CN102559911A - Method for assisting in identifying powdery mildew resistant plants and special primers for method - Google Patents
Method for assisting in identifying powdery mildew resistant plants and special primers for method Download PDFInfo
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Abstract
The invention discloses a method for assisting in identifying powdery mildew resistant plants and special primers for the method. The special primers comprises an Xcfd50 primer pair consisting of DNA shown in a sequence 1 and a sequence 2 in a sequence table, and also can comprise an XMag633 primer pair consisting of DNA shown in sequences 3 and 4. The special primers can be used for assisting in identifying the powdery mildew resistant plants. The primer composition and the method can assist in identifying the powdery mildew resistant plants, and have the advantages of simple operation, low cost and capacity of realizing early breeding. The primer composition and the method can be used for genetic breeding of plants, and have significance for breeding new varieties of identifying powdery mildew resistant plants.
Description
Technical field
The present invention relates to method and the primer special thereof of a kind of assistant identification mildew-resistance plant.
Background technology
Wheat is the important food crop of China, with the raising of national food safety, the national economic development, social stability and living standards of the people confidential relation is arranged.Wheat powdery mildew is a kind of global wheat diseases.Along with the popularization of short bar and semi-dwarf mutant wheat breed and the improvement of water and fertilizer condition; The increasing of planting density; Cause the wheatland closing, make Powdery Mildew serious day by day in the harm that China causes, onset area and scope constantly enlarge; Rise to main disease by less important disease, the grain-production of China and safety have been caused serious threat.
Seed selection is also promoted disease-resistant variety and has been acknowledged as control wheat powdery mildew economical, effective and safe the most approach, and the key of breeding resistant variety is constantly to excavate and effectively utilize new anti-source.The wheat breed that cultivation contains a plurality of mildew-resistance genes is to widen to resist to compose, improve resistance and keep one of effective way of resistance.Therefore, excavate new disease-resistant gene and closely linked molecule marker thereof, in time carry out the location work of new disease-resistant gene, significant for marker assisted selection.
The SSR mark is claimed little satellite (Microsatellite) mark again, by one group of 1～6 Nucleotide be cascaded, the simple repeated sequence of multiplicity between 5～10.They are prevalent in the higher organism, and are randomly distributed in the whole genome, owing to the different polymorphums that form dna sequence dna of its multiplicity.The both sides at each SSR seat generally are conservative relatively single-copy sequences, and therefore the polymorphum of this tandem repetitive sequence can easily be carried out the pcr amplification detection through the conserved sequence design special primer at two ends by ten minutes.
The SSR molecular marking technique has following characteristics: (1) site is many, quantity is abundant, covers whole genome; (2) all there are many equipotential forms in each site, and polymorphum is high; (3) majority is the codominant marker, can distinguish to isozygoty and the heterozygous genes type, thereby genetic information more complete on the single site is provided; (4) specification of quality to template DNA is not high, and the DNA consumption is few, is easy to utilize the round pcr analysis; (5) simple to operate, good reproducibility as a result; (6) most SSR marker chromosomes location awares can be applied in the researchs such as the assignment of genes gene mapping, exogenous genetic material evaluation easily.Wheat SSR marker has between wheat sibling species genus, belongs to interior transferability, and promptly most of SSR primer that designs according to the sequence information of common wheat also can play a role in the sibling species genus of wheat, amplifies normal dna fragmentation.
Li etc. (2009) utilize the SSR mark to analyzing from the mildew-resistance gene of wild emmer material IW2; Find that this mildew-resistance gene is between SSR mark Xbarc84 and Xwmc687; Genetic distance is respectively 3.2cM and 2.0cM; Utilize China spring nullisomic-limbs system, both-end system, disappearance based material with this assignment of genes gene mapping on karyomit(e) 3BL, with mildew-resistance gene called after Pm41.Hua etc. (2009) utilize the SSR mark to analyzing among the common wheat mildew-resistance strain P63 that comes from wild emmer; Find that this mildew-resistance gene is between SSR mark Xwmc257 and Xgwm148; Genetic distance is respectively 16cM and 6.7cM; And with this assignment of genes gene mapping on karyomit(e) 2BS, this mildew-resistance gene is named as Pm42.
