# Patent

## Title
Inbred maize line PHB6V

## Abstract

An inbred maize line, designated PHB6V, the seeds and plants of inbred maize line PHB6V, methods for producing a maize plant, either inbred or hybrid, produced by crossing the inbred maize line PHB6V with another maize plant, and seed and plants produced therefrom. The invention also relates to methods for producing a modified PHB6V maize plant that comprises in its genetic material one or more transgenes or backcross conversion genes and to the transgenic and backcross conversion maize plants produced by these methods. This invention also relates to methods for producing other inbred and hybrid maize lines derived from inbred maize line PHB6V and to the inbred and hybrid maize lines so produced.

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a continuation of U.S. patent application Ser. No. 10/355,622, now U.S. Pat. No. 6,967,268, filed on Jan. 31, 2003, the contents of which are hereby incorporated by reference in their entirety.


FIELD OF THE INVENTION
This invention relates generally to the field of maize breeding, specifically relating to an inbred maize line designated PHB6V.
BACKGROUND OF THE INVENTION
The goal of plant breeding is to combine, in a single variety or hybrid, various desirable traits. For field crops, these traits may include resistance to diseases and insects, tolerance to heat and drought, reducing the time to crop maturity, greater yield, and better agronomic quality. With mechanical harvesting of many crops, uniformity of plant characteristics such as germination and stand establishment, growth rate, maturity, and plant and ear height, is important. Traditional plant breeding is an important tool in developing new and improved commercial crops.
SUMMARY OF THE INVENTION
According to the invention, there is provided a novel inbred maize line, designated PHB6V. This invention thus relates to the seeds of inbred maize line PHB6V, to the plants of inbred maize line PHB6V, to plant parts of inbred maize line PHB6V, to methods for producing a maize plant produced by crossing the inbred maize line PHB6V with another maize plant, including a plant that is part of a synthetic or natural population, and to methods for producing a maize plant containing in its genetic material one or more backcross conversion trait or transgene and to the backcross conversion maize plants and plant parts produced by those methods. This invention also relates to inbred maize lines and plant parts derived from inbred maize line PHB6V, to methods for producing other inbred maize lines derived from inbred maize line PHB6V and to the inbred maize lines and their parts derived by the use of those methods. This invention further relates to hybrid maize seeds, plants and plant parts produced by crossing the inbred line PHB6V or a backcross conversion of PHB6V with another maize line.
Definitions
Certain definitions used in the specification are provided below. Also in the examples that follow, a number of terms are used herein. In order to provide a clear and consistent understanding of the specification and claims, including the scope to be given such terms, the following definitions are provided. NOTE: ABS is in absolute terms and % MN is percent of the mean for the experiments in which the inbred or hybrid was grown. PCT designates that the trait is calculated as a percentage. % NOT designates the percentage of plants that did not exhibit a trait. For example, STKLDG % NOT is the percentage of plants in a plot that were not stalk lodged. These designators will follow the descriptors to denote how the values are to be interpreted.
ABTSTK=ARTIFICIAL BRITTLE STALK. A count of the number of “snapped” plants per plot following machine snapping. A snapped plant has its stalk completely snapped at a node between the base of the plant and the node above the ear. Expressed as percent of plants that did not snap.
ALLELE. Any of one or more alternative forms of a genetic sequence. Typically, in a diploid cell or organism, the two alleles of a given sequence occupy corresponding loci on a pair of homologous chromosomes.
ANT ROT=ANTHRACNOSE STALK ROT (Colletotrichum graminicola). A 1 to 9 visual rating indicating the resistance to Anthracnose Stalk Rot. A higher score indicates a higher resistance.
BACKCROSSING. Process in which a breeder crosses a progeny line back to one of the parental genotypes one or more times.
BARPLT=BARREN PLANTS. The percent of plants per plot that were not barren (lack ears).
BREEDING. The genetic manipulation of living organisms.
BREEDING CROSS. A cross to introduce new genetic material into a plant for the development of a new variety. For example, one could cross plant A with plant B, wherein plant B would be genetically different from plant A. After the breeding cross, the resulting F1 plants could then be selfed or sibbed for one, two, three or more times (F1, F2, F3, etc.) until a new inbred variety is developed. For clarification, such new inbred varieties would be within a pedigree distance of one breeding cross of plants A and B.
BRTSTK=BRITTLE STALKS. This is a measure of the stalk breakage near the time of pollination, and is an indication of whether a hybrid or inbred would snap or break near the time of flowering under severe winds. Data are presented as percentage of plants that did not snap.
CLDTST=COLD TEST. The percent of plants that germinate under cold test conditions.
CLN=CORN LETHAL NECROSIS. Synergistic interaction of maize chlorotic mottle virus (MCMV) in combination with either maize dwarf mosaic virus (MDMV-A or MDMV-B) or wheat streak mosaic virus (WSMV). A 1 to 9 visual rating indicating the resistance to Corn Lethal Necrosis. A higher score indicates a higher resistance.
COMRST=COMMON RUST (Puccinia sorghi). A 1 to 9 visual rating indicating the resistance to Common Rust. A higher score indicates a higher resistance.
D/D=DRYDOWN. This represents the relative rate at which a hybrid will reach acceptable harvest moisture compared to other hybrids on a 1 to 9 rating scale. A high score indicates a hybrid that dries relatively fast while a low score indicates a hybrid that dries slowly.
DIPERS=DIPLODIA EAR MOLD SCORES (Diplodia maydis and Diplodia macrospora). A 1 to 9 visual rating indicating the resistance to Diplodia Ear Mold. A higher score indicates a higher resistance.
DIPROT=DIPLODIA STALK ROT SCORE. Score of stalk rot severity due to Diplodia (Diplodia maydis). Expressed as a 1 to 9 score with 9 being highly resistant.
DRPEAR=DROPPED EARS. A measure of the number of dropped ears per plot and represents the percentage of plants that did not drop ears prior to harvest.
D/T=DROUGHT TOLERANCE. This represents a 1 to 9 rating for drought tolerance, and is based on data obtained under stress conditions. A high score indicates good drought tolerance and a low score indicates poor drought tolerance.
EARHT=EAR HEIGHT. The ear height is a measure from the ground to the highest placed developed ear node attachment and is measured in centimeters.
EARMLD=GENERAL EAR MOLD. Visual rating (1 to 9 score) where a “1” is very susceptible and a “9” is very resistant. This is based on overall rating for ear mold of mature ears without determining the specific mold organism, and may not be predictive for a specific ear mold.
EARSZ=EAR SIZE. A 1 to 9 visual rating of ear size. The higher the rating the larger the ear size.
EBTSTK=EARLY BRITTLE STALK. A count of the number of “snapped” plants per plot following severe winds when the corn plant is experiencing very rapid vegetative growth in the V5-V8 stage. Expressed as percent of plants that did not snap.
ECB1 LF=EUROPEAN CORN BORER FIRST GENERATION LEAF FEEDING (Ostrinia nubilalis). A 1 to 9 visual rating indicating the resistance to preflowering leaf feeding by first generation European Corn Borer. A higher score indicates a higher resistance.
ECB2IT=EUROPEAN CORN BORER SECOND GENERATION INCHES OF TUNNELING (Ostrinia nubilalis). Average inches of tunneling per plant in the stalk.
ECB2SC=EUROPEAN CORN BORER SECOND GENERATION (Ostrinia nubilalis). A 1 to 9 visual rating indicating post flowering degree of stalk breakage and other evidence of feeding by European Corn Borer, Second Generation. A higher score indicates a higher resistance.
ECBDPE=EUROPEAN CORN BORER DROPPED EARS (Ostrinia nubilalis). Dropped ears due to European Corn Borer. Percentage of plants that did not drop ears under second generation corn borer infestation.
EGRWTH=EARLY GROWTH. This is a measure of the relative height and size of a corn seedling at the 2-4 leaf stage of growth. This is a visual rating (1 to 9), with 1 being weak or slow growth, 5 being average growth and 9 being strong growth. Taller plants, wider leaves, more green mass and darker color constitute higher score.
ELITE INBRED. An inbred that contributed desirable qualities when used to produce commercial hybrids. An elite inbred may also be used in further breeding for the purpose of developing further improved varieties.
ERTLDG=EARLY ROOT LODGING. Early root lodging is the percentage of plants that do not root lodge prior to or around anthesis; plants that lean from the vertical axis at an approximately 300 angle or greater would be counted as root lodged.
ERTLPN=EARLY ROOT LODGING. An estimate of the percentage of plants that do not root lodge prior to or around anthesis; plants that lean from the vertical axis at an approximately 30° angle or greater would be considered as root lodged.
ERTLSC=EARLY ROOT LODGING SCORE. Score for severity of plants that lean from a vertical axis at an approximate 30 degree angle or greater which typically results from strong winds prior to or around flowering recorded within 2 weeks of a wind event. Expressed as a 1 to 9 score with 9 being no lodging.
ESTCNT=EARLY STAND COUNT. This is a measure of the stand establishment in the spring and represents the number of plants that emerge on per plot basis for the inbred or hybrid.
EYESPT=EYE SPOT (Kabatiella zeae or Aureobasidium zeae). A 1 to 9 visual rating indicating the resistance to Eye Spot. A higher score indicates a higher resistance.
FUSERS=FUSARIUM EAR ROT SCORE (Fusarium moniliforme or Fusarium subglutinans). A 1 to 9 visual rating indicating the resistance to Fusarium ear rot. A higher score indicates a higher resistance.
GDU=GROWING DEGREE UNITS. Using the Barger Heat Unit Theory, which assumes that maize growth occurs in the temperature range 50° F.-86° F. and that temperatures outside this range slow down growth; the maximum daily heat unit accumulation is 36 and the minimum daily heat unit accumulation is 0. The seasonal accumulation of GDU is a major factor in determining maturity zones.
GDUSHD=GDU TO SHED. The number of growing degree units (GDUs) or heat units required for an inbred line or hybrid to have approximately 50 percent of the plants shedding pollen and is measured from the time of planting. Growing degree units are calculated by the Barger Method, where the heat units for a 24-hour period are:




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The highest maximum temperature used is 86° F. and the lowest minimum temperature used is 50° F. For each inbred or hybrid it takes a certain number of GDUs to reach various stages of plant development.
GDUSLK=GDU TO SILK. The number of growing degree units required for an inbred line or hybrid to have approximately 50 percent of the plants with silk emergence from time of planting. Growing degree units are calculated by the Barger Method as given in GDU SHD definition.
GENOTYPE. Refers to the genetic constitution of a cell or organism.
GIBERS=GIBBERELLA EAR ROT (PINK MOLD) (Gibberella zeae). A 1 to 9 visual rating indicating the resistance to Gibberella Ear Rot. A higher score indicates a higher resistance.
GIBROT=GIBBERELLA STALK ROT SCORE. Score of stalk rot severity due to Gibberella (Gibberella zeae). Expressed as a 1 to 9 score with 9 being highly resistant.
GLFSPT=GRAY LEAF SPOT (Cercospora zeae-maydis). A 1 to 9 visual rating indicating the resistance to Gray Leaf Spot. A higher score indicates a higher resistance.
GOSWLT=GOSS' WILT (Corynebacterium nebraskense). A 1 to 9 visual rating indicating the resistance to Goss' Wilt. A higher score indicates a higher resistance.
GRNAPP=GRAIN APPEARANCE. This is a 1 to 9 rating for the general appearance of the shelled grain as it is harvested based on such factors as the color of harvested grain, any mold on the grain, and any cracked grain. High scores indicate good grain quality.
HCBLT=HELMINTHOSPORIUM CARBONUM LEAF BLIGHT (Helminthosporium carbonum). A 1 to 9 visual rating indicating the resistance to Helminthosporium infection. A higher score indicates a higher resistance.
HD SMT=HEAD SMUT (Sphacelotheca reiliana). This score indicates the percentage of plants not infected.
HSKCVR=HUSK COVER. A 1 to 9 score based on performance relative to key checks, with a score of 1 indicating very short husks, tip of ear and kernels showing; 5 is intermediate coverage of the ear under most conditions, sometimes with thin husk; and a 9 has husks extending and closed beyond the tip of the ear. Scoring can best be done near physiological maturity stage or any time during dry down until harvested.
INC D/A=GROSS INCOME (DOLLARS PER ACRE). Relative income per acre assuming drying costs of two cents per point above 15.5 percent harvest moisture and current market price per bushel.
INCOME/ACRE. Income advantage of hybrid to be patented over other hybrid on per acre basis.
INC ADV=GROSS INCOME ADVANTAGE. GROSS INCOME advantage of variety #1 over variety #2.
KSZDCD=KERNEL SIZE DISCARD. The percent of discard seed; calculated as the sum of discarded tip kernels and extra large kernels.
LINKAGE. Refers to a phenomenon wherein alleles on the same chromosome tend to segregate together more often than expected by chance if their transmission was independent.
LINKAGE DISEQUILIBRIUM. Refers to a phenomenon wherein alleles tend to remain together in linkage groups when segregating from parents to offspring, with a greater frequency than expected from their individual frequencies.
L/POP=YIELD AT LOW DENSITY. Yield ability at relatively low plant densities on a 1 to 9 relative system with a higher number indicating the hybrid responds well to low plant densities for yield relative to other hybrids. A 1, 5, and 9 would represent very poor, average, and very good yield response, respectively, to low plant density.
LRTLDG=LATE ROOT LODGING. Late root lodging is the percentage of plants that do not root lodge after anthesis through harvest; plants that lean from the vertical axis at an approximately 30° angle or greater would be counted as root lodged.
LRTLPN=LATE ROOT LODGING. Late root lodging is an estimate of the percentage of plants that do not root lodge after anthesis through harvest; plants that lean from the vertical axis at an approximately 30° angle or greater would be considered as root lodged.
LRTLSC=LATE ROOT LODGING SCORE. Score for severity of plants that lean from a vertical axis at an approximate 30 degree angle or greater which typically results from strong winds after flowering. Recorded prior to harvest when a root-lodging event has occurred. This lodging results in plants that are leaned or “lodged” over at the base of the plant and do not straighten or “goose-neck” back to a vertical position. Expressed as a 1 to 9 score with 9 being no lodging.
MDMCPX=MAIZE DWARF MOSAIC COMPLEX (MDMV=Maize Dwarf Mosaic Virus and MCDV=Maize Chlorotic Dwarf Virus). A 1 to 9 visual rating indicating the resistance to Maize Dwarf Mosaic Complex. A higher score indicates a higher resistance.
MST=HARVEST MOISTURE. The moisture is the actual percentage moisture of the grain at harvest.
MSTADV=MOISTURE ADVANTAGE. The moisture advantage of variety #1 over variety #2 as calculated by: MOISTURE of variety #2−MOISTURE of variety #1=MOISTURE ADVANTAGE of variety #1.
NLFBLT=NORTHERN LEAF BLIGHT (Helminthosporium turcicum or Exserohilum turcicum). A 1 to 9 visual rating indicating the resistance to Northern Leaf Blight. A higher score indicates a higher resistance.
OILT=GRAIN OIL. Absolute value of oil content of the kernel as predicted by Near-infrared Transmittance and expressed as a percent of dry matter.
PEDIGREE DISTANCE. Relationship among generations based on their ancestral links as evidenced in pedigrees. May be measured by the distance of the pedigree from a given starting point in the ancestry.
PLANT. As used herein, the term “plant” includes reference to an immature or mature whole plant, including a plant that has been detasseled or from which seed or grain has been removed. Seed or embryo that will produce the plant is also considered to be the plant.
PLANT PARTS. As used herein, the term “plant parts” includes leaves, stems, roots, seed, grain, embryo, pollen, ovules, flowers, ears, cobs, husks, stalks, root tips, anthers, silk, tissue, cells and the like.
PLANT CELL. Plant cell, as used herein includes, plant cells whether isolated, in tissue culture or incorporated in a plant or plant part.
PLTHT=PLANT HEIGHT. This is a measure of the height of the plant from the ground to the tip of the tassel in centimeters.
POLSC=POLLEN SCORE. A 1 to 9 visual rating indicating the amount of pollen shed. The higher the score the more pollen shed.
POLWT=POLLEN WEIGHT. This is calculated by dry weight of tassels collected as shedding commences minus dry weight from similar tassels harvested after shedding is complete.
POP K/A=PLANT POPULATIONS. Measured as 1000s per acre.
POP ADV=PLANT POPULATION ADVANTAGE. The plant population advantage of variety #1 over variety #2 as calculated by PLANT POPULATION of variety #2−PLANT POPULATION of variety #1=PLANT POPULATION ADVANTAGE of variety #1.
PRM=PREDICTED RELATIVE MATURITY. This trait, predicted relative maturity, is based on the harvest moisture of the grain. The relative maturity rating is based on a known set of checks and utilizes standard linear regression analyses and is also referred to as the Comparative Relative Maturity Rating System that is similar to the Minnesota Relative Maturity Rating System.
PRMSHD=A relative measure of the growing degree units (GDU) required to reach 50% pollen shed. Relative values are predicted values from the linear regression of observed GDU's on relative maturity of commercial checks.
PROT=GRAIN PROTEIN. Absolute value of protein content of the kernel as predicted by Near-Infrared Transmittance and expressed as a percent of dry matter.
RTLDG=ROOT LODGING. Root lodging is the percentage of plants that do not root lodge; plants that lean from the vertical axis at an approximately 30° angle or greater would be counted as root lodged.
RTLADV=ROOT LODGING ADVANTAGE. The root lodging advantage of variety #1 over variety #2.
SCTGRN=SCATTER GRAIN. A 1 to 9 visual rating indicating the amount of scatter grain (lack of pollination or kernel abortion) on the ear. The higher the score the less scatter grain.
SDGVGR=SEEDLING VIGOR. This is the visual rating (1 to 9) of the amount of vegetative growth after emergence at the seedling stage (approximately five leaves). A higher score indicates better vigor.
SEL IND=SELECTION INDEX. The selection index gives a single measure of the hybrid's worth based on information for up to five traits. A maize breeder may utilize his or her own set of traits for the selection index. One of the traits that is almost always included is yield. The selection index data presented in the tables represent the mean value averaged across testing stations.
SLFBLT=SOUTHERN LEAF BLIGHT (Helminthosporium maydis or Bipolaris maydis). A 1 to 9 visual rating indicating the resistance to Southern Leaf Blight. A higher score indicates a higher resistance.
SOURST=SOUTHERN RUST (Puccinia polysora). A 1 to 9 visual rating indicating the resistance to Southern Rust. A higher score indicates a higher resistance.
STAGRN=STAY GREEN. Stay green is the measure of plant health near the time of black layer formation (physiological maturity). A high score indicates better late-season plant health.
STDADV=STALK STANDING ADVANTAGE. The advantage of variety #1 over variety #2 for the trait STK CNT.
STKCNT=NUMBER OF PLANTS. This is the final stand or number of plants per plot.
STKLDG=STALK LODGING REGULAR. This is the percentage of plants that did not stalk lodge (stalk breakage) at regular harvest (when grain moisture is between about 20 and 30%) as measured by either natural lodging or pushing the stalks and determining the percentage of plants that break below the ear.
STKLDL=LATE STALK LODGING. This is the percentage of plants that did not stalk lodge (stalk breakage) at or around late season harvest (when grain moisture is between about 15 and 18%) as measured by either natural lodging or pushing the stalks and determining the percentage of plants that break below the ear.
STKLDS=STALK LODGING SCORE. A plant is considered as stalk lodged if the stalk is broken or crimped between the ear and the ground. This can be caused by any or a combination of the following: strong winds late in the season, disease pressure within the stalks, ECB damage or genetically weak stalks. This trait should be taken just prior to or at harvest. Expressed on a 1 to 9 scale with 9 being no lodging.
STLLPN=LATE STALK LODGING. This is the percent of plants that did not stalk lodge (stalk breakage or crimping) at or around late season harvest (when grain moisture is below 20%) as measure by either natural lodging or pushing the stalks and determining the percentage of plants that break or crimp below the ear.
STLPCN=STALK LODGING REGULAR. This is an estimate of the percentage of plants that did not stalk lodge (stalk breakage) at regular harvest (when grain moisture is between about 20 and 30%) as measured by either natural lodging or pushing the stalks and determining the percentage of plants that break below the ear.
STRT=GRAIN STARCH. Absolute value of starch content of the kernel as predicted by Near-Infrared Transmittance and expressed as a percent of dry matter.
STWWLT=Stewart's Wilt (Erwinia stewartii). A 1 to 9 visual rating indicating the resistance to Stewart's Wilt. A higher score indicates a higher resistance.
TASBLS=TASSEL BLAST. A 1 to 9 visual rating was used to measure the degree of blasting (necrosis due to heat stress) of the tassel at the time of flowering. A 1 would indicate a very high level of blasting at time of flowering, while a 9 would have no tassel blasting.
TASSZ=TASSEL SIZE. A 1 to 9 visual rating was used to indicate the relative size of the tassel. The higher the rating the larger the tassel.
TAS WT=TASSEL WEIGHT. This is the average weight of a tassel (grams) just prior to pollen shed.
TEXEAR=EAR TEXTURE. A 1 to 9 visual rating was used to indicate the relative hardness (smoothness of crown) of mature grain. A 1 would be very soft (extreme dent) while a 9 would be very hard (flinty or very smooth crown).
TILLER=TILLERS. A count of the number of tillers per plot that could possibly shed pollen was taken. Data are given as a percentage of tillers: number of tillers per plot divided by number of plants per plot.
TSTWT=TEST WEIGHT (UNADJUSTED). The measure of the weight of the grain in pounds for a given volume (bushel).
TSWADV=TEST WEIGHT ADVANTAGE. The test weight advantage of variety #1 over variety #2.
WIN M %=PERCENT MOISTURE WINS.
WIN Y %=PERCENT YIELD WINS.
YIELD BU/A=YIELD (BUSHELS/ACRE). Yield of the grain at harvest in bushels per acre adjusted to 15% moisture.
YLDADV=YIELD ADVANTAGE. The yield advantage of variety #1 over variety #2 as calculated by: YIELD of variety #1−YIELD variety #2=yield advantage of variety #1.
YLDSC=YIELD SCORE. A 1 to 9 visual rating was used to give a relative rating for yield based on plot ear piles. The higher the rating the greater visual yield appearance.
Definitions for Area of Adaptability
When referring to area of adaptability, such term is used to describe the location with the environmental conditions that would be well suited for this maize line. Area of adaptability is based on a number of factors, for example: days to maturity, insect resistance, disease resistance, and drought resistance. Area of adaptability does not indicate that the maize line will grow in every location within the area of adaptability or that it will not grow outside the area.
Central Corn Belt: Iowa, Illinois, Indiana
Drylands: non-irrigated areas of North Dakota, South Dakota, Nebraska, Kansas, Colorado and Oklahoma
Eastern U.S.: Ohio, Pennsylvania, Delaware, Maryland, Virginia, and West Virginia North central U.S.: Minnesota and Wisconsin
Northeast: Michigan, New York, Vermont, and Ontario and Quebec Canada
Northwest U.S.: North Dakota, South Dakota, Wyoming, Washington, Oregon, Montana, Utah, and Idaho
South central U.S.: Missouri, Tennessee, Kentucky, Arkansas
Southeast U.S.: North Carolina, South Carolina, Georgia, Florida, Alabama, Mississippi, and Louisiana
Southwest U.S.: Texas, Oklahoma, New Mexico, Arizona
Western U.S.: Nebraska, Kansas, Colorado, and California
Maritime Europe: France, Germany, Belgium and Austria


