US009693531B1
`
`a2) United States Patent
`US 9,693,531 Bl
`(10) Patent No.:
`
` Odland (45) Date of Patent: Jul. 4, 2017
`
`
`(54) MAIZE INBRED PH25S9
`
`(56)
`
`References Cited
`
`.
`
`(71) Applicant: PIONEER HI-BRED
`INTERNATIONAL,INC., Johnston,
`IA (US)
`Inventor: MN(US) Odland, CannonFalls,
`(72)
`(73) Assignee: PIONEER HI-BRED
`INTERNATIONAL,INC., Johnston,
`1A (US)
`
`(*) Notice:
`,
`
`Subject to any disclaimer, the term ofthis
`:
`.
`rec 7sab)byOda under 35
`(21) Appl. No.: 15/193,403
`(22)
`Filed:
`Jun. 27, 2016
`
`(51)
`
`Int. CL
`AOLH 5/00
`CI2N 15/82
`AOLH 5/10
`AOIH 1/02
`(52) U.S. Cl.
`CPC vee AOLH 5/10 (2013.01); AOIH 1/02.
`(2013.01); C12N 15/8241 (2013.01); C12N
`15/8243 (2013.01); C12N 15/8245 (2013.01):
`C12N 15/8247 (2013.01); C12N 15/8251
`(2013.01); C12N 15/8271 (2013.01); C12N
`15/8274 (2013.01); C12N 15/8279 (2013.01);
`C12N 15/8286 (2013.01); C12N 15/8289
`(2013.01)
`
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`
`(58) Field of Classification Search
`None
`
`U.S. PATENT DOCUMENTS
`
`7,138,571 B2* 11/2006 Barker wesc AOLH 5/10
`435/412
`7,820,892 Bl
`10/2010 Noble
`8044.282 Bl
`10/2011 Smalley
`OTHER PUBLICATIONS
`.
`.
`.
`.
`US Plant Variety Protection Certificate No. PH12K5 for Maize
`Variety PH12K5; issued Jul. 13, 2013.
`US Plant Variety Protection Certificate No. 200800243 for Maize
`Variety PHFOD;issued Jan. 18, 2013.
`Breeding History for Maize Inbred PH25S9.
`ee
`.
`cited by examiner
`Primary Examiner — Vinod Kumar
`(74) Attorney, Agent, or Firm — Pioneer Hi Bred Int’l,
`Inc.
`
`(57)
`
`ABSTRACT
`
`A novel maize variety designated PH25S9 and seed, plants
`and plant parts thereof are provided. Methods for producing
`a maize plant comprise crossing maize variety PH25S9 with
`another maizeplant are provided. Methods for producing a
`maize plant containing in its genetic material one or more
`traits introgressed into PH25S89 through backcross conver-
`sion and/or transformation, and to the maize seed, plant and
`Plant part produced thereby are provided. Hybrid maize
`seed, plants or plant parts are produced by crossing the
`variety PH25S9 or a locus conversion of PH2589 with
`another maize variety.
`
`See application file for complete search history.
`
`20 Claims, No Drawings
`
`Inari Exhibit 1107
`Inari Exhibit 1107
`Inari v. Pioneer
`Inari v. Pioneer
`
`

`

`1
`MAIZE INBRED PH2589
`
`US 9,693,531 Bl
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`SUMMARY
`
`Processes are provided for making a maize plant contain-
`ing in its genetic material one or moretraits introgressed into
`PH25S9 through one or more of backcross conversion,
`genetic manipulation and transformation, and to the maize
`seed, plant and plant parts produced thereby. Hybrid maize
`seed, plants or plant parts produced by crossing the variety
`PH25S9 or a locus conversion of PH25S9 with another
`
`2
`certain embodiments, the plant seed growth mediais a soil
`or synthetic cultivation medium. In specific embodiments,
`BACKGROUND
`the growth medium may be comprised in a container or may,
`for example, be soil inafield.
