CN1539263A - Method of cultivating breed of wheat collecting multiple genes of against disease - Google Patents
Method of cultivating breed of wheat collecting multiple genes of against disease Download PDFInfo
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Abstract
A method for culturing the wheat variety containing multiple disease-resistant genes includes hybridizing between PP9-1 as yellow dwarf-resistant female parent and excellent wheat as male parent, backcrossing, choosing the yellow dwarf-resistant female parent for next bybrid, hybridizing between the Pm4b gene contained tetraploid wheat as male parent and the stripe rust-resistant wheat as female parent, backcrossing, choosing the powdery mildew-resistant male parent for next hybrid, hydridizing between said chosen male parent and female parent, self-crossing, choosing the yellow dwarf and powdery mildew resistant plant, anther culture, and natural reduplication.
Description
Technical field
The present invention relates to the breeding method of anti-disease wheat kind in the crop disease-resistant breeding field, particularly relate to the breeding method of the anti-disease wheat kind of multiple disease-resistant gene polymerization.
Background technology
Yellow stunt of wheat is one of main disease of world wheat, and all there is generation in the country of nearly all production wheat.Yellow stunt of wheat is caused that by luteovirus this virus is the line-up of delegates that yellow syndrome virus belongs to, and is a kind of monomer positive chain RNA virus, and the plastochondria form is icosahedron (T=3), hexagon, and diameter 25～30nm, no coating, kernel is made of geneome RNA.Genome is 5.6～5.9kb, 3 ' end no poly (A) tail.Luteovirus in China, causes the yellow dwarf virus of wheat morbidity to have 4 kinds of strain set types at least by aphis propagation, is respectively GPV, GAV, PAGV and RMV.PAGV, GPV and GAV's is pathogenic stronger, and RMV's is pathogenic the most weak.4 kinds of strains system passes the one-tenth of virus mediators, if aphid can both transmitted virus, greenbug is yellow dwarf virus GPV, effective biography virus mediator of system such as pathogenic strains such as 3 kinds strong of GAV and PAGV etc.Virus infects from the screen casing of the plant that catches an illness, causes phloem necorsis, hinders transportation, retarding of growing, and chlorophyll is reduced.Blade yellow (orange colour) can appear in the wheat after catching an illness, even plant downgrades final symptom such as withered, causes the output heavy losses.
The resistance that couchgrass is belonged to yellow dwarf is studied, in succession at middle couchgrass (Elytrigia intermedia (Nost) Nevski.), overgrow and found some disease-resistant materials in couchgrass (Ely trigiarepens (L) Desv) and the couchgrass (Ag.pulcherrimum), and, bred a collection of resistance to yellow dwarf germplasm that can on wheat breeding, utilize by the chromosome segment that will contain resistant gene with the common wheat hybridization common wheat of successfully having led the people.
(AABBDD is 2n=42) with middle couchgrass (E with common wheat Vilmorin27 for Cauderon
1E
1E
2E
2XX, 2n=42) disome alien addition line L1 (2n=44=21II+1II) (Cauderon Y, B Saigne, M Dauge, et al.In:Proc.4th.Wheat Genet Symp, Missouri.Columbia, 1973,401～407) is bred in hybridization.Studies show that, the additional dyad of L1 is middle couchgrass chromosome 7X, anti-BYDV gene be positioned at 7X long-armed on, be the dominance single-gene, 2 chromosomes (EE) in the couchgrass in the middle of containing among the L1, these 2 chromosomes are carrying several genes such as anti-yellow stunt of wheat, rust.Xin Zhiyong, Xu Huijun, old filial piety etc. are induced the offspring of homoeologous chromosomes reorganization by China spring Ph gene cryptic mutant from being anti-source with L1, select high anti-yellow stunt of wheat new germ plasm PP9-1 (Chinese science B collects, 1991, (1): 36～42).
