CN101213943A

CN101213943A - Hybrid corn plant and seed

Info

Publication number: CN101213943A
Application number: CNA2007101025241A
Authority: CN
Inventors: B·J·纳格尔
Original assignee: National Starch and Chemical Investment Holding Corp
Priority date: 2006-05-16
Filing date: 2007-05-14
Publication date: 2008-07-09
Anticipated expiration: 2027-05-14
Also published as: ZA200703824B; CN101213943B

Abstract

This invention provides hybrid maize plant designated PP59302. This invention further provides hybrid seed of PP59302, hybrid plants produced from such seed, and variants, mutants, and trivial modifications to hybrid PP59302, as well as methods of using the hybrid and products produced from the hybrid.

Description

Hybrid corn plant and seed

Technical field

The present invention is maize breeding field, is specifically related to the amylose hybrid corn of called after PP59302.

Background technology

With all publications of this paper and patent application for all purposes are incorporated herein by reference, just look like each independent publication or patent application by specifically with point out separately to be incorporated herein by reference the same.

Following description comprises can be used to understand information of the present invention.But be not to recognize that any information provided herein all is prior art or relevant with current claimed invention, any publication perhaps specifically any or that in the dark quote all is a prior art.

The purpose of plant breeding is the various required proterties of combination in single kind or hybrid, perhaps provides required proterties and to the not significantly infringement of other important properties.For the land for growing field crops crops, required proterties can comprise the resistance to disease and insect, to heat, the arid tolerance of cold-peace, shortens the crops maturation time, higher yield and better agronomy quality.About the mechanical harvest of many crops, plant characteristics is important as the uniformity of sprouting and plant associations establishment, growth rate, maturing stage and plant and fringe height.Other required proterties can be and the direct or indirect relevant proterties of the specific nutritions of crops and industrial type.The example of this type of special kind or hybrid comprise have higher oil content, different oily feature (profile), higher protein content, better those kinds or the hybrid of protein quality or higher amylose content.Also need to produce the plant that is particularly suited for given agricultural region.New hybrid be control the cost of raw material effort in pith.

Maize (Zea mays L.) is at the so-called corn of the U.S., and these two terms are used interchangeably in this application.Maize has male flower and female flower separately on same plant, they lay respectively on male flower fringe and the fringe.Thereby, by with self hybridization (self-pollination or selfing) or with mutually equal, be or another plant hybridization of kind (sib-pollination or compatriot's hybridization) or with not equal, be or another plant hybridization (cutcross or cross pollination) of kind carries out breeding.

The selection that the self-pollination repeatedly of plant is combined in required type in many generations obtains inbred line, thereby it is at offspring's the homogeneous group that isozygotys on nearly all locus and when carrying out further self-pollination generation isozygotied.Hybridization between two kinds of differences are isozygotied and are produces the homogeneous group of heterozygosis hybrid plant.The hybridization of two plant species of every kind of heterozygosis on many locus will produce xenogenesis plant population, and they are different and will not be uniform in heredity.

The maize kind that can hybridize by the following method, this method comprise that (1) select plant to be used for initial breeding cross from various germplasms storehouse; (2) will be from the selected plant selfing of breeding cross several generations to produce as above-mentioned a series of inbred line; (3) with selected inbred line and different inbred line cross with the offspring that hybridizes (F1).Preferably, inbred line will be included in about 95% or higher its locus on the allelomorph that isozygotys.

Pedigree breeding and recurrent selection are two examples that are used to develop the method for inbred line.

Pedigree breeding begins with two or more genotypic hybridization, and every kind of genotype can have one or more required features.With the superior progeny selfing and select in the successive generation, in this process, the homozygosity level improves.Being suitable for inbred line that hybrid produces can be after many generation selfing and selecting, for example 4,5,6 or more generations after produce.

The double haploid method can reduce to obtain the required generation number of inbred line.These methods relate to from female parent or the haploid of male parent gamete and doubling.Can identify monoploid with genetic marker, and the dliploid that doubling monoploids is isozygotied with formation is.

Recurrent selection makes the mutual cross pollination of independent plant to form the offspring, and it is grown then.Select superior progeny by many methods then, it comprises the offspring and the topcross of independent plant, half sibs offspring, full sibs offspring, selfing.With the mutual cross pollination of offspring selected to form the offspring of another colony.Plant this colony and select good plant once more with mutual cross pollination.The target of the method for described repetition is the proterties of improvement colony.The colony of improvement is used as the source of breeding material then to obtain and will be used for the inbred line of hybrid.

Backcross and to be used to improve the hybrid of inbred line and those inbred line preparations of use.Backcross and can be used for from a kind of system---supplier parent's specific required proterties is transferred to the inbred line that is called recurrent parent, and but this inbred line has overall good agronomy feature lacks this required proterties.This transfer can be by at first with recurrent parent and supplier parent hybridization, and backcross then (wherein offspring and recurrent parent mating) realizes.Can select described required proterties to the gained offspring then, and use selected individuality further to backcross.Usually, four or more backcross from generation to generation after, select described required proterties in wherein per generation, the offspring will be contained all genes basically of recurrent parent except the gene of controlling this required proterties.Then with the last selfing from generation to generation of backcrossing to obtain offspring for the gene pure breeding of being shifted.

Other plant breeding technique as known in the art as the selection of restriction fragment length polymorphism enhancing, selection and the conversion that genetic marker strengthens, also can be used to produce inbred line.For example, the selection in the breeding method can based on from the accumulation of the mark of the positive-effect allele linkage of Plant Genome and/or with removal from the mark of the negative effect allele linkage of Plant Genome.Usually, use the combination of these technology.

For the summary of the known plant breeding method of those skilled in the art, for example see Sprague and Dudley (eds.), Corn and Corn Improvement, ThirdEdition, American Society of Agronomy, Inc.986 pages, 1988; Fehr and Hadley (eds.), Hybridization of Crop Plants, American Society ofAgronomy, Inc.765 pages, 1980; Allard, Principles of Plant Breeding, John Wiley﹠amp; Sons, Inc.485 pages, 1960; Jensen, Plant BreedingMethodology, John Wiley﹠amp; Sons, Inc.676 pages, 1988; Simmonds, Principles of Plant Breeding, Longman Group Limited, 408 pages, 1979; With Hallauer and Miranda, Quantitative Genetics in MaizeBreeding, Iowa State University Press, 468 pages, 1981.

In the strain that hybridizes by two kinds of different inbred line crosses, advantageously make the minimizing possibility of self-pollination.Make self-pollination minimize the amount that minimizes and increase hybrid seed (cross pollination obtains) with the ratio of the essentially identical gained seed of inbred line (self-pollination obtains) with making.For this reason, commercial maize hybrid produces and uses the male sterility system so that female parent's male sterile.There is several method can be used for operating the maize plant, thereby makes that it is male sterile.These methods comprise make by hand or mechanical castration (perhaps eunuchs fringe (detasseling)), cytoplasmic genetic male sterility, nuclear genetic male sterility or gametocide (influence is to the chemical reagent of the cell of male fertile key, for example, as Carlson, Glenn R. U.S. Patent number 4, describe in 936,904).

In the eunuchs fringe, alternately being with of zeistic two kinds of inbred variety is planted in the field, and before pollen discharges, removes the male flower fringe that has pollen from a kind of inbred line (female).If have enough separating with external maize pollen source, the fringe of the inbred line of eunuchs fringe will only be fertilized from another inbred line (male) so, and therefore the gained seed is hybrid and will forms hybrid plant.

