CN102260669A - Molecular marking method for anti-wheat yellow mosaic virus active quantitative trait locus (QTL) in Xifeng wheat - Google Patents
Molecular marking method for anti-wheat yellow mosaic virus active quantitative trait locus (QTL) in Xifeng wheat Download PDFInfo
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Abstract
The invention discloses a molecular marking method for anti-wheat yellow mosaic virus active QTLQYm.nau-5A.1 in 'Xifeng wheat' and belongs to the field of biological technology. The invention first discloses the anti-wheat yellow mosaic virus active quantitative trait locus (QTL) in Xifeng wheat and then discloses two molecular markers tightly interlocked with the QTL, namely CINAU152 and CINAU153. In a material containing the QTLQYm.nau-5A.1, a product which is as long as about 1,100bps is obtained by amplification by using marker primers CINAU152F/CINAU152R and is tightly interlocked with the QYm.nau-5A.1, and the genetic distance is 0.0cM; and a product which is as long as about 900bps is obtained by amplification by using marker primers CINAU153F/CINAU153R and is tightly interlocked with the QYm.nau-5A.1, and the genetic distance is 0.1cM. The QYm.nau-5A.1 disclosed by the invention can be used as a new gene resource for breeding wheat yellow mosaic virus; and the primers which are tightly interlocked with the QYm.nau-5A.1 for marking can be used for molecular marker assisted seed breeding.
Description
One, technical field
The invention discloses ' west wind wheat ' anti-Wheat yellow mosaic virus main effect QTL
QYm.nau-5A.1Molecule marking method, belong to biological technical field.Can be the breeding of wheat anti yellow flower leaf disease poison new genetic resources is provided; With
QYm.nau-5A.1The primer of close linkage mark can be used for the molecular marker assisted selection breeding.
Two, background technology
Common wheat (
Triticum aestivumL.) 21 pairs of karyomit(e)s are arranged, every pair comprises two the same karyomit(e)s, these 21 karyomit(e)s are respectively 1A, 1B, 1D, 2A, 2B, 2D, 3A, 3B, 3D, 4A, 4B, 4D, 5A, 5B, 5D, 6A, 6B, 6D, 7A, 7B, 7D, and common wheat is represented (Fig. 1) with AABBDD usually.Every karyomit(e) comprises long-armed and galianconism again, represents with L and S respectively, comprises long-armed 5AL and galianconism 5AS such as 5A karyomit(e).
Wheat (
Triticum aestivumL) as global important food crop, in agriculture production, have critical role, but by the many slime moulds of fungi-cereal (
Polymyxa graminis) (document sees reference: Jie Dao Wheat yellow mosaic virus (Wheat yellow mosaic bymovirus Inouye T (1969) Viral pathogen of the wheat yellow mosaic disease. Nogaku Kenkyu 53:61-68), WYMV) the wheat yellow mosaic disease that causes (Wheat yellow mosaic, WYM) to wheat grow and output has caused serious harm, (document sees reference: 1 to become one of the severe diseases of Asian countries's Wheat Production such as harm China and Japan just day by day, Tao Jiafeng, Qin Jiazhong, Xiao Jiheng, Shen Yanzhang, Zhao Fuzhen, the Li Tian family dependant, Xie Yiyuan, He Daifu, after being full of, Huang Xianhua (1980) Sichuan soil passes the research of wheat yellow mosaic. Plant Pathology 10:5-25; 2, Yu Shanqian, Chen Zhongyi, the liter that blows gently, Zhang Ruoping, Wang Mingqi, Luo Ruiwu (1986) occurs in the Wheat yellow mosaic virus disease of China. plant protection journal 13:217-219; 3, Li Dawei, Han Chenggui, Xing Yiming, Tian Zhaofeng, Yu Jialin, Cai Zhunan, the RT-PCR that the Chinese Wheat yellow mosaic virus of Liu Yi (1997) (WYMV) distributes identifies. Plant Pathology 27:303-307).Resistance main effect QTL/gene is introduced the wheat cultivation kind, and cultivating and promote antiviral kind is to produce at present to go up the most economical effective means of control wheat yellow mosaic disease.