Wheat powdery mildew (Erysiphe graminis f.sp.tritici) is a kind of worldwide fungal disease, germ passes through air borne, past mainly occurs in gentle, the moist rainy some areas of weather, because water and fertilizer condition improves constantly, planting density strengthens, for the popular good field environment that provides takes place in powdery mildew, cause wheat powdery mildew harm area and occurrence frequency to enlarge year by year, occupy first of the four big diseases such as rust, banded sclerotial blight, head blight.For a long time, people pay attention to the research work of wheat powdery mildew always, cultivating the Basic Ways of disease-resistant variety as the control wheat powdery mildew.From nineteen thirty Australia scholar Water house reported first since wheat breed Thew carries a dominance mildew-resistance gene, the research of disease-resistant gene obtains various countries scientist's extensive concern.Mainly by key-gene control, its gene symbol is Pm (Powdery mildew) to the powder mildew resistance of having reported so far.Identified 30 disease-resistant genes, divided gene to be numbered Pm1, be distributed on 24 sites, comprising 8 multiple alleles to Pm30.Other has 1 gene (Mld) formally not name as yet.Wherein resistant gene is from 15 of common wheat, 10 of tetraploids, and 1 of dliploid wheat, 4 in rye, 3 of goatweeds, 1 of haynaldia villosa is originated uncertain 3.Identify according to the phytopathologist: Pm10,11,14,15 anti-grassy weed powdery mildews, not resist powdery mildew of wheat bacterium.Be Pm2,4b, 2+6,12,13,16,20,21,22,30 etc. to China's wheat powdery mildew resistance efficient gene at present.Wherein the Pm4b gene comes from the tetraploid of common wheat sibling species.
Stripe rust of wheat (puccinia striiformisf.sp.tritic) is a kind of pandemic worldwide disease that is caused by bar shaped handle rest fungus.To China North China, the northwest, southwest, the Yellow River-Huai River region Wheat Production causes regular threat, and other Mai Qu also often have popular.Find 23 anti-bars rust gene locis up to now altogether, called after Yr1-Yr23 (Yellow rest) respectively, comprising 3 multiple alleles, i.e. 26 major gene resistances.Yr1-Yr10, Yr15-Yr23 have been positioned on chromosome or the chromosome arm.These genes can be divided into two types on resistance, Yr1-Yr10, Yr15, Yr17-Yr23 are seedling stage and become strain phase microspecies specialization gene; Yr11-Yr14 and Yr16 are the strain-forming period resistance gene.Can be divided into two classes source from gene source, a class is from common wheat; Another kind of nearly source kind from wheat.Yr5, Yr8, Yr9 and Yr15 be respectively from spelt, tip awn goatweed, rye and wild emmer.All the other genes are all from common wheat.In the Stripe Rust Resistance Gene Yr1-Yr18 that has named, most of genes show as dominant inheritance, but the apparent recessiveness of anti-bar rust gene is according to biological strain in some kinds, hybrid combination (genetic background), the interaction of kind and microspecies and infection period different and show different.Yr2 for example, Yr6, Yr9 shows as recessive inheritance in some hybrid combination.Rich anti-13 is the recommended variety preferably of China's stripe rust resisting.
Wheat leaf rust also is one of main fungal disease of wheat, the end of the seventies, on the south the Shijiazhuang City, Hebei Province leaf rust burst popular, involve Henan and Shandong Mai Qu, output is subjected to heavy losses.Between year, the state of an illness slightly rises and falls later on.During " six or five ", the leaf rust resistance kind accounts for 17.3%, and the enforcement period of the seventh five-year plan is 10.5%, and during " eight or five ", in 2524 parts of kinds (material) that 30 breeding units provide, disease-resistant has only 5.4%.Should strengthen the screening and the utilization in the anti-source of leaf rust.
In recent years, the popular district of stem rust of wheat turns to southwestern Mai Qu from southeastern coast, Yangtze river basin Mai Qu.Black stem rust generally takes place in Mai Qu such as Yunnan, Sichuan, Hubei, Hunan, southwest, Henan, serious plot underproduction 20-30%, and minority is retransmitted the time even has been caused total crop failure.The morbidity plot is in when harvesting, the face of land even be covered with the conspicuous bronzing fungal spore of one deck powder.Puccinia graminis can form from the body circulation, and constantly outwards exports initial bacterium source.
The attribute of above-mentioned 5 kinds of diseases, circulation way difference, onset condition is variant, but can be in the popular harm in same wheat district, even takes place jointly in the same time.Therefore, select for use the kind of the anti-multiple diseases of holding concurrently not only can control the popular of disease, multiple diseases is carried out the comprehensive regulation, the kind that also can improve kind prolongs the service life of kind with being worth.Facts have proved: because source, the background difference of each gene, the synergy of a plurality of genes often is better than term single gene.For example, contain one section chromosomes of rye fragment (1B/1R) in " Lip river class " series of products, this fragment contains Pm8 gene and 3 rust-proofing ospc genes (Yr9, Sr31, Lr26) of mildew-resistance simultaneously, and " Lip river class " kind once was the main anti-source and the parent of China 20th century 80-90 age recommended variety.At present the disease-resistant variety (strain) of report mostly is anti-same disease, or the adding up of the different resistant genes of same disease, and holding concurrently resists the rarely seen report of kind (strain) of two or more disease.