Alternatively, because the hereditary factor in the plasmon group, female series can be a cytoplasmic male sterility.This feature is only by the female parent's heredity in the maize plant, because only female seed for fertilization provides cytoplasm.The CMS plant is used from the pollen fertilization that is not male sterile another inbred line.Pollen from second kind of inbred line can be with or without contribution to causing the male sterile gene of hybrid plant.The part that identical hybrid seed produces from the educated maize of eunuchs fringe and use part that the CMS system produces can fusion to guarantee that when crossbreed plant enough pollen loads can be used for being fertilized.

Can give hereditary male sterility by one of several available methods, described method is that it gives male sterility such as the locational a plurality of mutator genes that separate in genome, U.S. Patent number 4 as people such as Brar, 654,465 and 4,727, disclosed in 219, with Patterson at U.S. Patent number 3,861,709 and 3, the chromosome translocation of describing in 710,511.Described a kind of system in people's such as Albertsen U.S. Patent number 5,432,068, wherein male fertile gene is expressed under inducible promoter control.Additive method comprises the gene of sending the coding cytotoxic substance relevant with the male tissue specificity promoter in plant, perhaps antisense system, wherein identify to be inserted in the plant and (see Fabinjanski to the gene of fertility key and with the antisense thing of this gene, Deng people EPO 89/3010153.8, publication number 329,308 and PCT application PCT/CA90/00037, publication number WO 90/08828).

The hybridization by two kinds of different parent's inbred strains obtains required hybrid strain, may keep the even supply of hybrid seed.The colony that can keep mother plant by self-pollination repeatedly.In addition, because the parent isozygotys, so the hybrid that produces in the hybridization is always identical.Thereby,, just can provide hybrid seed continuously with same nature in case identified required hybrid.

The purpose that commercial maize hybrid is exploitation comprises the corn hybrid that exploitation is new; it can produce the grain of high yield; it needs less time or resource input; its more anti-environmental stress (for example; especially some growth area stress), its easier results and/or grain or other products that is particularly suited for required commercial object is provided.In order to obtain new hybrid, the good inbreeding parent system that the corn breeding person selects and exploitation is used to hybridize.Consider the number of isolated genes and the fact that the breeder does not know the details of required parent genotype usually, this categorical anything but method.Then, the breeder must identify the specific cross combination of the inbred line that produces required hybrid.Even obtained two kinds of good inbred line, can not guarantee that these combination will produce required hybrid F1 plant.Situation is so really, because many selection proterties (for example, output) depend on the effect of interactional numerous genes.Thereby, the selection of two kinds of parents system or combination results and the different unique hybrid of hybrid that as any parent and different inbreeding parent is gained when hybridizing.

Summary of the invention

The present invention relates to the exploitation of the new amylose maize hybrid of called after PP59302.PP59302 is than the higher output of amylose maize hybrid of the current growth with similar maturing stage, type and adaptation.For example, when inherent 4 places were tested together when 1 year, PP59302 was than every acre high 27.9 bushels of the average yields of two kinds of current commercial hybrids.Output difference is 39.7 bushels every acre in the test in 5 places of next year.PP59302 has the results water content suitable to other commercial amylose hybrids in similar maturing stage, has the stem quality of raising, and is the soft grain type that is suitable for wet-milling.PP59302 also provides the new amylose maize hybrid with high agronomy output for the maize culture person, and it is suitable for the corn growth area, center of the U.S..

According to the present invention, the new corn hybrid that produces by vA207 and vBL251 hybridization is provided, it is called PP59302.These two kinds of proprietary inbred line are by the exploitation of pedigree breeding method.Inbred line vA207 and vBL251 are respectively the female and male parents of hybrid PP59302.The representative sample of the seed of the hybrid plant of generation PP59302 is deposited in American type culture collection (" ATCC ") when plantation, and preserving number is PTA-7428.

On the one hand, the invention provides hybrid seed, its representative sample is deposited in ATCC with preserving number PTA-7428.The present invention also provides the colony of corn seed, at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% of wherein said seed is a hybrid seed, and its representative sample is deposited in ATCC with preserving number PTA-7428.

On the other hand, the present invention relates to by the growth seed hybrid plant that can obtain or obtain, the representative sample of described seed is deposited in ATCC with preserving number PTA-7428.

The invention still further relates to mutation, mutant and the slight modification of hybrid seed or plant.

Can have the substantially the same genotype of seed ATCC PTA-7428 with preservation according to seed of the present invention, plant, plant part, somatic tissue or cell, and/or can provide to produce and the substantially the same genotypic plant of hybrid seed as the tissue culture source with ATCC preserving number PTA-7428 preservation.

On the other hand, the invention provides the corn plant (perhaps its seed) of required proterties with hybrid PP59302.This corn plant can have all of hybrid PP59302 or all morphology or physiologic character basically.Randomly, plant can have one or more additional features, for example, owing to exist by technology well known by persons skilled in the art, the feature that produces as one or more nucleotide sequences of the importing of backcrossing as transgenic technology or conventional breeding method.In other words, hybrid corn plant of the present invention comprises the hybrid corn plant of PP59302, and it also comprises a kind of, two kinds, three kinds or the more kinds of external or heterologous gene that imports PP59302.This type of external or heterologous gene can be from the corn plant different with the inbred line that is used to produce PP59302 (promptly, corn inbred line, corn hybrid, corn monoploid, or the like), and/or (for example from the plant species that is different from maize (Zea mays), clover, soybean, low erucic acid mustard seed, tomato, potato, yew tree, Aztec marigold or the like), and/or from non-plant species (for example, bacterium, fungi, insect, mammal, jellyfish, or the like).

The invention still further relates to corn plant and seed from hybrid corn PP59302.These Plants and Seeds can be the kinds of deriving basically as definition in 41 (3) joints of plant variety protection bill (Plant Variety ProtectionAct), promptly such kind:

(i) mainly from hybrid PP59302 or from main kind from hybrid PP59302, and the expression that keeps essential characteristic, described essential characteristic is from genotype or the genotypic combination of hybrid PP59302;

(ii) obviously different with hybrid PP59302; With

(iii) except the difference that produces owing to the effect of deriving, meeting initial kind aspect the essential characteristic expression, described essential characteristic is from the genotype or the genotypic combination of initial kind.

The kind that can followingly be derived basically: by selecting mutant or somatic cell clone mutation natural or that induce, the variation individuality of the plant by selecting hybrid PP59302 is backcrossed, and transforms by gene engineering, perhaps by any other method.

Essential characteristic can be one or more required proterties that this paper provides.

In other embodiments, corn plant and seed from hybrid corn PP59302 can be from the tissue culture regeneration that produces from hybrid PP59302 plant, perhaps can be with plant or the seed of hybrid PP59302, as discussed further below as ancestors.

The present invention also provides from the tissue culture of the regenerable cell of hybrid plant PP59302 generation, and wherein said tissue culture can produce the plant of the required proterties with the hybrid PP59302 that provides as mentioned.This tissue culture can be directly or indirectly from hybrid PP59302.Preferably, tissue culture can produce the plant with the whole of hybrid PP59302 or whole basically morphology and physiologic character.Randomly, plant can have one or more extra features, for example, and the feature of giving by the nucleotide sequence that uses transgenosis or traditional breeding method to introduce.In some embodiments, plant can have the genetic complement thing of hybrid PP59302, randomly comprises one or more extra nucleotide sequences of the phenotype that when expressing (for example, as RNA or protein) can modified plant.Culture can for example, can be selected from leaf, pollen, embryo, root, the tip of a root, flower pesticide, fringe silk, flower, grain, fringe, cob, shell, stem, cell or protoplast from any tissue that can somatic embryo takes place.