`A plant of maize variety PH25S9 comprising an added
`There are numerous steps in the development of any
`heritable trait is provided. The heritable trait may comprise
`novel, desirable maize variety. Plant breeding begins with
`a genetic locus that is a dominantor recessive allele. In one
`the analysis and definition of problems and weaknesses of
`embodiment, a plant of maize variety PH25S9 comprising a
`the current germplasm, the establishment of program goals,
`single locus conversion in particular is provided. An added
`and the definition of specific breeding objectives. The next
`genetic locus which confers one or more traits such as, for
`step is selection of germplasm that possess the traits to meet
`example, male sterility, herbicide tolerance,
`insect resis-
`the program goals. The breeder’s goal is to combine in a
`tance, disease resistance, waxy starch, modified fatty acid
`single variety or hybrid, various desirable traits. For field
`metabolism, modified phytic acid metabolism, modified
`crops, these traits may include resistance to diseases and
`carbohydrate metabolism and modified protein metabolism
`insects, resistance to heat and drought, reducing the time to
`is provided. The trait may be, for example, conferred by a
`crop maturity, greateryield, altered fatty acid profile, abiotic
`naturally occurring maize gene introduced into the genome
`stress tolerance, improvements in compositionaltraits, and
`of the variety by backcrossing, a natural or induced muta-
`better agronomic characteristics and quality.
`tion, or a transgene introduced through genetic transforma-
`These product development processes, which lead to the
`tion techniques. When introduced through transformation, a
`final step of marketing and distribution, can take from six to
`genetic locus may comprise one or more transgenes inte-
`twelve years from the time the first cross is made until the
`grated at a single chromosomallocation.
`finished seed is delivered to the farmer for planting. There-
`An inbred maize plant of the variety designated PH25S9
`fore, development of new varieties and hybrids is a time-
`is provided, wherein a cytoplasmically-inherited trait has
`consuming process that requires precise planning, efficient
`been introduced into said inbred plant. Such cytoplasmi-
`use of resources, and a minimum of changesin direction. A
`cally-inherited traits are passed to progeny through the
`continuing goal of maize breeders is to develop stable, high
`female parent in a particular cross. An exemplary cytoplas-
`yielding maize varieties and hybrids that are agronomically
`mically-inherited trait is the male sterility trait. Cytoplas-
`sound with maximal yield over one or more different con-
`ditions and environments.
`mic-male sterility (CMS) is a pollen abortion phenomenon
`determined by the interaction between the genes in the
`cytoplasm and the nucleus. Alteration in the mitochondrial
`genomeand the lack of restorer genes in the nucleus will
`lead to pollen abortion. With either a normal cytoplasm or
`the presence of restorer gene(s) in the nucleus, the plant will
`produce pollen normally. A CMSplant can be pollinated by
`a maintainer version of the same variety, which has a normal
`cytoplasm but lacks the restorer gene(s) in the nucleus, and
`continue to be malesterile in the next generation. The male
`fertility of a CMSplantcan be restored by a restorer version
`of the same variety, which must have the restorer gene(s) in
`the nucleus. With the restorer gene(s) in the nucleus, the
`offspring of the male-sterile plant can produce normalpollen
`grains and propagate. A cytoplasmically inherited trait may
`be a naturally occurring maize trait or a trait introduced
`through genetic transformation techniques.
`A tissue culture of regenerable cells of a plant of variety
`PH25S9 is provided. The tissue culture can be capable of
`regenerating plants capable of expressing all of the physi-
`ological and morphological or phenotypic characteristics of
`maize variety are also provided.
`the variety, and of regenerating plants having substantially
`The inbred maize plant may further comprise a cytoplas-
`the same genotypeas other plants of the variety. Examples
`mic or nuclear factor capable of conferring malesterility or
`of some of the physiological and morphological character-
`otherwise preventing self-pollination, such as by self-incom-
`istics that may be assessed include characteristics related to
`patibility. Parts of the maize plant of the present invention
`yield, maturity, and kernel quality. The regenerable cells in
`are also provided, for example, pollen obtained from an
`such tissue cultures can be derived,
`for example, from
`inbred plant and an ovule of the inbred plant.