DETAILED DESCRIPTION OF THE INVENTION AND FURTHER EMBODIMENTS
All tables discussed in the Detailed Description of the Invention and Further Embodiments section can be found at the end of the section.
Morphological and Physiological Characteristics of PHB6V
Inbred maize line PHB6V is a yellow, dent-flint maize inbred that is best suited to be used as a female in the production of the first generation F1 maize hybrids. Inbred maize line PHB6V is best adapted to the Northcentral and Northwestern areas of the United States, and can be used to produce hybrids with approximately 90 maturity based on the Comparative Relative Maturity Rating System for harvest moisture of grain. Inbred maize line PHB6V demonstrates above average grain yield, above average resistance to ear mold and good emergence as an inbred per se. In hybrid combination, inbred PHB6V demonstrates high grain yield, fast drydown, good early root lodging resistance, and good resistance to head smut.
The inbred has shown uniformity and stability within the limits of environmental influence for all the traits as described in the Variety Description Information (Table 1, found at the end of the section). The inbred has been self-pollinated and ear-rowed a sufficient number of generations with careful attention paid to uniformity of plant type to ensure the homozygosity and phenotypic stability necessary for use in commercial hybrid seed production. The line has been increased both by hand and in isolated fields with continued observation for uniformity. No variant traits have been observed or are expected in PHB6V.
Inbred maize line PHB6V, being substantially homozygous, can be reproduced by planting seeds of the line, growing the resulting maize plants under self-pollinating or sib-pollinating conditions with adequate isolation, and harvesting the resulting seed using techniques familiar to the agricultural arts.
Genetic Marker Profile
To select and develop superior inbred parental lines for producing hybrids, it is necessary to identify and select genetically unique individuals that occur in a segregating population. The segregating population is the result of a combination of crossover events plus the independent assortment of specific combinations of alleles at many gene loci that results in specific and unique genotypes. The genetic variation among individual progeny of a breeding cross allows for the identification of rare and valuable new genotypes. Once identified, it is possible to utilize routine and predictable breeding methods to develop progeny that retain the rare and valuable new genotypes developed by the initial breeder.
Even if the entire genotypes of the parents of the breeding cross were characterized and a desired genotype known, only a few if any individuals having the desired genotype may be found in a large segregating F2 population. It would be very unlikely that a breeder of ordinary skill in the art would be able to recreate the same line twice from the very same original parents as the breeder is unable to direct how the genomes combine or how they will interact with the environmental conditions. This unpredictability results in the expenditure of large amounts of research resources in the development of a superior new maize inbred line. Once such a line is developed its value to society is substantial since it is important to advance the germplasm base as a whole in order to maintain or improve traits such as yield, disease resistance, pest resistance and plant performance in extreme weather conditions. Backcross trait conversions are routinely used to add or modify one or a few traits of such a line and this further enhances its value and usefulness to society.
Phenotypic traits exhibited by PHB6V can be used to characterize the genetic contribution of PHB6V to progeny lines developed through the use of PHB6V. Quantitative traits including, but not limited to, yield, maturity, stay green, root lodging, stalk lodging, and early growth are typically governed by multiple genes at multiple loci. PHB6V progeny plants that retain the same degree of phenotypic expression of these quantitative traits as PHB6V have received significant genotypic and phenotypic contribution from PHB6V. This characterization is enhanced when such quantitative trait is not exhibited in non-PHB6V breeding material used to developed the PHB6V progeny.
In addition to phenotypic observations, a plant can also be identified by its genotype. The genotype of a plant can be characterized through a genetic marker profile, which can identify plants of the same variety or a related variety or be used to determine or validate a pedigree. Genetic marker profiles can be obtained by techniques such as Restriction Fragment Length Polymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Amplified Fragment Length Polymorphisms (AFLPs), Simple Sequence Repeats (SSRs) which are also referred to as Microsatellites, and Single Nucleotide Polymorphisms (SNPs). For example, see Berry, Don, et al., “Assessing Probability of Ancestry Using Simple Sequence
Repeat Profiles: Applications to Maize Hybrids and Inbreds”, Genetics, 2002, 161:813-824, which is incorporated by reference herein.
Particular markers used for these purposes are not limited to the set of markers disclosed herein, but are envisioned to include any type of marker and marker profile which provides a means of distinguishing varieties. In addition to being used for identification of Inbred Line PHB6V, a hybrid produced through the use of PHB6V, and the identification or verification of pedigree for progeny plants produced through the use of PHB6V, the genetic marker profile is also useful in further breeding and in developing backcross conversions.
Means of performing genetic marker profiles using SSR polymorphisms are well known in the art. SSRs are genetic markers based on polymorphisms in repeated nucleotide sequences, such as microsatellites. A marker system based on SSRs can be highly informative in linkage analysis relative to other marker systems in that multiple alleles may be present. Another advantage of this type of marker is that, through use of flanking primers, detection of SSRs can be achieved, for example, by the polymerase chain reaction (PCR), thereby eliminating the need for labor-intensive Southern hybridization. The PCRTM detection is done by use of two oligonucleotide primers flanking the polymorphic segment of repetitive DNA. Repeated cycles of heat denaturation of the DNA followed by annealing of the primers to their complementary sequences at low temperatures, and extension of the annealed primers with DNA polymerase, comprise the major part of the methodology.
Following amplification, markers can be scored by gel electrophoresis of the amplification products. Scoring of marker genotype is based on the size of the amplified fragment as measured by molecular weight (MW) rounded to the nearest integer. While variation in the primer used or in laboratory procedures can affect the reported molecular weight, relative values should remain constant regardless of the specific primer or laboratory used. When comparing lines it is preferable if all SSR profiles are performed in the same lab. The SSR analyses reported herein were conducted in-house at Pioneer Hi-Bred. An SSR service is available to the public on a contractual basis by Paragen (formerly Celera AgGen) in Research Triangle Park, North Carolina.
Primers used for the SSRs reported herein are publicly available and may be found in the Maize GDB on the World Wide Web prefix followed by maizegdb.org (maintained by the USDA Agricultural Research Service), in Sharopova et al. (Plant Mol. Biol. 48(5-6):463-481), Lee et al (Plant Mol. Biol. 48(5-6); 453-461), or may be constructed from sequences if reported herein. Some marker information may be available from Paragen.
Map information is provided by bin number as reported in the Maize GDB. The bin number digits to the left of decimal point represent the chromosome on which such marker is located, and the digits to the right of the decimal represent the location on such chromosome. Any bin numbers reported in parenthesis represent other bin locations for such marker that have been reported in the literature or on the Maize GDB. A bin number.xx designation indicates that the bin location on that chromosome is not known. Map positions are also available on the Maize GDB for a variety of different mapping populations.
The SSR profile of Inbred PHB6V can be found in Table 2 found at the end of this section. The profile can be used to identify hybrids comprising PHB6V as a parent, since such hybrids will comprise two sets of allele, one set of which will be the same alleles as PHB6V. Because an inbred is essentially homozygous at all relevant loci, an inbred should, in almost all cases, have only one allele at each locus. In contrast, a genetic marker profile of a hybrid should be the sum of those parents, e.g., if one inbred parent had allele x at a particular locus, and the other inbred parent had allele y the hybrid is x.y (heterozygous) by inference. Subsequent generations of progeny produced by selection and breeding are expected to be of genotype x (homozygous), y (homozygous), or x.y (heterozygous) for that locus position. When the F1 plant is used to produce an inbred, the inbred should be either x or y for that allele.
Plants and plant parts substantially benefiting from the use of PHB6V in their development such as PHB6V comprising a backcross conversion, transgene, or genetic sterility factor, may be identified by having a molecular marker profile with a high percent identity to PHB6V.
The SSR profile of PHB6V also can be used to identify essentially derived varieties and other progeny lines developed from the use of PHB6V, as well as cells and other plant parts thereof. Progeny plants and plant parts produced using PHB6V may be identified by having a molecular marker profile of at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5% genetic contribution from inbred line PHB6V.
Unique SSR Profiles
While determining the SSR genetic marker profile of the inbreds described supra, several unique SSR profile loci sets were or may be identified through further analysis which did not appear in either parent of such inbred.
Such unique SSR profiles may arise during the breeding process from recombination or mutation. A combination of several unique alleles provides a means of identifying an inbred, a hybrid produced from such inbred, and progeny produced from such inbred. Such progeny may be further characterized as being within a pedigree distance of PHB6V, such as within 1,2,3,4 or 5 or less breeding crosses to a maize plant other than PHB6V or a plant that has PHB6V as a parent or other progenitor. Further unique molecular profiles may be identified with other molecular tools such as SNPs and RFLPs.
Each genetic marker profile represents a novel grouping of alleles unique to an inbred. Each of the unique SSR profiles that may be found are referred to as a “loci set”.
Comparing PHB6V to Other Inbreds
A breeder uses various methods to help determine which plants should be selected from the segregating populations and ultimately which inbred lines will be used to develop hybrids for commercialization. In addition to the knowledge of the germplasm and other skills the breeder uses, a part of the selection process is dependent on experimental design coupled with the use of statistical analysis. Experimental design and statistical analysis are used to help determine which plants, which family of plants, and finally which inbred lines and hybrid combinations are significantly better or different for one or more traits of interest. Experimental design methods are used to assess error so that differences between two inbred lines or two hybrid lines can be more accurately determined. Statistical analysis includes the calculation of mean values, determination of the statistical significance of the sources of variation, and the calculation of the appropriate variance components. Either a five or a one percent significance level is customarily used to determine whether a difference that occurs for a given trait is real or due to the environment or experimental error. One of ordinary skill in the art of plant breeding would know how to evaluate the traits of two plant varieties to determine if there is no significant difference between the two traits expressed by those varieties. For example, see Fehr, Walt, Principles of Cultivar Development, p. 261-286 (1987) which is incorporated herein by reference. Mean trait values may be used to determine whether trait differences are significant, and preferably the traits are measured on plants grown under the same environmental conditions.
In Tables 3A-3F, data from traits and characteristics of inbred maize line PHB6V per se are given and compared to other maize inbred lines.
Inbred by Tester comparisons are also used to evaluate two inbreds. Combined inbred by tester comparison results can be used to distinguish two inbred lines while also providing information about the combining ability of the inbreds used. Tables 4A-4C compare PHB6V to other inbreds, when each inbred is crossed to the same tester lines.
Methods of Introducing a New Gene or Trait into PHB6V.
Field crops are bred through techniques that take advantage of the plant's method of pollination. A plant is self-pollinated if pollen from one flower is transferred to the same or another flower of the same plant. A plant is sib-pollinated when individuals within the same family or line are used for pollination. A plant is cross-pollinated if the pollen comes from a flower on a different plant from a different family or line. The terms “cross-pollination” and “out-cross” as used herein do not include self-pollination or sib-pollination.
Maize (zea mays L.), often referred to as corn in the United States, can be bred by both self-pollination and cross-pollination techniques. Maize has separate male and female flowers on the same plant, located on the tassel and the ear, respectively. Natural or open pollination occurs in maize when wind blows pollen from the tassels to the silks that protrude from the tops of the ears.
Plants that have been self-pollinated and selected for type for many generations become homozygous at almost all gene loci and produce a uniform population of true breeding progeny. A cross between two different homozygous lines produces a uniform population of hybrid plants that may be heterozygous for many gene loci. A cross of two plants; each heterozygous at a number of gene loci, will produce a population of hybrid plants that differ genetically and will not be uniform.
One possible method of introducing a new gene or trait into PHB6V is producing a backcross conversion of PHB6V. A backcross conversion occurs when DNA sequences are introduced through traditional (non-transformation) breeding techniques, such as backcrossing (Hallauer et al., 1988). DNA sequences, whether naturally occurring or transgenes, may be introduced using these traditional breeding techniques. A backcross conversion may produce a plant with a trait conversion in at least one or more crosses, including at least 2 crosses, at least 3 crosses, at least 4 crosses, and the like. Molecular marker assisted breeding or selection may be utilized to reduce the number of backcrosses necessary to achieve the backcross conversion. The term backcross conversion is also referred to in the art as a single locus conversion. Reference is made to US 2002/0062506A1 for a detailed discussion of single locus conversions and traits that may be incorporated into PHB6V through backcross conversion. Desired traits transferred through this process include, but are not limited to, waxy starch, grain color, nutritional enhancements, altered fatty acid profile, increased digestibility, low phytate, industrial enhancements, disease resistance, insect resistance, herbicide resistance and yield enhancements. The trait of interest is transferred from the donor parent to the recurrent parent, in this case, the maize plant disclosed herein. The backcross conversion may result from either the transfer of a dominant allele or a recessive allele. Selection of progeny containing the trait of interest is accomplished by direct selection for a trait associated with a dominant allele. Selection of progeny for a trait that is transferred via a recessive allele, such as the waxy starch characteristic, requires growing and selfing the first backcross generation to determine which plants carry the recessive alleles. Recessive traits may require additional progeny testing in successive backcross generations to determine the presence of the gene of interest. Along with selection for the trait of interest, progeny are selected for the phenotype of the recurrent parent. While occasionally additional polynucleotide sequences or genes are transferred along with the backcross conversion, this is an insubstantial change to the variety, and backcross conversion lines are easily developed without undue experimentation. Hallauer et al. (1998, page 472) states that “For single gene traits that are relatively easy to classify, the backcross method is effective and relatively easy to manage.” Poehlman et al. (1995, page 334) states that “A backcross-derived inbred line fits into the same hybrid combination as the recurrent parent inbred line and contributes the effect of the additional gene added through the backcross.”
The advent of new molecular biological techniques has allowed the isolation and characterization of genetic elements with specific functions, such as encoding specific protein products. Scientists in the field of plant biology developed a strong interest in engineering the genome of plants to contain and express foreign genetic elements, or additional, or modified versions of native or endogenous genetic elements in order to alter the traits of a plant in a specific manner. Any DNA sequences, whether from a different species or from the same species, that are inserted into the genome using transformation are referred to herein collectively as “transgenes”. A transformed plant may contain at least one transgene but could contain at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 and/or no more than 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2. Over the last fifteen to twenty years several methods for producing transgenic plants have been developed, and the present invention, in particular embodiments, also relates to transformed versions of the claimed inbred maize line PHB6V as well as hybrid combinations thereof.
Numerous methods for plant transformation have been developed, including biological and physical plant transformation protocols. See, for example, Miki et al., “Procedures for Introducing Foreign DNA into Plants” in Methods in Plant Molecular Biology and Biotechnology, Glick, B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages 67-88 and Armstrong, “The First Decade of Maize Transformation: A Review and Future Perspective” (Maydica 44:101-109, 1999). In addition, expression vectors and in vitro culture methods for plant cell or tissue transformation and regeneration of plants are available. See, for example, Gruber et al., “Vectors for Plant Transformation” in Methods in Plant Molecular Biology and Biotechnology, Glick, B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages 89-119.
The most prevalent types of plant transformation involve the construction of an expression vector. Such a vector comprises a DNA sequence that contains a gene under the control of or operatively linked to a regulatory element, for example a promoter. The vector may contain one or more genes and one or more regulatory elements.
A genetic trait which has been engineered into a particular maize plant using transformation techniques, could be moved into another line using traditional breeding techniques that are well known in the plant breeding arts. For example, a backcrossing approach is commonly used to move a transgene from a transformed maize plant to an elite inbred line, and the resulting progeny would then comprise the transgene(s). Also, if an inbred line was used for the transformation then the transgenic plants could be crossed to a different inbred in order to produce a transgenic hybrid maize plant. As used herein, “crossing” can refer to a simple X by Y cross, or the process of backcrossing, depending on the context.
Various genetic elements can be introduced into the plant genome using transformation. These elements include but are not limited to genes; coding sequences; inducible, constitutive, and tissue specific promoters; enhancing sequences; and signal and targeting sequences. See the traits, genes and transforming methods listed in U.S. Pat. No. 6,118,055, which are herein incorporated by reference.
With transgenic plants according to the present invention, a foreign protein can be produced in commercial quantities. Thus, techniques for the selection and propagation of transformed plants, which are well understood in the art, yield a plurality of transgenic plants which are harvested in a conventional manner, and a foreign protein then can be extracted from a tissue of interest or from total biomass. Protein extraction from plant biomass can be accomplished by known methods which are discussed, for example, by Heney and Orr, Anal. Biochem. 114: 92-6 (1981). One embodiment, the biomass of interest is seed.
A genetic map can be generated, primarily via conventional Restriction Fragment Length Polymorphisms (RFLP), Polymerase Chain Reaction (PCR) analysis, and Simple Sequence Repeats (SSR) and Single Nucleotide Polymorphisms (SNP) which identifies the approximate chromosomal location of the integrated DNA molecule. For exemplary methodologies in this regard, see Glick and Thompson, METHODS IN PLANT MOLECULAR BIOLOGY AND BIOTECHNOLOGY 269-284 (CRC Press, Boca Raton, 1993).
Wang et al. discuss “Large Scale Identification, Mapping and Genotyping of Single-Nucleotide Polymorphisms in the Human Genome”, Science, 280:1077-1082, 1998, and similar capabilities are becoming increasingly available for the corn genome. Map information concerning chromosomal location is useful for proprietary protection of a subject transgenic plant. If unauthorized propagation is undertaken and crosses made with other germplasm, the map of the integration region can be compared to similar maps for suspect plants, to determine if the latter have a common parentage with the subject plant. Map comparisons would involve hybridizations, RFLP, PCR, SSR and sequencing, all of which are conventional techniques. SNP's may also be used alone or in combination with other techniques.
Likewise, by means of the present invention, plants can be genetically engineered to express various phenotypes of agronomic interest. Through the transformation of maize the expression of genes can be modulated to enhance disease resistance, insect resistance, herbicide resistance, agronomic traits as well as grain quality traits. Transformation can also be used to insert DNA sequences which control or help control male-sterility. DNA sequences native to maize as well as non-native DNA sequences can be transformed into maize and used to modulate levels of native or non-native proteins. Anti-sense technology, various promoters, targeting sequences, enhancing sequences, and other DNA sequences can be inserted into the maize genome for the purpose of modulating the expression of proteins. Exemplary transgenes implicated in this regard include, but are not limited to, those categorized below.
1. Transgenes that Confer Resistance to Pests or Disease and that Encode.
(A) Plant disease resistance genes. Plant defenses are often activated by specific interaction between the product of a disease resistance gene (R) in the plant and the product of a corresponding avirulence (Avr) gene in the pathogen. A plant variety can be transformed with cloned resistance gene to engineer plants that are resistant to specific pathogen strains. See, for example Jones et al., Science 266: 789 (1994) (cloning of the tomato Cf-9 gene for resistance to Cladosporium fulvum); Martin et al., Science 262: 1432 (1993) (tomato Pto gene for resistance to Pseudomonas syringae pv. tomato encodes a protein kinase); Mindrinos et al., Cell 78: 1089 (1994) (Arabidopsis RSP2 gene for resistance to Pseudomonas syringae). A plant resistant to a disease is one that is more resistant to a pathogen as compared to the wild type plant.
(B) A Bacillus thuringiensis protein, a derivative thereof or a synthetic polypeptide modeled thereon. See, for example, Geiser et al., Gene 48: 109 (1986), who disclose the cloning and nucleotide sequence of a Bt δ-endotoxin gene. Moreover, DNA molecules encoding δ-endotoxin genes can be purchased from American Type Culture Collection (Rockville, Md.), for example, under ATCC Accession Nos. 40098, 67136, 31995 and 31998. Other examples of Bacillus thuringiensis transgenes being genetically engineered are given in the following patents and hereby are incorporated by reference for this purpose: U.S. Pat. Nos. 5,188,960; 5,689,052; 5,880,275; and WO 97/40162.
(C) A lectin. See, for example, the disclosure by Van Damme et al., Plant Molec. Biol. 24: 25 (1994), who disclose the nucleotide sequences of several Clivia miniata mannose-binding lectin genes.
(D) A vitamin-binding protein such as avidin. See PCT Application US93/06487 the contents of which are hereby incorporated by reference for this purpose. The application teaches the use of avidin and avidin homologues as larvicides against insect pests.
(E) An enzyme inhibitor, for example, a protease inhibitor or an amylase inhibitor. See, for example, Abe et al., J. Biol. Chem. 262: 16793 (1987) (nucleotide sequence of rice cysteine proteinase inhibitor), Huub et al., Plant Molec. Biol. 21: 985 (1993) (nucleotide sequence of cDNA encoding tobacco proteinase inhibitor I), and Sumitani et al., Biosci. Biotech. Biochem. 57: 1243 (1993) (nucleotide sequence of Streptomyces nitrosporeus α-amylase inhibitor) and U.S. Pat. No. 5,494,813.
(F) An insect-specific hormone or pheromone such as an ecdysteroid and juvenile hormone, a variant thereof, a mimetic based thereon, or an antagonist or agonist thereof. See, for example, the disclosure by Hammock et al., Nature 344: 458 (1990), of baculovirus expression of cloned juvenile hormone esterase, an inactivator of juvenile hormone.
(G) An insect-specific peptide or neuropeptide which, upon expression, disrupts the physiology of the affected pest. For example, see the disclosures of Regan, J. Biol. Chem. 269: 9 (1994) (expression cloning yields DNA coding for insect diuretic hormone receptor), and Pratt et al., Biochem. Biophys. Res. Comm.163: 1243 (1989) (an allostatin is identified in Diploptera puntata). See also U.S. Pat. No. 5,266,317 to Tomalski et al., who disclose genes encoding insect-specific, paralytic neurotoxins.
(H) An insect-specific venom produced in nature by a snake, a wasp, etc. For example, see Pang et al., Gene 116: 165 (1992), for disclosure of heterologous expression in plants of a gene coding for a scorpion insectotoxic peptide.
(I) An enzyme responsible for an hyperaccumulation of a monterpene, a sesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivative or another non-protein molecule with insecticidal activity.
(J) An enzyme involved in the modification, including the post-translational modification, of a biologically active molecule; for example, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme, a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, a phosphatase, a kinase, a phosphorylase, a polymerase, an elastase, a chitinase and a glucanase, whether natural or synthetic. See PCT Application WO 93/02197 in the name of Scott et al., which discloses the nucleotide sequence of a callase gene. DNA molecules which contain chitinase-encoding sequences can be obtained, for example, from the ATCC under Accession Nos. 39637 and 67152. See also Kramer et al., Insect Biochem. Molec. Biol.23: 691 (1993), who teach the nucleotide sequence of a cDNA encoding tobacco hookworm chitinase, and Kawalleck et al., Plant Molec. Biol. 21: 673 (1993), who provide the nucleotide sequence of the parsley ubi-4-2 polyubiquitin gene.
(K) A molecule that stimulates signal transduction. For example, see the disclosure by Botella et al., Plant Molec. Biol. 24: 757 (1994), of nucleotide sequences for mung bean calmodulin cDNA clones, and Griess et al., Plant Physiol. 104: 1467 (1994), who provide the nucleotide sequence of a maize calmodulin cDNA clone.
(L) A hydrophobic moment peptide. See PCT Application WO95/16776 (disclosure of peptide derivatives of Tachyplesin which inhibit fungal plant pathogens) and PCT Application WO95/18855 (teaches synthetic antimicrobial peptides that confer disease resistance), the respective contents of which are hereby incorporated by reference for this purpose.
(M) A membrane permease, a channel former or a channel blocker. For example, see the disclosure by Jaynes et al., Plant Sci. 89: 43 (1993), of heterologous expression of a cecropin-β lytic peptide analog to render transgenic tobacco plants resistant to Pseudomonas solanacearum. 
(N) A viral-invasive protein or a complex toxin derived therefrom. For example, the accumulation of viral coat proteins in transformed plant cells imparts resistance to viral infection and/or disease development effected by the virus from which the coat protein gene is derived, as well as by related viruses. See Beachy et al., Ann. Rev. Phytopathol.28: 451 (1990). Coat protein-mediated resistance has been conferred upon transformed plants against alfalfa mosaic virus, cucumber mosaic virus, tobacco streak virus, potato virus X, potato virus Y, tobacco etch virus, tobacco rattle virus and tobacco mosaic virus. Id.
(O) An insect-specific antibody or an immunotoxin derived therefrom. Thus, an antibody targeted to a critical metabolic function in the insect gut would inactivate an affected enzyme, killing the insect. Cf. Taylor et al., Abstract #497, SEVENTH INT'L SYMPOSIUM ON MOLECULAR PLANT-MICROBE INTERACTIONS (Edinburgh, Scotland, 1994) (enzymatic inactivation in transgenic tobacco via production of single-chain antibody fragments).
(P) A virus-specific antibody. See, for example, Tavladoraki et al., Nature 366: 469 (1993), who show that transgenic plants expressing recombinant antibody genes are protected from virus attack.
(Q) A developmental-arrestive protein produced in nature by a pathogen or a parasite. Thus, fungal endo α-1,4-D-polygalacturonases facilitate fungal colonization and plant nutrient release by solubilizing plant cell wall homo-(x-1,4-D-galacturonase. See Lamb et al., Bio/Technology 10: 1436 (1992). The cloning and characterization of a gene which encodes a bean endopolygalacturonase-inhibiting protein is described by Toubart et al., Plant J. 2: 367 (1992).
(R) A developmental-arrestive protein produced in nature by a plant. For example, Logemann et al., Bio/Technology 10: 305 (1992), have shown that transgenic plants expressing the barley ribosome-inactivating gene have an increased resistance to fungal disease.
(S) Genes involved in the Systemic Acquired Resistance (SAR) Response and/or the pathogenesis related genes. Briggs, S., Current Biology, 5(2) (1995).
(T) Antifungal genes (Cornelissen and Melchers, PI. Physiol. 101:709-712, (1993) and Parijs et al., Planta 183:258-264, (1991) and Bushnell et al., Can. J. of Plant Path. 20(2): 137-149 (1998).
2. Transgenes that Confer Resistance to a Herbicide, For Example:
(A) A herbicide that inhibits the growing point or meristem, such as an imidazolinone or a sulfonylurea. Exemplary genes in this category code for mutant ALS and AHAS enzyme as described, for example, by Lee et al., EMBO J. 7: 1241 (1988), and Miki et al., Theor. Appl. Genet. 80: 449 (1990), respectively. See also, U.S. Pat. Nos. 5,605,011; 5,013,659; 5,141,870; 5,767,361; 5,731,180; 5,304,732; 4,761,373; 5,331,107; 5,928,937; and 5,378,824; and international publication WO 96/33270, which are incorporated herein by reference for this purpose.
(B) Glyphosate (resistance imparted by mutant 5-enolpyruvl-3-phosphikimate synthase (EPSP) and aroA genes, respectively) and other phosphono compounds such as glufosinate (phosphinothricin acetyl transferase (PAT) and Streptomyces hygroscopicus phosphinothricin acetyl transferase (bar) genes), and pyridinoxy or phenoxy proprionic acids and cycloshexones (ACCase inhibitor-encoding genes). See, for example, U.S. Pat. No. 4,940,835 to Shah et al., which discloses the nucleotide sequence of a form of EPSPS which can confer glyphosate resistance. U.S. Pat. No. 5,627,061 to Barry et al. also describes genes encoding EPSPS enzymes. See also U.S. Pat. Nos. 6,248,876 B1; 6,040,497; 5,804,425; 5,633,435; 5,145,783; 4,971,908; 5,312,910; 5,188,642; 4,940,835; 5,866,775; 6,225,114 B1; 6,130,366; 5,310,667; 4,535,060; 4,769,061; 5,633,448; 5,510,471; Re. 36,449; RE 37,287 E; and U.S. Pat. No. 5,491,288; and international publications WO 97/04103; WO 97/04114; WO 00/66746; WO 01/66704; WO 00/66747 and WO 00/66748, which are incorporated herein by reference for this purpose. Glyphosate resistance is also imparted to plants that express a gene that encodes a glyphosate oxido-reductase enzyme as described more fully in U.S. Pat. Nos. 5,776,760 and 5,463,175, which are incorporated herein by reference for this purpose. In addition glyphosate resistance can be imparted to plants by the over expression of genes encoding glyphosate N-acetyltransferase. See, for example, U.S. Application Ser. Nos. 60/244,385; 60/377,175 and 60/377,719. A DNA molecule encoding a mutant aroA gene can be obtained under ATCC accession No. 39256, and the nucleotide sequence of the mutant gene is disclosed in U.S. Pat. No. 4,769,061 to Comai. European Patent Application No. 0 333 033 to Kumada et al. and U.S. Pat. No. 4,975,374 to Goodman et al. disclose nucleotide sequences of glutamine synthetase genes which confer resistance to herbicides such as L-phosphinothricin. The nucleotide sequence of a phosphinothricin-acetyl-transferase gene is provided in European Patent No. 0 242 246 and 0 242 236 to Leemans et al. De Greef et al., Bio/Technology 7: 61 (1989), describe the production of transgenic plants that express chimeric bar genes coding for phosphinothricin acetyl transferase activity. See also, U.S. Pat. Nos. 5,969,213; 5,489,520; 5,550,318; 5,874,265; 5,919,675; 5,561,236; 5,648,477; 5,646,024; 6,177,616 B1; and 5,879,903, which are incorporated herein by reference for this purpose. Exemplary of genes conferring resistance to phenoxy proprionic acids and cycloshexones, such as sethoxydim and haloxyfop, are the Acc1-S1, Acc1-S2 and Acc1-S3 genes described by Marshall et al., Theor. Appl. Genet. 83: 435 (1992).
(C) A herbicide that inhibits photosynthesis, such as a triazine (psbA and gs+genes) and a benzonitrile (nitrilase gene). Przibilla et al., Plant Cell 3: 169 (1991), describe the transformation of Chlamydomonas with plasmids encoding mutant psbA genes. Nucleotide sequences for nitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker, and DNA molecules containing these genes are available under ATCC Accession Nos. 53435, 67441 and 67442. Cloning and expression of DNA coding for a glutathione S-transferase is described by Hayes et al., Biochem. J. 285:173 (1992).
(D) Acetohydroxy acid synthase, which has been found to make plants that express this enzyme resistant to multiple types of herbicides, has been introduced into a variety of plants (see, e.g., Hattori et al. (1995) Mol Gen Genet 246:419). Other genes that confer tolerance to herbicides include: a gene encoding a chimeric protein of rat cytochrome P4507A1 and yeast NADPH-cytochrome P450 oxidoreductase (Shiota et al. (1994) Plant Physiol 106:17), genes for glutathione reductase and superoxide dismutase (Aono et al. (1995) Plant Cell Physiol 36:1687, and genes for various phosphotransferases (Datta et al. (1992) Plant Mol Biol 20:619).
(E) Protoporphyrinogen oxidase (protox) is necessary for the production of chlorophyll, which is necessary for all plant survival. The protox enzyme serves as the target for a variety of herbicidal compounds. These herbicides also inhibit growth of all the different species of plants present, causing their total destruction. The development of plants containing altered protox activity which are resistant to these herbicides are described in U.S. Pat. Nos. 6,288,306 B1; 6,282,837 B1; and 5,767,373; and international publication WO 01/12825, which are incorporated herein by reference for this purpose.
3. Transgenes that Confer or Contribute to a Grain Trait, Such as:
(A) Modified fatty acid metabolism, for example, by transforming a plant with an antisense gene of stearoyl-ACP desaturase to increase stearic acid content of the plant. See Knultzon et al., Proc. Natl. Acad. Sci. USA 89: 2624 (1992).
(B) Decreased phytate content