The invention still further relates to the purposes of the complete plant of tissue culture deposits yields, relate to from the protoplast of described tissue culture deposits yields and the corn plant of regenerating from described tissue culture.From the method for the complete plant of tissue culture deposits yields can comprise following one or more: cultured cell in vitro forms up to observing the callus body during comprising the medium that promotes embryogenetic hormone; Organize body to form in the medium that promotes hormone to comprising cell transfer; Observe organize body to form after, with cell transfer in the medium that does not have described hormone to produce plantlet; And the described plantlet of growth, randomly be included in the described plantlet of growing on the minimal medium with sclerosis.

In another aspect of this invention, provide pollen or the ovule of hybrid plant PP59302, and by with the seed of described pollen or described ovule fertilization generation with from the plant of this seed growth.

Hybrid plant PP59302 can be hybridized with the corn plant of another kind system or kind, perhaps can compatriot hybridization or selfing to produce another kind of plant, (for example to be, inbred line) or the colony of plant (for example, the breeding population of plant), it is useful in plant breeding.

Thereby, on the other hand, the present invention relates to use hybrid PP59302 as plant or the seed of parent by procedure of breeding generation, wherein this plant or seed are described parent's offspring members from generation to generation, for example, the the the the the 1st, the 2nd, the 3rd, the 4th, the 5th, the 6th or more offsprings member from generation to generation.Thereby, the present invention includes the Plants and Seeds that use hybrid PP59302 to produce as ancestors.Can assess ancestors by the record that those of ordinary skills' routine is made.Can also for example, use molecular labeling, electrophoresis or the like assessment ancestors based on nucleic acid homogeneity.Plant of Chan Shenging or seed can have the required feature of hybrid PP59302 as discussed above like this, perhaps can have all morphology and the physiology proterties of hybrid PP59302.

On the other hand, the present invention relates to the purposes that hybrid PP59302 maize plant produces seed and/or offspring maize plant.The present invention also provides method, it comprises the plant that hybrid PP59302 is provided, with itself and self hybridization or with another maize plant (it can be another hybrid PP59302 plant or can be the plant of homology or kind not) hybridization so that produce seed, and gather in the crops described seed.This method also comprises the described seed of growth producing one or more offspring maize plants, and randomly, to produce progeny seed, can gather in the crops this seed from one or more described offspring maize plant breeding.The growth progeny seed and from the plant breeding of gained maize with the step that produces another kind of sub-group can repeat a generation or many generations (for example, 1,2,3,4,5,6 or more generations in).For example, the offspring can selfing, the compatriot hybridize, backcrosses, and colony hybridization or the like.From plant " breeding " refer to plant and self or with another plant hybridization of identical or different kind to produce the process of seed.Can select from generation to generation one or more offsprings.Selection can be one or more required proterties of selecting hybrid PP59302, for example, and one or more amylose contents of starch and agronomy output.Use visual inspection or use molecular labeling to select.

From then on thereby the plant that obtains of class methods will contain from the required proterties of hybrid PP59302 and will be benefited from the disclosure of this paper from the inventor's work neutralization.

For example, in one embodiment, method of the present invention can comprise that sib-pollination or self-pollination hybrid PP59302 are to produce first generation progeny plants.This method (for example can also be included in one or more further generations, 1,2,3,4,5,6 or the how follow-up generation) in compatriot hybridization or described offspring of selfing and/or double haploid breeding, basically isozygoty to produce, for example, greater than 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.95% or higher plant of isozygotying.This method can comprise that selection has the plant of one or more required proterties of mother plant.This selection can be carried out or can carry out with lower frequency from generation to generation each offspring, for example, in offspring's 1,2,3,4,5 or more generations, carry out (for example, first offspring from generation to generation and/or one or further the offspring from generation to generation in).

In another embodiment, as described herein the hybrid corn plant can also with the maize of different cultivars, as inbred line (for example, good inbred line) hybridization.The F1 offspring who produces from this hybridization 50% gene from generation to generation will be from hybrid PP59302.This method can also comprise from the self-pollinations of one or more plants of F1 colony to produce the F2 offspring from generation to generation.By accident, some F2 plants surpasses 50% gene from the parental generation hybrid plant.These genes can for example use molecular labeling to select or select one or more required proterties of hybrid PP59302 to select.Offspring's self-pollination can 1,2,3,4,5 or the how further generation in carry out to produce inbred line.Can be from generation to generation each offspring, perhaps with lower frequency, for example, 1,2,3,4,5 or more generations in select.

In some embodiments, this method comprises that also modification gained inbred line is to provide further one or more required proterties.For example, method can comprise gained inbred line and another plant variety hybridization with required proterties, and 1,2,3,4,5,6 or more generations in backcross described offspring so that described required proterties is inserted into genetic background, this genetic background is essentially the genetic background of inbred line.In another embodiment, this method can comprise the transgenosis modification of inbred line, and it can use the method for well known to a person skilled in the art to carry out.

In another embodiment, method comprises first kind of kind or the plant that is and second kind of different kind or the plant hybridization that is, wherein said first kind of kind or be to be hybrid PP59302.Second kind of kind or be can be inbred line and in some embodiments can be one of parent line of hybrid PP59302.This method can comprise described first offspring of growth from generation to generation.This method also comprises backcrosses described offspring one or more plants from generation to generation and second kind of kind or one or more plants of being to produce another offspring generation.Backcross can 1,2,3,4,5,6 or more generations in repeat.Last backcross from generation to generation can selfing to produce for required proterties pure breeding be.Can in one or more progeny populations, select, for example, have the plant of one or more required proterties of hybrid PP59302 with selection.

The present invention also is included in seed that arbitrary stage of above-mentioned breeding method produces or the colony of plant.In some embodiments, seed or plant can be inbred line seed or plant, for example, can be used for the further procedure of breeding or be used to develop other hybrids.

Corn is very useful crops, and many commercial products can provide by its different piece or from described part.Therefore, the invention provides plant as described herein and be used to produce purposes through the corn product of processing.

Following method also is provided, and it comprises one or more parts that plant as described herein is provided and processes described part to produce the corn product through processing.This method can also comprise the described plant of growth and/or gather in the crops described one or more parts.

Plant part can be above-mentioned any part, comprises stem, shell or cob, but will be fringe or grain in many embodiments.

The example of the corn product through processing is corn starch (corn starch ingredient that comprises separation is as amylose or amylopectin), flour, slag (grit), meal, corn syrup or glucose, corn oil, the corn grain products through processing (as grain canned, freezing or one-tenth mud), alcohol, paper, wallboard or charcoal.

For example, in one embodiment, the invention provides the method for producing corn starch, it comprises provides the grain of plant as described herein, and processing grain is to produce corn starch.Processing can comprise wet-milling.

In another embodiment, the invention provides the method for producing corn flour, it comprises provides the grain of plant as described herein, and processing grain is to produce corn flour.Processing can comprise dry grinding (dry-milling).

The present invention also provides following method, and corn product through processing that provides as above-mentioned is provided for it, uses described corn product through processing to produce manufactured goods.These can be any manufactured goods as described below.Example comprises food, packaging material, adhesive, paper or textile, medicine, cosmetics and residential care product.

The present invention also provides corn product or the manufactured goods through processing that produce by above-mentioned any method.Preferably the corn product through processing can be the starch or the flour of high amylose starches.

