CN102229986B - Method for assisted selection of wheat variety with stripe rust resistance and special PCR reagent used therein - Google Patents
Method for assisted selection of wheat variety with stripe rust resistance and special PCR reagent used therein Download PDFInfo
- Publication number
- CN102229986B CN102229986B CN 201110136905 CN201110136905A CN102229986B CN 102229986 B CN102229986 B CN 102229986B CN 201110136905 CN201110136905 CN 201110136905 CN 201110136905 A CN201110136905 A CN 201110136905A CN 102229986 B CN102229986 B CN 102229986B
- Authority
- CN
- China
- Prior art keywords
- wheat
- stripe rust
- primer pair
- sequence
- pcr
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for screening a wheat variety with stripe rust resistance assisted by molecular markers and special primers used therein. Sequences of the primer pairs provided in the invention for assisted selection of a wheat variety with stripe rust resistance are described in a primer sequence table (Table 1). The method for assisted selection of a wheat variety with stripe rust resistance provided in the invention is carried out on the basis of conventional breeding and causes no interruption to the breeding process of a breeder except that in the generations of F2 and F6, after the selection of the breeder, candidate individual plants are subjected to molecular marker detection and only those individual plants containing two stripe rust resistant genes are reserved, thereby helping the breeder to more effectively select excellent strains containing a plurality of stripe rust resistant genes and enhancing endurance of stripe rust resistance of the strains. According to the invention, a novel stripe rust resistant gene Yrcaas and its special primers are discovered in the wheat variety Zhou 8425B; the gene is on wheat 1B chromosome and is located between SSR mark Xbarc8 and SSR mark Xgwm582, wherein the two marks can be used for assisted breeding of stripe rust resistant plants. The special primers of the stripe rust resistance genes in the invention are important to the breeding of stripe rust resistant wheat.
Description
Technical field
The present invention relates to biological technical field, relate in particular to a kind of method and special-purpose PCR reagent thereof of assist-breeding stripe rust resisting wheat kind.
Background technology
Stripe rust of wheat is a kind of gas biography property leaf diseases that is caused by wheat stripe rust (Puccinia striiformis f.sp.tritici), after occuring, this disease will affect the grouting of plant development and wheat, and then having a strong impact on yield and quality of wheat, there are 4,300 ten thousand hm in the whole world
2Yi Faqu, account for the total cultivated area of wheat 19.4% (
Http:// www.cimmyt.org/Agricdb/fao/Default.aspx).China is maximum in the world Epidemics of Wheat Strip Rust district, approximately 2,000 ten thousand hm
2(Stubbs, 1988; Wan etc., 2004,2007), cool and the high altitude localities particularly serious.Since the 1950's, China 8 stripe rust successively occur and is very popular, wherein being very popular for 1950,1964,1990 and 2002 four times, the generation area is maximum, harm is the heaviest, front twice onset area reaches more than 200,000,000 mu, rear twice morbidity is more than 1.1 hundred million mu, lose respectively wheat 600,320,180 and 1,300,000 tons, bring massive losses (Li Zhenqi etc., 1989 to Wheat Production; Li Shimo, 2001; Li etc., 2000; Wan etc., 2004).Recent years is owing to the reasons such as generally use of climate change, triadimefon, its relative importance descends to some extent, long-pending approximately 0.63 hundred million mu of the every annual generating plane of 2004-09 China stripe rust, but also there are (Zhang Yuejin etc. about 100,000,000 mu in the maximum time, 2005-2009), and along with global warming, Climate Anomalies, stripe rust of wheat probably again rises to and is Major Diseases, thereby the stable high yield of serious threat China wheat.
Totally 51 of the wheat stripe rust resistance genes of at present in the world definite designation are distributed in 48 chromosomal focis, i.e. Yr1-Yr48 (Mcintosh RA etc., 2010).These Stripe Rust Resistance Gene belong to the disease-resistant gene in seedling stage with microspecies specialization mostly, seedling stage, disease-resistant gene was because its high resistance to the stripe rust evil is liked by breeding man deeply, the resistant gene of wheat population great majority are microspecies specialization resistance in recent decades, but the kind that the establishing in large scale disease-resistant gene is single, can accelerate the orthoselection of pathogenic bacteria microspecies, the result causes disease-resistant variety " forfeiture " original resistance after the large-area applications several years on producing, and loses using value (village ingeniously living 1996).In history, because " the Lip river class " and " numerous 6 " of establishing in large scale derivative system, cause in the new pathotype bar that No. 32 (CYR32) physiological strains occur and rise to rapidly being dominant races among No. 31 (CYR31) and bar, thereby caused that national stripe rust of wheat in 2002 is very popular.Recognize that after the potential hazard of single disease-resistant gene, breeding scholar and phytopathologist wish to prolong by methods such as multiple gene polymerization, gene layout and multiline varieties the resistance life-span of disease-resistant gene in seedling stage.If the disease-resistant gene of the different physiological strains of opposing can be aggregated in the kind, this kind just has the ability of the multiple physiological strain of opposing so, namely have multiresistance, lose resistance with regard to being not easy because of the variation of the microspecies of causing a disease like this, thereby the resistance time length is prolonged.Realize this purpose, need on the one hand abundant anti-source, will use on the other hand the easily more effective realization gene pyramiding of method.At present China still major gene of maintaining item rust resistance have Yr5, Yr10, Yr15, Yr24, Yr26 and YrZH84 etc. a few, therefore excavate new anti-source extremely urgent, simultaneously by the auxiliary breeding for disease resistance of molecule marker, thereby more effectively utilize anti-source also unusually important.
