CN102559911B - Method for assisting in identifying powdery mildew resistant plants and special primers for method - Google Patents
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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, close relationship 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 day by day serious in the harm that China causes, onset area and scope constantly enlarge, rise to Major Diseases by Minor diseases, 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 anti-spectrum, 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 claims again little satellite (Microsatellite) mark, by one group of 1～6 Nucleotide be cascaded, the simple repeated sequence of multiplicity between 5～10.They are prevalent in higher organism, and are randomly distributed in whole genome, due to the different polymorphisms that form DNA sequence dna of its multiplicity.The both sides at each SSR seat are generally the single-copy sequences of relatively guarding, therefore the polymorphism of this tandem repetitive sequence can be very easily conserved sequence design special primer by two ends carry out pcr amplification and detect.
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 polymorphism is high; (3) majority is the codominant marker, can distinguish to isozygoty and the heterozygous genes type, thereby more complete genetic information on Single locus 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 namely 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 nulli-tetrasomes 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 in 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 the application of primer pair first (Xcfd50 primer pair) in the test kit of preparation assistant identification powdery mildew disease-resistant plant; The primer pair that described primer pair first is comprised of single stranded DNA shown in the sequence 2 of single stranded DNA shown in the sequence 1 of sequence table and sequence table.Described plant is wheat.Described wheat specifically can be wheat lines 87-1, wheat lines IW132, wheat lines agricultural university 212, wheat lines agricultural university 211, filial generation and the self-bred progeny thereof of the offspring's (comprising filial generation, backcross progeny) who obtains as initial parents take wheat lines 87-1 and wheat lines IW132, described offspring and wheat lines agricultural university 212 (or wheat lines agricultural university 211).
The present invention also protects described primer pair first and the application of primer pair B (XMag633 primer pair) in the test kit of preparation assistant identification powdery mildew disease-resistant plant; The primer pair that described primer pair B is comprised of single stranded DNA shown in the sequence 4 of single stranded DNA shown in the sequence 3 of sequence table and sequence table.Described plant is wheat.Described wheat specifically can be wheat lines 87-1, wheat lines IW132, wheat lines agricultural university 212, wheat lines agricultural university 211, filial generation and the self-bred progeny thereof of the offspring's (comprising filial generation, backcross progeny) who obtains as initial parents take wheat lines 87-1 and wheat lines IW132, described offspring and wheat lines agricultural university 212 (or wheat lines agricultural university 211).
The present invention also protects the test kit of a kind of assistant identification powdery mildew disease-resistant plant, comprises described primer pair first.Described test kit also can comprise described primer pair B.
The present invention also protects the application of described primer pair first in assistant identification powdery mildew disease-resistant plant.
The present invention also protects described primer pair first and the application of described primer pair B in assistant identification powdery mildew disease-resistant plant.
The present invention also protects a kind of method of assistant identification powdery mildew disease-resistant plant, comprises the step first; Described step first comprises the steps: to treat that the genomic dna of measuring plants is template, carries out pcr amplification with described primer pair first; If have two specific fragments in pcr amplification product, treat that measuring plants is candidate's powdery mildew disease-resistant plant; If do not have described two specific fragments in pcr amplification product, treat that measuring plants is candidate's Powdery Mildew diseased plant; One in described two specific fragments is 505-515bp, and another is 485-495bp.
Described method also can comprise step second; Described step second comprises the steps: to treat that the genomic dna of measuring plants is template, carries out pcr amplification with described primer pair B; If have a specific fragment in pcr amplification product, treat that measuring plants is candidate's powdery mildew disease-resistant plant; If do not have a described specific fragment in pcr amplification product, treat that measuring plants is candidate's Powdery Mildew diseased plant; A described DNA fragmentation that specific fragment is 565-575bp.
Described plant is wheat.Described wheat specifically can be wheat lines 87-1, wheat lines IW132, wheat lines agricultural university 212, wheat lines agricultural university 211, filial generation and the self-bred progeny thereof of the offspring's (comprising filial generation, backcross progeny) who obtains as initial parents take wheat lines 87-1 and wheat lines IW132, described offspring and wheat lines agricultural university 212 (or wheat lines agricultural university 211).
The qualification result of described step first and described step second can be verified mutually.If samely describedly treat that measuring plants is consistent with the qualification result in described step second in described step first, this treats that measuring plants is the candidate's that further determines powdery mildew disease-resistant plant or the candidate's that further determines Powdery Mildew diseased plant.If samely describedly treat that the qualification result of measuring plants in described step first and described step second is inconsistent, can determine the anti-perception that this treats measuring plants dialogue powder disease in conjunction with other existing method.