    
    
        (1) Introduction of a phytase-encoding gene would enhance breakdown of phytate, adding more free phosphate to the transformed plant. For example, see Van Hartingsveldt et al., Gene 127: 87 (1993), for a disclosure of the nucleotide sequence of an Aspergillus niger phytase gene.
        (2) A gene could be introduced that reduces phytate content. In maize, this, for example, could be accomplished, by cloning and then reintroducing DNA associated with the single allele which is responsible for maize mutants characterized by low levels of phytic acid. See Raboy et al., Maydica 35: 383 (1990).
    
    


(C) Modified carbohydrate composition effected, for example, by transforming plants with a gene coding for an enzyme that alters the branching pattern of starch. See Shiroza et al., J. Bacteriol. 170: 810 (1988) (nucleotide sequence of Streptococcus mutans fructosyltransferase gene), Steinmetz et al., Mol. Gen. Genet. 200: 220 (1985) (nucleotide sequence of Bacillus subtilis levansucrase gene), Pen et al., Bio/Technology 10: 292 (1992) (production of transgenic plants that express Bacillus licheniformis α-amylase), Elliot et al., Plant Molec. Biol. 21: 515 (1993) (nucleotide sequences of tomato invertase genes), Sogaard et al., J. Biol. Chem. 268: 22480 (1993) (site-directed mutagenesis of barley α-amylase gene), and Fisher et al., Plant Physiol. 102: 1045 (1993) (maize endosperm starch branching enzyme II).
(D) Elevated oleic acid via FAD-2 gene modification and/or decreased linolenic acid via FAD-3 gene modification (see U.S. Pat. Nos. 6,063,947; 6,323,392; and WO 93/11245).
4. Genes that Control Male-Sterility
(A) Introduction of a deacetylase gene under the control of a tapetum-specific promoter and with the application of the chemical N-Ac-PPT (WO 01/29237).
(B) Introduction of various stamen-specific promoters (WO 92/13956, WO 92/13957).
(C) Introduction of the barnase and the barstar gene (Paul et al. Plant Mol. Biol. 19:611-622, 1992).
PHB6V Inbreds that are Male Sterile
Hybrid seed production requires elimination or inactivation of pollen produced by the female inbred parent. Incomplete removal or inactivation of the pollen provides the potential for self-pollination. A reliable method of controlling male fertility in plants offers the opportunity for improved plant breeding. This is especially true for development of maize hybrids, which relies upon some sort of male sterility system. It should be understood that PHB6V can be produced in a male-sterile form. There are several ways in which a maize plant can be manipulated so that it is male sterile. These include use of manual or mechanical emasculation (or detasseling), use of one or more genetic factors that confer male sterility, including cytoplasmic genetic and/or nuclear genetic male sterility, use of gametocides and the like. A male sterile inbred designated PHB6V may include one or more genetic factors, which result in cytoplasmic genetic and/or nuclear genetic male sterility. Such embodiments are also within the scope of the present claims. The term manipulated to be male sterile refers to the use of any available techniques to produce a male sterile version of maize line PHB6V. The male sterility may be either partial or complete male sterility.
Hybrid maize seed is often produced by a male sterility system incorporating manual or mechanical detasseling. Alternate strips of two maize inbreds are planted in a field, and the pollen-bearing tassels are removed from one of the inbreds (female). Provided that there is sufficient isolation from sources of foreign maize pollen, the ears of the detasseled inbred will be fertilized only from the other inbred (male), and the resulting seed is therefore hybrid and will form hybrid plants.
The laborious detasseling process can be avoided by using cytoplasmic male-sterile (CMS) inbreds. Plants of a CMS inbred are male sterile as a result of factors resulting from the cytoplasmic, as opposed to the nuclear, genome. Thus, this characteristic is inherited exclusively through the female parent in maize plants, since only the female provides cytoplasm to the fertilized seed. CMS plants are fertilized with pollen from another inbred that is not male-sterile. Pollen from the second inbred may or may not contribute genes that make the hybrid plants male-fertile, and either option may be preferred depending on the intended use of the hybrid. The same hybrid seed, a portion produced from detasseled fertile maize and a portion produced using the CMS system, can be blended to insure that adequate pollen loads are available for fertilization when the hybrid plants are grown.
There are several methods of conferring genetic male sterility available, such as multiple mutant genes at separate locations within the genome that confer male sterility, as disclosed in U.S. Pat. Nos. 4,654,465 and 4,727,219 to Brar et al. and chromosomal translocations as described by Patterson in U.S. Pat. Nos. 3,861,709 and 3,710,511. In addition to these methods, Albertsen et al., of Pioneer Hi-Bred, U.S. Pat. No. 5,432,068, have developed a system of nuclear male sterility which includes: identifying a gene which is critical to male fertility; silencing this native gene which is critical to male fertility; removing the native promoter from the essential male fertility gene and replacing it with an inducible promoter; inserting this genetically engineered gene back into the plant; and thus creating a plant that is male sterile because the inducible promoter is not “on” resulting in the male fertility gene not being transcribed. Fertility is restored by inducing, or turning “on”, the promoter, which in turn allows the gene that confers male fertility to be transcribed.
These, and the other methods of conferring genetic male sterility in the art, each possess their own benefits and drawbacks. Some other methods use a variety of approaches such as delivering into the plant a gene encoding a cytotoxic substance associated with a male tissue specific promoter or an antisense system in which a gene critical to fertility is identified and an antisense to that gene is inserted in the plant (see: Fabinjanski, et al. EPO 89/3010153.8 Publication No. 329,308 and PCT Application PCT/CA90/00037 published as WO 90/08828).
Another system for controlling male sterility makes use of gametocides. Gametocides are not a genetic system, but rather a topical application of chemicals. These chemicals affect cells that are critical to male fertility. The application of these chemicals affects fertility in the plants only for the growing season in which the gametocide is applied (see Carlson, Glenn R., U.S. Pat. No. 4,936,904). Application of the gametocide, timing of the application and genotype specificity often limit the usefulness of the approach and it is not appropriate in all situations.
Incomplete control over male fertility may result in self-pollinated seed being unintentionally harvested and packaged with hybrid seed. The male parent is grown next to the female parent in the field, so there is the very low probability that the male selfed seed could be unintentionally harvested and packaged with the hybrid seed. Once the seed from the hybrid bag is planted, it is possible to identify and select these self-pollinated plants. These self-pollinated plants will be genetically equivalent to one of the inbred lines used to produce the hybrid. Though the possibility of inbreds being included hybrid seed bags exists, the occurrence is very low because much care is taken to avoid such inclusions. It is worth noting that hybrid seed is sold to growers for the production of grain or forage and not for breeding or seed production.
These self-pollinated plants can be identified and selected by one skilled in the art due to their decreased vigor when compared to the hybrid. Inbreds are identified by their less vigorous appearance for vegetative and/or reproductive characteristics, including shorter plant height, small ear size, ear and kernel shape, cob color, or other characteristics.
Identification of these self-pollinated lines can also be accomplished through molecular marker analyses. See, “The Identification of Female Selfs in Hybrid Maize: A Comparison Using Electrophoresis and Morphology”, Smith, J. S. C. and Wych, R. D., Seed Science and Technology 14, pp. 1-8 (1995), the disclosure of which is expressly incorporated herein by reference. Through these technologies, the homozygosity of the self pollinated line can be verified by analyzing allelic composition at various loci along the genome. Those methods allow for rapid identification of the invention disclosed herein. See also, “Identification of Atypical Plants in Hybrid Maize Seed by Postcontrol and Electrophoresis” Sarca, V. et al., Probleme de Genetica Teoritica si Aplicata Vol. 20 (1) p. 29-42.
Development of Maize Hybrids Using PHB6V
A single cross maize hybrid results from the cross of two inbred lines, each of which has a genotype that complements the genotype of the other. The hybrid progeny of the first generation is designated F1. In the development of commercial hybrids in a maize plant breeding program, only the F1 hybrid plants are sought. F1 hybrids are more vigorous than their inbred parents. This hybrid vigor, or heterosis, can be manifested in many polygenic traits, including increased vegetative growth and increased yield.
PHB6V may be used to produce hybrid maize seed. One such embodiment is the method of crossing inbred maize line PHB6V with another maize plant, such as a different maize inbred line, to form a first generation F1 hybrid plant. The first generation F1 hybrid plant produced by this method is also an embodiment of the invention. This first generation F1 plant will comprise an essentially complete set of the alleles of inbred line PHB6V. One of ordinary skill in the art can utilize either breeder books or molecular methods to identify a particular F1 hybrid plant produced using inbred line PHB6V, and any such individual plant is also encompassed by this invention. These embodiments also cover use of these methods with transgenic, male sterile and/or backcross conversions of inbred line PHB6V.
The development of a maize hybrid in a maize plant breeding program involves three steps: (1) the selection of plants from various germplasm pools for initial breeding crosses; (2) the selfing of the selected plants from the breeding crosses for several generations to produce a series of inbred lines, which, although different from each other, breed true and are highly uniform; and (3) crossing the selected inbred lines with different inbred lines to produce the hybrids. During the inbreeding process in maize, the vigor of the lines decreases. Vigor is restored when two different inbred lines are crossed to produce the hybrid. An important consequence of the homozygosity and homogeneity of the inbred lines is that the hybrid between a defined pair of inbreds will always be the same. Once the inbreds that give a superior hybrid have been identified, the hybrid seed can be reproduced indefinitely as long as the homogeneity of the inbred parents is maintained.
PHB6V may be used to produce a single cross hybrid, a three-way hybrid or a double cross hybrid. A single cross hybrid is produced when two inbred lines are crossed to produce the F1 progeny. A double cross hybrid is produced from four inbred lines crossed in pairs (A×B and C×D) and then the two F1 hybrids are crossed again (A×B)×(C×D). A three-way cross hybrid is produced from three inbred lines where two of the inbred lines are crossed (A×B) and then the resulting F1 hybrid is crossed with the third inbred (A×B)×C. Much of the hybrid vigor and uniformity exhibited by F1 hybrids is lost in the next generation (F2). Consequently, seed produced from hybrids is not used for planting stock.
Inbred by Tester Comparisons
Combining ability of a line, as well as the performance of the line per se, is a factor in the selection of improved maize inbreds. Combining ability refers to a line's contribution as a parent when crossed with other lines to form hybrids. The hybrids formed for the purpose of selecting superior lines are designated as test crosses. One way of measuring combining ability is by using values based in part on the overall mean of a number of test crosses. This mean may be adjusted to remove environmental effects and it is adjusted for known genetic relationships among the lines.
Combining Ability Data
A general combining ability report for inbred PHB6V is provided in Table 5. Table 5 shows overall mean values of traits from numerous F1 lines produced by crosses between inbred PHB6V and other maize lines. Table 5 demonstrates that inbred PHB6V shows good general combining ability for hybrid production.
A specific combining ability report for inbred PHB6V is provided in Table 5A. Table 5A shows mean values of traits from numerous individual F1 lines produced by crosses between inbred PHB6V and other maize lines.
Hybrid Comparisons
The results in Tables 6A-6D compare a hybrid for which inbred PHB6V is a parent and 4 other hybrids.
Using PHB6V to Develop Other Maize Inbreds
Inbred maize lines are typically developed for use in the production of hybrid maize lines. However, PHB6V could also be used to derive new maize inbred lines. Inbred maize lines preferably should be highly homogeneous, substantially homozygous and reproducible to be useful as parents of commercial hybrids. Plant breeding techniques known in the art and used in a maize plant breeding program include, but are not limited to, recurrent selection, mass selection, bulk selection, mass selection, backcrossing, pedigree breeding, open pollination breeding, restriction fragment length polymorphism enhanced selection, genetic marker enhanced selection, making double haploids, and transformation. Often combinations of these techniques are used. The development of maize hybrids in a maize plant breeding program requires, in general, the development of homozygous inbred lines, the crossing of these lines, and the evaluation of the crosses. There are many analytical methods available to evaluate the result of a cross. The oldest and most traditional method of analysis is the observation of phenotypic traits. Alternatively, the genotype of a plant can be examined.
Using PHB6V in a Breeding Program
This invention is directed to methods for producing a maize plant by crossing a first parent maize plant with a second parent maize plant wherein either the first or second parent maize plant is an inbred maize plant of the line PHB6V. Thus, any such methods using the inbred maize line PHB6V are part of this invention: selfing, sibbing, backcrosses, mass selection, bulk selection, hybrid production, crosses to populations, and the like. All plants produced using inbred maize line PHB6V as a parent are within the scope of this invention, including plants essentially derived from inbred maize line PHB6V, as such term is defined in 7 U.S.C. § 2104(a)(3) of the Plant Variety Protection Act, which definition is hereby incorporated by reference. This also includes progeny plants and parts thereof with at least one ancestor that is PHB6V, and more specifically, where the pedigree of the progeny includes 1, 2, 3, 4, and/or 5 or less breeding crosses to a maize plant other than PHB6V or a plant that has PHB6V as a parent or other progenitor. All breeders of ordinary skill in the art maintain pedigree records of their breeding programs. These pedigree records contain a detailed description of the breeding process, including a listing of all parental lines used in the breeding process and information on how such line was used. Thus, a breeder would know if PHB6V were used in the development of a progeny line, and would also know how many crosses to a line other than PHB6V or line with PHB6V as a parent or other progenitor were made in the development of any progeny line. The inbred maize line may also be used in crosses with other, different, maize inbreds to produce first generation (F1) maize hybrid seeds and plants with superior characteristics.
Pedigree Breeding
Pedigree breeding starts with the crossing of two genotypes, such as PHB6V and one other elite inbred line having one or more desirable characteristics that is lacking or which complements PHB6V. If the two original parents do not provide all the desired characteristics, other sources can be included in the breeding population. In the pedigree method, superior plants are selfed and selected in successive filial generations. In the succeeding filial generations the heterozygous condition gives way to homogeneous lines as a result of self-pollination and selection. Typically in the pedigree method of breeding, five or more successive filial generations of selfing and selection is practiced: F1→F2; F2→F3; F3→F4; F4→F5, etc. After a sufficient amount of inbreeding, successive filial generations will serve to increase seed of the developed inbred. Preferably, an inbred line comprises homozygous alleles at about 95% or more of its loci.
Backcrossing
In addition to being used to create a backcross conversion, backcrossing can also be used to modify PHB6V and a hybrid that is made using the modified PHB6V. As discussed previously, backcrossing can be used to transfer a specific desirable trait from one line, the donor parent, to an inbred called the recurrent parent which has overall good agronomic characteristics yet lacks that desirable trait. This transfer of the desirable trait into an inbred with overall good agronomic characteristics can be accomplished by first crossing a recurrent parent and a donor parent (non-recurrent parent). The progeny of this cross is then mated back to the recurrent parent followed by selection in the resultant progeny for the desired trait to be transferred from the non-recurrent parent as well as selection for the characteristics of the recurrent parent. Typically after four or more backcross generations with selection for the desired trait and the characteristics of the recurrent parent, the progeny will contain essentially all genes of the recurrent parent except for the genes controlling the desired trait. However, the number of backcross generations can be less if molecular markers are used during selection or elite germplasm is used as the donor parent. The last backcross generation is then selfed to give pure breeding progeny for the gene(s) being transferred. In addition to the above described use, backcrossing can be used in conjunction with pedigree breeding to develop new inbred lines. For example, an F1 can be created that is backcrossed to one of its parent lines to create a BC1, BC2, BC3, etc. Progeny are selfed and selected so that the newly developed inbred has many of the attributes of the recurrent parent and some of the desired attributes of the non-recurrent parent. This approach leverages the value and strengths of the recurrent parent for use in new hybrids and breeding, and has very significant value for a breeder.
Recurrent Selection and Mass Selection
Recurrent selection is a method used in a plant breeding program to improve a population of plants. PHB6V can be used in a recurrent selection program to improve the population. The method entails individual plants cross pollinating with each other to form progeny which are then grown. The superior progeny are then selected by any number of methods, which include individual plant, half-sib progeny, full-sib progeny, selfed progeny and top crossing. The selected progeny are cross pollinated with each other to form progeny for another population. This population is planted and again superior plants are selected to cross pollinate with each other. Recurrent selection is a cyclical process and therefore can be repeated as many times as desired. The objective of recurrent selection is to improve the traits of a population. The improved population can then be used as a source of breeding material to obtain inbred lines to be used in hybrids or used as parents for a synthetic cultivar. A synthetic cultivar is the resultant progeny formed by the intercrossing of several selected inbreds. Mass selection is a useful technique when used in conjunction with molecular marker enhanced selection. In mass selection seeds from individuals are selected based on phenotype. These selected seeds are then bulked and used to grow the next generation. Similarly, bulk selection requires growing a population of plants in a bulk plot, allowing the plants to self-pollinate, harvesting the seed in bulk and then using a sample of the seed harvested in bulk to plant the next generation. Also, instead of directed pollination, open pollination could be used as part of the breeding program.
Mutation Breeding
Mutation breeding is one of many methods that could be used to introduce new traits into PHB6V. Mutations that occur spontaneously or are artificially induced can be useful sources of variability for a plant breeder. The goal of artificial mutagenesis is to increase the rate of mutation for a desired characteristic. Mutation rates can be increased by many different means including temperature, long-term seed storage, tissue culture conditions, radiation; such as X-rays, Gamma rays (e.g. cobalt 60 or cesium 137), neutrons, (product of nuclear fission by uranium 235 in an atomic reactor), Beta radiation (emitted from radioisotopes such as phosphorus 32 or carbon 14), or ultraviolet radiation (preferably from 2500 to 2900 nm), or chemical mutagens (such as base analogues (5-bromo-uracil), related compounds (8-ethoxy caffeine), antibiotics (streptonigrin), alkylating agents (sulfur mustards, nitrogen mustards, epoxides, ethylenamines, sulfates, sulfonates, sulfones, lactones), azide, hydroxylamine, nitrous acid, or acridines. Once a desired trait is observed through mutagenesis the trait may then be incorporated into existing germplasm by traditional breeding techniques. Details of mutation breeding can be found in “Principles of Cultivar Development” Fehr, 1993 Macmillan Publishing Company the disclosure of which is incorporated herein by reference.
Breeding with Molecular Markers
Molecular markers, which includes markers identified through the use of techniques such as Isozyme Electrophoresis, Restriction Fragment Length Polymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Amplified Fragment Length Polymorphisms (AFLPs), Simple Sequence Repeats (SSRs) and Single Nucleotide Polymorphisms (SNPs), may be used in plant breeding methods utilizing PHB6V.
Isozyme Electrophoresis and RFLPs as discussed in Lee, M., “Inbred Lines of Maize and Their Molecular Markers,” The Maize Handbook, (Springer-Verlag, New York, Inc. 1994, at 423-432) incorporated herein by reference for this purpose, have been widely used to determine genetic composition. Isozyme Electrophoresis has a relatively low number of available markers and a low number of allelic variants among maize inbreds. RFLPs allow more discrimination because they have a higher degree of allelic variation in maize and a larger number of markers can be found. Both of these methods have been eclipsed by SSRs as discussed in Smith et al., “An evaluation of the utility of SSR loci as molecular markers in maize (Zea mays L.): comparisons with data from RFLPs and pedigree”, Theoretical and Applied Genetics (1997) vol. 95 at 163-173 and by Pejic et al., “Comparative analysis of genetic similarity among maize inbreds detected by RFLPs, RAPDs, SSRs, and AFLPs,” Theoretical and Applied Genetics (1998) at 1248-1255 incorporated herein by reference. SSR technology is more efficient and practical to use than RFLPs; more marker loci can be routinely used and more alleles per marker locus can be found using SSRs in comparison to RFLPs. Single Nucleotide Polymorphisms may also be used to identify the unique genetic composition of the invention and progeny lines retaining that unique genetic composition. Various molecular marker techniques may be used in combination to enhance overall resolution.
Maize DNA molecular marker linkage maps have been rapidly constructed and widely implemented in genetic studies. One such study is described in Boppenmaier, et al., “Comparisons among strains of inbreds for RFLPs”, Maize Genetics Cooperative Newsletter, 65:1991, pg. 90, is incorporated herein by reference.
One use of molecular markers is Quantitative Trait Loci (QTL) mapping. QTL mapping is the use of markers, which are known to be closely linked to alleles that have measurable effects on a quantitative trait. Selection in the breeding process is based upon the accumulation of markers linked to the positive effecting alleles and/or the elimination of the markers linked to the negative effecting alleles from the plant's genome.
Molecular markers can also be used during the breeding process for the selection of qualitative traits. For example, markers closely linked to alleles or markers containing sequences within the actual alleles of interest can be used to select plants that contain the alleles of interest during a backorossing breeding program. The markers can also be used to select for the genome of the recurrent parent and against the genome of the donor parent. Using this procedure can minimize the amount of genome from the donor parent that remains in the selected plants. It can also be used to reduce the number of crosses back to the recurrent parent needed in a backcrossing program. The use of molecular markers in the selection process is often called genetic marker enhanced selection.
Production of Double Haploids
The production of double haploids can also be used for the development of inbreds in the breeding program. An F1 hybrid for which PHB6V is a parent can be used to produce double haploid plants. Double haploids are produced by the doubling of a set of chromosomes (1 N) from a heterozygous plant to produce a completely homozygous individual. For example, see Wan et al., “Efficient Production of Doubled Haploid Plants Through Colchicine Treatment of Anther-Derived Maize Callus”, Theoretical and Applied Genetics, 77:889-892,1989. This can be advantageous because the process omits the generations of selfing needed to obtain a homozygous plant from a heterozygous source.
Use of PHB6V in Tissue Culture
This invention is also directed to the use of PHB6V in tissue culture. As used herein, the term “tissue culture” includes plant protoplasts, plant cell tissue culture, cultured microspores, plant calli, plant clumps, and the like. As used herein, phrases such as “growing the seed” or “grown from the seed” include embryo rescue, isolation of cells from seed for use in tissue culture, as well as traditional growing methods.
Duncan, Williams, Zehr, and Widholm, Planta (1985) 165:322-332 reflects that 97% of the plants cultured that produced callus were capable of plant regeneration. Subsequent experiments with both inbreds and hybrids produced 91% regenerable callus that produced plants. In a further study in 1988, Songstad, Duncan & Widholm in Plant Cell Reports (1988), 7:262-265 reports several media additions that enhance regenerability of callus of two inbred lines. Other published reports also indicated that “nontraditional” tissues are capable of producing somatic embryogenesis and plant regeneration. K. P. Rao, et al., Maize Genetics Cooperation Newsletter, 60:64-65 (1986), refers to somatic embryogenesis from glume callus cultures and B. V. Conger, et al., Plant Cell Reports, 6:345-347 (1987) indicates somatic embryogenesis from the tissue cultures of maize leaf segments. Thus, it is clear from the literature that the state of the art is such that these methods of obtaining plants are, and were, “conventional” in the sense that they are routinely used and have a very high rate of success.
Tissue culture of maize, including tassel/anther culture, is described in U.S. 2002/0062506A1 and European Patent Application, publication 160,390, each of which are incorporated herein by reference for this purpose. Maize tissue culture procedures are also described in Green and Rhodes, “Plant Regeneration in Tissue Culture of Maize,” Maize for Biological Research (Plant Molecular Biology Association, Charlottesville, Virginia 1982, at 367-372) and in Duncan, et al., “The Production of Callus Capable of Plant Regeneration from Immature Embryos of Numerous Zea Mays Genotypes,” 165 Planta 322-332 (1985). Thus, another aspect of this invention is to provide cells which upon growth and differentiation produce maize plants having the genotype and/or physiological and morphological characteristics of inbred line PHB6V.
Progeny of the breeding methods described herein may be characterized in any number of ways, such as by traits retained in the progeny, pedigree and/or molecular markers. Using the breeding methods described herein, one can develop individual plants, plant cells, and populations of plants that retain at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5% genetic contribution from inbred line PHB6V. In pedigree analysis the percentage genetic contribution may not be actually known, but on average 50% of the starting germplasm would be expected to be passed to the progeny line after one cross to another line, 25% after another cross to a different line, and so on. With backcrossing, the expected contribution of PHB6V after 2, 3, 4 and 5 doses (or 1, 2, 3 and 4 backcrosses) would be 75%, 87.5%, 93.75% and 96.875% respectively. Actual genetic contribution may be much higher than the genetic contribution expected by pedigree, especially if molecular markers are used in selection. Molecular markers could also be used to confirm and/or determine the pedigree of the progeny line.
Specific methods and products produced using inbred line PHB6V in plant breeding are discussed in the following sections. The methods outlined are described in detail by way of illustration and example for purposes of clarity and understanding. However, it will be obvious that certain changes and modifications may be practiced within the scope of the invention.
One method for producing a line derived from inbred line PHB6V is as follows. One of ordinary skill in the art would produce or obtain a seed from the cross between inbred line PHB6V and another variety of maize, such as an elite inbred variety. The F1 seed derived from this cross would be grown to form a homogeneous population. The F1 seed would contain essentially all of the alleles from variety PHB6V and essentially all of the alleles from the other maize variety. The F1 nuclear genome would be made-up of 50% variety PHB6V and 50% of the other elite variety. The F1 seed would be grown and allowed to self, thereby forming F2 seed. On average the F2 seed would have derived 50% of its alleles from variety PHB6V and 50% from the other maize variety, but many individual plants from the population would have a greater percentage of their alleles derived from PHB6V (Wang J. and R. Bernardo, 2000, Crop Sci. 40:659-665 and Bernardo, R. and A. L. Kahler, 2001, Theor. Appl. Genet 102:986-992). The molecular markers of PHB6V could be used to select and retain those lines with high similarity to PHB6V. The F2 seed would be grown and selection of plants would be made based on visual observation, markers and/or measurement of traits. The traits used for selection may be any PHB6V trait described in this specification, including the inbred per se maize PHB6V traits described herein under the detailed description of inbred PHB6V. Such traits may also be the good general or specific combining ability of PHB6V, including its ability to produce hybrids with the approximate maturity and/or hybrid combination traits described herein in under the detailed description of inbred PHB6V. The PHB6V progeny plants that exhibit one or more of the desired PHB6V traits, such as those listed herein, would be selected and each plant would be harvested separately. This F3 seed from each plant would be grown in individual rows and allowed to self. Then selected rows or plants from the rows would be harvested individually. The selections would again be based on visual observation, markers and/or measurements for desirable traits of the plants, such as one or more of the desirable PHB6V traits listed herein. The process of growing and selection would be repeated any number of times until a PHB6V progeny inbred plant is obtained. The PHB6V progeny inbred plant would contain desirable traits derived from inbred plant PHB6V, some of which may not have been expressed by the other maize variety to which inbred line PHB6V was crossed and some of which may have been expressed by both maize varieties but now would be at a level equal to or greater than the level expressed in inbred variety PHB6V. However, in each case the resulting progeny line would benefit from the efforts of the inventor(s), and would not have existed but for the inventor(s) work in creating PHB6V. The PHB6V progeny inbred plants would have, on average, 50% of their nuclear genes derived from inbred line PHB6V, but many individual plants from the population would have a greater percentage of their alleles derived from PHB6V. This breeding cycle, of crossing and selfing, and optional selection, may be repeated to produce another population of PHB6V progeny maize plants with, on average, 25% of their nuclear genes derived from inbred line PHB6V, but, again, many individual plants from the population would have a greater percentage of their alleles derived from PHB6V. Another embodiment of the invention is a PHB6V progeny plant that has received the desirable PHB6V traits listed herein through the use of PHB6V, which traits were not exhibited by other plants used in the breeding process.
The previous example can be modified in numerous ways, for instance selection may or may not occur at every selfing generation, selection may occur before or after the actual self-pollination process occurs, or individual selections may be made by harvesting individual ears, plants, rows or plots at any point during the breeding process described. In addition, double haploid breeding methods may be used at any step in the process. The population of plants produced at each and any cycle of breeding is also an embodiment of the invention, and on average each such population would predictably consist of plants containing approximately 50% of its genes from inbred line PHB6V in the first breeding cycle, 25% of its genes from inbred line PHB6V in the second breeding cycle, 12.5% of its genes from inbred line PHB6V in the third breeding cycle and so on. However, in each case the use of PHB6V provides a substantial benefit. The linkage groups of PHB6V would be retained in the progeny lines, and since current estimates of the maize genome size is about 50,000-80,000 genes (Xiaowu, Gai et al., Nucleic Acids Research, 2000, Vol. 28, No. 1, 94-96), in addition to non-coding DNA that impacts gene expression, it provides a significant advantage to use PHB6V as starting material to produce a line that retains desired genetics or traits of PHB6V.
Another embodiment of this invention is the method of obtaining a substantially homozygous PHB6V progeny plant by obtaining a seed from the cross of PHB6V and another maize plant and applying double haploid methods to the F1 seed or F1 plant or to any successive filial generation. Such methods decrease the number of generations required to produce an inbred with similar genetics or characteristics to PHB6V. See Bernardo, R. and Kahler, A. L., Theor. Appl. Genet. 102:986-992, 2001.
The utility of inbred maize line PHB6V also extends to crosses with other species. Commonly, suitable species will be of the family Graminaceae, and especially of the genera Zea, Tripsacum, Coix, Schlerachne, Polytoca, Chionachne, and Trilobachne, of the tribe Maydeae. Potentially suitable for crosses with PHB6V may be the various varieties of grain sorghum, Sorghum bicolor (L.) Moench.
As is readily apparent to one skilled in the art, the foregoing are only some of the various ways by which the inbred can be obtained by those looking to use the germplasm. Other means are available, and the above examples are illustrative only.
INDUSTRIAL APPLICABILITY
Maize is used as human food, livestock feed, and as raw material in industry. The food uses of maize, in addition to human consumption of maize kernels, include both products of dry- and wet-milling industries. The principal products of maize dry milling are grits, meal and flour. The maize wet-milling industry can provide maize starch, maize syrups, and dextrose for food use. Maize oil is recovered from maize germ, which is a by-product of both dry- and wet-milling industries.
Maize, including both grain and non-grain portions of the plant, is also used extensively as livestock feed, primarily for beef cattle, dairy cattle, hogs, and poultry.
Industrial uses of maize include production of ethanol, maize starch in the wet-milling industry and maize flour in the dry-milling industry. The industrial applications of maize starch and flour are based on functional properties, such as viscosity, film formation, adhesive properties, and ability to suspend particles. The maize starch and flour have application in the paper and textile industries. Other industrial uses include applications in adhesives, building materials, foundry binders, laundry starches, explosives, oil-well muds, and other mining applications.
Plant parts other than the grain of maize are also used in industry: for example, stalks and husks are made into paper and wallboard and cobs are used for fuel and to make charcoal.
The seed of inbred maize line PHB6V, the plant produced from the inbred seed, the hybrid maize plant produced from the crossing of the inbred, hybrid seed, and various parts of the hybrid maize plant and transgenic versions of the foregoing, can be utilized for human food, livestock feed, and as a raw material in industry.
Deposits
Applicant has made a deposit of at least 2500 seeds of Inbred Maize Line PHB6V with the American Type Culture Collection (ATCC), Manassas, Va. 20110 USA, ATCC Deposit No. PTA-4874. The seeds deposited with the ATCC on Dec. 26, 2002 were taken from the deposit maintained by Pioneer Hi-Bred International, Inc., 7250 NW 62nd Avenue, Johnston, Iowa 50131-1000, since prior to the filing date of this application. Access to this deposit will be available during the pendency of the application to the Commissioner of Patents and Trademarks and persons determined by the Commissioner to be entitled thereto upon request. Upon allowance of any claims in the application, the Applicant will make the deposit available to the public pursuant to 37 C.F.R. § 1.808. This deposit of the Inbred Maize Line PHB6V will be maintained in the ATCC depository, which is a public depository, for a period of 30 years, or 5 years after the most recent request, or for the enforceable life of the patent, whichever is longer, and will be replaced if it becomes nonviable during that period. Additionally, Applicant has satisfied all of the requirements of 37 C.F.R. §§1.801-1.809, including providing an indication of the viability of the sample upon deposit. Applicant has no authority to waive any restrictions imposed by law on the transfer of biological material or its transportation in commerce. Applicant does not waive any infringement of his rights granted under this patent or under the Plant Variety Protection Act (7 USC 2321 et seq.). U.S. Plant Variety Protection of Inbred Maize Line PHB6V has been applied for under Application No. 200300226.