Molecular marker assisted selection be by with the objective trait close linkage or be divided into from molecule marker offspring's strain is carried out the screening of target gene or chromosome segment, and then early in generation, can obtain to contain the fine individual plant of target gene, improve breeding efficiency (Fang Xuanjun etc., 2001).Utilize the auxiliary breeding for disease resistance of molecule marker can deduct conventional disease-resistant evaluation work, and then overcome owing to falling ill insufficient or identifying the inaccurate selection mistake of bringing, simultaneously, this technology makes a plurality of disease-resistant gene importing work of detection become possibility, greatly improves the efficient of gene pyramiding.At present, molecular marker assisted selection has obtained some gratifying results in the breeding for disease resistance of world wheat main product state.Will be referred to the rust resistance gene at interior 42 proterties or gene such as the western australia, by Molecular Marker Assisted Selection Technology it has been carried out breed breeding and germplasm improvement (Caki etc., 2008); The Australia south utilizes molecule marker and DH technology, successfully changes the rust-proofing in the wheat breed Annuello and Fineness gene among the susceptible strain Stylet (Kuche etc., 2008); The U.S. is under the support of " wheat cdna is applied to put into practice " project, by the Soybean seed lipoxygenase selection-breeding method, 27 disease-resistant, anti insect genes and 20 high-quality allelotrope have been changed effectively over to (Sorrell etc. in 180 strains of U.S.'s wheat belt, 2007), and the University of California that is arranged in the Davis utilizes the molecule marker ancillary technique successfully to change stripe rust resistance genes Yr17 and brown leaf rust-resistance gene Lr37 over to wheat breed Patwin (Hospita etc., 2009); International corn wheat improvement center (CIMMTY) utilizes molecular marker assisted selection and the conventional breeding method that combines, 25 different Resistants, high-quality and economical character excellent genes are carried out polymerization, so that wheat breed is improved (Willia etc., 2007); Canada has obtained the molecule marker of the proterties such as some rust, smut, head blight, high-quality and fringe germination, at present, pass through molecular marker assisted selection, existing 2 high-quality wheat varieties are promoted, be Lillian and Goodeve (DePauw etc., 2005,2009), and the disease molecule marker utilize work to be in to carry out.In sum, molecule marker (comprising RAPD, RFLP, SSR and RGAP) assisted Selection has been able to widespread use in global wheat breeding for disease resistance, wherein the SSR mark is because the advantage such as its codominance, the high and technology of repeatability be simple is fit to molecular mark.
Though there are some correlative studys in China, the report of concrete Application effect is less.Li Zaifeng etc. (2006) utilize domestic stripe rust Epidemic Races CYR32 that all 8425B are carried out seedling resistance and identify, found that all 8425B carry dominant disease-resistant gene anti-in a pair of performance, and then utilize the SSR mark to be located on 7BL karyomit(e), name and be YrZH84, comprise with the closely linked molecule marker of this gene: Xgwm577, Xwmc276, Xwmc273, Xcfa2040, Xbarc32, Xbarc182 and Xwmc526, genetic distance is that 1.4cM is to 12.0cM, wherein be labeled as Xcfa2040-7B and Xbarc32-7B at the nearest SSR in YrZH84 both sides, genetic distance is respectively 1.4cM and 4.8cM.Yin Guihong etc. (2009) utilize the RGAP mark, by the segregating population fractional analysis method (BSA) based on marker genetype and Phenotypic Selection, obtained and the chain more closely RGA of wheat stripe rust resisting ospc gene YrZH84 mark Xrga-I, with the linkage distance of YrZH84 be 0.8cM.Ren Yan etc. (2010) utilize recently domestic stripe rust Epidemic Races CYR32 that all 8425B are carried out seedling resistance and identify, found that all 8425B carry the new dominant Stripe Rust Resistance Gene of a performance high resistance, and then utilize the SSR mark to be located on 1B karyomit(e), tentative Yrcaas by name, comprise with the closely linked molecule marker of this gene: H20, Xbarc8, Xgwm582, Xgwm131 and Xwmc216, genetic distance is that 0.6cM is to 16.8cM, wherein be labeled as Xbarc8 and Xgwm582 at the nearest SSR in YrZH84 both sides, genetic distance is respectively 0.6cM and 0.7cM, as shown in Figure 1.
Summary of the invention
An object of the present invention is to provide the primer of a kind of assist-breeding stripe rust resisting plant.
Primer provided by the invention is following 1)-4) in any one:
1) primer shown in is comprised of primer pair 1 (Xbarc32), primer pair 2 (Xcfa2040), primer pair 3 (Xbarc8), primer pair 4 (Xgwm582) and primer pair 5 (H20), and primer sequence sees table 1 for details;
2) primer shown in forms by any two kinds among primer pair 1, primer pair 2 and the following A-C: A: primer pair 3, B: primer pair 4 and C: primer pair 5;
3) primer shown in is comprised of primer pair 1, primer pair 2;
4) primer shown in forms by any two kinds among the following A-C: A: primer pair 3, B: primer pair 4 and C: primer pair 5;
The nucleotides sequence of a primer in the described primer pair 1 is classified the sequence 1 in the sequence table as, and the nucleotides sequence of another primer in the described primer pair is classified the sequence 2 in the sequence table as;
The nucleotides sequence of a primer in the described primer pair 2 is classified the sequence 3 in the sequence table as, and the nucleotides sequence of another primer in the described primer pair is classified the sequence 4 in the sequence table as;
The nucleotides sequence of a primer in the described primer pair 3 is classified the sequence 5 in the sequence table as, and the nucleotides sequence of another primer in the described primer pair is classified the sequence 6 in the sequence table as;
The nucleotides sequence of a primer in the described primer pair 4 is classified the sequence 7 in the sequence table as, and the nucleotides sequence of another primer in the described primer pair is classified the sequence 8 in the sequence table as;
The nucleotides sequence of a primer in the described primer pair 5 is classified the sequence 9 in the sequence table as, and the nucleotides sequence of another primer in the described primer pair is classified the sequence 10 in the sequence table as.
Another object of the present invention provides the PCR reagent of assist-breeding stripe rust resisting plant.
Provided by the invention following 1) the PCR reagent of the assist-breeding stripe rust resisting plant shown in arbitrary-4):
1) reagent of PCR shown in is comprised of PCR reagent 1, PCR reagent 2, PCR reagent 3, PCR reagent 4 and PCR reagent 5;
2) reagent of PCR shown in forms by any two kinds among PCR reagent 1, PCR reagent 2 and the following A-C: A:PCR reagent 3, B:PCR reagent 4 and C:PCR reagent 5;
3) reagent of PCR shown in is comprised of PCR reagent 1, PCR reagent 2;
4) reagent of PCR shown in forms by any two kinds among the following A-C: A:PCR reagent 3, B:PCR reagent 4 and C:PCR reagent 5;
Described PCR reagent 1 is comprised of described primer pair 1, dNTP, archaeal dna polymerase and pcr amplification damping fluid;
Described PCR reagent 2 is comprised of described primer pair 2, dNTP, archaeal dna polymerase and pcr amplification damping fluid;
Described PCR reagent 3 is comprised of described primer pair 3, dNTP, archaeal dna polymerase and pcr amplification damping fluid;
Described PCR reagent 4 is comprised of described primer pair 4, dNTP, archaeal dna polymerase and pcr amplification damping fluid;
Described PCR reagent 5 is comprised of described primer pair 5, dNTP, archaeal dna polymerase and pcr amplification damping fluid;
The final concentration of each primer in the PCR at its place reagent is 4pmol in all described primer pairs.