Definition

Unless otherwise defined, all technology used herein all have and the identical implication of one skilled in the art's common sense of the present invention with scientific terminology.Although may be used to practice or check the present invention to any method similar or of equal value described herein and material, describe preferable methods and material.

Term used herein " allelomorph " refers to any of several optional forms of gene.

" starch " used herein refers to the starch of nature or original form, also refers to the starch of modifying by physics, chemistry, enzymatic and biological method.

" amylose " used herein refers to starch polymer, and it gathers for the substantial linear of D-AGU, and described D-AGU is by α 1, and the 6-D-glycosidic bond connects.

" amylose content " used herein refers to the percentage of amylose type polymer about other starch polymers such as amylopectin.

" adaptation zone " used herein refers to have the zone of the particular combinations of environmental condition, and this corn hybrid is with well-grown under the described conditions.This term is not intended to and refers to that corn hybrid will be at this zone outgrowth, and particularly, it will not growth equally well in the zone of total identical or essentially identical condition combination.

" high amylose starches maize " used herein or " high amylose starches maize (amylomaize) " refers to have about 50% or the generic name of the corn of higher amylose content.Single recessive amylose replenishers (amylose-extender) genes (ae1) plus modifier has provided about 50% to about 94% amylose content scope.High amylose starches maize hybrid needs special management and cultivates more assurances that requirement is produced with the best grain that acceptable quality and purity are provided.The production field must isolate with normal horse profile of tooth corn.High amylose starches grain only is used for growing in order to supply wet-milling and dry grinding.The starch of amylose has complicated purposes in various industry.Be similar to output, the actual amylose content from the grain sample of concrete kind in any concrete test can be slightly different with its population mean or median amylose content, and this depends on the concrete environment of its growth.As plant in the zeistic field known to the skilledly, many factors participate in decision, and what forms the specific environment (for example, rainfall, temperature, soil types, the incidence of disease, cloud amount, or the like) of special test/seasons of growth.

" amylose maize inbred line " used herein refers to the maize inbred line, and it has about 50% or higher amylose content, wherein the amylose concentration by colorimetric method for determining grain.

" amylose maize hybrid " used herein refers to have about 50% or the maize hybrid of higher amylose content, wherein the amylose concentration by colorimetric method for determining grain.

Term used herein " hybridization " or " hybridization " or its grammer equivalent terms refer to transfer to the ovule of identical or different flower and cause fertilization from the pollen of a flower.With self hybridization plant be self-pollination or selfing; With the plant of same breed, section or another plant hybridization of being be sib-pollination or compatriot's hybridization, and with the plant of different cultivars, section or another plant hybridization of being be pollination cutcross or special-shaped.

Term used herein " cross pollination " or " crossbreeding " refer to that the pollen with a flower on the strain plant is applied to the method for ovule (column cap) of the flower of (manually or natively) another strain plant.

Term used herein " cultivated species " refers to kind, strain or the race of the plant that produces and do not find usually by gardening or agronomy technology in wild type colony.

Term used herein " good inbred line " refers to demonstrate the inbred line of the required quality of contribution when being used to produce commercial hybrid.

Term used herein " female " refers to produce the plant of ovule.Female generally produces seed at after fertilization.The plant that is called " female " can contain male and female sexual organ.Alternatively, " female " (for example, at the dioecism species) or because castration (for example, by the eunuchs fringe) and only contain female sexual organ natively.

Term used herein " filial generation " refers to cell, tissue or the biological any generation of specific parental generation after the generation.Parent's mating gained is that F1 (is called " F1 " or " F from generation to generation ₁"), and the filial generation of the individual hybridization of F1 is that second filial generation (is called " F2 " or " F ₂").

Term used herein " gamete " refers to reproductive cell, and but its cell nucleus (with common cytoplasm) and similar source another Gamete Fusion of opposite sex are with the formation zygote, and described zygote has the potentiality that develop into new individuality.Gamete is monoploid and is divided into male and female.

Term used herein " gene " refers to arbitrary fragment of the DNA relevant with biological function.Thereby gene includes but not limited to, the adjusting sequence that coded sequence and/or their expression are required.Gene can also comprise the dna fragmentation of non-expression, and it for example forms the fragment of the recognition sequence of other protein.Gene can obtain from multiple source, comprises from purpose source clone or synthetic from sequence information known or prediction, and can comprise the sequence that is designed to have desired parameters.Thereby the present invention also comprises maize plant of the present invention and its part, thereby it has been transformed and makes its genetic stocks contain one or more transgenosiss that are operably connected to one or more regulating elements.In addition, maize plant of the present invention or its part also comprise this type of maize plant or its part that contains the individual gene transformation.

The allelic close set that term used herein " genetic complement thing " phalangeal cell has.In plant or other somatic tissues or cell, complement will be diplontic-promptly, two allelomorph (identical or different) are arranged on each locus.

Term used herein " genotype " refers to the genetic constitution of individual cells, cell culture, tissue, plant or plant group.

Term used herein " grain " refers to that commercial grower is in order to be different from the one-tenth cooked maize grain that purpose produced of growth or breeding species.

Term used herein " heterologous polynucleotide " or " heterologous nucleic acids " or " exogenous dna fragment " refer to polynucleotides, nucleic acid or dna fragmentation, it is from the source of particular host cell outside, if perhaps from identical source, then modify from its original form.Thereby, but the heterologous gene in the host cell comprises the endogenous modified gene of particular host cell.Thereby this term refers to dna fragmentation, and it is external or allos for cell, is homology for cell perhaps, but in the position in host cell nucleic acid, does not find this fragment usually on this position.Exogenous dna fragment is expressed to produce allogenic polypeptide.

Term used herein " allos proterties " refers to give by the phenotype of transformed host cells or genetically modified organism by exogenous dna fragment, heterologous polynucleotide or heterologous nucleic acids.

Term used herein " heterozygote " refers to have dliploid or the polyploid individual cells or the plant of the not iso-allele (form of given gene) that is present at least one locus.

Term used herein " heterozygosis " refers to exist different allelomorph (form of given gene) on the specific gene seat.

Term used herein " homologue " refers to nucleic acid or peptide sequence, and it has common source or has similar function to nucleic acid or peptide sequence from another species.

Term used herein " homozygote " refers to have mutually homoallelic individual cells or plant on one or more locus.

Term used herein " isozygotys " and refer to exist on one or more locus identical allelomorph in the homologous chromosome fragment.

Term used herein " hybrid " refers to any individual cells, tissue or the plant that hybridization obtains between the parents different in one or more genes.

Term used herein " inbreeding " or " inbred line " refer to the strain of relative pure breeding.

Term used herein " grain " refers to comprise the corn caryopsis of mature embryo and endosperm, and it is the product of double fertilization.

Term used herein " is " to be used in a broad sense include but not limited to, by tissue culture technique from the vegetative one group of plant of single mother plant or since from common parent heredity and in heredity closely similar one group of inbreeding plant." belong to " specific system if following situation is then said plant: it (a) is elementary transformant (T0) plant from this material regeneration that is; (b) has the pedigree that comprises this T0 plant that is; Perhaps (c) is because common ancestors' (for example, by inbreeding or selfing) and closely similar in heredity.In this context, term " pedigree " for example refers to the pedigree of the plant of realizing according to sexual hybridization, thereby the combination of gene or gene is given required proterties to plant under the heterozygosis (hemizygous) or the condition of isozygotying.

Term used herein " locus " refers to upward any site of definition of heredity.Locus can be the part of gene or gene, and the dna sequence dna that perhaps has some regulating actions perhaps can be occupied by identical or different sequence.