Summary of the invention
The method and the primer special thereof that the purpose of this invention is to provide a kind of assistant identification mildew-resistance plant.
The invention provides primer to the application of first (Xcfd50 primer to) in the test kit of preparation assistant identification powdery mildew disease-resistant plant; The primer that said primer is made up of single stranded DNA shown in the sequence 2 of single stranded DNA shown in the sequence 1 of sequence table and sequence table first is right.Said plant is a wheat.Said wheat specifically can be wheat lines 87-1, wheat lines IW132, wheat lines agricultural university 212, wheat lines agricultural university 211, is the filial generation and the self-bred progeny thereof of the initial parents offspring's (comprising filial generation, backcross progeny), said offspring and the wheat lines agricultural university 212 (or wheat lines agricultural university 211) that obtain with wheat lines 87-1 and wheat lines IW132.
The present invention also protects said primer to first and primer pair B (the XMag633 primer to) application in the test kit of preparation assistant identification powdery mildew disease-resistant plant; Said primer pair B is right by the primer that single stranded DNA shown in the sequence 4 of single stranded DNA shown in the sequence 3 of sequence table and sequence table is formed.Said plant is a wheat.Said wheat specifically can be wheat lines 87-1, wheat lines IW132, wheat lines agricultural university 212, wheat lines agricultural university 211, is the filial generation and the self-bred progeny thereof of the initial parents offspring's (comprising filial generation, backcross progeny), said offspring and the wheat lines agricultural university 212 (or wheat lines agricultural university 211) that obtain with wheat lines 87-1 and wheat lines IW132.
The present invention also protects the test kit of a kind of assistant identification powdery mildew disease-resistant plant, comprises that said primer is to first.Said test kit also can comprise said primer pair B.
The present invention also protects said primer to the application of first in assistant identification powdery mildew disease-resistant plant.
The present invention also protects said primer to first and the application of said primer pair B in assistant identification powdery mildew disease-resistant plant.
The present invention also protects the method for a kind of assistant identification powdery mildew disease-resistant plant, comprises the step first; Said step first comprises the steps: that with the genomic dna of treating measuring plants be template, with said primer first is carried out pcr amplification; If have two specific fragments in the pcr amplification product, treat that measuring plants is candidate's a powdery mildew disease-resistant plant; If do not have said two specific fragments in the pcr amplification product, treat that measuring plants is candidate's the susceptible plant of Powdery Mildew; One in said two specific fragments is 505-515bp, and another is 485-495bp.
Said method also can comprise step second; Said step second comprises the steps: that with the genomic dna of treating measuring plants be template, carries out pcr amplification with said primer pair B; If have a specific fragment in the pcr amplification product, treat that measuring plants is candidate's a powdery mildew disease-resistant plant; If do not have a said specific fragment in the pcr amplification product, treat that measuring plants is candidate's the susceptible plant of Powdery Mildew; A said dna fragmentation that specific fragment is 565-575bp.
Said plant is a wheat.Said wheat specifically can be wheat lines 87-1, wheat lines IW132, wheat lines agricultural university 212, wheat lines agricultural university 211, is the filial generation and the self-bred progeny thereof of the initial parents offspring's (comprising filial generation, backcross progeny), said offspring and the wheat lines agricultural university 212 (or wheat lines agricultural university 211) that obtain with wheat lines 87-1 and wheat lines IW132.
The qualification result of said step first and said step second can be verified each other.If the same said measuring plants of treating is consistent with the qualification result in the said step second in said step first, this treats that measuring plants is the further candidate's who confirms powdery mildew disease-resistant plant or the further candidate's who confirms the susceptible plant of Powdery Mildew.If samely saidly treat that the qualification result of measuring plants in said step first and said step second is inconsistent, can combine other existing method to confirm this and treat the anti-perception that measuring plants dialogue powder is sick.