Each final concentration in the PCR at its place reagent described in described PCR reagent 1, described PCR reagent 2, described PCR reagent 3 and the described PCR reagent 4 among the dNTP is 0.2mM;
The final concentration of each among the dNTP in the described PCR reagent 5 is 0.15 μ M;
The final concentration of all described archaeal dna polymerases in the PCR at its place reagent is 0.067U/ μ L.
The pathogenic bacteria of described stripe rust is wheat stripe rust (Puccinia striifirmis f.sp.tritici), and described wheat stripe rust (Puccinia striiformis f.sp.tritici) is specially in wheat stripe rust (Puccinia striiformis f.sp.tritici) the microspecies bar No. 32.
The 3rd purpose of the present invention is the test kit of assist-breeding stripe rust resisting plant.
Provided by the invention following 1) test kit of assist-breeding stripe rust resisting plant shown in arbitrary-4):
1) test kit shown in is for containing described 1) shown in the test kit of PCR reagent;
2) test kit shown in is for containing described 2) shown in the test kit of PCR reagent;
3) test kit shown in is for containing described 3) shown in the test kit of PCR reagent;
4) test kit shown in is for containing described 4) shown in the test kit of PCR reagent.
Described primer or described PCR reagent or the application of described test kit in Resistant breeding also are the scope of protection of the invention;
Or described primer or described PCR reagent or the application of described test kit in assist-breeding stripe rust resisting plant also be the scope of protection of the invention, and described plant is specially wheat; The parent of described wheat is all wheats 22 and all wheats 24.
The pathogenic bacteria of described stripe rust is wheat stripe rust (Puccinia striiformis f.sp.tritici), and described wheat stripe rust (Puccinia striiformis f.sp.tritici) is specially No. 32 physiological strains in the bar.
The 4th purpose of the present invention provides a kind of the discriminating or the auxiliary method of differentiating stripe rust resisting wheat.
Method provided by the invention comprises the steps:
With described 1) shown in primer or described 1) shown in PCR reagent or described 1) shown in described primer pair 1, described primer pair 2, described primer pair 3, described primer pair 4 and described primer pair 5 in the test kit respectively wheat to be measured is carried out pcr amplification, detect pcr amplification product;
If the product of pcr amplification is following A)-in E) 5 kinds, then described wheat to be measured is or the candidate is stripe rust resisting wheat;
If the product of pcr amplification is not following A)-in E) 5 kinds, then described wheat to be measured is or the candidate is non-stripe rust resisting wheat:
A) described primer pair 1 amplification obtains the PCR product of 200bp;
B) described primer pair 2 amplifications obtain the PCR product of 150bp;
C) described primer pair 3 amplifications obtain the PCR product of 560bp;
D) described primer pair 4 amplifications obtain the PCR product of 350bp;
E) described primer pair 5 amplifications obtain the PCR product of 1598bp.
Annealing temperature when described primer pair 1, described primer pair 2, described primer pair 3 and described primer pair 4 carry out pcr amplification is 55 ℃, and annealing time is 1min;
Annealing temperature when described primer pair 5 carries out pcr amplification is 60 ℃, and annealing time is 45S;
Agarose gel electrophoresis is adopted in described detection.
The 5th purpose of the present invention provides the method for a kind of assist-breeding high yield and/or stripe rust resisting wheat.
Method provided by the invention comprises the steps:
1) hybridizes with all wheats 24 and all wheats 22, obtain F
1Generation;
2) with step 1) F that obtains
1Carry out selfing for wheat, obtain F
2For wheat;
3) with described 1) shown in primer or described 1) shown in PCR reagent or 1) shown in described primer pair 1, described primer pair 2, described primer pair 3, described primer pair 4 and described primer pair 5 in the test kit respectively to step 2) F that obtains
2Carry out pcr amplification for wheat, detect pcr amplification product;
If the product of pcr amplification is following A)-in E) 5 kinds, then described wheat to be measured is or the candidate is stripe rust resisting wheat;
If the product of pcr amplification is not following A)-in E) 5 kinds, then described wheat to be measured is or the candidate is non-stripe rust resisting wheat:
A) described primer pair 1 amplification obtains the PCR product of 200bp;
B) described primer pair 2 amplifications obtain the PCR product of 150bp;
C) described primer pair 3 amplifications obtain the PCR product of 560bp;
D) described primer pair 4 amplifications obtain the PCR product of 350bp;
E) described primer pair 5 amplifications obtain the PCR product of 1598bp.
In described step 3) after, also comprise the steps:
4) with step 3) candidate that obtains is stripe rust resisting F
2Carry out selfing for wheat and obtain F
3For wheat, with described F
3Obtain F for the wheat selfing
4For wheat, with described F
4Obtain F for the wheat selfing
5For wheat, with described F
5Obtain F for the wheat selfing
6For wheat;
5) with 1) shown in primer or 1) shown in PCR reagent or 1) shown in test kit to step 4) F that obtains
6Carry out pcr amplification for wheat, obtain the PCR product, detect pcr amplification product,
If the product of pcr amplification is following A)-E) shown in, described F then
2For wheat be or the candidate is stripe rust resisting F
6For wheat;
If the product of pcr amplification is not following A)-E) shown in, described F then
2For wheat be or the candidate is non-stripe rust resisting F
6For wheat:
A) described primer pair 1 amplification obtains the PCR product of 200bp;
B) described primer pair 2 amplifications obtain the PCR product of 150bp;
C) described primer pair 3 amplifications obtain the PCR product of 560bp;
D) described primer pair 4 amplifications obtain the PCR product of 350bp;
E) described primer pair 5 amplifications obtain the PCR product of 1598bp;
6) be stripe rust resisting F with described candidate
6Carry out selfing for wheat, obtain F
7For wheat, screening output is higher than the F of all wheats 22
7For wheat, be high yield, stripe rust resisting wheat;
Step 3) the described pcr amplification in is all with F
2Genomic dna for wheat is template;
Step 5) the described pcr amplification in is all with F
6Genomic dna for wheat is template;
The pathogenic bacteria of described stripe rust is wheat stripe rust (Puccinia striiformis f.sp.tritici), and described wheat stripe rust (Puccinia striiformis f.sp.tritici) is specially in wheat stripe rust (Puccinia striiformis f.sp.tritici) the microspecies bar No. 32;
Agarose gel electrophoresis is all adopted in described detection.
The output of described high yield, stripe rust resisting wheat is compared with all wheats 22, and the ratio of raising is not less than 5%.