TABLE 1





VARIETY DESCRIPTION INFORMATION


PHB6V


















1. TYPE: (Describe intermediate types in comments
Avg.
Sta.
Sam.












section)

Dev
Size



1 = Sweet, 2 = Dent, 3 = Flint, 4 = Flour,
2



5 = Pop and 6 = Ornamental.



Comments: Dent-Flint










2. MATURITY: DAYS HEAT UNITS
Days
H.Units













From emergence to 50% of plants in silk
58
1,292




From emergence to 50% of plants in pollen
57
1,245



From 10% to 90% pollen shed
2
61



From 50% Silk to harvest at 25% moisture







3. PLANT:












Plant Height (to tassel tip) (cm)
199.7
10.15
30



Ear Height (to base of top ear node) (cm)
73.7
11.24
30



Length of Top Ear Internode (cm)
13.6
1.73
30



Average Number of Tillers per Plant
0.0
0.01
6



Average Number of Ears per Stalk
1.1
0.17
6



Anthocyanin of Brace Roots: 1 = Absent, 2 =
3



Faint, 3 = Moderate, 4 = Dark







4. LEAF:












Width of Ear Node Leaf (cm)
8.3
0.76
30



Length of Ear Node Leaf (cm)
76.5
3.81
30



Number of Leaves above Top Ear
6.3
0.74
30



Leaf Angle: (measure from 2nd leaf above
22.0
4.11
30



ear at anthesis to stalk above leaf)(Degrees)










*
Leaf color: Dark Green Munsell code: 5GY36





Leaf Sheath Pubescence: 1 = none to 9 = like
2



peach fuzz







5. TASSEL:












Number of Primary Lateral Branches
4.6
1.38
30



Branch Angle from Central Spike
24.1
8.42
30



Tassel Length (from top leaf collar to tassel
55.7
3.27
30



tip)(cm):



Pollen Shed: 0 = male sterile, 9 = heavy shed
4










*
Anther Color:
Purple
Munsell code: 7.5RP36


*
Glume Color:
Light Green
Munsell code: 7.5GY56











*
Bar Glumes (glume bands): 1 = absent, 2 =
1





present



Peduncle Length (from top leaf to basal
20.4
3.32
30



branches) (cm):










6a. EAR (Unhusked ear)
Avg.
St.
Sam.











*
Silk color:
Red
Munsell c.
2.5R48









(3 days after silk emergence)











*
Fresh husk color:
Dark Green
Munsell c.
5GY56









(25 days after 50% silking)











*
Dry husk color:
Buff
Munsell c.
5Y92









(65 days after 50% silking)











Ear position at dry husk stage: 1 = upright , 2 =
3




horizontal, 3 = pendant



Husk tightness: (1 = very loose, 9 = very tight)
4



Husk extension (at harvest): 1 = short (ears
2



exposed), 2 = medium (<8 cm), 3 = long (8-10



cm), 4 = v. long (>10 cm)







6b. EAR (Husked ear data)












Ear length (cm):
16.4
1.30
30



Ear diameter at mid-point (mm)
35.6
2.22
30



Ear weight (gm):
95.7
23.65
30



Number of Kernel Rows:
13.1
1.14
30



Kernel Rows: 1 = indistinct, 2 = distinct
2



Row alignment: 1 = straight, 2 = slightly
2



curved, 3 = spiral



Shank Length (cm):
11.3
3.59
30



Ear Taper: 1 = slight cylind., 2 = average,
2



3 = extreme conic.







7. KERNEL (Dried):












Kernel Length (mm):
10.4
0.77
30



Kernel Width (mm):
7.7
0.69
30



Kernel Thickness (mm):
5.0
0.85
30



Round Kernels (shape grade) (%)
40.6
5.37
6



Aleurone Color pattern: 1 = homozygous,
1



2 = segregating










*
Aleurone Color:
Yellow
Munsell c.: 2.5Y814


*
Hard Endo. color:
Yellow
Munsell c.: 1.25Y714












Endosperm Type:
3





1 = sweet (su1), 2 = extra sweet (sh2), 3 =



normal starch, 4 = high amylose starch, 5 =



waxy starch, 6 = high protein, 7 = high lysine,



8 = super sweet (se), 9 = high oil, 10 = other



Weight per 100 kernels (unsized sample) (gm):
25.8
2.64
6







8. COB:











*
Cob Diameter at mid-point (mm):
20.0
1.02
30










*
Cob Color:
Red
Munsell c.: 5R46







10. DISEASE RESISTANCE:









(Rate from 1 = most-susceptable to 9 = most-resistant. Leave



blank if not tested, leave race or strain options blank if



polygenic.)







A. LEAF BLIGHTS, WILTS, AND LOCAL INFECTION DISEASES











Anthracnose Leaf Blight (Colletotrichum graminicola)



5
Common Rust (Puccinia sorghi)




Common Smut (Ustilago maydis)



8
Eyespot (Kabatiella zeae)




Gross's Wilt (Clavibacter michiganense spp. nebraskense)



4
Gray Leaf Spot (Cercospora zeae-maydis)




Helminthosporium Leaf Spot (Bipolaris zeicola) Race:



4
Northern Leaf Blight (Exserohilum turcicum) Race:




Southern Leaf Blight (Bipolaris maydis) Race:




Southern Rust (Puccinia polysora)



7
Stewart's Wilt (Erwinia stewartii)




Other (Specify):—————







B. SYSTEMIC DISEASES











Corn Lethal Necrosis (MCMV and MDMV)



9
Head Smut (Sphacelotheca reiliana)




Maize Chlorotic Dwarf Virus (MDV)




Maize Chlorotic Mottle Virus (MCMV)




Maize Dwarf Mosaic Virus (MDMV)




Sorghum Downy Mildew of Corn (Peronosclerospora sorghi)




Other (Specify):—————







C. STALK ROTS










8
Anthracnose Stalk Rot (Colletotrichum graminicola)




Diploidia Stalk Rot (Stenocarpella maydis)




Fusarium Stalk Rot (Fusarium moniliforme)




Gibberella Stalk Rot (Gibberella zeae)




Other (Specify):—————







D. EAR AND KERNEL ROTS











Aspergillus Ear and Kernel Rot (Aspergillus flavus)




Diplodia Ear Rot (Stenocarpella maydis)



7
Fusarium Ear and Kernel Rot (Fusarium moniliforme)




Gibberella Ear Rot (Gibberella zeae)




Other (Specify):—————







11. INSECT RESISTANCE:









(Rate from 1 = most-suscept. to 9 = most-resist., leave blank if



not tested.)



Corn Worm (Helicoverpa zea)



Leaf Feeding



Silk Feeding



Ear Damage



Corn Leaf Aphid (Rophalosiphum maydis)



Corn Sap Beetle (Capophilus dimidiatus)



European Corn Borer (Ostrinia nubilalis)



1st. Generation (Typically whorl leaf feeding)



2nd. Generation (Typically leaf sheath-collar feeding)



Stalk Tunneling



cm tunneled/plant



Fall armyworm (Spodoptera fruqiperda)



Leaf Feeding



Silk Feeding



mg larval wt.



Maize Weevil (Sitophilus zeamaize)



Northern Rootworm (Diabrotica barberi)



Southern Rootworm (Diabrotica undecimpunctata)



Southwestern Corn Borer (Diatreaea grandiosella)



Leaf Feeding



Stalk Tunneling



cm tunneled/plant



Two-spotted Spider Mite (Tetranychus utricae)



Western Rootworm (Diabrotica virgifrea virgifrera)



Other (Specify):—————







12. AGRONOMIC TRAITS:










3
Staygreen (at 65 days after anthesis; rate from 1-worst to




9-excellent)



0
% Dropped Ears (at 65 days after anthesis)




% Pre-anthesis Brittle Snapping




% Pre-anthesis Root Lodging




% Post-anthesis Root Lodging (at 65 days after anthesis)




% Post-anthesis Stalk Lodging









4,860.0 Kg/ha Yield of inbred per se (at 12-13% grain moisture)







*Munsell Glossy Book of Color, a standard color reference Kollmorgen Inst. Corp. New Windsor NY.