`embryos, meristematic cells, immature tassels, microspores,
`Seed of the inbred maize variety PH25S9 is provided. The
`pollen, leaves, anthers,roots, roottips, silk, flowers, kernels,
`inbred maize seed may be an essentially homogeneous
`ears, cobs, husks, or stalks, or from callus or protoplasts
`population of inbred maize seed of the variety designated
`derived from those tissues. Maize plants regenerated from
`PH25S9. Essentially homogeneous populations of inbred
`the tissue cultures of the invention, the plants having all the
`seed are generally free from substantial numbers of other
`physiological and morphological characteristics of variety
`seed. Therefore, inbred seed generally forms at least about
`PH25S9 are also provided.
`97% of the total seed. The population of inbred maize seed
`Processes are provided for producing maize seeds or
`of the invention may be particularly defined as being essen-
`
`tially free from hybrid seed. The inbred seed population may plants, which processes generally comprise crossingafirst
`be separately grownto provide an essentially homogeneous
`parent maize plant as a male or female parent with a second
`population of inbred maize plants designated PH25S9.
`parent maize plant, wherein at least one ofthefirst or second
`Compositions are provided comprising a seed of maize
`parent maize plants is a plant of the variety designated
`variety PH25S9 comprised in plant seed growth media. In
`PH25S9. These processes may be further exemplified as
`
`Provided is a novel maize, Zea mays L., variety, desig-
`nated PH25S9 and processes for making PH25S9. Seed of
`maize variety PH25S9, plants of maize variety PH25S9,
`plant parts and cells of maize variety PH25S9, and processes
`for making a maize plant that comprise crossing maize
`variety PH25S9 with another maize plant are provided. Also
`provided are maize plants having all the physiological and
`morphological characteristics of the inbred maize variety
`PH25S9.
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`US 9,693,531 Bl
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`3
`processes for preparing hybrid maize seed or plants, wherein
`a first inbred maize plant is crossed with a second maize
`plant of a different, distinct variety to provide a hybrid that
`has, as one of its parents, the inbred maize plant variety
`PH25S9.
`In these processes, crossing will result
`in the
`production of seed. The seed production occurs regardless of
`whether the seed is collected or not.
`
`In some embodiments, the first step in “crossing” com-
`prises planting, such as in pollinating proximity, seeds of a
`first and second parent maize plant, and such as, seeds of a
`first inbred maize plant and a second, distinct inbred maize
`plant. Where the plants are not in pollinating proximity,
`pollination can nevertheless be accomplishedby transferring
`a pollen or tassel bag from oneplant to the other as described
`below.
`Asecond step comprises cultivating or growing the seeds
`of said first and second parent maize plants into plants that
`bear flowers (maize bears both male flowers (tassels) and
`female flowers (silks) in separate anatomical structures on
`the sameplant).
`A third step comprises preventing self-pollination of the
`plants,
`i.e., preventing the silks of a plant from being
`fertilized by any plant of the samevariety, including the
`same plant. This can be done by emasculating the male
`flowers of the first or second parent maize plant,
`(i.e.,
`treating or manipulating the flowers so as to prevent pollen
`production, in order to produce an emasculated parent maize
`plant). Self-incompatibility systems may also be used in
`some hybrid crops for the same purpose. Self-incompatible
`plants still shed viable pollen and can pollinate plants of
`othervarieties but are incapable of pollinating themselves or
`other plants of the same variety.
`A fourth step may comprise allowing cross-pollination to
`occur between the first and second parent maize plants.
`Whenthe plants are not in pollinating proximity, this is done
`by placing a bag, usually paper or glassine, over the tassels
`of the first plant and another bag over the silks of the
`incipient ear on the secondplant. The bags are left in place
`for at least 24 hours. Since pollen is viable for less than 24
`hours, this assures that the silks are not pollinated from other
`pollen sources, that any stray pollen on thetassels of thefirst
`plant is dead, and that the only pollen transferred comes
`from thefirst plant. The pollen bag overthe tassel ofthefirst
`plant is then shaken vigorously to enhancerelease of pollen
`from the tassels, and the shoot bag is removed from thesilks
`of the incipient ear on the secondplant. Finally, the pollen
`bag is removed from thetassel of the first plant and is placed
`overthe silks of the incipient ear of the second plant, shaken
`again and left in place. Yet another step comprises harvest-
`ing the seeds from at least one of the parent maize plants.