The molecule marker that is used for assistantly screening anti-stripe rust wheat that provides that experiment showed, of the present invention has 5, is respectively: Xcfa2040, Xbarc32, Xbarc8, Xgwm582 and H20.The invention provides the method for the auxiliary breeding for disease resistance of a kind of molecule marker, the method is consistent with the process of conventional breeding, and different is that after parents' apolegamy, it carries out Molecular Detection at first to utilize above-mentioned primer pair, and then determines which combination can utilize the method; To these applicable combinations, at F
2The time, candidate's individual plant of breeding man being selected with described 2 SSR marks carries out respectively pcr amplification, detect amplified production, when the purpose fragment that above-mentioned all primer pairs answer is arranged in the amplified production, this individual plant is the candidate's individual plant that contains above-mentioned two Stripe Rust Resistance Gene, only these individual plants is used for next step breeding work; At F
6The time, after the selection of breeding man, again utilize the strain that described 5 pairs of primer pairs isozygoty substantially to carry out Molecular Detection, thereby verify whether selected strain still contains above-mentioned two Stripe Rust Resistance Gene, keep the strain that contains anti-rust gene, further by comprehensive agronomy proterties comparation and assessment, and then obtain the seed materials of the good and high stripe rust resisting of economical character, finally obtain kind.
Description of drawings
Fig. 1 is the linkage map of 5 SSR marks and Yrcaas gene
Fig. 2 is the pedigree that wheat breed is deposited wheat No. 7
Fig. 3 is SSR primer pair F
2For plant with to F
6The amplification of strain
Embodiment
Employed experimental technique is ordinary method if no special instructions among the following embodiment.
Used material, reagent etc. if no special instructions, all can obtain from commercial channels among the following embodiment.
The sequence of used primer is as shown in table 1 below among the following embodiment, is synthetic:
The discovery of a new anti-rust gene Yrcaas and the acquisition of SSR mark thereof among embodiment 1, the wheat lines week 8425B:
One, phenotypic acquisition and colony's wheat seeding situation thereof
With all 8425B and Avocet S hybridization, obtain F
1Colony is with F
1Colony's selfing obtains F
2Colony, F
2Colony is 586 individual plants.With No. 32 physiological strains in the present stripe rust Epidemic Races bar of China experiment material is carried out inoculated identification, the result shows: all 8425B wheat seedings are high resistance (response type IT=0; ) to immunity (IT=0), the AvocetS wheat seeding is high sense (IT=4), F
1Colony all shows as high resistance (IT=0; ), F
2439 strains are disease-resistant individual plant in the colony, and 147 strains are susceptible individual plant, show the dominant gene that carries 1 pair of high stripe rust resisting among all 8425B.According to seedling stage qualification result select 10 extremely disease-resistant individual plants and 10 extremely susceptible individual plants respectively its DNA to be carried out balanced mix to form disease-resistant pond and susceptible pond.
Two, the right acquisition of primer special
Select 655 couples of SSR primer pair parents to carry out the polymorphism screening, all primers are synthetic to be finished by Beijing AudioCodes biotech firm.The result shows at 3 mark H20, Xbarc8 on the 1B karyomit(e) and Xgwm582 all has polymorphism between parent and anti-sense pond.Show that tentatively these marks and Stripe Rust Resistance Gene (Yrcaas) are chain.
Three, the acquisition of linkage map and source thereof
With 3 mark H20, Xbarc8 on the 1B and Xgwm582 to 586 F
2Individual plant increases respectively, obtain colony's genotype, the data obtained utilizes mapping software MapMaker 3.0 and Map Manager QTX20 to carry out linkage analysis, the result shows that these 5 marks are all chain with Stripe Rust Resistance Gene Yrcaas, nearest Xbarc8 and the Xgwm582 of being labeled as apart from the Yrcaas both sides wherein, genetic distance is respectively 9.5cM and 9.6cM, as shown in Figure 1.
The PCR reaction system of 2 SSR marks (being Xbarc8 and Xgwm582) is 15 μ l, that is: 8.8 μ l sterilized waters, 1.5 μ l10 * PCR buffer (remittance day east), 0.3 μ l 10mM dNTPs (remittance day east), every primer 4pmol, Taq archaeal dna polymerase 0.067U/ μ L (remittance day east), the 20ng template DNA; Response procedures is: 94 ℃ of denaturations 5 minutes, and each 94 ℃ of sex change 1min that circulate, 55 ℃ of annealing 1min, 72 ℃ are extended 1min, 35 circulations, last 72 ℃ are extended 10min; The PCR product is stored in 4 ℃; Every part of amplified production adds 3 μ l sex change load sample indicator [98% without the ion methane amide, 10mM EDTA (pH 8.0), 0.1% dimethylbenzene is blue or green, 0.1% tetrabromophenol sulfonphthalein], 95 ℃ of sex change 10 minutes.Point sample 5 μ l carry out electrophoretic separation in 6% denaturing polyacrylamide gel.
The reaction system that silver dyes development STS mark H20 is 20 μ l, contains 10 * PCR buffer, 2 μ l, each 200 μ molL of dNTP (A, T, C, G)
-1, every primer 4pmol, Taq archaeal dna polymerase (TaKaRa) 0.067U/ μ L, template DNA 50ng; The PCR program is 94 ℃ of denaturation 5min; 94 ℃ of sex change 45s, 60 ℃ of annealing 45s, 72 ℃ are extended 1.5min, 38 circulations; 72 ℃ are extended 10min; Pcr amplification product is with 1.5% agarose gel electrophoresis separation detection, and buffering liquid is 1 * TAE solution, 220V voltage electrophoresis 10min, ethidium bromide staining.
3 are labeled as H20, Xbarc8 and Xgwm582, and the primer pair that above-mentioned 3 marks are corresponding is as shown in table 1.
Mark Xbarc32 and Xcfa2040 are for detecting the special marker of Stripe Rust Resistance Gene YrZH84.
Table 1 detection Stripe Rust Resistance Gene YrZH84 in seedling stage and the required primer sequence table of Yrcaas
Annotate: mark Xbarc32 and Xcfa2040 are the primer specials of screening Stripe Rust Resistance Gene YrZH84, and mark Xbarc8, Xgwm582 and H20 are the primer specials of screening Stripe Rust Resistance Gene Yrcaas.
Embodiment 2, deposit the breeding process of No. 7 wheats of wheat
Depositing wheat is for No. 7 semi-winterness, the middle dwarf varietiy of wheat that forms with all wheat 24/ all wheat 22 selection cross, and its pedigree sees Fig. 2 for details.This Varieties In The Seedling Stage is to stripe rust mixed culture performance high resistance, and the strain phase is high stripe rust resisting not only, and high leaf rust resistance, has participated at present 2010-2011 year national the Yellow River and Huai He River sheet prerun.