Common wheat kind ' west wind wheat ' is (public, document sees reference: Qian Cunming, fly the Zhou Dynasty, Yao Guocai, Yao Jinbao, Sheng Peiying, seed selection and application that the peaceful wheat of Yang Xueming (1999) new variety of wheat is No. 9. Jiangsu agricultural sciences 3:19-20) have excellent proterties such as resistant to lodging, Powdery Mildew, head blight, stripe rust, fringe germination, disease-resistant proterties is outstanding, and comprehensive proterties is good.Agricultural University Of Nanjing's cytogenetics identifies that to ' west wind wheat ' anti yellow flower leaf disease it is high anti-that the result shows that ' west wind wheat ' shows the wheat yellow mosaic disease, ' the high sense of town's 9523 ' performance.In order further to study ' west wind wheat ' genetics of resistance mechanism and location resistance QTL to the wheat yellow mosaic disease, ' town's 9523 ' (authorization kind) made male parent, (Single-seed descent SSD) has made up F to pass method by single seed so ' the west wind wheat ' of the high anti-wheat yellow mosaic disease of Agricultural University Of Nanjing's cytogenetics makes female parent, high sense wheat anti yellow flower leaf disease
6Recombinant inbred lines (Recombinant inbred line, RIL), and carried out utilize this RIL colony carry out resistance QTL location and with the screening operation of main effect QTL compact linkage molecule mark, and carried out the secondary F of utilization " RILV-6 * town 9523 "
2Segregating population is further estimated disease-resistant main effect QTL
QYm.nau-5A.1Disease-resistant Study on Effect, ' west wind wheat ' derived varieties checking of having carried out utilizing high anti-wheat yellow mosaic disease at last with
QYm.nau-5A.The researchs such as validity of compact linkage molecule mark.
Three,
Summary of the invention
Technical problemThe objective of the invention is to open anti-wheat yellow mosaic disease main effect QTL from common wheat kind ' west wind wheat '
QYm.nau-5A.1, this QTL can be the breeding of disease-resistant wheat poison new genetic resources is provided; Simultaneously, provide with
QYm.nau-5A.1The compact linkage molecule mark
CINAU152With
CINAU153Primer and usage, can be used for the molecular marker assisted selection breeding.
Technical scheme
1, the main effect QTL among the present invention
QYm.nau-5A.1From anti-wheat yellow mosaic disease wheat breed ' west wind wheat ',
QYm.nau-5A.1Resistance to the wheat yellow mosaic disease is stable, and (2007 in six directions test point, Nanjing in four environment; Continuous 3 years of 2008-2010 is in test point, Yangzhou, Jiangsu) all can detect; Contribution rate is big, the phenotypic variation of soluble 25.9-53.7%.
2, among the present invention with anti-wheat yellow mosaic disease main effect QTL
QYm.nau-5A.1The primer of closely linked two marks is respectively with wheat EST(BE426712 and BE497829) sequence is that stencil design obtains (Fig. 3), its sequence is,
Mark
CINAU152Primer:
CINAU152F：CTTGGTTTCGGTGTGTGTAT
CINAU152R：CCATTCTGATGGAAGCAATA；
Or mark
CINAU153Primer:
CINAU153F：GCAAAAATGTAATGCACCAT
CINAU153R：GTTGCTATTGCCTTCAGTTG。
Wherein, use mark
CINAU152Primer from containing
QYm.nau-5A.1Disease-resistant plant in amplify the specific band of about 1100bp, perhaps use mark
CINAU153Primer from containing
QYm.nau-5A.1Disease-resistant plant in amplify the specific band of about 900bp; Mark
CINAU152With
CINAU153With
QYm.nau-5A.1Between genetic distance be respectively 0.0 and 0.1cM.