The different gene pyramiding in multiple source or be accumulated in together, general employing is the method for artificial hybridization traditionally, carry out one or many hybridization with carrying heterogeneic material more than 2 or 2 respectively,, breed into colony then from separating the individual plant that the offspring selects to contain gene pyramiding.This method make polymerization or gene time (from generation to generation) in the process of isozygotying of adding up longer, the gene of polymerization is many more, the time is long more, and the kind of breeding always exists heterozygous genes, population resistance poor stability more or less.
Summary of the invention
The breeding method that the purpose of this invention is to provide the anti-disease wheat kind of one kind of multiple disease-resistant gene polymerizations, this method breeding time is short, and the strain resistance of acquisition is stable.
The breeding method of the anti-disease wheat kind of one kind of multiple disease-resistant gene polymerizations may further comprise the steps:
1) the individual plant PP9-1 that selects resistance to yellow dwarf is hybridized, is backcrossed as male parent to promote wheat breed as female parent, selects the female parent of the individual plant of resistance to yellow dwarf as the polymerization of next round disease-resistant gene in backcross progeny;
3) being male parent with the tetraploid that contains the Pm4b gene, is that female parent is hybridized, backcrossed with the popularization wheat breed of stripe rust resisting, selects the male parent of the individual plant of mildew-resistance as the polymerization of next round disease-resistant gene in backcross progeny;
4) the above-mentioned female parent and the male parent that are used for the polymerization of next round disease-resistant gene that obtains hybridized;
5) plant have yellow dwarf and powder mildew resistance is simultaneously selected in hybrid generation's selfing in the self progeny, and the flower pesticide of this plant is cultivated;
6) in the anther culture process through doubling naturally, the common wheat strain of the anti-multiple diseases that obtains holding concurrently.
Popularization wheat breed in the described first step is preferably Shan 7859 or rich resisting No. 8.
Popularization wheat breed in described the 3rd step is preferably rich anti-13, and described backcrossing carries out 3 on behalf of benefit.
Popularization wheat breed with stripe rust resisting in described the 3rd step is after female parent is hybridized, backcrossed, and carries out selfing again, selects the male parent of the individual plant of mildew-resistance as the polymerization of next round disease-resistant gene in the self progeny.Carry out selfing and can improve the economical character and the homozygosity of next round disease-resistant gene polymerization male parent.Described selfing carries out 4 on behalf of benefit.
The described F that is used for anther cultural plant with selfing
2Be advisable for plant.
Because the contained resistant gene resistance in long-term use of common wheat itself descends, and some disease do not have resistant gene in common wheat, and the disease-resistant gene of selecting the wheat sibling species to belong to is that the anti-source in the wheat breeding is the important channel that addresses the above problem.The present inventor selects the filial generation PP9-1 of common wheat-middle couchgrass disome alien addition line L1 dexterously for use and contains the donor of the tetraploid of Pm4b gene as the external source disease-resistant gene through a large amount of experiments.Make the anti-disease wheat that obtains the several genes polymerization become possibility.
Anther cultural purpose is quick homozygous gene, shortens the seed selection time limit.From hybrid F
2Choose suitable flower pesticide in the plant of resistance to yellow dwarf and the powdery mildew held concurrently after identify in the field and cultivate, through doubling naturally, produce regeneration plant in the anther culture process.The genotype of these regeneration plants is all isozygotied, and the offspring is difficult for taking place to show recessive gene segregation phenomenon, genetic stability.Experiment confirm is chosen in F
2Carry out anther culture, the probability that occurs the several genes polymerization in the regeneration plant of anther culture descendant is much higher than F
1Shi Huapei.
The method that the present invention utilizes biotechnology and routine techniques to combine makes a plurality of resistant gene polymerizations, and has occurred indivedual genes generation variations, the phenomenon that resistance strengthens in the seed selection process.With the resistant variety that this method obtains, help limiting the popular of multiple diseases and expansion, increase disease-resistant persistence and the regional adaptability of kind.Have far-reaching theory and practice significance.