TABLE 2







SSR PROFILE OF PHB6V











Locus
Bin #
PHB6V mwt















bnlg1014
1.01
128



phi427913
1.01
131



phi056
1.01
255



bnlg1127
1.02
117



bnlg1627
1.02
205



bnlg1429
1.02
207



bnlg1083
1.02
225



bnlg1953
1.02
252



bnlg439
1.03
236



phi109275
1.03
136



bnlg1484
1.03
138



phi339017
1.03
148



bnlg1203
1.03
307



bnlg2238
1.04
203



bnlg2086
1.04
228



bnlg1016
1.04
255



bnlg1886
1.05
150



bnlg1832
1.05
236



bnlg1615
1.06
223



bnlg1057
1.06
251



bnlg1556
1.07
211



phi423298
1.08
129



phi323065
1.08
331



phi002
1.08
72



phi335539
1.08
91



bnlg1331
1.09
145



phi011
1.09
230



bnlg1720
1.09(1.10)
240



phi308707
1.10
134



phi265454
1.11
238



phi227562
1.11
322



phi064
1.11
97



phi402893
2.00
209



phi96100
2.01
303



bnlg1017
2.02
178



bnlg2277
2.02
296



phi109642
2.03
152



bnlg1064
2.03
204



phi083
2.04
132



bnlg1018
2.04
137



bnlg1909
2.05
312



bnlg1396
2.06
136



bnlg1831
2.06
197



bnlg1138
2.06
219



phi251315
2.07
127



phi427434
2.08
123



phi328189
2.08
124



phi127
2.08
124



phi090
2.08
141



bnlg1141
2.08
183



bnlg1940
2.08
211



phi435417
2.08
219



bnlg2237
2.08
220



bnlg1520
2.09
288



phi101049
2.10
234



bnlg1690
2.XX
114



phi453121
3.00
213



phi104127
3.01
165



phi404206
3.01
305



phi193225
3.02
137



bnlg1647
3.02
154



phi374118
3.02
220



bnlg1523
3.03
307



bnlg1452
3.04
128



bnlg1019
3.04
158



phi029
3.04
161



bnlg1035
3.05
106



phi053
3.05
169



phi102228
3.06
131



bnlg1951
3.06
134



bnlg1160
3.06
223



phi072
4.00(4.01)
143



phi213984
4.01
304



bnlg1162
4.03
98



phi308090
4.04
223



phi096
4.04
238



phi079
4.05
190



bnlg1265
4.05
196



phi438301
4.05
214



bnlg1755
4.05
241



bnlg1937
4.05(4.06)
241



bnlg1189
4.07
141



bnlg2244
4.08
219



phi093
4.08
291



phi076
4.11
161



phi396160
5.02
303



phi109188
5.03
164



bnlg653
5.04
154



bnlg1208
5.04
121



phi331888
5.04
131



phi330507
5.04
144



bnlg1892
5.04
156



phi333597
5.05
219



phi085
5.06
262



bnlg1711
5.07
178



bnlg1118
5.07
86



phi159819
6.00(6.08)
126



phi423796
6.01
131



phi389203
6.03
307



phi452693
6.04
126



bnlg1174
6.05
222



phi445613
6.05
99



phi299852
6.07
123



phi364545
6.07
134



bnlg1740
6.07
212



phi070
6.07
83



phi338882
6.XX
172



phi034
7.02
122



bnlg2271
7.03
237



phi328175
7.04
124



phi051
7.05
142



phi069
7.05
196



phi116
7.06
168



phi420701
8.00
294



bnlg1194
8.02
144



phi100175
8.03
140



bnlg2082
8.03
174



phi115
8.03
303



bnlg2046
8.04
327



bnlg1176
8.05
223



bnlg1152
8.06
151



bnlg1065
8.07
243



phi015
8.08
96



bnlg1056
8.08
97



phi233376
8.09
142



bnlg2122
9.01
226



phi033
9.01
252



bnlg1159
9.04
149



bnlg1012
9.04
164



phi032
9.04
240



phi236654
9.05
126



phi108411
9.05
129



phi448880
9.06(9.07)
187



bnlg619
9.07
275



bnlg1375
9.07
178



bnlg1129
9.08
303



phi041
10.00
202



phi059
10.02
156



phi96342
10.02
249



bnlg1037
10.03
121



bnlg1655
10.03
150



bnlg1079
10.03
175



phi050
10.03
80



phi301654
10.04
132



phi062
10.04
161



phi323152
10.05
144



bnlg1074
10.05
174



bnlg1450
10.07
187



bnlg1185
10.07
191

















TABLE 3A





PAIRED INBRED COMPARISON REPORT


Variety #1: PHB6V


Variety #2: PH185



























YIELD
YIELD
MST
TSTWT
EGRWTH
ESTCNT
TILLER
GDUSHD
GDUSLK
POLWT
POLWT



BU/A 56#
BU/A 56#
PCT
LB/BU
SCORE
COUNT
PCT
GDU
GDU
VALUE
VALUE


Stat
ABS
% MN
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
% MN





Mean1
77.4
117.8
15.9
54.2
5.7
23.5
5.3
126.4
130.3
81.0
66.8


Mean2
58.4
91.2
17.2
56.3
5.6
19.7
1.6
117.2
119.5
116.3
95.4


Locs
31
31
35
6
21
13
13
42
42
10
10


Reps
35
35
39
6
21
13
13
42
42
19
19


Diff
19.0
26.6
1.3
−2.2
0.1
3.8
−3.7
9.2
10.8
−35.3
−28.6


Prob
0.000
0.000
0.015
0.085
0.561
0.003
0.223
0.000
0.000
0.000
0.000





















TASBLS
TASSZ
PLTHT
EARHT
STAGRN
STKLDG
BRTSTK
SCTGRN
EARSZ
TEXEAR



SCORE
SCORE
CM
CM
SCORE
% NOT
% NOT
SCORE
SCORE
SCORE


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS





Mean1
9.0
4.1
208.8
82.7
4.3
78.3
97.8
6.6
4.0
5.5


Mean2
9.0
4.0
182.1
65.1
4.5
92.9
100.0
7.3
3.0
5.5


Locs
3
34
25
11
4
2
2
11
1
2


Reps
3
34
25
11
4
2
2
11
1
2


Diff
0.0
0.1
26.7
17.7
−0.3
−14.6
−2.2
−0.6
1.0
0.0


Prob
1.000
0.751
0.000
0.006
0.873
0.500
0.500
0.132

1.000






EARMLD
BARPLT
DRPEAR
GLFSPT
NLFBLT
STWWLT
ANTROT
FUSERS
GIBERS
EYESPT



SCORE
% NOT
% NOT
SCORE
SCORE
SCORE
SCORE
SCORE
SCORE
SCORE


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS





Mean1
7.5
97.7
100.0
4.5
3.5
6.8
8.5
6.8
4.5
7.5


Mean2
7.3
94.1
100.0
5.0
5.5
6.8
9.0
7.0
5.8
5.5


Locs
6
15
1
1
2
4
2
5
2
1


Reps
6
15
1
2
2
4
4
5
3
2


Diff
0.2
3.6
0.0
−0.5
−2.0
0.0
−0.5
−0.2
−1.3
2.0


Prob
0.741
0.136


1.000
1.000
0.500
0.749
0.500






COMRST
CLDTST
CLDTST
KSZDCD
HD SMT
ERTLDG
LRTLDG
ERTLPN
LRTLPN
STLLPN



SCORE
PCT
PCT
PCT
% NOT
% NOT
% NOT
% NOT
% NOT
% NOT


Stat
ABS
ABS
% MN
ABS
ABS
ABS
ABS
ABS
ABS
ABS





Mean1
4.8
87.1
101.6
11.1
100.0
100.0
100.0
83.3
89.0
20.4


Mean2
5.3
82.3
95.2
5.1
98.9
1.8
100.0
56.7
87.0
96.2


Locs
4
12
12
13
2
1
1
3
5
1


Reps
4
12
12
13
4
1
1
3
6
1


Diff
−0.5
4.8
6.4
6.0
1.1
98.2
0.0
26.7
2.0
−75.8


Prob
0.182
0.229
0.211
0.032
0.500


0.270
0.772
















TABLE 3B





PAIRED INBRED COMPARISON REPORT


Variety #1: PHB6V


Variety #2: PHAA0



























YIELD
YIELD
MST
TSTWT
EGRWTH
ESTCNT
TILLER
GDUSHD
GDUSLK
POLWT
POLWT



BU/A 56#
BU/A 56#
PCT
LB/BU
SCORE
COUNT
PCT
GDU
GDU
VALUE
VALUE


Stat
ABS
% MN
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
% MN





Mean1
78.1
117.8
16.3
54.0
5.7
23.3
5.3
126.5
130.2
83.1
68.0


Mean2
77.6
117.3
16.5
55.5
6.1
20.4
1.2
120.2
122.2
88.8
70.7


Locs
28
28
31
4
23
15
13
51
51
9
9


Reps
32
32
35
4
23
15
13
51
51
17
17


Diff
0.6
0.5
0.2
−1.5
−0.4
2.9
−4.2
6.3
8.0
−5.7
−2.7


Prob
0.872
0.927
0.719
0.297
0.119
0.029
0.245
0.000
0.000
0.586
0.735





















TASBLS
TASSZ
PLTHT
EARHT
STAGRN
STKLDG
BRTSTK
SCTGRN
EARSZ
TEXEAR



SCORE
SCORE
CM
CM
SCORE
% NOT
% NOT
SCORE
SCORE
SCORE


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS





Mean1
9.0
4.2
206.9
83.6
3.4
55.9
97.8
6.9
4.0
5.5


Mean2
9.0
2.9
188.0
76.6
3.3
58.9
100.0
7.6
4.0
6.0


Locs
3
43
28
13
7
3
2
15
1
2


Reps
3
43
28
13
7
3
2
15
1
2


Diff
0.0
1.3
18.9
7.0
0.1
−3.1
−2.2
−0.7
0.0
−0.5


Prob
1.000
0.000
0.000
0.084
0.838
0.848
0.500
0.076

0.874






EARMLD
BARPLT
DRPEAR
GLFSPT
NLFBLT
STWWLT
ANTROT
FUSERS
GIBERS
EYESPT



SCORE
% NOT
% NOT
SCORE
SCORE
SCORE
SCORE
SCORE
SCORE
SCORE


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS





Mean1
7.5
96.8
100.0
3.5
3.5
7.0
8.5
6.8
4.5
7.5


Mean2
7.5
98.2
100.0
3.3
5.5
7.0
8.5
8.0
5.0
5.5


Locs
6
18
1
2
2
3
2
5
2
1


Reps
6
18
1
3
2
3
4
5
3
2


Diff
0.0
−1.4
0.0
0.3
−2.0
0.0
0.0
−1.2
−0.5
2.0


Prob
1.000
0.360

0.795
0.295
1.000
1.000
0.324
0.500






COMRST
CLDTST
CLDTST
KSZDCD
HD SMT
ERTLDG
LRTLDG
ERTLPN
LRTLPN
STLLPN



SCORE
PCT
PCT
PCT
% NOT
% NOT
% NOT
% NOT
% NOT
% NOT


Stat
ABS
ABS
% MN
ABS
ABS
ABS
ABS
ABS
ABS
ABS





Mean1
4.8
87.2
101.4
10.0
100.0
100.0
100.0
83.3
90.8
20.4


Mean2
4.0
89.2
103.5
5.0
100.0
84.1
100.0
66.7
97.5
14.0


Locs
4
9
9
12
2
1
1
3
6
1


Reps
4
9
9
12
4
1
1
3
7
1


Diff
0.8
−2.0
−2.1
5.0
0.0
15.9
0.0
16.7
−6.7
6.4


Prob
0.058
0.396
0.453
0.042
1.000


0.300
0.235
















TABLE 3C





PAIRED INBRED COMPARISON REPORT


Variety #1: PHB6V


Variety #2: PH5TG



























YIELD
YIELD
MST
TSTWT
EGRWTH
ESTCNT
TILLER
GDUSHD
GDUSLK
POLWT
POLWT



BU/A 56#
BU/A 56#
PCT
LB/BU
SCORE
COUNT
PCT
GDU
GDU
VALUE
VALUE


Stat
ABS
% MN
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
% MN





Mean1
77.4
117.8
15.9
54.5
5.7
23.4
5.3
126.5
130.2
81.0
66.8


Mean2
81.1
124.3
16.8
55.9
6.2
21.4
10.2
125.2
129.2
116.4
96.6


Locs
31
31
35
5
24
14
13
50
50
10
10


Reps
35
35
39
5
24
14
13
50
50
19
19


Diff
−3.6
−6.5
0.9
−1.3
−0.5
2.0
4.8
1.3
1.0
−35.4
−29.8


Prob
0.377
0.317
0.123
0.253
0.037
0.031
0.211
0.021
0.119
0.000
0.000





















TASBLS
TASSZ
PLTHT
EARHT
STAGRN
STKLDG
BRTSTK
SCTGRN
EARSZ
TEXEAR



SCORE
SCORE
CM
CM
SCORE
% NOT
% NOT
SCORE
SCORE
SCORE


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS





Mean1
9.0
4.2
206.9
83.6
3.4
55.9
97.8
6.9
4.0
5.5


Mean2
9.0
4.4
201.3
80.6
3.1
85.4
90.9
6.3
4.0
5.5


Locs
3
43
28
13
7
3
2
15
1
2


Reps
3
43
28
13
7
3
2
15
1
2


Diff
0.0
−0.2
5.6
2.9
0.3
−29.5
6.9
0.7
0.0
0.0


Prob
1.000
0.193
0.062
0.301
0.457
0.261
0.406
0.126

1.000






EARMLD
BARPLT
DRPEAR
GLFSPT
NLFBLT
STWWLT
ANTROT
FUSERS
GIBERS
EYESPT



SCORE
% NOT
% NOT
SCORE
SCORE
SCORE
SCORE
SCORE
SCORE
SCORE


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS





Mean1
7.5
96.8
100.0
4.5
3.5
6.8
8.5
6.8
4.5
7.5


Mean2
8.2
95.0
100.0
4.5
7.5
7.3
8.5
8.2
6.0
5.5


Locs
6
18
1
1
2
4
2
5
2
1


Reps
6
18
1
2
2
4
4
5
3
2


Diff
−0.7
1.9
0.0
0.0
−4.0
−0.5
0.0
−1.4
−1.5
2.0


Prob
0.444
0.036


0.156
0.391
1.000
0.184
0.205






COMRST
CLDTST
CLDTST
KSZDCD
HD SMT
ERTLDG
LRTLDG
ERTLPN
LRTLPN
STLLPN



SCORE
PCT
PCT
PCT
% NOT
% NOT
% NOT
% NOT
% NOT
% NOT


Stat
ABS
ABS
% MN
ABS
ABS
ABS
ABS
ABS
ABS
ABS





Mean1
4.8
87.1
101.6
11.1
100.0
100.0
100.0
83.3
90.8
20.4


Mean2
4.8
88.4
102.7
6.6
100.0
46.4
100.0
80.0
90.8
61.1


Locs
4
12
12
13
2
1
1
3
6
1


Reps
4
12
12
13
4
1
1
3
7
1


Diff
0.0
−1.3
−1.1
4.5
0.0
53.6
0.0
3.3
0.0
−40.7


Prob
1.000
0.578
0.693
0.019
1.000


0.885
1.000
















TABLE 3D





PAIRED INBRED COMPARISON REPORT


Variety #1: PHB6V


Variety #2: PH75K

























YIELD
YIELD
MST
TSTWT
EGRWTH
ESTCNT
TILLER
GDUSHD
GDUSLK



BU/A 56#
BU/A 56#
PCT
LB/BU
SCORE
COUNT
PCT
GDU
GDU


Stat
ABS
% MN
ABS
ABS
ABS
ABS
ABS
ABS
ABS



















Mean1
77.4
117.8
15.9
54.5
5.7
23.5
5.3
126.4
130.4


Mean2
68.3
103.9
17.4
54.1
6.0
21.2
1.9
122.3
122.3


Locs
31
31
35
5
21
13
13
41
41


Reps
35
35
39
5
21
13
13
41
41


Diff
9.1
13.9
1.4
0.5
−0.3
2.4
−3.4
4.1
8.1


Prob
0.011
0.012
0.009
0.834
0.167
0.023
0.336
0.000
0.000




















POLWT
POLWT
TASBLS
TASSZ
PLTHT
EARHT
STAGRN
STKLDG
BRTSTK



VALUE
VALUE
SCORE
SCORE
CM
CM
SCORE
% NOT
% NOT


Stat
ABS
% MN
ABS
ABS
ABS
ABS
ABS
ABS
ABS



















Mean1
71.8
67.0
9.0
4.1
208.8
82.7
4.3
78.3
97.8


Mean2
70.4
65.3
9.0
4.4
188.7
76.8
4.3
92.5
96.3


Locs
2
2
3
33
25
11
4
2
2


Reps
3
3
3
33
25
11
4
2
2


Diff
1.4
1.7
0.0
−0.3
20.1
6.0
0.0
−14.2
1.5


Prob
0.921
0.898
1.000
0.194
0.000
0.090
1.000
0.500
0.500




















SCTGRN
EARSZ
TEXEAR
EARMLD
BARPLT
DRPEAR
GLFSPT
NLFBLT
STWWLT



SCORE
SCORE
SCORE
SCORE
% NOT
% NOT
SCORE
SCORE
SCORE


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS



















Mean1
6.6
4.0
5.5
7.2
97.7
100.0
4.5
3.5
6.8


Mean2
7.3
4.0
7.0
7.0
92.3
98.1
3.0
6.0
7.3


Locs
11
1
2
5
15
1
1
2
4


Reps
11
1
2
5
15
1
2
2
4


Diff
−0.6
0.0
−1.5
0.2
5.3
1.9
1.5
−2.5
−0.5


Prob
0.387

0.742
0.847
0.025


0.126
0.391




























HD



ANTROT
FUSERS
GIBERS
EYESPT
COMRST
CLDTST
CLDTST
KSZDCD
SMT



SCORE
SCORE
SCORE
SCORE
SCORE
PCT
PCT
PCT
% NOT


Stat
ABS
ABS
ABS
ABS
ABS
ABS
% MN
ABS
ABS



















Mean1
8.5
6.8
4.5
7.5
4.8
87.1
101.6
11.1
100.0


Mean2
8.8
7.6
3.8
5.5
4.0
87.0
101.2
4.2
100.0


Locs
2
5
2
1
4
12
12
13
2


Reps
4
5
3
2
4
12
12
13
4


Diff
−0.3
−0.8
0.8
2.0
0.8
0.1
0.4
6.9
0.0


Prob
0.500
0.477
0.795

0.058
0.973
0.882
0.012
1.000


















ERTLDG
LRTLDG
ERTLPN
LRTLPN
STLLPN




% NOT
% NOT
% NOT
% NOT
% NOT



Stat
ABS
ABS
ABS
ABS
ABS


















Mean1
100.0
100.0
83.3
86.3
20.4



Mean2
89.3
100.0
66.7
72.5
40.7



Locs
1
1
3
4
1



Reps
1
1
3
4
1



Diff
10.7
0.0
16.7
13.8
−20.4



Prob


0.199
0.241

















TABLE 3E





PAIRED INBRED COMPARISON REPORT


Variety #1: PHB6V


Variety #2: PHJ40
























EGRWTH
ESTCNT
TILLER
GDUSHD
GDUSLK
TASBLS
TASSZ
PLTHT



SCORE
COUNT
PCT
GDU
GDU
SCORE
SCORE
CM


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS


















Mean1
4.7
23.8
1.8
128.7
132.2
9.0
4.6
205.6


Mean2
5.6
19.4
2.5
122.0
124.1
9.0
4.0
173.1


Locs
7
5
7
22
22
1
18
12


Reps
7
5
7
22
22
1
18
12


Diff
−0.9
4.4
0.8
6.8
8.1
0.0
0.6
32.5


Prob
0.045
0.108
0.622
0.000
0.000

0.145
0.000



















EARHT
STAGRN
STKLDG
SCTGRN
BARPLT
GLFSPT
NLFBLT
STWWLT



CM
SCORE
% NOT
SCORE
% NOT
SCORE
SCORE
SCORE


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS


















Mean1
91.3
4.5
100.0
4.0
98.4
3.5
3.5
7.0


Mean2
80.2
4.5
100.0
4.0
87.8
3.5
6.5
7.0


Locs
5
2
1
1
6
2
2
1


Reps
5
2
1
1
6
3
2
1


Diff
11.1
0.0
0.0
0.0
10.6
0.0
−3.0
0.0


Prob
0.111
1.000


0.403
1.000
1.000
























HD






ANTROT
GIBERS
EYESPT
COMRST
SMT
ERTLPN
LRTLPN




SCORE
SCORE
SCORE
SCORE
% NOT
% NOT
% NOT



Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS




















Mean1
8.5
1.0
7.5
6.0
100.0
100.0
100.0



Mean2
8.8
1.5
6.5
5.5
100.0
100.0
100.0



Locs
2
1
1
2
2
1
1



Reps
4
2
2
2
4
1
1



Diff
−0.3
−0.5
1.0
0.5
0.0
0.0
0.0



Prob
0.500


0.500
1.000

















TABLE 3F





PAIRED INBRED COMPARISON REPORT


Variety #1: PHB6V


Variety #2: PHJ40

























EGRWTH
ESTCNT
TILLER
GDUSHD
GDUSLK
TASBLS
TASSZ
PLTHT
EARHT



SCORE
COUNT
PCT
GDU
GDU
SCORE
SCORE
CM
CM


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS



















Mean1
4.7
23.8
1.8
128.7
132.2
9.0
4.6
205.6
91.3


Mean2
5.6
19.4
2.5
122.0
124.1
9.0
4.0
173.1
80.2


Locs
7
5
7
22
22
1
18
12
5


Reps
7
5
7
22
22
1
18
12
5


Diff
−0.9
4.4
0.8
6.8
8.1
0.0
0.6
32.5
11.1


Prob
0.045
0.108
0.622
0.000
0.000

0.145
0.000
0.111



















STAGRN
STKLDG
SCTGRN
BARPLT
GLFSPT
NLFBLT
STWWLT
ANTROT



SCORE
% NOT
SCORE
% NOT
SCORE
SCORE
SCORE
SCORE


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS


















Mean1
4.5
100.0
4.0
98.4
3.5
3.5
7.0
8.5


Mean2
4.5
100.0
4.0
87.8
3.5
6.5
7.0
8.8


Locs
2
1
1
6
2
2
1
2


Reps
2
1
1
6
3
2
1
4


Diff
0.0
0.0
0.0
10.6
0.0
−3.0
0.0
−0.3


Prob
1.000


0.403
1.000
1.000

0.500






















HD






GIBERS
EYESPT
COMRST
SMT
ERTLPN
LRTLPN




SCORE
SCORE
SCORE
% NOT
% NOT
% NOT



Stat
ABS
ABS
ABS
ABS
ABS
ABS



















Mean1
1.0
7.5
6.0
100.0
100.0
100.0



Mean2
1.5
6.5
5.5
100.0
100.0
100.0



Locs
1
1
2
2
1
1



Reps
2
2
2
4
1
1



Diff
−0.5
1.0
0.5
0.0
0.0
0.0



Prob


0.500
1.000

















TABLE 4A





Average Inbred By Tester Performance Comparing PHB6V To PHAA0


Crossed To The Same Inbred Testers And Grown In The


Same Experiments.





