`The harvested seed can be grown to produce a maize plant
`or hybrid maize plant.
`Also provided are maize seed and plants produced by a
`process that comprises crossing a first parent maize plant
`with a second parent maize plant, whereinat least one of the
`first or second parent maize plants is a plant of the variety
`designated PH25S9. In one embodiment of the invention,
`maize seed and plants produced by the process are first
`generation (F1) hybrid maize seed and plants produced by
`crossing an inbred in accordance with the invention with
`another, distinct inbred. Seed of an F1 hybrid maize plantis
`contemplated and an F1 hybrid maize plant and seed thereof
`are provided.
`The genetic complement of the maize plant variety des-
`ignated PH25S9 is provided. The phrase “genetic comple-
`ment” is used to refer to the aggregate of nucleotide
`sequences, the expression of which sequences defines the
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`phenotypeof, in the present case, a maize plant, or a cell or
`tissue of that plant. A genetic complement thus represents
`the genetic make-up of an inbredcell, tissue or plant, and a
`hybrid genetic complement represents the genetic make-up
`of a hybrid cell, tissue or plant. Maize plant cells that have
`a genetic complementin accordance with the inbred maize
`plant cells disclosed herein, and plants, seeds and diploid
`plants containing such cells are provided.
`Plant genetic complements may be assessed by genetic
`marker profiles, and by the expression of phenotypic traits
`that are characteristic of the expression of the genetic
`complement, e.g., isozyme typing profiles. It is understood
`that variety PH25S9 could be identified by any of the many
`well-known techniques used for genetic profiling disclosed
`herein.
`
`In still yet another aspect, the present invention provides
`hybrid genetic complements, as represented by maize plant
`cells, tissues, plants, and seeds, formed by the combination
`of a haploid genetic complementof an inbred maize plant of
`the invention with a haploid genetic complementof a second
`maizeplant, such as, another, distinct inbred maize plant. In
`another aspect, the present invention provides a maize plant
`regenerated from a tissue culture that comprises a hybrid
`genetic complementof this invention.
`Methods of producing an inbred maize plant derived from
`the maize variety PH25S9 are provided, the method com-
`prising the steps of: (a) preparing a progeny plant derived
`from maize variety PH25S9, wherein said preparing com-
`prises crossing a plant of the maize variety PH25S9 with a
`second maize plant;
`(b) crossing the progeny plant with
`itself or a second plant to produce a seed of a progeny plant
`of a subsequent generation; (c) repeating steps (a) and (b)
`with sufficient inbreeding until a seed of an inbred maize
`plant derived from the variety PH25S9 is produced. In the
`method,
`it may be desirable to select particular plants
`resulting from step (c) for continued crossing according to
`steps (b) and (c). By selecting plants having one or more
`desirable traits, an inbred maize plant derived from the
`maize variety PH25S9 is obtained which possesses some of
`the desirable traits of maize variety PH25S9 as well as
`potentially other selected traits.
`
`DETAILED DESCRIPTION
`
`A new and distinctive maize inbred variety designated
`PH25S9, which has been the result of years of careful
`breeding and selection in a comprehensive maize breeding
`program is provided.
`
`Definitions
`
`Maize (Zea mays) can be referred to as maize or corn.
`Certain definitions used in the specification are provided
`below. Also in the examples that follow, a numberof terms
`are used herein. In order to provide a clear and consistent
`understanding ofthe 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 percent-
`age of plants that did not exhibit a trait. For example,
`STKLDG % NOTis 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.
`Below are the descriptors used in the data tables included
`herein.