(Triticum dicoccoides, 2n=4 *=28 are the secondary GENE SOURCES of common wheat AABB) to wild emmer, and wheat leaf rust, stem rust and Powdery Mildew are had good resistance, in the improvement of disease-resistant wheat sex-controlled inheritance, huge application potential are arranged.Wild emmer is the tetraploid ancestors kind of common wheat, and with common wheat hybridization success easily, hybrid F1 partially or completely can educate, and its disease resistance can be transferred in the common wheat through hybridization and method such as backcross easily.Therefore, fully excavate the anti-source of Powdery Mildew of wild emmer, and it is imported in the common wheat, add up, have important practical significance for the variety and the gene that improve wheat resistance genes.The discovery of mildew-resistance gene Ml7K65 has also further confirmed the effect of wild emmer on the common wheat mildew-resistance, also for the seed selection of new variety of wheat foundation is provided.The present invention has screened 2 and the closely linked molecule marker of mildew-resistance gene Ml7K65: Xcfd50 and Xmag633.These two marks can be used for the marker assisted selection of M17K65 gene effectively.Be labeled as road sign with this, can carry out the Fine Mapping of Ml7K65 gene or through the chromosome walking method near the Ml7K65 gene, thereby lay the first stone for the clone of Ml7K65 gene.
Use primer sets compound provided by the invention and method, can assistant identification powder mildew resistance plant, have advantage easy and simple to handle, with low cost, as can to realize early stage seed selection.Primer sets compound of the present invention and method can be used for plant genetics and breeding, have great value for cultivating the mildew-resistance new variety of plant.
Description of drawings
Fig. 1 identifies the result of the 7K65/ 211-F2 of agricultural university for plant for using the Xcfd50 primer to PCR.
Fig. 2 identifies the result of the 7K65/ 212-F2 of agricultural university for plant for using the Xcfd50 primer to PCR.
Fig. 3 identifies the result of the 7K65/ 211-F2 of agricultural university for plant for using the XMag633 primer to PCR.
Fig. 4 identifies the result of the 7K65/ 212-F2 of agricultural university for plant for using the XMag633 primer to PCR.
Embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique among the following embodiment like no specified otherwise, is ordinary method.Used test materials among the following embodiment like no specified otherwise, is to buy from routine biochemistry reagent shop and obtains.Quantitative test in following examples all is provided with repeated experiments three times, results averaged.
Wheat lines 87-1 (common wheat of easy infection wheat powdery mildew): academy of agricultural sciences, Handan, Hebei province; The public can obtain from China Agricultural University; Reference: Zhang Liansong, Huawei, Guan Haiying, Li Genqiao, Zhang Hongtao, Xie Chaojie, Yang Zuomin, Sun Qixin, the brave wild emmer of Liu Zhi imports the mildew-resistance gene MlWE29 molecule marker location of common wheat, Acta Agronomica Sinica 2009,35 (6): 998-1005.
Wheat lines IW132 (wild emmer of high mildew-resistance): Haifa, Israel university; The public can obtain from China Agricultural University; Reference: Xie Chaojie, Sun Qixin, poplar makes people Israel wild emmer disease resistance in seedling stage and identifies.Wheat Lou Acta Agronomica Sinica 2003,23 (2): 39～42.
Wheat lines Morocco (common wheat of easy infection wheat powdery mildew): the public can obtain from China Agricultural University; Reference: Liu Huiyuan, K Suenaga, He Zhonghu, Wang Zhulin, beam sparkles, and horse is equal, M Bernard, PSourdille, summer elder generation's spring common wheat Powdery Mildew becomes the qtl analysis of strain resistance.Acta Agronomica Sinica 2006,32 (2): 197-202.
Wheat lines agricultural university 212 (common wheat of easy infection wheat powdery mildew): China Agricultural University's agronomy and biotechnology institute wheat group are cultivated new variety of wheat, and the public can buy from Tongzhou District, Beijing City seeds company.