Above-mentioned primer is used for ' west wind wheat ' anti-wheat yellow mosaic disease main effect QTL
QYm.nau-5A.1Molecule marking method, comprising:
(1) with the DNA of material to be identified as template, use
CINAU152Or
CINAU153Carry out pcr amplification as primer;
The PCR reagent set becomes: contain the 1.0 μ L dna profilings of 20-100ng DNA, 1.0 μ L, 10 * PCR buffer, 0.8 μ L MgCl
2, 0.8 μ L dNTP, each 0.2 μ L of left and right sides primer, 0.15 μ L
TaqDNA polymerase, 5.85 μ L ddH
2O;
The PCR program is: 94 ℃ of pre-sex change 3 minutes; 94 ℃ of sex change 30 seconds, 55 ℃ of annealing 50 seconds, 72 ℃ were extended 35 circulations 1 minute and 10 seconds; 72 ℃ were extended 10 minutes; 10 ℃ of preservations;
The PCR product detects: the PCR product is electrophoretic separation on the non-denaturing polyacrylamide gel of 39:1 at acrylamide and methene acrylamide mass ratio, detects with argentation again;
(2) two pairs of labeled primers are identified
QYm.nau-5A.1The result be:
Use mark
CINAU152Primer carry out pcr amplification, the plant that can amplify about 1100bp specific band is for containing
QYm.nau-5A.1Plant;
Perhaps use mark
CINAU153Primer carry out pcr amplification, the plant that can amplify about 900bp specific band is for containing
QYm.nau-5A.1Plant.
Beneficial effect:
1, disclosed anti-wheat yellow mosaic disease main effect QTL among the present invention from ' west wind wheat '
QYm.nau-5A.1Resistance to virus is stable, and (2007 in six directions test point, Nanjing in four environment; Continuous 3 years of 2008-2010 is in test point, Yangzhou, Jiangsu) all can detect; Contribution rate is big, the phenotypic variation of soluble 25.9-53.7%.Anti-wheat yellow mosaic disease main effect QTL from ' west wind wheat '
QYm.nau-5A.1In wheat breeding for disease resistance, has higher utility value.
2, anti-wheat yellow mosaic disease main effect QTL in disclosed and ' the west wind wheat ' among the present invention
QYm.nau-5A.1The primer of closely linked two pairs of molecule markers (CINAU152F/CINAU152R and CINAU153F/CINAU153R) is based on the wheat est sequence and designs, and is used for identifying in the plant whether contain
QYm.nau-5A.1, simple, convenient and swift, amplification is stablized.
3, labeled primer CINAU152F/CINAU152R and CINAU153F/CINAU153R the amplification product with
QYm.nau-5A.1Between genetic distance very near, be respectively 0.0 and 0.1cM, so carry out pcr amplification, utilize molecular marker assisted selection with these two pairs of labeled primers
QYm.nau-5A.1The accuracy height.
Four, description of drawings
Fig. 1: common wheat karyomit(e) diagrammatic sketch
Fig. 2: wheat anti yellow flower leaf disease individual plant resistance is identified grade scale
Fig. 3: the two couples of labeled primer CINAU152F/CINAU152R and pairing wheat EST(BE426712 of CINAU153F/CINAU153R and BE497829) sequence and primer sequence position
Fig. 4: anti-wheat yellow mosaic disease main effect QTL in ' west wind wheat '
QYm.nau-5A.1Distribution on 5A karyomit(e)
Annotate: left-hand digit is represented synergetic genetic distance (cM), and the position of four environment and the detected QTL peak value of population mean thereof marks with different triangles.
Fig. 5: anti-with " west wind wheat * town 9523 " parents of RIL colony and part, the DNA of sense family is as template, use two couples of labeled primer CINAU152F/CINAU152R (a) and CINAU153F/CINAU153R (b) among the present invention to carry out pcr amplification respectively, the result shows that labeled primer CINAU152F/CINAU152R is at disease-resistant parent ' west wind wheat ' (swimming lane 1) and disease-resistant family ( swimming lane 3,4,7,8,10,14,17,18,20 and 21) all amplify the specific band of about 1100 bp in, labeled primer CINAU153F/CINAU153R all amplifies the specific band of about 900 bp; And these two pairs of labeled primers all do not amplify corresponding specific band in susceptible parent and family.Arrow is depicted as specific band.