Its Breeding Process is as follows:
1, F
1Acquisition for wheat
With all wheat 24 (academy of agricultural sciences, Zhoukou City improved variety; obtained kind power protection (CNA002834E) in 2006; 2007 careful by state; all No. 24 features of wheat and cultivation technique; Wang Shigang plants the industry guide, the 05th phase in 2010; the 29th page, the public can deposit the acquisition of wheat improving technology institute from Institute of Crop Science, Chinese Academy of Agricultural Science and sky, Henan Province.) be maternal, all wheats 22 (improved variety of academy of agricultural sciences, Zhoukou City, all wheat 22 high-yield culture techniques, Gu Sumei plants the industry guide, the 1st phase in 2011, the 25-27 page or leaf, the public can deposit the acquisition of wheat improving technology institute from Institute of Crop Science, Chinese Academy of Agricultural Science and sky, Henan Province.) be male parent, hybridize, obtain F
1Generation, F
1In generation, do not eliminate, all F of results after the impurity elimination
1Plant;
2, F
2Acquisition and evaluation for wheat
With F
1Carry out selfing for wheat and obtain F
2For wheat, about program request 1200 strains, in its process of growth, breeding man selects 54 strain F by individual characters (in good, the fringe of snappiness of short stem, the stem stalk large, thousand seed weight 48g) and field performance (fringe is many, filling speed is fast, the Huang that falls good)
2For individual plant.
Extract above-mentioned 54 strain F
2Genomic dna for individual plant, adopt respectively Xbarc32-F in the table 1 of embodiment 1 and Xbarc32-R, Xcfa2040-F and these 2 pairs of primers of Xcfa2040-R to carry out pcr amplification, primer Xbarc32 and Xcfa2040 are for detecting the primer special of Stripe Rust Resistance Gene YrZH84.With all 8425B (Molecular tagging of stripe rust resistance gene YrZH84 in Chinese wheat line Zhou 8425B, Li etc, Theor Appl Genet (2006) 112:1098-1103; The public can deposit the acquisition of wheat improving technology institute from Institute of Crop Science, Chinese Academy of Agricultural Science and sky, Henan Province; Above-mentioned studies show that, all 8425B carry Stripe Rust Resistance Gene YrZH84), (32 make the become rusty present situation in anti-source of its China's wheat bar, Yang Zuomin etc., Acta Agronomica Sinica, the 29th volume, the 2nd phase, in March, 2003,161-168 page or leaf to Avocet S in the rear bar; The susceptible strain Avocet S of Australia does not contain Stripe Rust Resistance Gene YrZH84; The public can deposit the acquisition of wheat improving technology institute from Institute of Crop Science, Chinese Academy of Agricultural Science and sky, Henan Province; ) for contrasting.
The PCR reaction system is 15 μ l, 8.8 μ l sterilized water, 1.5 μ l 10 * PCR buffer (remittance day east), 0.3 μ l 10mM (final concentration is 0.2mM) dNTPs (remittance day east), Taq archaeal dna polymerase 0.067U/ μ L (remittance day east), the 20ng template DNA, every primer 4pmol; The program of described pcr amplification is: 94 ℃ of denaturations 5 minutes; 94 ℃ of sex change 1min, 55 ℃ of annealing 1min, 72 ℃ are extended 1min, 35 circulations; Extend at last 10min.
Every part of pcr amplification product adds 3 μ l sex change load sample damping fluids, mixing, 95 ℃ of sex change 8 minutes.With 6% denaturing polyacrylamide gel electrophoresis separation detection, buffering liquid is: upper groove 0.3 * TBE, lower groove 0.5 * TBE, and power 80W, electrophoresis 50-70min, silver dyes development.
The result is shown in Fig. 3 a and 3b, and wherein 3a is that SSR primer Xcfa2040 is to F
2The amplification of individual plant, Fig. 3 b are that SSR primer Xbarc32 is to F
2The amplification of individual plant, M is Marker, 1-30 is F
2For individual plant.
Can find out from Fig. 3 a, take Xbarc32-F and Xbarc32-R as primer (primer sequence is respectively sequence 1 and sequence 2 in the sequence table), the 1-20 plant obtains the PCR fragment 1 (200bp) shown in the arrow, and the 21-30 plant obtains the PCR fragment 2 (170bp) shown in the arrow; With above-mentioned primer amplification, week, 8425B obtained PCR fragment 1, Avocet S obtains PCR fragment 2, and (specific fragment of disease-resistant plant is PCR fragment 1, the specific fragment of disease plant is PCR fragment 2, consistent with disease-resistant parent and Susceptible parent respectively, thereby can distinguish disease-resistant plant and disease plant from clip size).
Can find out from Fig. 3 b, take Xcfa2040-F and Xcfa2040-R as primer, the 1-20 plant obtains the PCR fragment 3 (150bp) shown in the arrow, and the 21-30 plant obtains the PCR fragment 4 (165bp) shown in the arrow; With above-mentioned primer amplification, all 8425B obtain PCR fragment 3, and Avocet S obtains PCR fragment 4.
Extract above-mentioned 54 strain F
2For the genomic dna of individual plant, adopt respectively Xbarc8-F in the table 1 of embodiment 1 and Xbarc8-R, Xgwm582-F and Xgwm582-R, H20-F and these 3 pairs of primers of H20-R to carry out pcr amplification.Take all 8425B, Avocet S as contrast.
The result is as follows:
Take Xbarc8-F and Xbarc8-R as primer amplification, 1-20 plant F
2Obtain the fragment of PCR fragment 5 (560bp) for individual plant, all 8425B also obtain the specific fragment of 560bp, and Avocet S does not obtain the as above fragment of size.
Take Xgwm582-F and Xgwm582-R as primer amplification, 1-20 plant F
2Obtain the fragment of PCR fragment 6 (350bp) for individual plant, all 8425B also obtain the specific fragment of 350bp, and Avocet S does not obtain the as above fragment of size.
Take H20-F and H20-R as primer amplification, 1-20 plant F
2Obtain the fragment of the fragment of PCR fragment 7 (1598bp) for individual plant, all 8425B also obtain the specific fragment of 1598bp, and Avocet S does not obtain the as above fragment of size.