`
`

`

`US 9,693,531 Bl
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`5
`ABIOTIC STRESS TOLERANCE: resistance to non-
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`BRLPNE=ARTIFICIAL ROOT LODGING EARLY
`
`biological sources of stress conferred by traits such as
`nitrogen utilization efficiency, altered nitrogen responsive-
`ness, drought resistance, cold, and salt resistance
`ABTSTK=ARTIFICIAL BRITTLE STALK: A count of
`
`the numberof “snapped”plants per plot following machine
`snapping. A snappedplanthasits 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. In a diploid cell or organism, the two
`alleles of a given sequence typically occupy corresponding
`loci on a pair of homologous chromosomes.
`ALTER: The utilization of up-regulation, down-regula-
`tion, or gene silencing.
`ANTHESIS: The time of a flower’s opening.
`ANTIOXIDANT: A chemical compound or substance
`that inhibits oxidation, including but not limited to tocoph-
`erol or tocotrienols.
`
`ANT ROT=ANTHRACNOSE STALK ROT (Colletotri-
`chum graminicola): A 1
`to 9 visual rating indicating the
`resistance to Anthracnose Stalk Rot. A higherscore indicates
`a higher resistance. Data are collected only when sufficient
`selection pressure exists in the experiment measured.
`BACKCROSSING:Process in which a breeder crosses a
`
`hybrid progenyvariety back to one of the parental genotypes
`one or more times.
`
`BACKCROSS PROGENY:Progeny plants produced by
`crossing one maize line (recurrent parent) with plants of
`another maize line (donor) that comprise a desired trait or
`locus, selecting progeny plants that comprise the desired
`trait or locus, and crossing them with the recurrent parent
`one or moretimes to produce backcross progeny plants that
`comprise said trait or locus.
`BARPLT=BARREN PLANTS: Thepercent of plants per
`plot that were not barren (lack ears).
`BLUP=BEST LINEAR UNBIASED PREDICTION.The
`
`BLUPvalues are determined from a mixed model analysis
`of hybrid performance observationsat various locations and
`replications. BLUP values for inbred maize plants, breeding
`values, are estimated from the same analysis using pedigree
`information.
`BORBMN=ARTIFICIAL BRITTLE STALK MEAN:
`
`The mean percentofplants not “snapped”in a plot following
`artificial selection pressure. A snapped plant has its stalk
`completely snapped at a node between the base of the plant
`and the node abovethe ear. Expressed as percent of plants
`that did not snap. A higher numberindicates better tolerance
`to brittle snapping.
`BRENGMN=BRITTLE STALK ENERGY MEAN: The
`mean amount of energy per unit area neededto artificially
`brittle snap a corn stalk. A high number indicates better
`tolerance to brittle snapping.
`BREEDING:The genetic manipulation of living organ-
`isms.
`
`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.
`BREEDING VALUE: A relative value determined by
`evaluating the progeny of the parent. For corn the progeny
`is often the Fl generation and the parent is often an inbred
`variety.
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`SEASON: Thepercent of plants not root lodged in a plot
`followingartificial selection pressure applied prior to flow-
`ering. A plant is considered root lodgedif it leans from the
`vertical axis at an approximately 30 degree angle or greater.
`Expressedas percentof plants that did not root lodge. A high
`numberindicates higher tolerance to root lodging.
`BRLPNL=ARTIFICIAL ROOT LODGING LATE SEA-
`
`lodged in a plot
`SON: The percent of plants not root
`following artificial selection pressure during grain fill. A
`plant is considered root lodged if it leans from the vertical
`axis at an approximately 30 degree angle or greater.
`Expressedas percentof plants that did not root lodge. A high
`numberindicates higher tolerance to root lodging.
`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 snapor break
`near the time of flowering under severe winds. Data are
`presented as percentage of plants that did not snap. Data are
`collected only when sufficient selection pressure exists in the
`experiment measured.
`BRTPCN=BRITTLE STALKS: This is an estimate of the
`stalk breakage near the time of pollination, and is an
`indication of whether a hybrid or inbred would snapor break
`near the time of flowering under severe winds. Data are
`presented as percentage of plants that did not snap. Data are
`collected only when sufficient selection pressure exists in the
`experiment measured.
