921, sl922, sl925, sl931, sl933, sl939, sl946, sl949, s2054, s2055, s2057, s2058, s2097, s2122, s2125, s2150, s2156 and s2175, and b) identify maize plants or maize lineages which, based on their genotype, are estimated to produce transgressive segregants for oil concentration in the grain

4. A trait site that controls the concentration of oil in the grain, Terized because the site map is plotted by a genetic marker selected from the group consisting of s2054, sl647, sl500, S1545, S1774 and s2097.

5. The trait site that controls the concentration of oil in the grain, characterized in that the site map is plotted by a genetic marker selected from the group consisting of sl817 and s2057.

6. A trait site that controls the concentration of oil in the grain, characterized in that the site map is 'traced by a' genetic marker selected from the group consisting of sl860, sl931, s2150 and sl925 7.

A trait site - which controls the concentration of oil in the grain, characterized in that the site map is drawn by a genetic marker selected from the group consisting of sl457, s2055, sl757, s2125, S1780, sl375, sl797, sl416, sl432 and sl921.

A trait site that controls the concentration of oil in the grain, characterized in that the site map is traced by a genetic marker selected from the group consisting of sl544, sl633, sl384, S1813, S1767, s2058, sl933, sl513 and sl484

9. A trait site that controls the concentration of oil in the grain, characterized in that the site map is mapped by a genetic marker selected from the group consisting of sl476, sl772, sl816, S2122 and S1836.

10. A trait site that controls the concentration of oil in the grain, characterized in that the site map is plotted by a genetic marker selected from the group consisting of sl939 and sl946.

11. A trait site that controls the concentration of oil in the grain, characterized in that the site map is 'traced by a' genetic marker selected from the group consisting of sl478, sl853 and sl949

12. A trait site that controls the concentration of oil in the grain, characterized in that the site map is plotted by a genetic marker selected from the group consisting of sl630, sl422 and s2156.

13. A trait site that controls the oil concentration in the grain, characterized because the site map is plotted by the sl756 genetic marker.

14. A trait site that controls the oil concentration in the grain, characterized because the site map is plotted by the sl922 genetic marker.

15. A trait site that controls the concentration of oil in the grain, characterized in that the site map is mapped by the sl529 genetic marker.

16. A trait site that controls the oil concentration in the grain, characterized in that the site map is plotted by the sl394 genetic marker.

17. A trait site that controls the concentration of oil in the grain, characterized in that the site map is mapped by the genetic marker S1750.

18. A trait site that controls the oil concentration in the grain, characterized because the site map is plotted by the sl870 genetic marker.

19. A "trait site that controls the concentration of oil in the grain, characterized in that the site map is mapped by the genetic marker 's2175

20. Corn plants that have a higher concentration of oil in the grain, characterized by produced by the method of claim 1.