Term used herein " male " refers to produce the plant of pollen grain." androecy " refers generally to produce the sex of the gamete that is used to fertilize an egg.The plant that is called " androecy " can contain male and female sexual organ.Alternatively, " androecy " (for example at the dioecism species) or because castration (for example, by removing ovary) and only contain male sexual organ natively.

Term used herein " mix select " refers to a kind of selection of form, wherein selects bion and of future generation from their the aggregation breeding of seed.

Term used herein " open pollination " refers to freely be exposed to the plant population of some gene flows, and this is with opposite in sealing pollination, and the latter is to the effective barrier of gene flow.

Term used herein " colony of open pollination " or " kind of open pollination " refer to can carry out at least usually the plant of some cross pollinations, the certain Standard Selection of basis, but it can demonstrate variation also have one or more genotype or phenotypic characteristic, and this colony or kind are distinguished with other colonies or kind by described feature.It is the colony of open pollination or the kind of open pollination that cross pollination is not had the hybrid of barrier.

Term used herein " ovule " refers to the female gamete body, and term " pollen " refers to male gametophyte.

Term used herein " phenotype " refers to the observable feature of individual cells, cell culture, plant or plant group, and it is produced by the interaction between this individual genetic constitution (being genotype) and the environment.

Term used herein " a kind of recombinant " or " multiple recombinant " refer to experience recombinant DNA cell transformed, tissue or biology.Initial recombinant is called " R0 " or " R ₀".Selfing R ₀Generation is called " R1 " or " R ₁" the generation that transforms first.

Term used herein " various plants " or " plant species " or its grammer equivalent terms (are plant with broad understanding for both being included in the complete biology of their arbitrary developmental stage, sometimes be also referred to as complete plant), also comprise plant cell, plant protoplast, tissue culture, plant callus, plant embryos or plant part are as root, the tip of a root, stem, leaf, flower, flower pesticide, fringe, cob, shell, fringe silk and grain.

Term used herein " seed " refers to the one-tenth cooked maize grain for the purpose generation of breeding species.

Term used herein " self-pollination " or " self-pollination " refer to the pollen of a flower on the strain plant is used (manually or natively) on identical on the identical plant or the ovule (column cap) that do not suit.

" MST PCT " used herein actual water content of grain when referring to gather in the crops.

" fringe that percentage falls (PER CENT DROPPED EARS) " used herein refers to break away from and fall from plant the percentage of corncob on the ground.

" PLTPOP " used herein refers to compare the percentage of the plant that has emerged after the plantation with the average percent of all hybrids in the conventional test.

" evergreen (staygreen) " used herein refers to the tolerance of plant health, and its percentage by the chlorenchyma compared with dry brown tissue on the physiology maturing stage plant is recently measured.

" dehydration (drydown) " used herein or " dehydration (dry down) " refer to the forfeiture as time passes of grain moisture.

" STKLOD PCT " used herein refers to the percentage of the plant that the stem under the joint of fringe wherein breaks.

" TST/WT LB/BU " used herein refer to for given bushel volume, the measurement of the grain weight of representing with pound.

Term used herein " synthetic " refers to one group of offspring that the mutual friendship that is by the clone of particular group or seminal propagation obtains.The synthetic mixture that can contain the seed that allogamy, self-fertilization and/or compatriot's fertilization obtain.

Term used herein " conversion " refers to that nucleic acid (being nucleotide polymer) transfers in the cell.Term used herein " genetic transformation " refers to DNA, and especially recombinant DNA shifts and is incorporated in the cell.

Term used herein " transformant " refers to experience cell transformed, tissue or organ.Initial transformant is called " T0 " or " T ₀".Selfing T0 produces and is called " T1 " or " T ₁" the generation that transforms first.

Term used herein " genetically modified " refers to accept by one of various method for transformation cell, cell culture, biology, plant and the offspring of the plant of external or modified gene, wherein said external or modified gene from the plant of accepting external or modified gene or the biological identical or different species of species.

Term used herein " kind " refers to the segmentation of species, and it is made up of group of individuals in the species, described group of individuals on shape or function with other similar group individual different.

Detailed Description Of The Invention

PP59302 is the Hybrid between female inbred line vA207 and the male inbred line vBL251.Exploitation inbred line vA207 and vBL251, thus make each inbred line all have the content of starch of 55% to 65% amylose.

The feature of hybrid PP59302 is high agronomy output, average drydown and strong excrescence.Its strong excrescence is especially noticeable proterties.Amylose maize hybrid generally on agronomy inferior to the maize hybrid of the other types that are used for commodity production, as (waxy) of horse profile of tooth (dent) and wax.According to more relative maturing stage system of grain results water content, hybrid PP59302 has about 112 days relative maturing stage.It is suitable for the east center corn field zone of Indiana and Ohio.According to the data of collecting from the sub-district, field of the Lebanon that is positioned at Indiana, this hybrid has following feature.

Table 1

The mutation of hybrid seed or plant PP59302, mutant and small modification are within the scope of the invention.Small modification can be the modification of the genetic coding of hybrid plant, and it causes having the required proterties of aforesaid hybrid PP59302, and preferably has all of hybrid PP59302 or the plant of all morphology or physiologic character basically.

Preferably according to seed of the present invention or plant, for example, the genome of genome of mutation seed or plant and hybrid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% genetic identity.

The progeny plants of hybrid PP59302 (in any generation) or from the plant of hybrid PP59302 can be preferably and hybrid corn plant PP59302 have at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or 100% genetic identity.

Use the conventional plant breeder's record preserved of those skilled in the art can assess plant genotype and with the genetic identity degree of hybrid PP59302.Use molecular marking technique, for example, can be additionally or alternatively assess genotype by the genetic marker signature analysis.

Can obtain the genetic marker feature by following technology, amplification region (SCARs), amplified fragment length polymorphism (AFLPs), simple sequence as restriction fragment length polymorphism (RFLP), randomly amplified polymorphic DNA (RAPD), arbitrarily primed PCR (AP-PCR), DNA cloning fingerprint technique (DAF), sequence characterization repeat (SSRs) (being also referred to as little satellite), and single nucleotide polymorphism (SNPs).For example, see Berry, people such as Don " AssessingProbability of Ancestry Using Simple Sequence Repeat Profiles:Applications to Maize Hybrids and Inbreds " Genetics 2002,161:813-824.

SSRs is through being usually used in the purpose of mapping.This method is based on repetitive sequence, and it can repeat different number of times at any given locus, thereby produces polymorphism, has a plurality of allelic possibilities.Comprise that by several different methods PCR can realize the detection of SSR.Two primers (as the primer that can openly obtain) with the regional flank that contains repetitive sequence carry out the PCR detection.After the amplification, mark is given a mark by the gel electrophoresis of amplified production.The marking of marker gene type is rounded up to nearest integer based on the size of the amplified fragments that passes through molecular weight measurement.Relative value should be regardless of concrete primer that uses or accurate technology and all keep constant.

Thereby can quote percentage molecular labeling feature homogeneity to quoting of percentage genetic identity.The molecular labeling feature can be the SSR feature.Percentage can refer to contribute from the heredity in the molecular labeling feature of hybrid PP59302.