Description of drawings
Fig. 1 analyzes collection of illustrative plates for STS.
Embodiment
The cultivation of the anti-disease wheat kind of embodiment 1, multiple disease-resistant gene polymerization
The cultivation of the anti-disease wheat kind of multiple disease-resistant gene polymerization may further comprise the steps:
1, the filial generation PP9-1 with common wheat-middle couchgrass disome alien addition line L1 is a female parent, and Shan 7859 is male parent, hybridizes, and hybrid inoculation in 1 seedling stage in generation carries the aphid of yellow dwarf virus, to identify yellow dwarf resistance;
2, the individual plant of selecting high resistance to yellow dwarf is as female parent, and recommended variety is hybridized as male parent for rich anti-No. 8, from hybrid select in 1 generation that economical character is better, the individual plant of high resistance to yellow dwarf is as the female parent of next round disease-resistant gene polymerization;
3, being male parent with the tetraploid Persian wheat that contains the Pm4b gene, is that female parent is hybridized with the popularization wheat breed of stripe rust resisting rich anti-13;
4,1 generation of hybrid is carried out the seedling stage powder mildew resistance and identify that selecting the individual plant of mildew-resistance is male parent, rich anti-13 backcross for female parent, backcross continuous 3 generations;
5, select the individual plant of anti-hundred powder diseases to carry out 4 generations of selfing, in per generation, carries out powdery mildew and identifies, keeps the plant of mildew-resistance, selects F
4Economical character is better, the individual plant of high mildew-resistance is as the male parent of next round disease-resistant gene polymerization;
6, the above-mentioned female parent and the male parent that are used for the polymerization of next round disease-resistant gene that obtains hybridized;
7, hybrid generation's selfing, F
2Identify yellow dwarf and the powder mildew resistance of plant seedling stage simultaneously;
8, choose suitable flower pesticide in the plant of double certainly resistance to yellow dwarf and powdery mildew, utilize conventional method to carry out anther culture, through doubling naturally, produce regeneration plant in the incubation, concrete grammar is, flower pesticide is inoculated on the C17 medium, produce callus after 30 days, forward on the differential medium about 10 days, differentiate green seedling, forward on the strong seedling culture base in good time, cultivate between 30 days transplant fields.
9, from some regeneration plants through yellow dwarf, powdery mildew, yellow rust, black stem rust with leaf rust is identified and the selection of economical character, select the hold concurrently strain of anti-multiple diseases of common wheat.
The hold concurrently common wheat strain of anti-multiple diseases of embodiment 2, the present invention is identified the resistance of yellow dwarf
1, field resistance is identified
In the field poison (GPV and GAV strain system) aphid is raised in hold concurrently common wheat strain, recommended variety capital, Beijing area 411 and No. 9 each the 50 strain inoculations of middle wheat of anti-multiple diseases of the present invention, 10 of every strain inoculations, observe incidence after 40 days, the hold concurrently common wheat strain of anti-multiple diseases of the present invention is not found susceptible strain, No. 9 50 strains of capital 411 and middle wheat are all susceptible, and susceptible rate all reaches 100%.Illustrate that hold concurrently existing 2 the popular BYDV strains of the high anti-China of common wheat strain of anti-multiple diseases of the present invention are GPV and GAV.
2, genomic in situ hybridization (GISH) is analyzed
In situ hybridization print middle couchgrass chromosome hybridization signal under fluorescence microscope blueness (B) fluorescence excitation is yellow, and the chromosome of wheat background is red.The result of GISH shows: the present invention holds concurrently has 2 chromosome ends yellow hybridization signal to occur in common wheat strain root-tip cells chromosome mitosis metaphase of anti-multiple diseases, remainder chromosome takes on a red color, show these 2 chromosome ends contain in the middle of the dna fragmentation of couchgrass, and this chromosome segment is less, the chromosome configuration 2n=21 II of the common wheat strain of anti-multiple diseases because the present invention holds concurrently, explanation is the wheat-middle couchgrass small fragment translocation line that isozygotys.