Wheat lines agricultural university 211 (common wheat of easy infection wheat powdery mildew): China Agricultural University's agronomy and biotechnology institute wheat group are cultivated new variety of wheat, and the public can buy from Tongzhou District, Beijing City seeds company.
Wheat lines Xue is (common wheat of easy infection wheat powdery mildew) early: academy of agricultural sciences, Beijing crop institute; The public can obtain from China Agricultural University; Reference: Zhang Liansong, Huawei, Guan Haiying, Li Genqiao, Zhang Hongtao, Xie Chaojie, Yang Zuomin, Sun Qixin, the brave wild emmer of Liu Zhi imports the mildew-resistance gene MlWE29 molecule marker location of common wheat.Acta Agronomica Sinica 2009,35 (6): 998-1005.
E09 physiological strain: the Chinese Academy of Agricultural Sciences Institute of Plant Protection; The public can obtain from China Agricultural University; Reference: Zhang Liansong, Huawei, Guan Haiying, Li Genqiao, Zhang Hongtao, Xie Chaojie, Yang Zuomin, Sun Qixin, the brave wild emmer of Liu Zhi imports the mildew-resistance gene MlWE29 molecule marker location of common wheat.Acta Agronomica Sinica 2009,35 (6): 998-1005.
The division of resistance of plants on wheat Powdery Mildew pathogenic bacterium and perception is dependent on the response type in seedling stage of plant behind the inoculation wheat powdery mildew pathogenic bacterium, and grade scale is seen table 1.
The response type grade scale in seedling stage (Liu et al.1999) of plant behind the table 1 inoculation wheat powdery mildew pathogenic bacterium
The acquisition of embodiment 1, vegetable material
One, the acquisition of disease-resistant parent 7K65
1, with 1 strain wheat lines 87-1 (as male parent) and 1 strain wheat lines IW132 (as female parent) hybridization, obtains 10 seeds, be F1 generation.
2,10 strain F1 are hybridized with 1 strain wheat lines Morocco for wheat (as female parent), obtain 50 seeds, be BC1 generation.
3, with 50 BC1 for planting seed; Inoculate wheat powdery mildew pathogenic bacterium (E09 physiological strain) in seedling stage; Select the inoculation pathogenic bacterium to show as disease-resistant seedling (totally 2 strains) after 15 days; Wait to ear back and 1 strain wheat lines 87-1 hybridization (resistance seedling as maternal, wheat lines 87-1 as male parent) obtain 23 seeds, are BC2 generation.
4, sow 23 BC2 for seed, and gather in the crops selfed seed respectively.
5,23 groups of selfed seeds that step 4 obtained are planted in China Agricultural University's research park experimental field; Every group of seed 1 row; Inoculate wheat powdery mildew pathogenic bacterium (E09 physiological strain) respectively in seedling stage; Wherein capable number the plant for " 7K65 plant " all shows as the resist powdery mildew of wheat that isozygotys, and keeps this row plant (the mildew-resistance material isozygotys) and is used for follow-up study, is disease-resistant parent 7K65.The pedigree of disease-resistant parent 7K65 is: IW132/87-1//Morocco/3/87-1.
Two, the 212-F2 of 7K65/ agricultural university is for the acquisition of disease-resistant segregating population
1, disease-resistant parent 7K65 of 1 strain (as female parent) and 1 strain wheat lines agricultural university 212 (as male parent) are hybridized, obtain 18 seeds, be F1 generation.
2,18 strain F1 are carried out selfing for wheat, obtain 247 seeds, be the 212-F2 of 7K65/ agricultural university generation.
Three, the 211-F2 of 7K65/ agricultural university is for the acquisition of disease-resistant segregating population
1, disease-resistant parent 7K65 of 1 strain (as female parent) and 1 strain wheat lines agricultural university 211 (as male parent) are hybridized, gather in the crops 20 seeds, be F1 generation.
2,20 strain F1 are carried out selfing for wheat, obtain 268 seeds, be the 211-F2 of 7K65/ agricultural university generation.