As can be seen from the above, above-mentioned 1-20 plant obtains PCR fragment 1, PCR fragment 3, PCR fragment 5, PCR fragment 6, PCR fragment 7, this class F
2Be disease-resistant plant for the wheat individual plant, obtain altogether 30 stripe rust resisting F
2For the wheat individual plant; 24 are stripe rust resisting F not
2For the wheat individual plant.
Be stripe rust resisting F with 30 candidates
2Be stripe rust resisting F not for wheat and 24 candidates
2All carry out stripe rust resistance evaluation in seedling stage for wheat, method is as follows: material to be detected is planted in the polypots of 9*9*9,1 part of material to be identified of every alms bowl kind (that is, candidate F
2Seed for harvesting wheat, 20) and the susceptible contrast of 3 strains engrave virtuous 169 (Quantitative Trait Loci Mapping for Adult-Plant Resistance to Stripe Rust in Chinese Landrace Wheat Cultivar Pingyuan 50, Lan CX etc., Phytopathology (2010), 100:313-318; The public can deposit the acquisition of wheat improving technology institute from Institute of Crop Science, Chinese Academy of Agricultural Science and sky, Henan Province.Engrave and virtuous 169 in whole breeding time all strip rust bacteria physiological strains are all shown as high sense, think at present and do not contain any disease-resistant gene), when wheat seeding grows to wholeheartedly a leaf, with sweeping No. 32 ((Puccinaia striiformis f.sp.tritici in wiping manipulation inoculation wheat stripe rust (the Puccinaia striiformis f.sp.tritici) bar, Wan An people, Wu Li people, Jin Shelin, Yao Ge, Wang Baotong. No. 32 name and feature thereof in the Wheat in China Stripe Rust bar. the plant protection journal, 2003,30 (4): 347-352., the public can deposit the acquisition of wheat improving technology institute from Institute of Crop Science, Chinese Academy of Agricultural Science and sky, Henan Province), inoculate rear about 15 days, the documented response type infects the type grade scale and adopts 6 grade standards when treating that susceptible contrast is fully fallen ill, and namely 0,0; , 1,2,3 and 4, and with+and-expression is strong and weak, wherein, 0～2+ type is disease-resistant, 3-～4 types are susceptible (Bariana and McIntosh, 1993).The result proves that 30 candidates are stripe rust resisting F
2Be disease-resistant plant for wheat, 24 candidates are stripe rust resisting F not
2For the equal disease plant of wheat.
The checking of proof present method is correct.
3, F
6Acquisition and evaluation for wheat
Be stripe rust resisting F with above-mentioned 30 candidates
2Obtain F for the equal selfing of wheat
3For wheat, with described F
3Obtain F for the wheat selfing
4For wheat, with described F
4Obtain F for the wheat selfing
5For wheat, with described F
5Obtain F for the wheat selfing
6For wheat;
Extract respectively 35 strain F
6Genomic dna for individual plant, the 5 couples of primer Xbarc32-F that adopt respectively that embodiment 1 obtains and Xbarc32-R, Xcfa2040-F and Xcfa2040-R, Xbarc8-F and Xbarc8-R, Xgwm582-F and Xgwm582-R, H20-F and H20-R carry out pcr amplification, be respectively equipped with contrast, specifically see the result.
The PCR reaction system is the same, and annealing temperature sees Table 1.
The result is shown in Fig. 3 c-3g, and M is Marker, and 1-11 is F
6For individual plant, the red arrow indication is the corresponding fragment of disease-resistant gene among the figure;
3c is that SSR primer pair Xbarc32-F and Xbarc32-R are to F
6The amplification of strain; Take all 8425B, AvocetS, all wheats 22, all wheats 24 as contrast, the 1-7 plant obtains the PCR fragment 1 (200bp) of arrow indication; The 8-11 plant obtains the PCR fragment 2 (170bp) shown in the arrow; With above-mentioned primer amplification, all 8425B and all wheats 22 obtain PCR fragment 1, Avocet S and all wheats 24 and obtain PCR fragment 2.
Fig. 3 d is that SSR primer pair Xcfa2040-F and Xcfa2040-R are to F
6The amplification of strain; Take all 8425B, Avocet S, all wheats 22, all wheats 24 as contrast, the 1-7 plant obtains the PCR fragment 3 (150bp) of arrow indication; The 8-11 plant obtains the PCR fragment 4 (165bp) shown in the arrow; With above-mentioned primer amplification, all 8425B and all wheats 22 obtain PCR fragment 3, Avocet S and all wheats 24 and obtain PCR fragment 4.
Fig. 3 e is that SSR primer pair Xbarc8-F and Xbarc8-R are to F
6The amplification of strain; Take all 8425B, Avocet S, all wheats 22, all wheats 24 as contrast, the 1-5 plant obtains the PCR fragment 5 (560bp) of arrow indication; The 8-10 plant does not amplify above-mentioned specific fragment; With above-mentioned primer amplification, all 8425B and all wheats 22 obtain PCR fragment 5, and Avocet S and all wheats 24 do not amplify this fragment.
Fig. 3 f is that SSR primer pair Xgwm582-F and Xgwm582-R are to F
6The amplification of strain; Take all 8425B, Avocet S, all wheats 22, all wheats 24 as contrast, the 1-7 plant obtains the PCR fragment 6 (350bp) of arrow indication; The 8-11 plant does not amplify above-mentioned specific fragment; With above-mentioned primer amplification, all 8425B and all wheats 22 obtain PCR fragment 6, Avocet S and all wheats 24 and do not amplify this fragment.
Fig. 3 g (: STS primer H20 is to F
6The amplification of strain; Take all 8425B, Avocet S, all wheats 22, all wheats 24 as contrast, the 1-6 plant obtains the PCR fragment 6 (1598bp) of arrow indication; The 8-11 plant does not amplify above-mentioned specific fragment; With above-mentioned primer amplification, all 8425B and all wheats 22 obtain PCR fragment 6, and AvocetS and all wheats 24 do not amplify this fragment.
Can find out, above-mentioned 1-7 plant can amplify specific PCR fragment 1,3,5,6 and 7, therefore thinks that it keeps above-mentioned 2 Stripe Rust Resistance Gene, is the stripe rust resisting plant, and its candidate is stripe rust resisting F
6For wheat, obtaining altogether 11 candidates is stripe rust resisting F
6For wheat; 24 candidates are stripe rust resisting F not
6For wheat.
Be stripe rust resisting F with 11 candidates
6For wheat and 24 candidates stripe rust resisting F not
6All carry out stripe rust resistance evaluation in seedling stage for wheat, method is the same.
The result shows that 11 candidates are stripe rust resisting F
6Be the stripe rust resisting plant for wheat; 24 candidates are stripe rust resisting F not
6All feel the stripe rust plant for wheat.