3, RFLP Markers for Detection
Select for use and be positioned at the long-armed end of wheat the 7th homoeologous group chromosome, (Devos K M apart from about kinetochore 104cM and the 115cM respectively, M D Atkinson, C N Chinoy, et al.Theor Appl Genet, 1992,83:931～949) 2 probe Psr687 and Wg380, restriction enzyme EcoR I and Dra I carry out the RFLP Markers for Detection.Finding that Psr687 and EcoR I enzyme are cut is combined in the hold concurrently DNA of common wheat strain of anti-multiple diseases of L1 and the present invention and hybridizes the characteristic strip that all shows a 7XL in the banding pattern in the 2.1kb place, and the hold concurrently common wheat strain of anti-multiple diseases of the present invention lacks the 7DL characteristic strip at 8.6kb place; Wg380 and Dra I enzyme are cut and are combined in the present invention and hold concurrently in the DNA hybridization banding pattern of common wheat strain of anti-multiple diseases in the characteristic strip of a 7XL of 10.0kb place demonstration, and lack the 7DL characteristic strip at 6.3kb place.Illustrate that the 7DL chromosome segment of holding concurrently the present invention the common wheat strain of anti-multiple diseases containing Xpsr687 site and Xwg380 site is replaced by the 7X chromosome segment.
The hold concurrently common wheat strain of anti-multiple diseases of embodiment 3, the present invention is identified the resistance of powdery mildew
1, the resistance of wheat powdery mildew is identified
Respectively with the hold concurrently common wheat strain of anti-multiple diseases of the present invention in powdery mildew fungus strain inoculation seedling stage of the different toxicity of 28 biological strains numbering between 11-715, the reaction of investigation resistance, the result shows the high anti-biological strain below No. 213 of common wheat strain that the present invention holds concurrently anti-multiple diseases, and is susceptible to No. 335 later biological strains.Because at present the white powder germ found of China is mainly based on No. 177 following microspecies, toxicity frequency accounts for more than 90%, so the hold concurrently resistance of common wheat strain of anti-multiple diseases of the present invention is still effective.
2, resistant gene genetic analysis
No. 15 microspecies inoculations of wheat powdery mildew qualification result shows the high mildew-resistance of common wheat strain that the present invention holds concurrently anti-multiple diseases.The hold concurrently hybrid F of the common wheat strain of anti-multiple diseases and susceptible variety capital 771, China spring of the present invention
1In generation, is all disease-resistant, hybrid F
2In generation,, anti-, sense strain ratio was respectively 185: 52,123: 47, and anti-sense separates and meets 1 pair of dominant gene segregation ratio of Mendel 3: 1, illustrated that the hold concurrently common wheat strain of anti-multiple diseases of the present invention contains 1 pair of dominance mildew-resistance gene.
Hold concurrently common wheat strain and the notice of invitation for examination of anti-multiple diseases of the present invention shows the F of gene line Pm2, Pm4a, Pm6, Pm12, Pm13, Pm16 test cross
2Anti-sense of generation individual plant ratio is 15: 1, and illustrate has 2 pairs of genes in the hybrid, has the hold concurrently resistant gene of common wheat strain of anti-multiple diseases of this deduction the present invention different with said gene.F with the Pm4b test cross
2In generation, do not found susceptible individual plant, illustrates that the hold concurrently contained gene of common wheat strain of anti-multiple diseases of the present invention is identical with Pm4b.
3, the molecular labeling of mildew-resistance gene
1) structure of anti-sense gene pool (BSA)
From the hold concurrently F of the common wheat strain of anti-multiple diseases and China spring hybridization of the present invention
2In the anti-sense segregation population of No. 15 microspecies of pickup kind powdery mildew, DNA is extracted in disease-resistant and each 10 strain of disease plant of picked at random respectively, and anti-, sense individual plant DNA constitutes disease-resistant pond and susceptible pond in the mixing of equal-volume ratio.