The powder mildew resistance of embodiment 2, vegetable material is identified
Respectively the 212-F2 of 7K65/ agricultural university is carried out the disease resistance evaluation for plant (247 strain) and the 211-F2 of 7K65/ agricultural university for plant (268 strain), early as susceptible contrast (3 strain), concrete grammar (carries out in the greenhouse) as follows with wheat lines Xue:
1, method inoculation wheat lines Xue who utilizes natural propagation and manual work to stroke Powdery Mildew pathogenic bacterium E09 physiological strain to brush lightly (stroke and brush lightly 1 time every day during growth of seedling) early; Repeat repeatedly to inoculate; Thicker subiculum appears on wheat leaf blade; Big volume production spore and scab are linked to be sheet, promptly can be used as numerous bacterium basin.
2, seed to be measured (seed) is planted in plastic seeding culturing plate, about 15 of every cave plantations, seed is sprouted also and is grown to a leaf during one heart stage, and numerous bacterium basin is placed around the seedling pan, and Powdery Mildew pathogenic bacterium spore is inoculated the seedling of plant to be measured through natural propagation.
3, inoculation (placing numerous bacterium basin) is back 15 days, and susceptible contrast this moment is fully fallen ill, and according to the disease resistance of observed and recorded plant to be measured, after 3 days, checks once its disease resistance of accurate recording.
The result shows that the result of twice observed and recorded plant disease resistance to be measured is consistent.The 247 strain 7K65/ 212-F2 of agricultural university are disease-resistant for there being 174 individual plants to show as in the plant, and 73 individual plant performances are susceptible, and the Chi-square test result shows χ
2 3: 1=2.73, (χ
2 0.05, 1=3.840) (P＜0.05) meets 3: 1 single-gene segregation ratio, shows that the mildew-resistance proterties of this combination is controlled by the dominance single-gene.The 268 strain 7K65/ 211-F2 of agricultural university are disease-resistant for there being 196 individual plants to show as in the plant, and 72 individual plant performances are susceptible, and the Chi-square test result shows χ
2 3: 1=0.497, (χ
2 0.05,1=3.84) (P＜0.05) meets 3: 1 single-gene segregation ratio, shows that also the mildew-resistance proterties of this combination is controlled by the dominance single-gene.Temporarily this disease-resistant gene is named and be Ml7K65.
The PCR of embodiment 3, vegetable material identifies
One, the association analysis of SSR mark and plant powdery mildew resistance
Utilize cluster compartment analysis (Bulked Segregant Analysis; BSA) method makes up anti-, sense pond: from F2 for 10 strains of picking the individual plant isozygoty disease-resistant individual plant and 10 strains isozygoty susceptible individual plant DNA respectively balanced mix form disease-resistant pond of DNA and the susceptible pond of DNA, be used to screen the molecule marker chain with disease-resistant gene.Final find the Xcfd50 primer to XMag633 primer pair and Ml7K65 gene close linkage, thereby can adopt the powder mildew resistance of these two pairs of primer assistant identification plants.
Two, the preparation of primer sets compound
The primer sets compound by the Xcfd50 primer to the XMag633 primer to forming.
The Xcfd50 primer is following to the sequence of the upstream and downstream primer of (annealing temperature that is applied to PCR is 60.0 ℃):
Upstream primer (sequence 1 of sequence table): 5 '-TTCTGCAACATTTTGTCCCA-3 ';
Downstream primer (sequence 2 of sequence table): 5 '-CGTATGATCCTAACGAGGGC-3 '.
The XMag633 primer is following to the sequence of the upstream and downstream primer of (annealing temperature that is applied to PCR is 52.0 ℃):
Upstream primer (sequence 3 of sequence table): 5 '-CGCCAAAACTCTGGGACG-3 ';
Downstream primer (sequence 4 of sequence table): 5 '-GAAAGAAGGACCGTGCTACAAG-3 '.
Synthetic respectively above four primers.