Wild emmer (Triticum dicoccoides, 2n=4 *=28, AABB) be the secondary GENE SOURCES of common wheat, 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 is arranged.Wild emmer is the tetraploid ancestors kind of common wheat, and with easily success of common wheat hybridization, hybrid F1 partially or completely can educate, and its disease resistance can be transferred in common wheat easily by hybridization and the method such as backcross.Therefore, fully excavate the anti-source of Powdery Mildew of wild emmer, and it is imported in common wheat, cumulative for the diversity that improves wheat resistance genes and gene, have important practical significance.The discovery of mildew-resistance gene Ml7K65 has also further confirmed the effect of wild emmer on the common wheat mildew-resistance, and also the seed selection for new variety of wheat provides foundation.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 by the chromosome walking method near the Ml7K65 gene, thereby lay the first stone for the Ml7K65 gene cloning.
Use primer sets compound provided by the invention and method, can assistant identification powder mildew resistance plant, have advantages of easy and simple to handle, with low cost, can 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 that for using Xcfd50 primer pair PCR the 7K65/ 211-F2 of agricultural university is for the result of plant.
Fig. 2 identifies that for using Xcfd50 primer pair PCR the 7K65/ 212-F2 of agricultural university is for the result of plant.
Fig. 3 identifies that for using XMag633 primer pair PCR the 7K65/ 211-F2 of agricultural university is for the result of plant.
Fig. 4 identifies that for using XMag633 primer pair PCR the 7K65/ 212-F2 of agricultural university is for the result of plant.
Embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique in following embodiment if no special instructions, is ordinary method.Test materials used in following embodiment if no special instructions, is and purchases available from routine biochemistry reagent shop.Quantitative test in following examples all arranges 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 Identification in seedling stage.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 is glittering, and horse is equal, M Bernard, PSourdille, the qtl analysis of first spring common wheat Powdery Mildew strain resistance of summer.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 the resistance of plants on wheat Powdery Mildew pathogenic bacterium and perception is dependent on the response type in seedling stage of plant after inoculation wheat powdery mildew pathogenic bacterium, and grade scale sees Table 1.
The response type grade scale in seedling stage (Liu et al.1999) of plant after 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, obtain 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, Seedling Inoculation wheat powdery mildew pathogenic bacterium (E09 physiological strain), select the inoculation pathogenic bacterium to show as disease-resistant seedling (totally 2 strains) after 15 days, after ear with 1 strain wheat lines 87-1 hybridization (resistance seedling as maternal, wheat lines 87-1 as male parent), obtain 23 seeds, be BC2 generation.
4, sow 23 BC2 for seed, and gather in the crops respectively selfed seed.
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, respectively Seedling Inoculation wheat powdery mildew pathogenic bacterium (E09 physiological strain), wherein line number all shows as for the plant of " 7K65 plant " resist powdery mildew of wheat that isozygotys, keep this row plant (the mildew-resistance material isozygotys) and be used for follow-up study, be disease-resistant parent 7K65.The pedigree of disease-resistant parent 7K65 is: IW132/87-1//Morocco/3/87-1.
Two, the 7K65/ 212-F2 of agricultural university is for the acquisition of disease-resistant segregating population
1, the 1 disease-resistant parent 7K65 of strain (as female parent) and 1 strain wheat lines agricultural university 212 (as male parents) are hybridized, obtain 18 seeds, be F1 generation.
2,18 strain F1 are carried out selfing for wheat, obtain 247 seeds, be the 7K65/ 212-F2 of agricultural university generation.
Three, the 7K65/ 211-F2 of agricultural university is for the acquisition of disease-resistant segregating population
1, the 1 disease-resistant parent 7K65 of strain (as female parent) and 1 strain wheat lines agricultural university 211 (as male parents) 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 7K65/ 211-F2 of agricultural university generation.
The powder mildew resistance of embodiment 2, vegetable material is identified
Respectively the 7K65/ 212-F2 of agricultural university is carried out Disease Resistance Identification for plant (247 strain) and the 7K65/ 211-F2 of agricultural university for plant (268 strain), with wheat lines Xue early as susceptible contrast (3 strain), concrete grammar following (carrying out in the greenhouse):
1, Powdery Mildew pathogenic bacterium E09 physiological strain utilized natural propagation and manually stroke method inoculation wheat lines Xue morning of brushing lightly (stroke and brush lightly 1 time every day during growth of seedling), repeatedly inoculation, until occur thicker subiculum on wheat leaf blade, large volume production spore and scab are linked to be sheet, namely can be used as numerous bacterium basin.