YIELD
MST
EGRWTH
ESTCNT
GDUSHD


—
BU/A 56#
PCT
SCORE
COUNT
GDU





Reps
28
28
5
7
17


Locs
28
28
5
7
17


PHB6V
163.0
23.6
5.6
55.1
116.8


PHAA0
155.0
23.7
5.4
57.3
114.3


Diff
8.0
0.0
0.2
2.1
2.5


Pr > T
0.009
0.991
0.704
0.216
0.025










TSTWT



GDUSLK
PLTHT
EARHT
STAGRN
LB/BU


—
GDU
CM
CM
SCORE
ABS





Reps
6
13
13
14
17


Locs
6
13
13
14
17


PHB6V
117.3
288.9
122.4
5.6
52.9


PHAA0
113.2
263.7
125.0
4.4
54.0


Diff
4.1
25.2
2.6
1.2
1.0


Pr > T
0.077
0.001
0.771
0.028
0.020

















EYESPT
COMRST
STLLPN
EBTSTK




SCORE
SCORE
% NOT
% NOT



—
ABS
ABS
ABS
ABS







Reps
1
4
1
5



Locs
1
4
1
5



PHB6V
6.0
5.8
100.0
93.4



PHAA0
6.0
5.0
100.0
80.1



Diff
0.0
0.8
0.0
13.3



Pr > T

0.215

0.471







*Pr > T values are valid only for comparisons with Locs >= 10













TABLE 4B





Average Inbred By Tester Performance Comparing PHB6V To PH185


Crossed To The Same Inbred Testers And Grown In The Same


Experiments.





















YIELD
MST
EGRWTH
ESTCNT
GDUSHD


—
BU/A 56#
PCT
SCORE
COUNT
GDU





Reps
17
17
5
5
9


Locs
17
17
5
5
9


PHB6V
157.2
22.6
5.6
52.0
114.9


PH185
141.6
21.7
5.6
55.2
110.9


Diff
15.6
0.9
0.0
3.2
4.0


Pr > T
0.001
0.208
1.000
0.150
0.004




















TSTWT



GDUSLK
PLTHT
EARHT
STAGRN
LB/BU


—
GDU
CM
CM
SCORE
ABS





Reps
2
9
9
8
11


Locs
2
9
9
8
11


PHB6V
115.0
298.0
122.2
5.4
53.1


PH185
110.0
279.1
131.2
3.5
55.1


Diff
5.0
18.9
9.0
1.9
2.0


Pr > T
0.344
0.002
0.575
0.011
0.000
















EYESPT
COMRST
EBTSTK




SCORE
SCORE
% NOT



—
ABS
ABS
ABS







Reps
1
4
3



Locs
1
4
3



PHB6V
6.0
5.8
92.3



PH185
6.0
6.0
67.0



Diff
0.0
0.3
25.3



Pr > T

0.391
0.352







*Pr > T values are valid only for comparisons with Locs >= 10













TABLE 4C





Average Inbred By Tester Performance Comparing PHB6V To PH5TG


Crossed To The Same Inbred Testers And Grown In The Same


Experiments.





