May be preferably, according to seed of the present invention or plant when relatively the time, having one or more extra required proterties and/or give the nucleotide sequence of one or more insertions of required proterties with hybrid PP59302.Nucleotide sequence can be inserted among any ancestors of seed or plant or its by any method well known by persons skilled in the art, and described method is for example transgenic technology or traditional breeding method, as backcrosses.Required proterties includes but not limited to, insect, insect or disease resistance, to the modification (for example carbohydrate of the phytic acid content of the fatty acid metabolism of Xiu Shiing, reduction, modification composition or the like) of the character of the arid of the resistance of weed killer herbicide, enhancing or cold resistance, male sterility and corn grain.The source of nucleic acid can be the plant of identical or different plant species, perhaps can be that any other is biological as animal (for example, insect), prokaryotes, fungi or virus.Nucleic acid can also be artificial nucleic acid, i.e. the nucleic acid of natural appearance not.

The particular instance of this genoid is well known to a person skilled in the art, but more operable genes comprise EPSP or aroA gene, antisense ACP gene or the phytic acid enzyme coding gene of bacillus thuringiensis (Bacillus thuringiensis) protein, disease resistance in plants gene, agglutinin, vitamin-binding protein such as avidin, protease inhibitors or amylase inhibitor, sudden change.Nucleic acid can be can modified plant when in plant, expressing phenotype, for example, give or improve any genetic stocks of required proterties, comprise the nucleotide sequence of antisensenucleic acids, siRNA or the like and coded protein.Nucleic acid can also be or comprise the enhancer of promotor.The example of suitable nucleic acid can be seen US 6,777,598, and its disclosure clearly is incorporated herein by reference.

Transgenic method is well known to a person skilled in the art.The physics of Plant Transformation and biological method all are well known in the artly (for example to see, people such as Miki, " Procedures forIntroducing Foreign DNA into Plants ", in Methods in Plant MolecularBiology and Biotechnology, Glick, B.R. and Thompson, J.E.Eds (CRCPress, Inc, Boca Raton, 1993) pages 67-88).Also can utilize the extracorporeal culturing method of expression vector and plant cell and metaplasia and the regeneration of plant.For example see, people such as Gruber " Vectors for Plant Transformation ", in Methods in PlantMolecular Biology and Biotechnology, Glick, B.R. and Thompson, J.E.Eds (CRC Press, Inc, Boca Raton, 1993) pages 89-119, with U.S. Patent number 6,118,055.

The present invention also relates to tissue culture in some respects with in the embodiment, relates to the purposes of these cultures and produces the methods of plants from these cultures.

Duncan, Williams, Zehr and Widholm, Planta, (1985) 165:322-332 have reflected that 97% the plant of being cultivated that produces callus can carry out plant regeneration.The experiment of carrying out subsequently with inbred line and hybrid has produced 91% renewable callus, and it produces plant.In another research in 1988, Songstad, Duncan﹠amp; Widholm has reported several culture medium additives at Plant Cell Reports (1988) among the 7:262-265, it has strengthened the recyclability of the callus of two kinds of inbred line.Other disclosed reports show that also " unconventional " tissue can produce somatic embryo and take place and plant regeneration.K.P.Rao, Deng people .Maize Genetics Cooperation Newsletter, 60:64-65 (1986) relates to and carries out somatic embryo from lepicena callus culture thing and take place, and B.V.Conger, Deng people .Plant Cell Reports, 6:345-347 (1987) points out that carrying out somatic embryo from the tissue culture of maize leaf fragment takes place.Thereby, from document can know the technology status of finding out this area be these methods that obtain plant now and the past be normally used and have very that the meaning of high success rate is " routine " at them.

Disclose in 160,390 (being introduced into this paper as a reference) at european patent application and to have described zeistic tissue culture.At Green and Rhodes, " Plant Regeneration inTissue Culture of Maize; " Maize for Biological Research (PlantMolecular Biology Association, Charlottesville, Va.1982, at 367-372) and Duncan, Deng the people. among " The Production of Callus Capable of PlantRegeneration from Immature Embryos of Numerous Zea MaysGenotypes, " 165 Planta 322-332 (1985) the maize method for tissue culture has been described also.

In the hybrid seed production process, make great efforts to prevent the self-pollination of inbreeding parent system.This can give one of parent male sterility by the technology (comprising technology discussed above) that it will be apparent to those skilled in the art and carry out.Yet, in the field, female parent's holandry sterility is difficult to realize very much, and therefore in the hybrid seed of packing, possibility that has the female parent comprise a small amount of selfing, though at processing or treated female seed so that when making male sterile.And, because male parent's adjacent growth with female parent, so seed that might male selfing is gathered in the crops unintentionally and packed with hybrid seed in the field.

Therefore, most of seeds that can comprise two kinds of parents' hybridization generation according to the colony of seed of the present invention, and the seed (being equal to the male and female parent of inbreeding is) that produces of the selfing parental line that comprises certain level, can expect that it obtains from the method that hybridizes normally.For example, plant sub-group and can comprise at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or 100% the seed that obtains from two kinds of parents' hybridization.The amount of female inbred line (i.e. the seed that produces from the female parent of selfing) can be less than 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1% or 0.05%.The amount of male inbred line (i.e. the seed that produces from the male parent of selfing) can be less than 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1% or 0.05%.

The plant of self-pollination can be identified and be distinguished with hybrid seed, because the plant of self-pollination will be equal to one of inbred line that is used to hybridize in heredity.Because the homozygosity level, when comparing with hybrid, they will demonstrate lower vigor.For example, can identify inbred line by their nutrition and/or the lower vigor outward appearance of reproduction feature, described outward appearance comprises lower plant height, small ear size, fringe and grain shape, cob color or other features.

Also can finish the evaluation that is of these self-pollinations by molecular marker analysis.See " TheIdentification of Female Selfs in Hybrid Maize:A Comparison UsingElectrophoresis and Morphology ", Smith, J.S.C. and Wych, R.D.Seed Science and Technology 14, pp.1-8 (1995) clearly is incorporated herein by reference its disclosure.Inbred line can be accredited as on one or more locus and isozygoty.Also referring to " Identification of Atypical Plants in Hybrid Maize Seed byPostcontrol and Electrophoresis " Sarca, people .Probleme deGenetica Teoritica si Aplicata Vol.20 (1) such as V. p.29-42.

Industrial usability

Corn has purposes widely, can be used as animal feed, is used to provide human consumption's food and is used to provide industrial raw materials.

Corn comprises that grain and non-grain partly are widely used as the feed of domestic animal such as pig, ox and poultry.Grain also is used for the human consumption.In addition, corn grain can wet-milling to produce corn starch, corn syrup and glucose, perhaps can dry grind to produce corn flour, slag and meal.Reclaim corn oil from maize germ, maize germ is the accessory substance of wet-milling and dry grinding industry.

The purposes of corn starch is based on functional character, as the ability of viscosity, film formation, sticking property and suspended particulate.Can in industry, be used for papermaking, textile and adhesive in the corn starch.It can also be used for construction material, foundry binders, laundry starch, explosive, oil well mud, bore flow of oil (oil-drilling fluid) and other mining application.Because their biodegradable and renewable character, starch is used to many other products day by day, comprises packing, plastics, detergent, medicinal tablet, insecticide and cosmetics.Starch can also be fermented into alcohol and can be processed to corn syrup and sweetener, as high-fructose corn syrup and glucose.Starch can use (for example, the corn starch of the corn starch of sour modification, paste, oxidation, pregelatinised starch and chemically derived starch) with modification or unmodified form.

Corn starch is made up of two kinds of component amyloses and amylopectin.Amylose mainly by passing through 1, form by the glucose monomer linear chain that the 4-glycosidic bond connects.In amylopectin, chain is by adding 1,6-glycosidic bond branch.Have the amylose of different proportion and the starch of amylopectin and be particularly suited for different industrial purposes with flour.