Annotate: M. Marker; 1. west wind wheat; 2. press down 9523; 3-24. part is anti-, the sense family; R. it is disease-resistant to isozygoty; S. it is susceptible to isozygoty
Fig. 6: with " RILV-6 * town 9523 " secondary F
2Segregating population parents and anti-, sense pond and part are anti-, the DNA of sense individual plant is as template, use labeled primer CINAU152F/CINAU152R (a) and CINAU153F/CINAU153R (b) among the present invention to carry out pcr amplification respectively, the result shows that labeled primer CINAU152F/CINAU152R is at disease-resistant parent ' RILV-6 ' (swimming lane 1), anti-pond (swimming lane 3), disease-resistant individual plant (swimming lane 6 isozygotys, 11 and 12), the disease-resistant individual plant of heterozygosis (swimming lane 7,9-10,13-14,16,19-22 and 24) all amplify the specific band of about 1100 bp in, labeled primer CINAU153F/CINAU153R all amplifies the specific band of about 900 bp; And these two pairs of labeled primers all do not amplify corresponding specific band in susceptible parent and individual plant.Arrow is depicted as specific band.
Annotate: M. Marker; 1. RIL5-6; 2. press down 9523; 3. anti-pond; 4. sense pond; 5-24. part is anti-, the sense individual plant; R. it is disease-resistant to isozygoty; H. heterozygosis is disease-resistant; S. it is susceptible to isozygoty.
Fig. 7: with the DNA of the derived varieties of anti-wheat yellow mosaic disease ' west wind wheat ' and sense wheat yellow mosaic disease kind as template, use labeled primer CINAU152F/CINAU152R (a) and CINAU153F/CINAU153R (b) among the present invention to carry out pcr amplification respectively, the result shows that labeled primer CINAU152F/CINAU152R all amplifies the specific band of about 1100 bp in ' west wind wheat ' (swimming lane 1) and deutero-disease-resistant variety (swimming lane 2-4) thereof, and labeled primer CINAU153F/CINAU153R all amplifies the specific band of about 900 bp; And these two pairs of labeled primers all do not amplify corresponding specific band in susceptible variety (swimming lane 5-16).Arrow is depicted as specific band.
Annotate: M. Marker; 1. ' west wind wheat '; 2-4. ' west wind wheat ' derived varieties of high anti-wheat yellow mosaic disease ' peaceful wheat No. 9 ', ' peaceful wheat No. 16 ' and ' raising wheat No. 18 '; 5-16. high sense wheat yellow mosaic disease kind ' town 9523 ', ' Anhui wheat 36 ', ' Zheng wheat 9094 ', ' all wheats 18 ', ' Shan wheat 139 ', ' locust wheat ', ' are raised wheat 10 ', ' No. 5, Jinan ', ' Beijing 11 ', ' awns being arranged red No. 18 ', ' and see 0089 ' and ' face farming 11 '; R. it is disease-resistant to isozygoty; S. it is susceptible to isozygoty.
Five, embodiment
1, the excavation of anti-wheat yellow mosaic disease main effect QTL in ' west wind wheat '
The resistance standard of perfection of wheat anti yellow flower leaf disease is divided into 0-5 level (Fig. 2) (reference: Liu Weihua, He Zhentian, Geng Bo, Hou Mingsheng, Zhang Min, Nie Huan etc. (2004) wheat is to the resistance evaluation of yellow mosaic disease and the genetic analysis of typical species. Plant Pathology 34:542-547): 0 grade is that plant is asymptomatic; 1 grade is stunt not obviously for plant, and slight floral leaf does not generally produce tangible streaked necrosis spot, not yellow of blade or minority yellow; 2 grades are stunt not obviously for plant, and slight floral leaf produces tangible streaked necrosis spot, and the blade yellow is obvious; 3 grades are slightly stunt for plant, and floral leaf is obvious, and the streaked necrosis spot accounts for leaf area about 1/2, the partial blade yellow, and minority is withered, and the yellow of tillering is stunt; 4 grades are obviously stunt for plant, serious floral leaf, and the striped spot accounts for leaf area about 3/4, the thin and delicate distortion of lobus cardiacus or be the top shape that contracts, partial blade and tiller withered; 5 grades are seriously stunt for plant, most of blade and tiller or whole strain withered.' west wind wheat ' is high anti-to the performance of wheat yellow mosaic disease, and resistance level is 0 grade; ' the then high sense of performance of town 9523 ', anti-level level is 5 grades.(2007 in six directions test point, Nanjing in four environment; Continuous 3 years of 2008-2010 is in test point, Yangzhou, Jiangsu) the RIL colony (recombinant inbred lines) of " west wind wheat * town 9523 " has been carried out wheat yellow mosaic disease resistance identified, locate the QTL of anti-wheat yellow mosaic disease in ' west wind wheat ' in conjunction with the molecular marker linkage maps that makes up, wherein be positioned at the main effect QTL on ' west wind wheat ' 5AL karyomit(e)
QYm.nau-5A.1In four environment, all be detected the phenotypic variation of soluble 25.9-53.7% (Fig. 4).Newly detected
QYm.nau-5A.1Be that former studies was never reported, can be used as the new genetic resources of wheat anti yellow flower leaf disease breeding and be used.