`CARBOHYDRATE: Organic compounds comprising
`carbon, oxygen and hydrogen,
`including sugars, starches
`and cellulose.
`
`CELL: Cell as used herein includes a plant cell, whether
`isolated, in tissue culture or incorporated in a plant or plant
`part.
`CLDTST=COLD TEST: The percent of plants that ger-
`minate under cold test conditions.
`
`CLN=CORN LETHAL NECROSIS: Synergistic interac-
`tion of maize chlorotic mottle virus (MCMV)in combina-
`tion 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 Necro-
`sis. A higher score indicates a higher resistance. Data are
`collected only when sufficient selection pressure exists in the
`experiment measured.
`CMSMT=COMMON SMUT: This is the percentage of
`plants not infected with Common Smut. Data are collected
`only when sufficient selection pressure exists in the experi-
`ment measured.
`COMRST=COMMONRUST(Puccinia sorghi): A 1 to 9
`visual rating indicating the resistance to Common Rust. A
`higher score indicates a higher resistance. Data are collected
`only when sufficient selection pressure exists in the experi-
`ment measured.
`CROSS POLLINATION:Fertilization by the union of
`two gametes from different plants.
`CROSSING: The combination of genetic material by
`traditional methods such as a breeding cross or backcross,
`but also including protoplast fusion and other molecular
`biology methods of combining genetic material from two
`sources.
`
`D and D1-Dn:represents the generation of doubled hap-
`loid.
`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 thatdriesrelatively fast while a low
`score indicates a hybrid that dries slowly.
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`US 9,693,531 Bl
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`DIGENG=DIGESTIBLE ENERGY:Near-infrared trans-
`mission spectroscopy, NIT, prediction of digestible energy.
`DIPERS=D/PLODIA 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. Data are collected only
`when sufficient selection pressure exists in the experiment
`measured.
`DIPLOID PLANTPART:Refers to a plant part or cell that
`has a same diploid genotype.
`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.
`Data are collected only when sufficient selection pressure
`exists in the experiment measured.
`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. Data are
`collected only whensufficient selection pressure exists in the
`experiment measured.
`D/T=DROUGHT TOLERANCE:This represents a 1 to 9
`rating for drought tolerance, and is based on data obtained
`understress conditions. A high score indicates good drought
`tolerance and a low score indicates poor droughttolerance.
`Data are collected only when sufficient selection pressure
`exists in the experiment measured.
`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 notbe predictive for a specific ear mold. Data are
`collected only whensufficient selection pressure exists in the
`experiment measured.
`EARHT=EDEARHT=EAR HEIGHT:Theear height is a
`measure fromthe groundto the highest placed developed ear
`node attachment and is measured in inches (EARHT) or
`centimeters (EDEARHT).
`EARSZ=EARSIZE: A 1 to 9 visualrating of ear size. The
`higher the rating the larger the ear size.
`EBTSTK=EARLY BRITTLE STALK: A count of the
`
`numberof “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. Data are collected only when sufficient
`selection pressure exists in the experiment measured.
`ECB1 LF=EUROPEAN CORN BORER FIRST GEN-
`
`FRATION LEAF FERDING (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. Data are collected only
`when sufficient selection pressure exists in the experiment
`measured.
`
`ECB2IT=EUROPEAN CORN BORER SECOND GEN-
`ERATION INCHES OF TUNNELING(Ostrinia nubilalis):
`Average inches of tunneling per plant in the stalk. Data are
`collected only whensufficient selection pressure exists in the
`experiment measured.
`ECB2SC=EUROPEAN CORN BORER SECOND GEN-
`
`ERATION (Ostrinia nubilalis): A 1 to 9 visual rating indi-
`cating post flowering degree of stalk breakage and other
`evidence of feeding by second generation European Corn
`Borer. A higher score indicates a higher resistance. Data are
`collected only whensufficient selection pressure exists in the
`experiment measured.