Can be by the starch of the property identification high amylose starches below one or more.Particle is two kinds of different types, and is spherical and irregular, and littler than normal starch granules.Birefringence end point temperature (Birefringence End Point Temperature) (" BEPT ") is reported as 97 ℃.BEPT is the temperature of starch molecule when losing organized structure.Some particles even after boiling for a long time, also do not lose all birefringences; Under 95 ℃, swelling power only is that about 1/4 and solubility of ordinary maize starch is the pact half (Corn and Corn Improvement, third edition, Ed.Sprague andDudley) of ordinary maize starch.

The starch of high amylose starches is particularly useful for confectionery, as the candy (because their quick thickenings) of gluing, fried snack (because infiltration of their anti-cooking oil) and photographic film (because their toughness and transparency), and purposes discussed above (for example, textile, biodegradable packaging material, be used to produce the adhesive of corrugation cardboard, or the like).Also propose by using high amylose starches flour can improve the anti-aging character of bread.Other purposes are included in the classification of braiding front glass fiber, and preparation is used for transparent, the dispersible edible film of hot water of sheathing foodstuff, dyestuff and other soluble materials and be used to reduce the coated paper of water and fat absorption.Attention: be intended to add nutrition aspect (because these mainly is that we develop with the starch of high amylose starches), especially high fiber, to the height resistance that digests, low in calories, and the control of blood glucose response (glycemic response).

Amylopectin especially can be used for papermaking and adhesive (because its branched chain makes it have bigger binding ability), with the food that is used for producing easily, as freezing and canned food (because it strengthens stability and pot-life) and fruit pie filling (wherein it is as thickener).It can be used to form semi-transparent film, and it can easily dissolve again, and purposes discussed above.

Other purposes of corn comprise that purposes and cob that stem and shell are used for paper and wallboard are used for fuel, with preparation charcoal and the purposes that is used to produce furfural (fufural).

Embodiment

To understand embodiment described herein and embodiment and only be used to illustrate purpose, and prompting be had its various modifications or change for those skilled in the art, and described modification or change will be included in the application's the spirit and scope.

The agronomy character of embodiment 1 hybrid relatively

The agronomy character that carries out A5414 and A5515 and PP59302 compares, and A5414 is business-like amylose hybrid with A5515 and has similar maturing stage and adaptation.In table 2 and 3, provide relatively.

The hybrid yield summary data of table 2.A5515 and PP59302.The mean value of the yield trials that the comparing data representative is carried out in 3 year sections in 21 places of Indiana.

(BU/A#56=is with every bushel of 56lbs, the bushel number of every acre of grain output; PCT=percentage)

The hybrid yield summary data of table 3.A5414 and PP59302.The mean value of the yield trials that the comparing data representative is carried out in 3 year sections in 11 places of Indiana.(BU/A#56=is with every bushel of 56lbs, the bushel number of every acre of grain output; PCT=percentage)

As shown in table 2 and 3, PP59302 has obviously higher agronomy output than A5414 and A5515.Grain water content when results is suitable, thereby shows that three kinds of hybrids have the suitable maturing stage.PP59302 demonstrates the remarkable advantage that early excrescence (data do not provide) and plant associations are established, and plant associations are established and pass through PLTPOP PCT data representation.PP59302 also has than A5414 and the better stem quality of A5515, as pointing out by STKLOD PCT data.

Table 4 provides with many different commerce and test corn hybrid and has compared the summary of the average agronomy character data of PP59302.

Table 4. hybrid yield summary data.The mean value of the farm test that the comparing data representative is carried out in 15 places of Indiana (Beck 5422 is horse profile of tooth kinds, calculates % horse profile of tooth value from it).

Hybrid	Water content	Output (Bu/A)	% horse profile of tooth
Hybrid	Water content	Output (Bu/A)	% horse profile of tooth	A5414	17.5	140.9	59.7
A5515	17.4	149.8	63.4	A5414	17.5	140.9	59.7
A5515	17.4	149.8	63.4	PP59302	18.2	162.1	68.7
PP59401	17.4	141.4	59.9	PP59302	18.2	162.1	68.7
PP59401	17.4	141.4	59.9	A7701	17.9	160.2	67.8
A7504	17.0	157.8	24.5	A7701	17.9	160.2	67.8
A7504	17.0	157.8	24.5	PP79311	17.5	163.9	69.4
PP79411	18.5	150.6	63.8	PP79311	17.5	163.9	69.4
PP79411	18.5	150.6	63.8	PP79412	17.3	157.5	66.7
BECK 5422	9.3	236.1		PP79412	17.3	157.5	66.7

As shown above, PP59302 has high yield when comparing with other hybrids in suitable maturing stage.

Embodiment 2. amylose contents

Amylose concentration by colorimetric method for determining grain.The amylose selectivity absorbs iodine, and producing highly colored amylose-Surgidine, and the amount of the amylose of the intensity of this color and existence is proportional.Use spectrophotometer to measure percent transmittance in 610 nanometers.Obtain amylose percentage from calibration curve.This calibration curve is drawn from the percent transmittance value that obtains with the starch with known percentage amylose content.Colorimetric method used herein is following to be provided:

Measure amylose content by colorimetric analysis

Equipment:

1.Tecator Cemotec ^TMSample grinding mill or equivalent apparatus

2. four cap nut teat glasses that have lid, 20 * 125mm

3. four cap nut teat glasses that have lid, 20 * 150mm

4.4 individual solid PTFE (polytetrafluoroethylene (PTFE) or teflon) splash bar, long 8 "

5. boiling water bath

6. the centrifuge that can hold 20 * 125mm test tube

7.4 individual porcelain Buchner funnel, the 43mm board diameter

8. glass microfiber filter, 4.25cm diameter, 1-1.2 μ m porosity (Whatman

#1821-042, VWR#28333-141, or be equal to filter)

9. automation diluter, double syringe (Hamilton Microlab Series 500

Or equivalent)

10. the glass syringe of diluter, 10mL

11. the glass syringe of diluter, 500 μ L

12. the flow injection spectrophotometer of automation, the 590nm wavelength is as FossTecator FIAStar ^TMThe flow injection analyzer system, it has Tecator 5042Detector ^TM, Tecator 5012 Analyzer ^TM, Tecator 5027 Sampler ^TM

13. projection polariscope, the 589nm wavelength

14. vavuum pump

15. filter flask, 500mL

Reagent:

Dense calcium chloride solution

3.5kg reagent grade calcium chloride dihydrate is dissolved in the water of purifying, cool to room temperature, and regulate proportion to 1.3 with calcium chloride or purified water, use the pH to 2.0 of the careful regulator solution of reagent grade glacial acetic acid then; Before use the cellular glass funnel of solution by medium porosity filtered

Rare calcium chloride solution

With purified water the dense calcium chloride solution of 600mL is adjusted to 2L

The deposit iodine solution

8.00g reagent grade potassium iodide and 4.16g reagent grade iodine are dissolved in about 10mL purified water, and are adjusted to 1L with rare calcium chloride solution; Solution should be kept in the amber bottle

The work iodine solution

25mL is laid in iodine solution add to 200mL with the dilution calcium chloride solution

The maize grain sample of known amylose content is as calibration criterion

Program:

1. use Cemotec ^TMThe sample grinding mill with 3-4g calibration standard sample fine gtinding in suitable containers.

2. to laboratory sample repeating step 1, thereby guarantee that grinding mill is clean between each sample is milled.