2, with
QYm.nau-5A.1The design of compact linkage molecule labeled primer
Make up and anti-wheat yellow mosaic disease main effect QTL at above-mentioned molecular marker linkage maps
QYm.nau-5A.1In the research of excavating, two pairs of molecule markers
CINAU152With
CINAU153With
QYm.nau-5A.1Close linkage (Fig. 4), and be respectively 0.0 and 0.4cM with the genetic distance of its LOD peak value (52.1cM).The primer of these two pairs of marks is (public according to two est sequences (BE426712 and BE497829) on wheat the 5th homology group, document Qi LL sees reference, Echalier B, Chao S, Lazo GR, Butler GE, Anderson OD et al (2004) A chromosome bin map of 16,000 expressed sequence tag loci and distribution of genes among the three genomes of polyploid wheat. Genetics 168:701-712), adopt online primer-design software Primer3 V0.4.0 to design (http://frodo.wi.mit.edu/primer3/).
Mark
CINAU152Primer be CINAU152F:CTTGGTTTCGGTGTGTGTAT and CINAU152R:CCATTCTGATGGAAGCAATA; (Fig. 3)
Mark
CINAU153Primer be CINAU153F:GCAAAAATGTAATGCACCAT and CINAU153R:GTTGCTATTGCCTTCAGTTG).(Fig. 3)
3, labeled primer CINAU152F/CINAU152R and the CINAU153F/CINAU153R Molecular Detection in " west wind wheat * town 9523 " RIL colony
Utilizing labeled primer CINAU152F/CINAU152R to carry out pcr amplification in 164 familys of RIL colony and parents thereof detects.The result shows: all amplify the specific band of about 1100bp in the anti-parent of height ' west wind wheat ' and 83 familys, and ' (Fig. 5 a) all not amplify this specific band in town 9523 ' and remaining 81 familys height sense parent.
Utilizing labeled primer CINAU153F/CINAU153R to carry out pcr amplification in 164 familys of RIL colony and parents thereof detects.The result shows: all amplify the specific band of about 900bp in the anti-parent of height ' west wind wheat ' and 84 familys, and ' do not amplify this specific band (Fig. 5 b) in town 9523 ' and remaining 80 familys height sense parent.
The PCR reagent set becomes: contain the 1.0 μ L dna profilings of 20-100ng DNA, 1.0 μ L10 * PCR buffer, 0.8 μ LMgCl
2, 0.8 μ LdNTP, each 0.2 μ L of left and right sides primer, 0.15 μ L
TaqDNA polymerase, 5.85 μ L dd H
2O.
The PCR program is: 94 ℃ of pre-sex change 3 minutes; 94 ℃ of sex change 30 seconds, 55 ℃ of annealing 50 seconds, 72 ℃ were extended 35 circulations 1 minute and 10 seconds; 72 ℃ were extended 10 minutes; 10 ℃ of preservations, PCR product are electrophoretic separation on the non-denaturing polyacrylamide gel of 39:1 at acrylamide and methene acrylamide mass ratio, dye with argentation again.