2) STS special primer PCR reaction
Upstream primer P1:5 '-ACGAGTGATGCTCCAGGATATGG-3 ';
Downstream primer P2:5 '-GATCCACCTTTTCCTTGACAAGC-3 ';
Amplification reaction system is 25 μ L, wherein contains template DNA 40ng, 10mmol/L Tris-HCl (pH8.3), 50mmol/L KCl, 2.0mmol/L MgCl
2, 4 kinds of each 100 μ mol/L of dNTP, Taq enzyme 1U, each 10 μ mol/L of primer.Reaction condition is 94 ℃, and 30 circulations are carried out in sex change 1 minute subsequently: 96 ℃ 1 minute, 56 ℃ 1 minute, 72 ℃ 1 minute, last 72 ℃ were extended 10 minutes.After the first round amplification, from amplified production, get 3.0 μ L, carry out second and take turns amplification as template DNA.Amplified production electrophoresis in 1.2% Ago-Gel dyeed in the ethidium bromide about 2 hours, observed on the ultraviolet scanner and photograph.The result as shown in Figure 1, hold concurrently common wheat strain, Pm4a, Pm4b and the disease-resistant pond of anti-multiple diseases, disease-resistant individual plant of the present invention all amplifies the special band of a 1.7kb, and (the 1st swimming lane is Marker among the figure and do not have this band in susceptible variety China spring and susceptible pond, the susceptible individual plant; The 2nd swimming lane is the hold concurrently common wheat strain of anti-multiple diseases of the present invention; The 3rd swimming lane is a China spring; The 4th swimming lane is Khapli/8
*Cc (Pm4a); The 5th swimming lane is VPM/ hundred farmings 3217 * 3 (Pm4b); The 6th swimming lane is disease-resistant pond; The 7th swimming lane is susceptible pond; 8th, 10,12,14,16 swimming lanes are disease-resistant individual plant; 9th, 11,13,15,17 swimming lanes are susceptible individual plant), illustrate that the hold concurrently resistant gene of common wheat strain of anti-multiple diseases of the present invention is Pm4.
The hold concurrently common wheat strain of anti-multiple diseases of embodiment 4, the present invention is identified the resistance of yellow rust
1, identify in the field
With main breed capital, Beijing area 411 is contrast, inoculates CY27, CY28, CY29, CY30, CY31 and the fresh bacterial classification of their mixing with injection, and each every kind is inoculated 25 strains.The 411 abundant anti-sense of investigation plant during morbidity in the capital, by having or not illness to be divided into anti-, sense secondary, check is once again after the heading.The result shows, contrast capital 411 is all susceptible, the present invention hold concurrently anti-multiple diseases the common wheat strain seedling phase with become the strain phase that CY27, CY28, CY29, CY30, CY31 and their mixing microspecies are all showed immunity, illustrate the present invention hold concurrently anti-multiple diseases the common wheat strain to the present popular microspecies of China all highly the opposing.
2, disease-resistant gene genetic analysis
With the present invention hold concurrently the common wheat strain of anti-multiple diseases and 771 hybridization of spring habit sense yellow rust kind capital, obtain F
2Colony.With semar technique to (common wheat strain/capital 771 that the present invention holds concurrently anti-multiple diseases) F
2And the fresh bacterial classification of parents' 2 leaf age seedlings inoculation CY31 microspecies, the 30 days anti-sense of " Invest, Then Investigate " strain numbers move into the greenhouse then, No. 15 biological strains of inoculation powdery mildew behind the slow seedling, the anti-sense of powdery mildew morbidity " Invest, Then Investigate " plant.
The result is: in 87 individual plants, and 66 strain stripe rust resistings, 21 strain sense yellow rust, anti-sense ratio are 3: 1 (as shown in table 1).In 64 strains of mildew-resistance, 48 strain stripe rust resistings, 16 strain sense yellow rust; In 23 strains of sense powdery mildew, 18 strains of stripe rust resisting, 5 strains of sense yellow rust, anti-sense are than being 3: 1, show that the hold concurrently common wheat strain of anti-multiple diseases of the present invention controlled by 1 pair of dominant gene the resistance of CY31.Can see also that from table the hold concurrently common wheat strain of anti-multiple diseases of the present invention is controlled by a pair of dominant gene the resistance of No. 15 microspecies of powdery mildew.The anti-bar of finding simultaneously to hold concurrently is white: anti-bar sense is white: the sense bar is anti-white: the white individual plant ratio of sense bar sense is 9: 3: 3: 1, the ratio that meets two pairs of gene independent allocation illustrates that there is not linkage relationship in the present invention's Stripe Rust Resistance Gene and mildew-resistance gene that contains in the common wheat strain of anti-multiple diseases of holding concurrently in heredity
Table 1, F
2Colony separates the resistance of No. 15 microspecies of yellow rust CY31 and powdery mildew
T
| Total strain number | Stripe rust resisting strain number | Sense yellow rust strain number | Anti-sense ratio R: S | ?????x 2c | |