Three, the PCR of vegetable material identifies (use Xcfd50 primer to)
Respectively the 212-F2 of 7K65/ agricultural university is carried out the PCR evaluation for plant (247 strain) and the 211-F2 of 7K65/ agricultural university for plant (268 strain); Adopt wheat lines 87-1, wheat lines agricultural university 212 and wheat lines agricultural university 211 as negative control; Wheat lines IW132 and disease-resistant parent 7K65 are as positive control, and concrete grammar is following:
1, the blade of each seedling among the clip embodiment 2 extracts genomic dna.
2, the genomic dna with step 1 is a template, to carrying out pcr amplification, obtains pcr amplification product with the Xcfd50 primer.
3, the pcr amplification product with step 2 carries out 8% polyacrylamide gel electrophoresis, utilizes argentation to dye then, last statistical experiment result.Partial results is seen Fig. 1 (on behalf of disease-resistant individual plant, S, R represent susceptible individual plant).
In 174 disease-resistant individual plants, two specific bands of 163 strain electrophoresis showed 510bp and 490bp are arranged.In 73 susceptible individual plants, have only two specific bands of 13 strain electrophoresis showed 510bp and 490bp.The result shows, uses the Xcfd50 primer genomic dna to plant to be measured is carried out pcr amplification, and the dna fragmentation that whether has 510bp and 490bp according to amplified production can assistant identification plant to be measured still be the Powdery Mildew disease plant for the powdery mildew disease-resistant plant.Each pcr amplification product is carried out sequence verification, consistent with electrophoresis result.
Four, the PCR of vegetable material identifies (use XMag633 primer to)
Respectively the 212-F2 of 7K65/ agricultural university is carried out the PCR evaluation for plant (247 strain) and the 211-F2 of 7K65/ agricultural university for plant (268 strain); Adopt wheat lines 87-1, wheat lines agricultural university 212 and wheat lines agricultural university 211 as negative control; Wheat lines IW132 and disease-resistant parent 7K65 are as positive control, and concrete grammar is following:
1, the blade of each seedling among the clip embodiment 2 extracts genomic dna.
2, the genomic dna with step 1 is a template, to carrying out pcr amplification, obtains pcr amplification product with the XMag633 primer.
3, the pcr amplification product with step 2 carries out 8% polyacrylamide gel electrophoresis, utilizes argentation to dye then, last statistical experiment result.Partial results is seen Fig. 2 (on behalf of disease-resistant individual plant, S, R represent susceptible individual plant).Each pcr amplification product is carried out sequence verification, consistent with electrophoresis result.
In 174 disease-resistant individual plants, the specific band of 171 strain electrophoresis showed 570bp is arranged.In 73 susceptible individual plants, has only the specific band of 9 strain electrophoresis showed 570bp.The result shows, uses the XMag633 primer genomic dna to plant to be measured is carried out pcr amplification, and the dna fragmentation that whether has 570bp according to amplified production can assistant identification plant to be measured still be the Powdery Mildew disease plant for the powdery mildew disease-resistant plant.
Claims (10)
1. primer is to the application of first in the test kit of preparation assistant identification powdery mildew disease-resistant plant; The primer that said primer is made up of single stranded DNA shown in the sequence 2 of single stranded DNA shown in the sequence 1 of sequence table and sequence table first is right.
2. primer is to first and the application of primer pair B in the test kit of preparation assistant identification powdery mildew disease-resistant plant; The primer that said primer is made up of single stranded DNA shown in the sequence 2 of single stranded DNA shown in the sequence 1 of sequence table and sequence table first is right; Said primer pair B is right by the primer that single stranded DNA shown in the sequence 4 of single stranded DNA shown in the sequence 3 of sequence table and sequence table is formed.
3. the test kit of an assistant identification powdery mildew disease-resistant plant comprises that primer is to first; The primer that said primer is made up of single stranded DNA shown in the sequence 2 of single stranded DNA shown in the sequence 1 of sequence table and sequence table first is right.