4, labeled primer CINAU152F/CINAU152R and CINAU153F/CINAU153R are at F
2Molecular Detection in the segregating population
For further evaluation
QYm.nau-5A.1Disease-resistant effect, this laboratory is again according to the amplification of molecule marker in RIL colony and parents in detected 3 QTL fiducial intervals, and in conjunction with the resistance qualification result of each family in four environment, identifies 12 altogether and only contain main effect QTL
QYm.nau-5A.1And do not contain other little effect QTL, yellow mosaic disease resistance level is 0 grade high anti-family, appoints and selects one of them high anti-family ' RILV-6 ', has made up " RILV-6 * town 9523 " secondary F
2Segregating population (280 individual plants).Genetic analysis shows,
QYm.nau-5A.1At secondary F
2Be the dominance monogenic inheritance in the segregating population.Utilize labeled primer CINAU152F/CINAU152R at F
2The parents of colony carry out pcr amplification detection (condition and method are the same) with resisting, feeling in pond and the colony's individual plant.The result shows: at 280 F
2In the individual plant, the 60 strains disease-resistant individual plant that isozygotys all amplifies the specific band consistent with high anti-parent ' RILV-6 ' (about 1100bp), the 77 strains susceptible individual plant that isozygotys amplifies all that ' specific band (about 850bp) that town 9523 ' is consistent, (Fig. 6 a) for two kinds of specific bands and the disease-resistant individual plant of 143 strain heterozygosis can increase simultaneously with high sense parent.Utilize labeled primer CINAU153F/CINAU153R at F
2The parents of colony carry out pcr amplification detection (condition and method are the same) with resisting, feeling in pond and the colony's individual plant.The result shows: at 280 F
2In the individual plant, the disease-resistant individual plant of 203 strains all amplifies the specific band consistent with high anti-parent ' RILV-6 ' (about 900bp), and the susceptible individual plant of 77 strains does not all amplify this specific band (Fig. 6 b).According to amplification and linkage analysis, the result shows mark
CINAU152With
CINAU153With
QYm.nau-5A.1Between genetic distance be respectively 0.0 and 0.1cM.
Secondary F
2The result of segregating population and RIL colony verifies mutually, promptly
QYm.nau-5A.1Being the main effect QTL of anti-wheat yellow mosaic disease in ' west wind wheat ', is again anti-wheat yellow mosaic disease gene in the high anti-family ' RILV-6 ', and mark
CINAU152With
CINAU153With
QYm.nau-5A.1Close linkage.
5, utilize the derived varieties of anti-wheat yellow mosaic disease ' west wind wheat ' and the validity of sense wheat yellow mosaic disease kind verification mark primer CINAU152F/CINAU152R and CINAU153F/CINAU153R
Domestic breeding work person directly or indirectly utilizes ' west wind wheat ' to breed the wheat breed of some high anti-wheat yellow mosaic diseases of extensively promoting by the conventional breeding method, (public as ' peaceful wheat No. 9 ', the authorization kind), ' peaceful wheat No. 16 ' is (public, authorize kind) and (public, authorization kinds) such as ' raising wheat No. 18 '.
And ' town 9523 ' is (public, the authorization kind), ' Anhui wheat 36 ' is (public, the authorization kind), ' Zheng wheat 9094 ' is (public, the authorization kind), ' all wheats 18 ' are (public, the authorization kind), ' Shan wheat 139 ' is (public, the authorization kind), ' locust wheat ' is (public, the authorization kind), ' it is (public to raise wheat 10 ', the authorization kind), ' No. 5, Jinan ' is (public, the authorization kind), ' Beijing 11 ' is (public, the authorization kind), ' awns being arranged red No. 18 ' is (public, the authorization kind), ' sight 0089 ' is (public, authorize kind) and ' face farming 11 ' (public, authorization kind) and then be sense wheat yellow mosaic disease kind.