`ECBDPE=EUROPEAN CORN BORER DROPPED
`
`EARS(Ostrinia nubilalis): Dropped ears due to European
`Corn Borer. Percentage of plants that did not drop ears under
`
`8
`second generation European Corn Borer infestation. Data
`are collected only whensufficient selection pressure exists in
`the experiment measured.
`ECBLSI-EUROPEAN CORN BORER LATE SEASON
`
`INTACT(Ostrinia nubilalis): A 1 to 9 visual rating indicat-
`ing late season intactness of the corn plant given damage
`(stalk breakage above and below the top ear) caused pri-
`marily by 2”“ and/or 3” generation ECB larval feeding
`before harvest. A higher score indicates more intact plants.
`Data are collected only when sufficient selection pressure
`exists in the experiment measured.
`EDANTCOLs=ANTHER COLOR: Rated on a 1
`
`to 7
`
`scale where 1 is green, 2 is yellow, 3 is pink, 5 is red, and
`7 is purple.
`EDantants=ANTHER
`
`=ANTHOCYANIN
`
`COLOR
`
`INTENSITY: A measure ofanther anthocyanin color inten-
`sity rated on a 1 to 9 scale where 1 is absent or very weak,
`3 is weak, 5 is medium, 7 is strong, and 9 is very strong.
`Observed in the middle third of the main branch on fresh
`anthers.
`EDbarants=GLUME ANTHOCYANIN COLORATION
`
`AT BASE (WHOLE PLANT, EAR INSERTION LEVEL):
`A measure of the color intensity at the base of the glume,
`rated on a 1 to 9 scale where 1 is absent or very weak, 3 is
`weak, 5 is medium, 7 is strong, and 9 is very strong.
`Observed in the middle third of the main branch ofthe tassel.
`EDBARCOLs=BAR GLUME COLOR INTENSITY: A
`
`measure of the bar glumecolorintensity. Bar glumeis a dark
`purple band that may occur on the bottom of a glume. Bar
`glumecolorintensity is measured on a scale of 1 to 7 where
`1 is absent, 2 is weak, 3 is medium,5 is strong, and 7 is very
`strong.
`EDBRROANTs=BRACE ROOTS ANTHOCYANIN
`
`COLORATION: A measure of the color intensity of the
`brace roots rated on a 1 to 9 scale where 1 is absent or very
`weak, 3 is weak, 5 is medium, 7 is strong, and 9 is very
`strong. Observed when well developed and fresh brace roots
`are present on 50% ofplants.
`ANTHOCYANIN
`EDCOBAINTs=COB
`GLUME
`COLOR INTENSITY: Rated on a 1 to 9 scale where 1 is
`
`absent or very weak, 3 is weak, 5 is medium,7 is strong, and
`9 is very strong. Anthocyanin coloration should be observed
`on the middle third of the uppermost cob, after the removal
`of some of the grains.
`EDCOBCOLs—COB COLOR:A measure ofthe intensity
`of pink or salmon coloration of the cob, rated on a 1 to 9
`scale where 1 is absent or white, 2 is light pink, 3 is pink,
`4 is medium red, 5 is red, 6 is medium red, 7 is dark red, 8
`is dark to very dark red, and 9 is present.
`EDCOBDIA=COB DIAMETER:Measured in mm.
`EDCOBICAs=COB ANTHOCYANIN COLOR INTEN-
`
`SITY: A measure of the intensity of pink or salmon color-
`ation of the cob, rated ona 1 to 9 scale where 1 is very weak,
`3 is weak, 5 is medium, 7 is strong, and 9 is very strong.
`EDEARDIA=EAR DIAMETER: Measured in mm.
`EDEARHULs=EAR HUSK LENGTH: A measure of ear
`
`husk length rated on a 1 to 9 scale where 1 is very short, 3
`is short, 5 is medium, 7 is long, and 9 is very long.
`EDEARLNG=EAR LENGTH: Measured in mm.
`EDEARROW=NUMBER OF ROWS OF GRAIN ON
`EAR.
`
`EDEARSHAs=EAR SHAPE (TAPER): Rated on a 1 to 3
`scale where 1
`is conical, 2 is conico-cylindrical, and 3 is
`cylindrical.