3. in calibration sample to three independent 20 * 125mm test tube that weighing 0.2g, 0.4g and 0.6g mill.

4. in weighing 0.4g laboratory sample to four 20 * 125mm test tube.

5. add the dense calcium chloride solution of 8mL to each 20 * 125mm test tube.

6. in each 20 * 125mm test tube, place the PTFE splash bar.Use described rod to disperse grain.

7. 4 20 * 125mm test tubes were placed in the boiling water bath 30 minutes.Use splash bar in preceding 5 minutes, to continue to stir the content of test tube.Stirred content about 1 minute in per then 5 minutes.

8. take out test tube from water-bath.Take out stirring rod immediately, do not wash, and allow the sample cool to room temperature.

9. in each sample cell, add the rare calcium chloride solution of 8mL.Covering each pipe also acutely shakes.

10. with 1, the centrifugal sample cell of 800RPM 5 minutes.

11. 20 * 150mm test tube is carefully put into filter flask (can place sponge in the bottom of bottle destroys to prevent test tube).

12. in 20 * 150mm test tube, insert the Buchner funnel handle.In Buchner funnel, place 1-1.2 μ M microfibre filter.Open vavuum pump.

13. pour out the solution of the centrifugal grain of milling to the bottom and pour on the microfibre filter from 1 20 * 125mm test tube.Allow sample filtering, entered 20 * 150mm test tube up to all solution, and dry filter.

14. cover 20 * 150mm test tube and inversion several times, with biased sample.The solution that filters should be clarification and do not have floating particle at this moment.If not, filtered sample once more.

15. to laboratory sample and each calibration standard sample completing steps 11-14.

16. use automation double syringe diluter, with rare calcium chloride solution with the diluted sample of 400 μ L filtration to 10mL.

17. use the solution of flow injection spectrophotometer analysis dilution.The work iodine solution should as will with the last reagent of sample mix of injection.Flow injection pump line with 0.38mm internal diameter can be used to send work iodine, and the flow injection pump line with 0.89mm internal diameter can be used to send sample.If necessary, other reagent bottles that are full of rare calcium chloride solution can use with further dilute sample together with flow injection analyzer, mix with the work iodine solution afterwards.In scouring stage, should use purified water between analyzing, to wash flow chamber.

18. the absorption peak of the solution of record iodinate.

19. use remainder, write down the optical activity of each sample from the filtering solution of step 14.

20. use result, draw the figure that absorbs (amylose content X optical activity) from three calibration standard samples.Determine the slope (m) and the y-intercept (b) of this line.

21. use the slope definite in the step 20 and the absorption peak and the optical activity value of y-values of intercept and laboratory sample, can use following equation to determine the amylose content of laboratory sample:

Three hybrid A5414 that do not carry out the same year and commercial that following table provides in six places of middle part Indiana and A5515 PP59302 amylose content percentage relatively.

Table 5. amylose content percentage.The mean value of the experiment that the comparing data representative is carried out in the triennium section in 6 places of Indiana.

As shown in table 5, PP59302 has higher amylose content than commercial hybrid A5414 and A5515, but is based on the average amylose content of hybrid in all tests, falls into the classification of amylose hybrid.

Preservation information

The applicant according to budapest treaty (Budapest Treaty) on March 8th, 2006 with corn hybrid Corn, Zea Maize, at least 2500 seeds of L.PP59302 (describing as this paper) are deposited in American type culture collection (ATCC), Rockville, Md.20852 USA, ATCC preserving number PTA-7428.In the application's submission a few days ago, the preservation thing of keeping from National Starch and Chemical Company takes out seed and is deposited in ATCC.This preservation thing of corn hybrid PP59302 will be in the public preservation ATCC of institute preservation institute 30 year of preservation section, back 5 years of perhaps nearest request perhaps in the implemented time limit of this patent, is as the criterion with elder more, if and the preservation thing can't be survived in this time period, then will be replaced.In addition, the applicant has satisfied all requirements of 37C.F.R. § § 1.801-1.809, comprises the proof of sampling survival, perhaps satisfies described requirement before the license based on this application.The applicant to the availability of this preserved material of ATCC without limits, yet the applicant does not abandon in any restriction that biomaterial shifts or it is applied by law during in commercial transportation.The applicant has the right to investigate any infringement of the right that this patent is awarded.

Although described the present invention in conjunction with specific embodiments of the present invention, but will understand it can further be revised and the application is intended to cover of the present invention any change, use or the reorganization of generally making according to principle of the present invention, and comprise and the departing from of the disclosure of invention, these depart from the known or habitual practice that drops on field of the present invention, and can be applied to above provide and essential characteristic within the scope of the appended claims.

Although purpose in order to be aware and understand, describe the invention of front to a certain extent in detail by explanation and embodiment, but obviously can make some change and modification within the scope of the invention, scope of the present invention is only limited by the scope of claims.

Claims

1. the seed of the hybrid corn of called after PP59302, its representative sample is deposited in ATCC with preserving number PTA-7428.

2. pass through the available maize plant of seed or its part of growth claim 1.

3. the maize plant of claim 2 or its part, wherein this plant or its part are transformed, thus its genetic stocks contains one or more transgenosiss of one or more regulating elements that are operably connected.

4. all or the maize plant of all morphology and physiologic character basically that have the plant of claim 2.

5. all or the plant part of all morphology and physiologic character basically that have the plant part of claim 2.

6. accessory rights requires the tissue culture of the regenerable cell that 2 plant or its part produce.

7. accessory rights requires the maize plant of the tissue culture regeneration of 2 plant or its part.

8. the ovule of the plant of claim 2.

9. the pollen of the plant of claim 2.

10. produce the method for maize seed, it comprises the maize plant of claim 2 and self or another kind of maize plant hybridization, and gathers in the crops the gained seed.

11. the method for claim 10, it also comprises growth gained seed producing one or more offspring maize plants, producing progeny seed, and gathers in the crops described progeny seed from the plant breeding of one or more described offspring maize.

12. the method for claim 11, its also be included in 1,2,3,4,5,6 or more generations in the described progeny seed of growth, producing seed, and gather in the crops described seed from the plant breeding of gained maize.

13. the maize seed that produces by claim 10,11 or 12 method.

14. the maize plant of the seed growth of accessory rights requirement 13.

15. seed, it produces the plant of claim 4 when growth.

16. produce the method through the corn product of processing, it comprises at least a plant part that claim 2 is provided, and described part is processed into corn product through processing.

17. the method for claim 16, wherein said part are one or more corn grain.

18. the method for claim 16, wherein said corn product through processing is a corn starch.

19. the method for claim 16, wherein said corn product through processing is a corn flour.

20. the corn product that produces by claim 16,17,18 or 19 method through processing.

21. method, it comprises the corn product through processing that claim 16 is provided, and comprises the described corn product production manufactured goods through processing of use.

22. the method for claim 21, wherein said manufactured goods are selected from confectionery, deep-fried food and bakery.

23. the method for the maize plant that generation PP59302 derives, it comprises:

A) with hybrid corn PP59302 and second kind of maize plant hybridization, and results gained maize seed, wherein the representative seed of PP59302 is deposited in ATCC with preserving number PTA-7428; With

B) the described maize seed that obtains of growth is to produce the maize plant that PP59302 derives.

In the plant breeding program, develop the method for maize plant 24. use the plant breeding technology, it comprises and uses maize plant or its part as the plant breeding material source, comprises that maize plant or its part of using claim 2 originate as breeding material.