4. test kit as claimed in claim 3 is characterized in that: said test kit also comprises primer pair B; Said primer pair B is right by the primer that single stranded DNA shown in the sequence 4 of single stranded DNA shown in the sequence 3 of sequence table and sequence table is formed.
5. primer is to the application of first in assistant identification powdery mildew disease-resistant plant; The primer that said primer is made up of single stranded DNA shown in the sequence 2 of single stranded DNA shown in the sequence 1 of sequence table and sequence table first is right.
Primer to first and with the application of primer pair B in assistant identification powdery mildew disease-resistant plant; The primer that said primer is made up of single stranded DNA shown in the sequence 2 of single stranded DNA shown in the sequence 1 of sequence table and sequence table first is right; Said primer pair B is right by the primer that single stranded DNA shown in the sequence 4 of single stranded DNA shown in the sequence 3 of sequence table and sequence table is formed.
7. the method for an assistant identification powdery mildew disease-resistant plant comprises the step first;
Said step first comprises the steps: that with the genomic dna of treating measuring plants be template, with primer first is carried out pcr amplification; If have two specific fragments in the pcr amplification product, treat that measuring plants is candidate's a powdery mildew disease-resistant plant; If do not have said two specific fragments in the pcr amplification product, treat that measuring plants is candidate's the susceptible plant of Powdery Mildew; One in said two specific fragments is 505-515bp, and another is 485-495bp; The primer that said primer is made up of single stranded DNA shown in the sequence 2 of single stranded DNA shown in the sequence 1 of sequence table and sequence table first is right.
8. method as claimed in claim 7 is characterized in that: said method also comprises step second;
Said step second comprises the steps: that with the genomic dna of treating measuring plants be template, carries out pcr amplification with primer pair B; If have a specific fragment in the pcr amplification product, treat that measuring plants is candidate's a powdery mildew disease-resistant plant; If do not have a said specific fragment in the pcr amplification product, treat that measuring plants is candidate's the susceptible plant of Powdery Mildew; A said dna fragmentation that specific fragment is 5635-575bp; Said primer pair B is right by the primer that single stranded DNA shown in the sequence 4 of single stranded DNA shown in the sequence 3 of sequence table and sequence table is formed.
9. method as claimed in claim 8; It is characterized in that: if the same said measuring plants of treating is consistent with the qualification result in the said step second in said step first, this treats that measuring plants is the further candidate's who confirms powdery mildew disease-resistant plant or the further candidate's who confirms the susceptible plant of Powdery Mildew.
10. like claim 1 or 2 or 5 or 6 described application, or like claim 3 or 4 described test kits, or like arbitrary described method in the claim 7 to 9, it is characterized in that: said plant is a wheat.
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| CN108342505A (en) * | 2018-04-25 | 2018-07-31 | 中国农业科学院作物科学研究所 | The relevant chromosome segment of leaf rust resistance and its application |
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| CN108165656A (en) * | 2017-11-08 | 2018-06-15 | 河南丰德康种业有限公司 | Wheat molecular marker and its application in wheat powdery mildew resistance is identified |
| CN108165656B (en) * | 2017-11-08 | 2021-07-27 | 河南丰德康种业有限公司 | Wheat molecular marker and application thereof in identification of wheat powdery mildew resistance |
| CN108342505A (en) * | 2018-04-25 | 2018-07-31 | 中国农业科学院作物科学研究所 | The relevant chromosome segment of leaf rust resistance and its application |
| CN108342505B (en) * | 2018-04-25 | 2021-06-22 | 中国农业科学院作物科学研究所 | Chromosome segment related to leaf rust resistance and application thereof |
| CN108690883A (en) * | 2018-08-17 | 2018-10-23 | 华南农业大学 | A kind of molecular labeling RMD7 of the soybean powder mildew resistance of auxiliary identification soybean to be measured |
| CN108690883B (en) * | 2018-08-17 | 2020-09-15 | 华南农业大学 | Molecular marker RMD7 for assisting in identifying soybean powdery mildew resistance of soybean to be detected |
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