The checking of proof present method is correct.
4, high yield, disease-resistant Screening and Identification of depositing wheat No. 7
Be stripe rust resisting F with 11 candidates
6Obtain F for the equal selfing of wheat
7For wheat, with candidate F
7Carry out the strain evaluation for the wheat strain and (namely under higher water and fertilizer condition, adopt randomized block design, three repetitions, every residential quarter 6.67m
2, the amount of broadcasting is consistent with production level.Selecting local one or two best kinds or product is contrast, and the contrast of this research is all wheats 22.
Screening obtains the F that output is higher than contrast
7For the wheat family, its called after is deposited wheat No. 7.Wherein, contrast 22 super high-yielding of all wheats, the output three elements are coordinated (400,000 fringe/mu * 36 * 45.0 restrain/thousand), and yielding ability is good.
And the output three elements of depositing wheat No. 7 are/thousand of 410,000 fringe/mu * 36 * 46.5 grams.
Improved seeds are deposited wheat utilizes for No. 7 above-mentioned special primer to carry out Markers for Detection and stripe rust resisting evaluation in seedling stage, prove that this kind contains anti-rust gene YrZH84 and Yrcaas, pedigree analysis shows, all (Markers for Detection shows the resistant gene of this kind from all wheats 22, all wheats contain above-mentioned two disease-resistant genes for No. 22, therefore and all wheats 24 do not contain above-mentioned two disease-resistant genes, infer that the disease-resistant gene of depositing wheat No. 7 is from parent week wheat 22).
Claims (1)
1. the method for a seed selection or assist-breeding stripe rust resisting and/or High-yield Wheat comprises the steps:
1) hybridizes with all wheat 24 wheats and all wheats 22, obtain F
1Generation;
2) F that step 1) is obtained
1Carry out selfing for wheat, obtain F
2For wheat;
3) primer pair 1, described primer pair 2, described primer pair 3, described primer pair 4 and described primer pair 5 are respectively to step 2) F that obtains
2Carry out pcr amplification for wheat, detect pcr amplification product;
If the product of pcr amplification is following A)-in E) 5 kinds, then described F
2For wheat be or the candidate is stripe rust resisting F
2For wheat;
If the product of pcr amplification is not following A)-in E) 5 kinds, then described F
2For wheat be or the candidate is non-stripe rust resisting F
2For wheat:
A) described primer pair 1 amplification obtains the PCR product of 200 bp;
B) described primer pair 2 amplifications obtain the PCR product of 150bp;
C) described primer pair 3 amplifications obtain the PCR product of 560bp;
D) described primer pair 4 amplifications obtain the PCR product of 350bp;
E) described primer pair 5 amplifications obtain the PCR product of 1598bp;
The nucleotides sequence of a primer in the described primer pair 1 is classified the sequence 1 in the sequence table as, and the nucleotides sequence of another primer in the described primer pair is classified the sequence 2 in the sequence table as;
The nucleotides sequence of a primer in the described primer pair 2 is classified the sequence 3 in the sequence table as, and the nucleotides sequence of another primer in the described primer pair is classified the sequence 4 in the sequence table as;
The nucleotides sequence of a primer in the described primer pair 3 is classified the sequence 5 in the sequence table as, and the nucleotides sequence of another primer in the described primer pair is classified the sequence 6 in the sequence table as;
The nucleotides sequence of a primer in the described primer pair 4 is classified the sequence 7 in the sequence table as, and the nucleotides sequence of another primer in the described primer pair is classified the sequence 8 in the sequence table as;
The nucleotides sequence of a primer in the described primer pair 5 is classified the sequence 9 in the sequence table as, and the nucleotides sequence of another primer in the described primer pair is classified the sequence 10 in the sequence table as;
4) candidate who step 3) is obtained is stripe rust resisting F
2Carry out selfing for wheat and obtain F
3For wheat, with described F
3Obtain F for the wheat selfing
4For wheat, with described F
4Obtain F for the wheat selfing
5For wheat, with described F
5Obtain F for the wheat selfing
6For wheat;
5) with the described primer pair 1 to 5 of the step 3) F to obtaining respectively
6Carry out pcr amplification for wheat, obtain the PCR product, detect pcr amplification product;
If the product of pcr amplification is following A)-in E) 5 kinds, then described F
6For wheat be or the candidate is stripe rust resisting F
6For wheat;
If the product of pcr amplification is not following A)-in E) 5 kinds, then described F
6For wheat be or the candidate is non-stripe rust resisting F
6For wheat:
A) described primer pair 1 amplification obtains the PCR product of 200 bp;
B) described primer pair 2 amplifications obtain the PCR product of 150bp;
C) described primer pair 3 amplifications obtain the PCR product of 560 bp;
D) described primer pair 4 amplifications obtain the PCR product of 350 bp;
E) described primer pair 5 amplifications obtain the PCR product of 1598 bp;
6) be stripe rust resisting F with described candidate
6Carry out selfing for wheat, obtain F
7For wheat, screening output is higher than the F of all wheats 22
7For wheat, be high yield, stripe rust resisting wheat;
Described pcr amplification in the step 3) is all with F
2Genomic dna for wheat is template;
Described pcr amplification in the step 5) is all with F
6Genomic dna for wheat is template;
The pathogenic bacteria of described stripe rust be wheat stripe rust (
Puccinia striiformisF. sp.