2, seed to be measured (seed) is planted in plastic seeding culturing plate, 15 left and right of plantation, every cave, seed is sprouted and is grown to a leaf during one heart stage, and numerous bacterium basin is placed in the seedling pan surrounding, and Powdery Mildew pathogenic bacterium spore is inoculated the seedling of plant to be measured by natural propagation.
3, inoculation (placing numerous bacterium basin) is rear 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, checks once afterwards its disease resistance of accurate recording after 3 days.
Result shows, 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 plant, and 73 individual plant performances are susceptible, and different sexes shows χ
2 3: 1=2.73, (χ
2 0.05, 1=3.840) (P＜0.05) meets the single-gene segregation ratio of 3: 1, shows that the mildew-resistance proterties of this combination is by dominant Dominant gene.The 268 strain 7K65/ 211-F2 of agricultural university are disease-resistant for there being 196 individual plants to show as in plant, and 72 individual plant performances are susceptible, and different sexes shows χ
2 3: 1=0.497, (χ
2 0.05,1=3.84) (P＜0.05) meets the single-gene segregation ratio of 3: 1, shows that also the mildew-resistance proterties of this combination is by dominant Dominant 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 builds anti-, sense pond: from F2 for 10 strains of picking individual plant isozygoty disease-resistant individual plant and 10 strains isozygoty susceptible individual plant DNA respectively balanced mix form the disease-resistant pond of DNA and DNA susceptible pond, be used for the screening molecule marker chain with disease-resistant gene.Final discovery Xcfd50 primer pair and XMag633 primer pair and Ml7K65 gene close linkage, thus the powder mildew resistance of these two pairs of primer assistant identification plants can be adopted.
Two, the preparation of primer sets compound
The primer sets compound is comprised of Xcfd50 primer pair and XMag633 primer pair.
The sequence of the upstream and downstream primer of Xcfd50 primer pair (annealing temperature that is applied to PCR is 60.0 ℃) is as follows:
Upstream primer (sequence 1 of sequence table): 5 '-TTCTGCAACATTTTGTCCCA-3 ';
Downstream primer (sequence 2 of sequence table): 5 '-CGTATGATCCTAACGAGGGC-3 '.
The sequence of the upstream and downstream primer of XMag633 primer pair (annealing temperature that is applied to PCR is 52.0 ℃) is as follows:
Upstream primer (sequence 3 of sequence table): 5 '-CGCCAAAACTCTGGGACG-3 ';
Downstream primer (sequence 4 of sequence table): 5 '-GAAAGAAGGACCGTGCTACAAG-3 '.
Synthesize respectively above four primers.
Three, the PCR of vegetable material identifies (using the Xcfd50 primer pair)
Respectively the 7K65/ 212-F2 of agricultural university is carried out the PCR evaluation for plant (247 strain) and the 7K65/ 211-F2 of 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 as follows:
1, the blade of each seedling in clip embodiment 2 extracts genomic dna.
2, take the genomic dna of step 1 as template, carry out pcr amplification with the Xcfd50 primer pair, obtain pcr amplification product.
3, the pcr amplification product with step 2 carries out 8% polyacrylamide gel electrophoresis, then utilizes argentation to dye, last statistical experiment result.Partial results is seen Fig. 1 (R represents that disease-resistant individual plant, S 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, only have two specific bands of 13 strain electrophoresis showed 510bp and 490bp.Result shows, uses the Xcfd50 primer pair genomic dna of 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 be powdery mildew disease-resistant plant or Powdery Mildew disease plant.Each pcr amplification product is carried out sequence verification, consistent with electrophoresis result.
Four, the PCR of vegetable material identifies (using the XMag633 primer pair)
Respectively the 7K65/ 212-F2 of agricultural university is carried out the PCR evaluation for plant (247 strain) and the 7K65/ 211-F2 of 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 as follows:
1, the blade of each seedling in clip embodiment 2 extracts genomic dna.
2, take the genomic dna of step 1 as template, carry out pcr amplification with the XMag633 primer pair, obtain pcr amplification product.