| Total strain number | ????87 | ????66 | ????21 | ????3∶1 | ?????0.034 |
| The anti-sense ratio of sense white powder diseased plant number R is counted in the mildew-resistance strain: S x 2c | ????64 ????23 ????3∶1 ????0.0345 | ????48 ????18 ????3∶1 ????0.08 | ????16 ????5 ????3∶1 ????0.016 | ????3∶1 ????3∶1 | ?0.0000 ?0.0145 |
X
2 0.05＝3.84
3, RAPD analyzes
Utilize (common wheat strain/capital 771 that the present invention holds concurrently anti-multiple diseases) F
2Anti-sense behind the seedling inoculation CY31 separates plant, and conventional method is extracted DNA, forms anti-sense pond.The resistant, susceptible pond is anti-by 20 strains, the DNA geometric ratio of sense individual plant mixes.Pcr amplification carries out in 25 μ l reactant liquors.Wherein template DNA 50ng, MgCl
21.5mmol/L, dNTP 0.1mmol/L, Tag archaeal dna polymerase 1U, random primer (Operon company product) 0.2 μ mol/L.Totally 45 circulations, the condition of each circulation is: 95 ℃ of 15s, 37 ℃ of 5s, 72 ℃ of 1min.Amplified production is electrophoresis on 1.5% Ago-Gel, has detected 520 primers altogether.From 520 primers, only filtering out a primer OPY08 (5 '-AGGCAGAGCA-3 ') has polymorphism in anti-sense pond.OPY08 can amplify the DNA product of 5 kinds of different lengths on the present invention holds concurrently the common wheat strain of anti-multiple diseases, disease-resistant pond DNA, and in susceptible pond, capital 771 and rich anti-No. 13, amplify the DNA product of 4 kinds of different lengths, compare with disease-resistant pond with the hold concurrently common wheat strain of anti-multiple diseases of the present invention, lack the DNA product of a 700bp length.Can think the present invention hold concurrently the common wheat strain of anti-multiple diseases and this special band OPY08 that disease-resistant pond has
700The entrained stripe rust resisting bacterium CY31 gene-correlation of common wheat strain of holding concurrently anti-multiple diseases with the present invention.
The hold concurrently common wheat strain leaf rust resistance of anti-multiple diseases of embodiment 5, the present invention is identified
With 5389 kinds is contrast, and with injection inoculation leaf rust PHT, THT, mixed bacterias such as PCR, FHP, each every kind is inoculated 25 strains.Investigate anti-sense plant when leaf rust is fully fallen ill, the disease reaction type is checked once after the heading by the record of 0,1,2,3,4 level Four again.The result shows, contrast 5389 is all susceptible, the present invention hold concurrently anti-multiple diseases the common wheat strain seedling phase with become the strain phase that mixed bacterias such as PHT, THT, PCR, FHP are all showed immunity and high anti-, illustrate that the hold concurrently common wheat strain of anti-multiple diseases of the present invention has highly resistivity to leaf rust.
The hold concurrently common wheat strain of anti-multiple diseases of embodiment 6, the present invention is identified the resistance of black rust
With 701 kinds is contrast, and with injection inoculation puccinia graminis bacterium No. 21 and No. 34 little populations, each every kind is inoculated 25 strains.Investigate anti-sense plant when black rust is fully fallen ill, the disease reaction type is checked once after the heading by the record of 0,1,2,3,4 level Four again.The result shows, check variety 701 is all susceptible, the present invention hold concurrently anti-multiple diseases the common wheat strain seedling phase with become the strain phase that puccinia graminis bacterium No. 21 and No. 34 little populations are all showed immunity and high anti-, illustrate that the hold concurrently common wheat strain of anti-multiple diseases of the present invention has highly resistivity to leaf rust.
Claims (6)
1, the breeding method of the anti-disease wheat kind of one kind of multiple disease-resistant gene polymerizations may further comprise the steps:
1) the individual plant PP9-1 that selects resistance to yellow dwarf is hybridized, is backcrossed as male parent to promote wheat breed as female parent, selects the female parent of the individual plant of resistance to yellow dwarf as the polymerization of next round disease-resistant gene in backcross progeny;
3) being male parent with the tetraploid that contains the Pm4b gene, is that female parent is hybridized, backcrossed with the popularization wheat breed of stripe rust resisting, selects the male parent of the individual plant of mildew-resistance as the polymerization of next round disease-resistant gene in backcross progeny;
4) the above-mentioned female parent and the male parent that are used for the polymerization of next round disease-resistant gene that obtains hybridized;
5) plant have yellow dwarf and powder mildew resistance is simultaneously selected in hybrid generation's selfing in the self progeny, and the flower pesticide of this plant is cultivated;
6) in the anther culture process through doubling naturally, the common wheat strain of the anti-multiple diseases that obtains holding concurrently.