Tritici);
Agarose gel electrophoresis is all adopted in described detection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110136905 CN102229986B (en) | 2011-05-24 | 2011-05-24 | Method for assisted selection of wheat variety with stripe rust resistance and special PCR reagent used therein |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110136905 CN102229986B (en) | 2011-05-24 | 2011-05-24 | Method for assisted selection of wheat variety with stripe rust resistance and special PCR reagent used therein |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102229986A CN102229986A (en) | 2011-11-02 |
| CN102229986B true CN102229986B (en) | 2013-04-17 |
Family
ID=44842591
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110136905 Expired - Fee Related CN102229986B (en) | 2011-05-24 | 2011-05-24 | Method for assisted selection of wheat variety with stripe rust resistance and special PCR reagent used therein |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102229986B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103789431B (en) * | 2014-01-26 | 2016-01-06 | 甘肃省农业科学院小麦研究所 | Assisting sifting and cultivate the primer special of stripe rust resisting wheat new lines, reagent and test kit and selection thereof |
| CN105200122B (en) * | 2014-11-17 | 2019-01-18 | 中国农业科学院植物保护研究所 | Quantitative detection kit for wheat stripe rust and application thereof |
| CN104774934B (en) * | 2015-04-01 | 2017-07-25 | 周口市农业科学院 | A kind of method for identifying No. 23 wheat breeds of all wheats |
| CN104975015A (en) * | 2015-07-13 | 2015-10-14 | 中国农业科学院作物科学研究所 | Method for assisting in screening stripe-rust-resistant wheat and special primers thereof |
| CN110663542A (en) * | 2019-11-19 | 2020-01-10 | 中国科学院西北高原生物研究所 | Breeding method for long-acting resistance of wheat stem rust Ug99 |
| CN113832249B (en) * | 2021-09-29 | 2023-07-07 | 河南农业大学 | Molecular marker for detecting specific chromosome fragment of wheat diaphysis germplasm 8425B and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1858256A (en) * | 2006-03-03 | 2006-11-08 | 中国农业科学院作物科学研究所 | Method for screening stripe rust resisting wheat and its special primer |
-
2011
- 2011-05-24 CN CN 201110136905 patent/CN102229986B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1858256A (en) * | 2006-03-03 | 2006-11-08 | 中国农业科学院作物科学研究所 | Method for screening stripe rust resisting wheat and its special primer |
Non-Patent Citations (7)
| Title |
|---|
| A Microsatellite Map of Wheat;Marion S. Roder et al.;《Genetics》;19980831(第149期);第2007-2023页 * |
| Marion S. Roder et al..A Microsatellite Map of Wheat.《Genetics》.1998,(第149期),第2007-2023页. |
| 小麦条锈菌鉴别寄主抗条锈病基因Yr9的微卫星标记;翁东旭等;《遗传学报》;20050930;第32卷(第9期);第937-941页 * |
| 抗条锈病小麦基因组中黑麦DNA插入片段的分子检测和序列分析;李春来;《中国优秀硕士学位论文全文数据库 农业科技辑》;20051215(第8期);摘要、第22页 * |
| 李峰奇等.黄淮麦区126 个小麦品种(系)抗条锈病基因的分子检测.《中国农业科学》.2008,第41卷(第10期),第3060-3069页. * |
| 李春来.抗条锈病小麦基因组中黑麦DNA插入片段的分子检测和序列分析.《中国优秀硕士学位论文全文数据库 农业科技辑》.2005,(第8期),摘要、第22页. |
| 翁东旭等.小麦条锈菌鉴别寄主抗条锈病基因Yr9的微卫星标记.《遗传学报》.2005,第32卷(第9期),第937-941页. |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102229986A (en) | 2011-11-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ratnaparkhe et al. | Inheritance of inter-simple-sequence-repeat polymorphisms and linkage with a fusarium wilt resistance gene in chickpea | |
| CN102229986B (en) | Method for assisted selection of wheat variety with stripe rust resistance and special PCR reagent used therein | |
| CN102220430B (en) | Auxiliary screening method for stripe rust-resistance wheat and its special primers | |
| CN103305510A (en) | Rice blast resistance gene Pi9 gene specificity molecular marker Pi9SNP as well as preparation and application thereof | |
| CN103320437A (en) | Gene-specific molecular marker Pi2SNP of rice blast-resistant gene Pi2 as well as preparation method and application thereof | |
| CN103146691A (en) | SNP loci linked with blight resistant gene Fon-1 in watermelon, and markers thereof | |
| CN104313016B (en) | The molecular labeling of the QTL/ major gene resistance relevant with cotton verticillium wilt resistance | |
| CN109321671A (en) | Wheat stripe rust resisting ospc gene QYr.sicau-1B-1 SSR molecular marker and application | |
| CN110512025A (en) | A kind of molecular labeling and its application with powdery mildew resistance gene in wheat PmJM23 close linkage | |
| CN101491212B (en) | Marker-assisted selection quick cultivation method of rice strain containing amylase of middle content | |
| CN114032235B (en) | SSR marker, primer pair, application of primer pair and screening method of SSR marker locus related to upland cotton early-maturing molecular breeding | |
| CN103952403B (en) | The closely linked molecular marker of rice bacterial blight resistance new gene Xa39 | |
| CN110468229A (en) | Rice wide spectrum high resistance to hoja blanca gene Xa45's (t) isolates molecular labeling Hxjy-1 | |
| CN104762298A (en) | Rice seedling-stage salt-tolerant gene qST11 and molecular marker method thereof | |
| CN101659989A (en) | Molecular markers of wheat powdery mildew resistance gene Pm2 and method for acquiring same | |
| CN110273022B (en) | Molecular marker, primer, detection method and application for detecting soybean phytophthora root rot resistance gene | |
| CN103275975A (en) | Wheat new few-tillering QTL (quantitative trait locus), primer pair, molecular marker, molecular marking method and application | |
| CN109576387B (en) | SNP molecular marker of fiber length major gene derived from Xinluzao 24 and Lumian 28 | |
| CN103114140B (en) | Zoysia salt tolerance major gene site qLF-2 molecular marking method | |
| CN111944920A (en) | InDel marker closely linked with melon epidemic disease resistance gene and application thereof | |
| CN103789431A (en) | Special primers, reagent and kit for auxiliary screening and cultivation of stripe rust-resistant new wheat strains, and breeding method of stripe rust-resistant novel wheat strains | |
| CN109355426B (en) | Diagnostic molecular marker for detecting powdery mildew resistance gene Pm2a of wheat and application thereof | |
| CN111073990B (en) | Dominant molecular marker of rice blast resistance gene Pi67(t) and application thereof | |
| CN108085407B (en) | SSR marker closely linked with recessive cotton bud yellow gene v1 and application thereof | |
| CN109554492B (en) | SNP molecular marker of fiber strength major gene from Xinluzao 24 and Lumian 28 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20111102 Assignee: Henan Fengdekang Seed Co., Ltd. Assignor: Institute of crop science of Chinese Academy of Agricultural Sciences|Henan Institute of Wheat Improvement Technology Contract record no.: 2014410000110 Denomination of invention: Method for assisted selection of wheat variety with stripe rust resistance and special PCR reagent used therein Granted publication date: 20130417 License type: Exclusive License Record date: 20141110 |
|
| LICC | Enforcement, change and cancellation of record of contracts on the licence for exploitation of a patent or utility model | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130417 Termination date: 20160524 |
|
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| EM01 | Change of recordation of patent licensing contract |
Change date: 20191029 Contract record no.: 2014410000110 Assignee after: Henan fengdekang Seed Industry Co., Ltd Assignee before: Henan Fengdekang Seed Co., Ltd. |
|
| EM01 | Change of recordation of patent licensing contract |