YIELD
MST
EGRWTH
ESTCNT
GDUSHD


—
BU/A 56#
PCT
SCORE
COUNT
GDU





Reps
93
93
14
13
63


Locs
93
93
14
13
63


PHB6V
169.7
25.6
6.0
61.9
119.7


PH5TG
161.8
25.5
6.1
62.4
118.4


Diff
7.9
0.1
0.1
0.5
1.3


Pr > T
0.000
0.633
0.807
0.687
0.000




















TSTWT



GDUSLK
PLTHT
EARHT
STAGRN
LB/BU


—
GDU
CM
CM
SCORE
ABS





Reps
29
38
38
53
54


Locs
29
38
38
53
54


PHB6V
120.1
285.8
131.0
5.3
52.1


PH5TG
118.4
273.5
125.0
5.1
51.8


Diff
1.7
12.3
5.9
0.2
0.3


Pr > T
0.009
0.000
0.006
0.106
0.354

















GIBERS
GIBROT
STLLPN
EBTSTK




SCORE
SCORE
% NOT
% NOT



—
ABS
ABS
ABS
ABS







Reps
2
4
8
20



Locs
2
4
8
20



PHB6V
7.0
6.3
95.0
83.0



PH5TG
7.5
8.0
97.4
87.4



Diff
0.5
1.8
2.3
4.3



Pr > T
0.795
0.326
0.503
0.187







*Pr > T values are valid only for comparisons with Locs >= 10













TABLE 5





General Combining Ability Report for Inbred PHB6V



















YIELD bu/a 56# ABS
Mean
157.8



YIELD bu/a 56# ABS
Locs
69



YIELD bu/a 56# ABS
Reps
402



YIELD bu/a 56# ABS
Years
3



MST pct ABS
Mean
23.2



MST pct ABS
Locs
70



MST pct ABS
Reps
403



MST pct ABS
Years
3



EGRWTH score ABS
Mean
5.3



EGRWTH score ABS
Locs
19



EGRWTH score ABS
Reps
74



EGRWTH score ABS
Years
3



ESTCNT count ABS
Mean
55.4



ESTCNT count ABS
Locs
34



ESTCNT count ABS
Reps
158



ESTCNT count ABS
Years
3



GDUSHD GDU ABS
Mean
118.6



GDUSHD GDU ABS
Locs
37



GDUSHD GDU ABS
Reps
302



GDUSHD GDU ABS
Years
3



GDUSLK GDU ABS
Mean
117.6



GDUSLK GDU ABS
Locs
16



GDUSLK GDU ABS
Reps
136



GDUSLK GDU ABS
Years
3



PLTHT cm ABS
Mean
291.2



PLTHT cm ABS
Locs
32



PLTHT cm ABS
Reps
197



PLTHT cm ABS
Years
3



EARHT cm ABS
Mean
131.5



EARHT cm ABS
Locs
32



EARHT cm ABS
Reps
197



EARHT cm ABS
Years
3



STAGRN score ABS
Mean
5.6



STAGRN score ABS
Locs
28



STAGRN score ABS
Reps
212



STAGRN score ABS
Years
3



TSTWT lb/bu ABS
Mean
53



TSTWT lb/bu ABS
Locs
63



TSTWT lb/bu ABS
Reps
349



TSTWT lb/bu ABS
Years
3



NLFBLT score ABS
Mean
4.5



NLFBLT score ABS
Locs
1



NLFBLT score ABS
Reps
2



NLFBLT score ABS
Years
1



ANTROT score ABS
Mean
5.5



ANTROT score ABS
Locs
3



ANTROT score ABS
Reps
6



ANTROT score ABS
Years
2



GIBERS score ABS
Mean
7



GIBERS score ABS
Locs
2



GIBERS score ABS
Reps
3



GIBERS score ABS
Years
2



EYESPT score ABS
Mean
6.1



EYESPT score ABS
Locs
2



EYESPT score ABS
Reps
7



EYESPT score ABS
Years
2



COMRST score ABS
Mean
5.6



COMRST score ABS
Locs
4



COMRST score ABS
Reps
19



COMRST score ABS
Years
1



ECB1LF score ABS
Mean
4.2



ECB1LF score ABS
Locs
2



ECB1LF score ABS
Reps
5



ECB1LF score ABS
Years
2



ECB2SC score ABS
Mean
4.5



ECB2SC score ABS
Locs
6



ECB2SC score ABS
Reps
15



ECB2SC score ABS
Years
2



BRTSTK % NOT ABS
Mean
94.9



BRTSTK % NOT ABS
Locs
3



BRTSTK % NOT ABS
Reps
32



BRTSTK % NOT ABS
Years
2



STLLPN % NOT ABS
Mean
82.9



STLLPN % NOT ABS
Locs
12



STLLPN % NOT ABS
Reps
67



STLLPN % NOT ABS
Years
3



ABTSTK % NOT ABS
Mean
50.9



ABTSTK % NOT ABS
Locs
2



ABTSTK % NOT ABS
Reps
9



ABTSTK % NOT ABS
Years
1



ERTLPN % NOT ABS
Mean
92.8



ERTLPN % NOT ABS
Locs
8



ERTLPN % NOT ABS
Reps
22



ERTLPN % NOT ABS
Years
2



LRTLPN % NOT ABS
Mean
86.5



LRTLPN % NOT ABS
Locs
11



LRTLPN % NOT ABS
Reps
43



LRTLPN % NOT ABS
Years
2

















TABLE 5A





Combining Ability Report for Inbred PHB6V


F1 data for Inbred Line PHB6V when crossed to various tester lines






















YIELD
YIELD
YIELD






bu/a 56#
bu/a 56#
bu/a 56#
MST pct
MST pct
MST pct



ABS
ABS
ABS
ABS
ABS
ABS


Tester
Mean
Locs
Reps
Mean
Locs
Reps
















48091F
148.3
11
11
21.6
11
11


61685C
164.9
12
12
23.5
12
12


61701F
163.2
12
12
22.2
12
12


214361J
150.5
8
10
22.4
8
10


51622B
159
7
8
22.5
7
8


53413B
141.9
14
17
26.8
15
18


57076F
148.4
8
9
23.3
8
9


57077H
145
7
9
23.8
7
9


57079G
153.6
11
11
23.3
11
11


57079H
162.5
41
73
22.1
41
73


57080A
152
8
9
21.9
8
9


61695K
153.4
7
8
23.7
7
8


61696B
166.7
8
9
23.8
8
9


62024C
148.4
8
9
24.3
8
9


76028E
160.9
22
28
22.2
22
28


76028F
164.6
8
9
24.3
8
9


76057H
159.4
8
10
22.6
8
10


76106J
141
9
11
23.7
9
11


76107K
160.8
7
9
21.8
7
9


77181K
172.8
12
12
24.8
12
12


214361K
168.6
8
10
24
8
10


214362A
162.8
7
9
23.8
7
9


214362B
159.3
7
9
23.5
7
9


214362C
161.7
7
9
21.8
7
9


214362D
141.8
7
9
25.8
7
9


214362E
170.7
7
9
21.5
7
9


214362F
155
8
10
23.5
8
10


61692H
160
12
12
24.2
12
12

















EGRWTH
EGRWTH
EGRWTH
ESTCNT
ESTCNT
ESTCNT



score
score
score
count
count
count



ABS
ABS
ABS
ABS
ABS
ABS


Tester
Mean
Locs
Reps
Mean
Locs
Reps
















48091F
5.3
4
4
50.3
4
4


61685C
4.8
5
5


61701F
5.2
5
5


214361J



63.5
2
2


51622B
4
1
1
51.2
12
13


53413B



54.6
10
11


57076F



63.5
2
2


57077H



62
3
3


57079G
6
4
4
47.4
5
5


57079H
4.9
17
30
56.4
24
49


57080A



63.5
2
2


61695K



62
2
2


61696B



64
2
2


62024C



61.5
2
2


76028E
6.5
6
6
52.7
13
14


76028F



64
2
2


76057H



66
2
2


76106J



64.7
3
3


76107K



65
2
2


77181K
5.6
5
5


214361K



64
2
2


214362A



64
2
2


214362B



59
2
2


214362C



58.5
2
2


214362D



63.3
3
3


214362E



62
2
2


214362F



64.5
2
2


61692H
6.2
5
5

















GDUSHD
GDUSHD
GDUSHD
GDUSLK
GDUSLK
GDUSLK



GDU ABS
GDU ABS
GDU ABS
GDU ABS
GDU ABS
GDU ABS


Tester
Mean
Locs
Reps
Mean
Locs
Reps
















48091F
117.7
7
7
115
2
2


61685C
123
8
8
123.5
2
2


61701F
123.6
8
8
120.5
2
2


214361J
118.7
7
9
115
4
5


51622B
110.8
4
5


53413B
108.6
6
8
119
1
1


57076F
119.1
7
8
117
4
5


57077H
110.7
6
7
116.3
3
3


57079G
121.6
8
8
115
2
2


57079H
117.6
29
48
120
15
24


57080A
118.1
7
8
115
4
5


61695K
117.9
6
7
116
4
5


61696B
118.8
7
8
116.6
4
5


62024C
120.8
7
8
117
4
5


76028E
120.8
14
20
117.9
4
7


76028F
121.6
7
8
117.8
4
5


76057H
121.7
7
9
117.6
4
5


76106J
113.3
6
7
118
3
3


76107K
118.5
6
8
113
3
4


77181K
124.1
8
8
124
2
2


214361K
121.1
6
8
120
4
5


214362A
119.9
7
9
117
4
5


214362B
120
7
9
119
4
5


214362C
120.4
7
9
117.4
4
5


214362D
111.9
6
7
120
3
3


214362E
120
7
9
115.6
4
5


214362F
119.4
7
9
114.8
4
5


61692H
123.8
8
8
121
2
2

















PLTHT
PLTHT
PLTHT
EARHT
EARHT
EARHT



cm
cm
cm
cm
cm
cm



ABS
ABS
ABS
ABS
ABS
ABS


Tester
Mean
Locs
Reps
Mean
Locs
Reps
















48091F
309.2
7
7
124.5
7
7


61685C
284.9
6
6
128.3
6
6


61701F
279
6
6
124.5
6
6


214361J
295.3
3
4
123.8
3
4


51622B
299.3
5
6
130.8
5
6


53413B
287
5
7
129.9
5
7


57076F
283.2
3
4
135.3
3
4


57077H
293.6
4
5
135.6
4
5


57079G
317.9
7
7
148.4
7
7


57079H
291.5
19
36
131.6
19
36


57080A
288.3
3
4
137.2
3
4


61695K
288.3
3
4
124.5
3
4


61696B
281.3
3
4
132.1
3
4


62024C
281.9
3
3
128.7
3
3


76028E
277.4
12
14
128.5
12
14


76028F
290.2
3
4
132.1
3
4


76057H
285.8
3
4
130.2
3
4


76106J
291.6
4
5
133.1
4
5


76107K
287
3
4
128.9
3
4


77181K
288.7
6
6
128.3
6
6


214361K
300.4
3
4
142.9
3
4


214362A
299.1
3
4
133.4
3
4


214362B
288.3
3
4
134
3
4


214362C
295.9
3
4
138.4
3
4


214362D
291.5
3
4
131.4
3
4


214362E
286.4
3
4
130.2
3
4


214362F
289.6
3
4
129.5
3
4


61692H
270.5
6
6
126.2
6
6

















STAGRN
STAGRN
STAGRN
TSTWT
TSTWT
TSTWT



score ABS
score ABS
score ABS
lb/bu ABS
lb/bu ABS
lb/bu ABS


Tester
Mean
Locs
Reps
Mean
Locs
Reps
















48091F
6
6
6
53.1
11
11


61685C
5.7
6
6
52.3
11
11


61701F
5.8
6
6
52.6
10
10


214361J
5
4
6
53.7
5
7


51622B
4
3
4
53.2
7
8


53413B
3.7
5
6
53.9
15
18


57076F
5.4
4
5
52.9
6
7


57077H
4
4
5
52.4
5
7


57079G
6.2
6
6
53.3
11
11


57079H
5.8
20
34
53.4
36
63


57080A
5.4
4
5
53.8
6
7


61695K
6
3
4
52.7
5
6


61696B
5.6
4
5
51.5
6
7


62024C
6.6
4
5
52.7
6
7


76028E
5
10
14
55.2
19
25


76028F
6
3
4
50.7
6
7


76057H
5.7
4
6
52.8
6
8


76106J
4.3
5
6
52
7
9


76107K
5.3
4
6
52.3
5
7


77181K
6.7
6
6
52.4
11
11


214361K
6.2
4
6
51.2
6
8


214362A
6
4
6
51.5
6
8


214362B
5.8
4
6
52.4
6
8


214362C
5.5
4
6
52.9
6
8


214362D
5.2
4
5
51.2
5
7


214362E
5.3
4
6
51.1
6
8


214362F
6.2
4
6
52.5
6
8


61692H
5.7
6
6
52.3
11
11

















BRTSTK
BRTSTK
BRTSTK
STLLPN
STLLPN
STLLPN



% NOT
% NOT
% NOT
% NOT
% NOT
% NOT



ABS
ABS
ABS
ABS
ABS
ABS


Tester
Mean
Locs
Reps
Mean
Locs
Reps
















48091F



88.5
2
2


61685C
70
1
1
47.2
2
2


61701F
95
1
1
64.4
2
2


214361J
100
1
1
96.7
1
1


51622B



90
1
1


53413B



100
2
2


57076F
98.3
1
1
96.7
1
1


57077H
100
1
1
86.7
1
1


57079G



98.4
3
3


57079H
94.1
3
6
84.1
8
18


57080A
94.8
1
1
100
1
1


61695K
96.6
1
1
100
1
1


61696B
96.6
1
1
96.7
1
1


62024C
100
1
1
100
1
1


76028E
90
3
3
72.7
7
8


76028F
100
1
1
100
1
1


76057H
96.6
1
1
90
1
1


76106J
98.4
1
1
90
1
1


76107K
100
1
1
96.7
1
1


77181K
66.7
1
1
61.5
2
2


214361K
100
1
1
96.7
1
1


214362A
96.9
1
1
93.3
1
1


214362B
100
1
1
96.7
1
1


214362C
98.4
1
1
93.1
1
1


214362D
100
1
1
70
1
1


214362E
100
1
1
70
1
1


214362F
96.6
1
1
90
1
1


61692H
98.3
1
1
61.5
2
2

















ERTLPN
ERTLPN
ERTLPN
LRTLPN
LRTLPN
LRTLPN



% NOT
% NOT
% NOT
% NOT
% NOT
% NOT



ABS
ABS
ABS
ABS
ABS
ABS


Tester
Mean
Locs
Reps
Mean
Locs
Reps
















48091F








61685C
100
1
1
91.8
5
5


61701F
100
1
1
96.6
5
5


214361J


51622B
100
1
1


53413B
100
1
1


57076F


57077H


57079G


57079H
89.9
6
12
78.4
8
17


57080A


61695K


61696B


62024C


76028E
87.7
3
3
89
5
5


76028F


76057H


76106J



100
1
1


76107K


77181K
100
1
1
86.8
5
5


214361K


214362A


214362B


214362C


214362D


214362E


214362F


61692H
100
1
1
93.4
5
5
















TABLE 6A





INBREDS IN HYBRID COMBINATION REPORT


Variety #1: HYBRID CONTAINING PHB6V - Variety #2: 3893

























YIELD
YIELD
MST
EGRWTH
ESTCNT
GDUSHD
GDUSLK
STKCNT
PLTHT



BU/A 56#
BU/A 56#
PCT
SCORE
COUNT
GDU
GDU
COUNT
CM


Stat
ABS
% MN
% MN
% MN
% MN
% MN
% MN
% MN
% MN



















Mean1
165.7
103.0
98.9
90.9
106.9
101.3
102.3
101.1
104.6


Mean2
154.5
96.0
97.3
95.8
104.9
98.2
98.1
100.1
95.3


Locs
64
64
64
19
29
39
22
118
29


Reps
111
111
111
33
52
64
38
251
49


Diff
11.2
7.0
−1.6
−4.8
1.9
3.1
4.2
1.0
9.3


Prob
0.000
0.000
0.208
0.382
0.336
0.000
0.000
0.115
0.000




















EARHT
STAGRN
ERTLSC
LRTLSC
STKLDS
STKLDG
ABTSTK
DRPEAR
TSTWT



CM
SCORE
SCORE
SCORE
SCORE
% NOT
% NOT
% NOT
LB/BU


Stat
% MN
% MN
ABS
ABS
ABS
% MN
% MN
% MN
ABS



















Mean1
102.1
108.5
8.7
7.7
8.4
100.5
98.0
99.3
53.3


Mean2
96.2
93.2
6.0
6.7
8.4
101.0
128.9
100.1
54.3


Locs
29
27
3
3
5
2
2
1
37


Reps
49
46
3
4
5
4
9
2
63


Diff
5.9
15.3
2.7
1.0
0.0
−0.5
−30.9
−0.8
−1.0


Prob
0.001
0.044
0.347
0.225
1.000
0.500
0.095

0.000



















NLFBLT
GOSWLT
ANTROT
FUSERS
GIBERS
EYESPT
COMRST
ECB1LF



SCORE
SCORE
SCORE
SCORE
SCORE
SCORE
SCORE
SCORE


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS


















Mean1
5.7
3.0
5.8
8.0
6.0
6.0
5.5
4.0


Mean2
5.5
5.5
6.0
8.1
6.0
5.8
5.0
3.9


Locs
3
1
2
2
1
2
4
3


Reps
6
2
4
5
2
3
4
8


Diff
0.2
−2.5
−0.3
−0.1
0.0
0.3
0.5
0.1


Prob
0.840

0.910
0.910

0.500
0.495
0.840






















HD






ECB2SC
HSKCVR
BRTSTK
SMT
ERTLPN
LRTLPN




SCORE
SCORE
% NOT
% NOT
% NOT
% NOT



Stat
ABS
ABS
ABS
ABS
ABS
ABS



















Mean1
4.8
5.6
94.1
95.8
89.9
81.9



Mean2
4.1
4.9
99.5
86.7
69.9
95.3



Locs
12
19
3
2
6
10



Reps
26
34
6
3
12
21



Diff
0.8
0.7
−5.3
9.1
20.0
−13.3



Prob
0.050
0.023
0.449
0.410
0.084
0.080

















TABLE 6B





INBREDS IN HYBRID COMBINATION REPORT


Variety #1: HYBRID CONTAINING PHB6V-Variety #2: 38R92
























YIELD
YIELD
MST
EGRWTH
ESTCNT
GDUSHD
GDUSLK
STKCNT



BU/A 56#
BU/A 56#
PCT
SCORE
COUNT
GDU
GDU
COUNT


Stat
ABS
% MN
% MN
% MN
% MN
% MN
% MN
% MN


















Mean1
165.1
101.3
98.2
84.6
109.4
100.2
101.1
101.3


Mean2
166.5
102.1
100.9
101.6
92.4
98.2
98.0
98.4


Locs
45
45
46
14
23
32
23
95


Reps
87
87
88
28
45
56
39
217


Diff
−1.4
−0.8
2.7
−17.0
17.1
2.1
3.1
2.9


Prob
0.399
0.457