2, breeding method according to claim 1 is characterized in that: the popularization wheat breed in the described first step is Shan 7859 or rich resisting No. 8.
3, breeding method according to claim 1 is characterized in that: the popularization wheat breed in described the 3rd step is rich anti-13, and described backcrossing carried out for 3 generations.
4, breeding method according to claim 1, it is characterized in that: the popularization wheat breed with stripe rust resisting in described the 3rd step is after female parent is hybridized, backcrossed, carry out selfing again, in the self progeny, select the male parent of the individual plant of mildew-resistance as the polymerization of next round disease-resistant gene.
5, breeding method according to claim 4 is characterized in that: described selfing carried out for 4 generations.
6, breeding method according to claim 1 is characterized in that: describedly be used for the F that anther cultural plant is selfing
2For plant.
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| CNA03109788XA CN1539263A (en) | 2003-04-21 | 2003-04-21 | Method of cultivating breed of wheat collecting multiple genes of against disease |
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| CNA03109788XA CN1539263A (en) | 2003-04-21 | 2003-04-21 | Method of cultivating breed of wheat collecting multiple genes of against disease |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101558735B (en) * | 2009-06-03 | 2011-09-07 | 西北农林科技大学 | New wheat stripe rust resisting polygene-polymerized flower-nurture breeding method |
| CN103931484A (en) * | 2014-04-04 | 2014-07-23 | 福建农林大学 | Breeding method of widely-resistive wheat variety |
| CN106258937A (en) * | 2016-08-12 | 2017-01-04 | 西北农林科技大学 | A kind of quickly breeding is disease-resistant, the method for good quality wheat new lines |
| CN109924124A (en) * | 2019-04-01 | 2019-06-25 | 江苏丘陵地区镇江农业科学研究所 | A method of lasting powdery-mildew-resistance wheat breeding element is formulated based on multiple gene polymerization |
| CN113179884A (en) * | 2021-04-29 | 2021-07-30 | 山东省农业科学院作物研究所 | Method for cultivating high-yield wheat with gibberellic disease resistance in Huanghuai wheat area based on polygene polymerization |
| CN114711048A (en) * | 2022-05-17 | 2022-07-08 | 北方民族大学 | High-efficiency and stable Chinese wolfberry powdery mildew inoculation method |
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2003
- 2003-04-21 CN CNA03109788XA patent/CN1539263A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101558735B (en) * | 2009-06-03 | 2011-09-07 | 西北农林科技大学 | New wheat stripe rust resisting polygene-polymerized flower-nurture breeding method |
| CN103931484A (en) * | 2014-04-04 | 2014-07-23 | 福建农林大学 | Breeding method of widely-resistive wheat variety |
| CN103931484B (en) * | 2014-04-04 | 2016-05-11 | 福建农林大学 | A kind of selection of wide anti-wheat breed |
| CN106258937A (en) * | 2016-08-12 | 2017-01-04 | 西北农林科技大学 | A kind of quickly breeding is disease-resistant, the method for good quality wheat new lines |
| CN106258937B (en) * | 2016-08-12 | 2018-03-13 | 西北农林科技大学 | A kind of quickly breeding is disease-resistant, the method for good quality wheat strain |
| CN109924124A (en) * | 2019-04-01 | 2019-06-25 | 江苏丘陵地区镇江农业科学研究所 | A method of lasting powdery-mildew-resistance wheat breeding element is formulated based on multiple gene polymerization |
| CN113179884A (en) * | 2021-04-29 | 2021-07-30 | 山东省农业科学院作物研究所 | Method for cultivating high-yield wheat with gibberellic disease resistance in Huanghuai wheat area based on polygene polymerization |
| CN113179884B (en) * | 2021-04-29 | 2022-05-10 | 山东省农业科学院作物研究所 | Method for cultivating high-yield wheat with gibberellic disease resistance in Huanghuai wheat area based on polygene polymerization |
| CN114711048A (en) * | 2022-05-17 | 2022-07-08 | 北方民族大学 | High-efficiency and stable Chinese wolfberry powdery mildew inoculation method |
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