`
`

`

`US 9,693,531 Bl
`
`9
`EDEARSHLs=EAR SHANK LENGTH SCALE: A mea-
`
`sure of the length of the ear shank or peduncle, rated on a 1
`to 9 scale where 1 is very short, 3 is short, 5 is medium, 7
`is long, 9 is very long.
`EDFILEANs=SHEATH ANTHOCYANIN COLOR
`INTENSITY AT FIRST
`
`LEAF STAGE: A measure of the anthocyanin color
`intensity of the sheath ofthe first leaf, rated on a 1 to 9 scale
`where 1 is absent or very weak, 3 is weak, 5 is medium, 7
`is strong, and 9 is very strong.
`EDFILECOs=FOLIAGE INTENSITY OF GREEN
`
`COLOR:A measure of the green coloration intensity in the
`leaves, rated on a 1 to 3 scale where 1 is light, 2 is medium,
`and 3 is dark.
`EDFILESHs=LEAF TIP SHAPE: An indication of the
`shape of the apex ofthe first leaf, rated on a 1 to 5 scale
`where 1 is pointed, 2 is pointed to rounded, 3 is rounded, 4
`is rounded to spatulate, and 5 is spatulate.
`EDGLUANTs=GLUME ANTHOCYANIN COLOR
`
`EXCLUDING BASE: A measure of the color intensity of
`the glume excludingthe base, rated on a 1 to 9 scale where
`1 is absent or very weak, 3 is weak, 5 is medium,7 is strong,
`and 9 is very strong. Observed in the middle third of the
`main branch of the tassel.
`EDGLUCOLs=GLUME COLOR:Rated on a 1 to 7 scale
`where 1 is green, 2 is yellow, 3 is pink, 5 is red, and 7 is
`purple.
`EDKERDOCs=DORSAL SIDE OF GRAIN COLOR:
`
`Rated on a 1 to 10 scale where 1 is white, 2 is yellowish
`white, 3 is yellow, 4 is yellow orange, 5 is orange, 6 is red
`orange, 7 is red, 8 is purple, 9 is brownish, and 10 is blue
`black. Observed in the middle third of the uppermost ear
`when well developed.
`EDKERSHAs=KERNEL SHAPE: Rated on a 1 to 3 scale
`where 1 is round, 2 is kidney-shaped, and 3 is cuneiform.
`EDKERTCOs=TOP OF GRAIN COLOR:Rated ona 1 to
`
`AND STEM:A measure of the angle formed between stem
`and leaf, rated on a 1 to 9 scale where 1 is very small (<5
`degrees), 3 is small (6 to 37 degrees), 5 is medium (38 to 62
`degrees), 7 is large (63 to 90 degrees), and 9 is very large
`(>90 degrees). Observed on the leafjust above the upper ear.
`FEDILEAATTs=ILEAF ATTITUDE OF ENTIRE PLANT:
`
`A measureofleaf curvature or attitude, rated on a 1 to 9 scale
`where 1
`is absent or very slightly recurved, 3 is slightly
`recurved, 5 is moderately recurved, 7 is strongly recurved,
`and 9 is very strongly recurved. Observed on the leaf just
`above the upperear.
`EDLEALNGs=LEAF LENGTH SCORE: A measure of
`leaf length rated on a 1 to 9 scale where 1 indicates<0.70 m,
`3 indicates 0.70 m to 0.80 m, 5 indicates 0.80 m to 0.90 m,
`7 indicates 0.90 m to 1 m, and 9 indicates>1.00 m.
`EDLEAWID=LEAF WIDTH OF BLADE:A measure of
`
`the average leaf width in centimeters.
`EDLELIANTs=LEAF LIMB ANTHOCYANIN COLOR
`INTENSITY OF ENTIRE PLANT: A measure of the leaf
`limb anthocyanin coloration, rated on a 1 to 9 scale with 1
`being absent or very weak, 3 being weak, 5 being medium,
`7 being strong, and 9 being very strong.
`EDNODANTS=NODES ANTHOCYANIN CO