3, the pcr amplification product with step 2 carries out 8% polyacrylamide gel electrophoresis, then utilizes argentation to dye, last statistical experiment result.Partial results is seen Fig. 2 (R represents that disease-resistant individual plant, S 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, only has the specific band of 9 strain electrophoresis showed 570bp.Result shows, uses the XMag633 primer pair genomic dna of 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 be powdery mildew disease-resistant plant or Powdery Mildew disease plant.
Claims (9)
1. the application of primer pair first in the test kit of preparation assistant identification powdery mildew disease-resistant wheat; The primer pair that described primer pair first is comprised of single stranded DNA shown in the sequence 2 of single stranded DNA shown in the sequence 1 of sequence table and sequence table.
2. primer pair first and the primer pair B application in the test kit of preparation assistant identification powdery mildew disease-resistant wheat; The primer pair that described primer pair first is comprised of single stranded DNA shown in the sequence 2 of single stranded DNA shown in the sequence 1 of sequence table and sequence table; The primer pair that described primer pair B is comprised of single stranded DNA shown in the sequence 4 of single stranded DNA shown in the sequence 3 of sequence table and sequence table.
3. the test kit of an assistant identification powdery mildew disease-resistant wheat, comprise the primer pair first; The primer pair that described primer pair first is comprised of single stranded DNA shown in the sequence 2 of single stranded DNA shown in the sequence 1 of sequence table and sequence table.
4. test kit as claimed in claim 3, it is characterized in that: described test kit also comprises primer pair B; The primer pair that described primer pair B is comprised of single stranded DNA shown in the sequence 4 of single stranded DNA shown in the sequence 3 of sequence table and sequence table.
5. the application of primer pair first in assistant identification powdery mildew disease-resistant wheat; The primer pair that described primer pair first is comprised of single stranded DNA shown in the sequence 2 of single stranded DNA shown in the sequence 1 of sequence table and sequence table.
6. primer pair first and the primer pair B application in assistant identification powdery mildew disease-resistant wheat; The primer pair that described primer pair first is comprised of single stranded DNA shown in the sequence 2 of single stranded DNA shown in the sequence 1 of sequence table and sequence table; The primer pair that described primer pair B is comprised of single stranded DNA shown in the sequence 4 of single stranded DNA shown in the sequence 3 of sequence table and sequence table.
7. the method for an assistant identification powdery mildew disease-resistant wheat, comprise the step first;
Described step first comprises the steps: that genomic dna take wheat to be measured as template, carries out pcr amplification with the primer pair first; If have two specific fragments in pcr amplification product, wheat to be measured is candidate's powdery mildew disease-resistant wheat; If do not have described two specific fragments in pcr amplification product, wheat to be measured is the susceptible wheat of candidate's Powdery Mildew; One in described two specific fragments is 505-515bp, and another is 485-495bp; The primer pair that described primer pair first is comprised of single stranded DNA shown in the sequence 2 of single stranded DNA shown in the sequence 1 of sequence table and sequence table.
8. method as claimed in claim 7, it is characterized in that: described method also comprises step second;
Described step second comprises the steps: that genomic dna take wheat to be measured as template, carries out pcr amplification with primer pair B; If have a specific fragment in pcr amplification product, wheat to be measured is candidate's powdery mildew disease-resistant wheat; If do not have a described specific fragment in pcr amplification product, wheat to be measured is the susceptible wheat of candidate's Powdery Mildew; A described DNA fragmentation that specific fragment is 565-575bp; The primer pair that described primer pair B is comprised of single stranded DNA shown in the sequence 4 of single stranded DNA shown in the sequence 3 of sequence table and sequence table.
9. method as claimed in claim 8, it is characterized in that: if same described wheat to be measured is consistent with the qualification result in described step second in described step first, this wheat to be measured is the further candidate's who determines powdery mildew disease-resistant wheat or the further candidate's who determines the susceptible wheat of Powdery Mildew.
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| 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 |
| 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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| CN101659989A (en) * | 2008-08-29 | 2010-03-03 | 李祥 | Molecular markers of wheat powdery mildew resistance gene Pm2 and method for acquiring same |
| CN101955986A (en) * | 2009-07-14 | 2011-01-26 | 河南农业大学 | PCR marker linked with wheat powdery mildew resistant genes PmLK906 and using method thereof |
| CN101988064A (en) * | 2010-12-03 | 2011-03-23 | 河南省农业科学院 | Marker primer interlocked with wheat powdery mildew resistance gene PmHNK54 and application thereof |
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