0.025
0.021
0.000
0.000
0.000
0.013



















PLTHT
EARHT
STAGRN
LRTLSC
STKLDG
ABTSTK
DRPEAR
TSTWT



CM
CM
SCORE
SCORE
% NOT
% NOT
% NOT
LB/BU


Stat
% MN
% MN
% MN
ABS
% MN
% MN
% MN
ABS


















Mean1
104.6
102.5
104.3
6.0
100.5
98.0
99.3
53.5


Mean2
96.0
98.4
109.6
4.5
100.5
99.1
100.1
54.2


Locs
19
19
19
1
2
2
1
25


Reps
39
39
35
2
4
9
2
47


Diff
8.6
4.1
−5.2
1.5
0.0
−1.1
−0.8
−0.7


Prob
0.000
0.014
0.463

1.000
0.965

0.005



















NLFBLT
GOSWLT
ANTROT
FUSERS
GIBERS
EYESPT
ECB1LF
ECB2SC



SCORE
SCORE
SCORE
SCORE
SCORE
SCORE
SCORE
SCORE


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS


















Mean1
5.7
3.0
5.8
8.0
6.0
6.0
4.0
4.7


Mean2
5.5
6.5
5.0
8.3
5.0
6.0
5.8
4.4


Locs
3
1
2
2
1
1
3
11


Reps
6
2
4
5
2
2
8
23


Diff
0.2
−3.5
0.8
−0.3
1.0
0.0
−1.8
0.3


Prob
0.840

0.656
0.910


0.047
0.464




















HD






HSKCVR
BRTSTK
SMT
ERTLPN
LRTLPN




SCORE
% NOT
% NOT
% NOT
% NOT



Stat
ABS
ABS
ABS
ABS
ABS


















Mean1
5.3
94.1
95.8
89.9
81.9



Mean2
5.2
99.1
95.0
79.6
95.3



Locs
15
3
2
6
10



Reps
29
6
3
11
19



Diff
0.2
−5.0
0.8
10.3
−13.4



Prob
0.596
0.459
0.942
0.214
0.085

















TABLE 6C





INBREDS IN HYBRID COMBINATION REPORT


Variety #1: HYBRID CONTAINING PHB6V-Variety #2: 38R69
























YIELD
YIELD
MST
EGRWTH
ESTCNT
GDUSHD
GDUSLK
STKCNT



BU/A 56#
BU/A 56#
PCT
SCORE
COUNT
GDU
GDU
COUNT


Stat
ABS
% MN
% MN
% MN
% MN
% MN
% MN
% MN


















Mean1
164.1
100.9
98.2
84.6
109.4
100.1
101.0
101.3


Mean2
162.9
100.3
100.2
108.0
99.5
98.3
97.1
99.5


Locs
48
48
48
14
23
31
22
96


Reps
92
92
93
28
46
56
38
225


Diff
1.2
0.6
2.0
−23.4
9.9
1.8
3.8
1.8


Prob
0.465
0.554
0.067
0.001
0.001
0.000
0.000
0.011



















PLTHT
EARHT
STAGRN
LRTLSC
STKLDG
ABTSTK
DRPEAR
TSTWT



CM
CM
SCORE
SCORE
% NOT
% NOT
% NOT
LB/BU



% MN
% MN
% MN
ABS
% MN
% MN
% MN
ABS


















Mean1
104.4
102.2
104.3
6.0
100.5
98.0
99.3
53.4


Mean2
96.4
102.0
104.2
3.0
100.5
84.4
100.1
54.0


Locs
20
20
19
1
2
2
1
27


Reps
41
41
38
2
4
9
2
52


Diff
8.0
0.2
0.2
3.0
0.0
13.7
−0.8
−0.6


Prob
0.000
0.863
0.985

1.000
0.514

0.015



















NLFBLT
GOSWLT
ANTROT
FUSERS
GIBERS
EYESPT
ECB1LF
ECB2SC



SCORE
SCORE
SCORE
SCORE
SCORE
SCORE
SCORE
SCORE


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS


















Mean1
5.7
3.0
5.8
8.0
6.0
6.0
4.0
4.8


Mean2
4.5
5.5
6.8
7.3
6.5
5.5
4.8
3.8


Locs
3
1
2
2
1
1
3
12


Reps
6
2
4
5
2
2
8
25


Diff
1.2
−2.5
−1.0
0.7
−0.5
0.5
−0.8
1.0


Prob
0.336

0.795
0.295


0.038
0.028




















HD






HSKCVR
BRTSTK
SMT
ERTLPN
LRTLPN




SCORE
% NOT
% NOT
% NOT
% NOT



Stat
ABS
ABS
ABS
ABS
ABS


















Mean1
5.3
94.1
95.8
89.9
81.9



Mean2
4.1
99.2
93.0
67.9
89.4



Locs
15
3
3
6
10



Reps
30
6
5
12
21



Diff
1.2
−5.0
2.8
22.0
−7.5



Prob
0.003
0.452
0.714
0.061
0.223

















TABLE 6D





INBREDS IN HYBRID COMBINATION REPORT


Variety #1: HYBRID CONTAINING PHB6V-Variety #2: 39K40
























YIELD
YIELD
MST
EGRWTH
ESTCNT
GDUSHD
GDUSLK
STKCNT



BU/A 56#
BU/A 56#
PCT
SCORE
COUNT
GDU
GDU
COUNT


Stat
ABS
% MN
% MN
% MN
% MN
% MN
% MN
% MN


















Mean1
163.8
100.8
98.3
84.6
109.4
101.3
102.5
101.4


Mean2
153.6
94.6
95.7
85.8
108.0
99.9
100.0
101.4


Locs
47
47
47
14
23
29
20
93


Reps
91
91
92
28
46
54
36
225


Diff
10.2
6.2
−2.6
−1.2
1.4
1.4
2.6
0.0


Prob
0.000
0.000
0.033
0.830
0.699
0.002
0.000
0.951



















PLTHT
EARHT
STAGRN
LRTLSC
STKLDG
ABTSTK
DRPEAR
TSTWT



CM
CM
SCORE
SCORE
% NOT
% NOT
% NOT
LB/BU


Stat
% MN
% MN
% MN
ABS
% MN
% MN
% MN
ABS


















Mean1
104.4
102.2
104.3
6.0
100.5
98.0
99.3
53.4


Mean2
99.7
95.0
93.3
1.5
101.0
104.7
100.1
54.2


Locs
20
20
19
1
2
2
1
27


Reps
41
41
37
2
4
9
2
52


Diff
4.7
7.2
11.1
4.5
−0.5
−6.7
−0.8
−0.8


Prob
0.000
0.004
0.054

0.794
0.501

0.002



















NLFBLT
GOSWLT
ANTROT
FUSERS
GIBERS
EYESPT
ECB1LF
ECB2SC



SCORE
SCORE
SCORE
SCORE
SCORE
SCORE
SCORE
SCORE


Stat
ABS
ABS
ABS
ABS
ABS
ABS
ABS
ABS


















Mean1
5.7
3.0
5.8
8.0
6.0
6.0
4.0
4.8


Mean2
5.5
4.5
7.3
8.0
6.0
5.0
5.0
4.1


Locs
3
1
2
2
1
1
3
12


Reps
6
2
4
5
2
2
8
26


Diff
0.2
−1.5
−1.5
0.0
0.0
1.0
−1.0
0.7


Prob
0.921

0.500
1.000


1.000
0.097




















HD






HSKCVR
BRTSTK
SMT
ERTLPN
LRTLPN




SCORE
% NOT
% NOT
% NOT
% NOT



Stat
ABS
ABS
ABS
ABS
ABS


















Mean1
5.3
94.1
95.8
89.9
81.9



Mean2
4.7
100.0
92.5
56.4
79.3



Locs
15
3
3
6
10



Reps
30
6
5
12
21



Diff
0.6
−5.9
3.2
33.5
2.6



Prob
0.036
0.393
0.423
0.064
0.548










All publications, patents and patent applications mentioned in the specification are indicative of the level of those skilled in the art to which this invention pertains. All such publications, patents and patent applications are incorporated by reference herein for the purpose cited to the same extent as if each was specifically and individually indicated to be incorporated by reference herein.
The foregoing invention has been described in detail by way of illustration and example for purposes of clarity and understanding. However, it is obvious that certain changes and modifications such as backcross conversions and mutations, somoclonal variants, variant individuals selected from large populations of the plants of the instant inbred and the like may be practiced within the scope of the invention, as limited only by the scope of the appended claims.

## Claims

1. A seed comprising at least one set of the chromosomes of maize inbred line PHB6V, representative seed of said line having been deposited under ATCC Accession No: PTA-4874.

2. A maize plant produced by growing the seed of claim 1.

3. A maize plant part of the maize plant of claim 2.

4. An F1 hybrid maize seed produced by crossing a plant of maize inbred line designated PHB6V, representative seed of said line having been deposited under ATCC Accession No: PTA-4874, with a different maize plant and harvesting the resultant F1 hybrid maize seed, wherein said F1 hybrid maize seed comprises two sets of chromosomes and one set of the chromosomes is the same as maize inbred line PHB6V.

5. A maize plant produced by growing the F1 hybrid maize seed of claim 4.

6. A maize plant part of the maize plant of claim 5.

7. An F1 hybrid maize seed comprising an inbred maize plant cell of inbred maize line PHB6V, representative seed of said line having been deposited under ATCC Accession No: PTA-4874.

8. A maize plant produced by growing the F1 hybrid maize seed of claim 7.

9. The F1 hybrid maize seed of claim 7 wherein the inbred maize plant cell comprises two sets of chromosomes of maize inbred line PHB6V.

10. A maize plant produced by growing the F1 hybrid maize seed of claim 9.

11. A process of introducing a desired trait into maize inbred line PHB6V comprising:
(a) crossing PHB6V plants grown from PHB6V seed, representative seed of which has been deposited under ATCC Accession No: PTA-4874, with plants of another maize line that comprise a desired trait to produce F1 progeny plants, wherein the desired trait is selected from the group consisting of waxy starch, male sterility, herbicide resistance, insect resistance, bacterial disease resistance, fungal disease resistance, and viral disease resistance;
(b) selecting F1 progeny plants that have the desired trait to produce selected F1 progeny plants;
(c) crossing the selected progeny plants with the PHB6V plants to produce backcross progeny plants;
(d) selecting for backcross progeny plants that have the desired trait and the alleles of inbred line PHB6V at the SSR loci listed in Table 2 to produce selected backcross progeny plants; and
(e) repeating steps (c) and (d) to produce backcross progeny plants that comprise the desired trait and comprise at least 95% of the alleles of inbred line PHB6V at the SSR loci listed in Table 2.

12. A plant produced by the process of claim 11, wherein the plant comprises at least 95% of the alleles of inbred line PHB6V at the SSR loci listed in Table 2.

13. A maize plant having all the physiological and morphological characteristics of inbred line PHB6V, wherein a sample of the seed of inbred line PHB6V was deposited under ATCC Accession No: PTA-4874.

14. A process of producing maize seed, comprising crossing a first parent maize plant with a second parent maize plant, wherein one or both of the first or the second parent maize plants is the plant of claim 13, wherein seed is allowed to form.

15. The maize seed produced by the process of claim 14.

16. The maize seed of claim 15, wherein the maize seed is hybrid seed.

17. A hybrid maize plant, or its parts, produced by growing said hybrid seed of claim 16.

18. The maize plant of claim 13, further comprising an SSR profile in accordance with the profile shown in Table 2.

19. A cell of the maize plant of claim 13.

20. The cell of claim 19, wherein said cell is further defined as having an SSR profile in accordance with the profile shown in Table 2.

21. A seed comprising the cell of claim 19.

22. The maize plant of claim 13, further comprising a genome having a single gene conversion.

23. The maize plant of claim 22, wherein the single gene was stably inserted into the maize genome by transformation.

24. The maize plant of claim 22, wherein the gene is selected from the group consisting of a dominant allele and a recessive allele.

25. The maize plant of claim 22, wherein the gene confers a trait selected from the group consisting of herbicide tolerance; insect resistance; resistance to bacterial, fungal, nematode or viral disease; waxy starch; male sterility and restoration of male fertility.

26. The maize plant of claim 13, wherein said plant further comprises a gene conferring male sterility.

27. The maize plant of claim 13, wherein said plant further comprises a transgene conferring a trait selected from the group consisting of male sterility, herbicide resistance, insect resistance and disease resistance.

28. A method of producing a maize plant comprising the steps of:
(a) growing a progeny plant produced by crossing the plant of claim 13 with a second maize plant;
(b) crossing the progeny plant with itself or a different plant to produce a seed of a progeny plant of a subsequent generation;
(c) growing a progeny plant of a subsequent generation from said seed and crossing the progeny plant of a subsequent generation with itself or a different plant; and
(d) repeating steps (b) and (c) for an additional 0-5 generations to produce a maize plant.

29. The method of claim 28 wherein the produced maize plant is an inbred maize plant.

30. The method of claim 29, further comprising the step of crossing the inbred maize plant with a second, distinct inbred maize plant to produce an F1 hybrid maize plant.

31. A method for developing a second maize plant in a maize plant breeding program comprising applying plant breeding techniques to a first maize plant, or its parts, wherein said first maize plant is the maize plant of claim 13 and wherein application of said techniques results in development of said second maize plant.

32. The method for developing a maize plant in a maize plant breeding program of claim 31 wherein plant breeding techniques are selected from the group consisting of recurrent selection, backcrossing, pedigree breeding, restriction fragment length polymorphism enhanced selection, genetic marker enhanced selection, and transformation.

33. A method of plant breeding comprising the steps of:
(a) obtaining a molecular marker profile of maize inbred line PHB6V, representative seed of said line having been deposited under ATCC Accession No. PTA-4874;
(b) obtaining an F1 hybrid seed for which the maize plant of claim 13 is a parent;
(c) crossing a plant grown from the F1 hybrid seed with a different maize plant; and
(d) selecting progeny that retain the molecular marker profile of PHB6V.