For certification mark
CINAU152With
CINAU153Follow the trail of the anti-wheat yellow mosaic disease of ' west wind wheat ' derived varieties QTL
QYm.nau-5A.1Validity, the derived varieties of the anti-wheat yellow mosaic disease of this research and utilization ' west wind wheat ' and sense wheat yellow mosaic disease kind are test material, found that mark
CINAU152With
CINAU153Primer in the derived varieties of all anti-wheat yellow mosaic diseases ' west wind wheat ', amplify about 1100bp(Fig. 7 respectively a) and 900bp specific band (Fig. 7 b), in sense yellow mosaic disease kind, do not amplify corresponding specific band.This experimental results show that mark
CINAU152With
CINAU153Primer can be used for the molecular marker assisted selection breeding, follow the trail of the anti-wheat yellow mosaic disease QTL on ' west wind wheat ' 5AL karyomit(e) specifically
QYm.nau-5A.1
SEQUENCE?LISTING
＜110〉Agricultural University Of Nanjing
＜120〉' west wind wheat ' anti-Wheat yellow mosaic virus main effect QTL
QYm.nau-5A.1Molecule marking method
<130> 000
<140> 000
<141> 2011-06-30
<160> 4
<170> PatentIn?version?3.1
<210> 1
<211> 20
<212> DNA
＜213〉artificial
<220>
<221> CINAU152F
<222> (1)..(20)
<223>
<400> 1
cttggtttcg?gtgtgtgtat 20
<210> 2
<211> 20
<212> DNA
＜213〉artificial
<220>
<221> CINAU152R
<222> (1)..(20)
<223>
<400> 2
ccattctgat?ggaagcaata 20
<210> 3
<211> 20
<212> DNA
＜213〉artificial
<220>
<221> CINAU153F
<222> (1)..(20)
<223>
<400> 3
gcaaaaatgt?aatgcaccat 20
<210> 4
<211> 20
<212> DNA
＜213〉artificial
<220>
<221> CINAU153R
<222> (1)..(20)
<223>
<400> 4
gttgctattg?ccttcagttg 20
Claims (3)
1. anti-wheat yellow mosaic disease main effect QTL in ' west wind wheat '
QYm.nau-5A.1The molecule marker primer, its sequence is,
Mark
CINAU152Primer:
CINAU152F：CTTGGTTTCGGTGTGTGTAT
CINAU152R：CCATTCTGATGGAAGCAATA；
Or mark
CINAU153Primer:
CINAU153F：GCAAAAATGTAATGCACCAT
CINAU153R：GTTGCTATTGCCTTCAGTTG。
2. according to anti-wheat yellow mosaic disease main effect QTL in the claim 1 described ' west wind wheat '
QYm.nau-5A.1The molecule marker primer is characterized in that, uses mark
CINAU152Primer from containing
QYm.nau-5A.1Disease-resistant plant in amplify the specific band of about 1100bp, perhaps use mark
CINAU153Primer from containing
QYm.nau-5A.1Disease-resistant plant in amplify the specific band of about 900bp; Mark
CINAU152With
CINAU153With
QYm.nau-5A.1Between genetic distance be respectively 0.0 and 0.1cM, QTL
QYm.nau-5A.1Resistance to the wheat yellow mosaic disease is stable, explains the phenotypic variation of 25.9-53.7%.
3. claim 1 or 2 described primers are used for ' west wind wheat ' anti-wheat yellow mosaic disease main effect QTL
QYm.nau-5A.1Molecule marking method, comprising:
(1) with the DNA of material to be identified as template, use
CINAU152Or
CINAU153Primer carry out pcr amplification;
The PCR reagent set becomes: contain the 1 μ L dna profiling of 20-100ngDNA, 1.0 μ L, 10 * PCR buffer, 0.8 μ L MgCl
2, 0.8 μ L dNTP, each 0.2 μ L of left and right sides primer, 0.15 μ L
TaqDNA polymerase, 5.85 μ L ddH
2O;
The PCR program is: 94 ℃ of pre-sex change 3 minutes; 94 ℃ of sex change 30 seconds, 55 ℃ of annealing 50 seconds, 72 ℃ were extended 35 circulations 1 minute and 10 seconds; 72 ℃ were extended 10 minutes; 10 ℃ of preservations;
The PCR product detects: the PCR product is electrophoretic separation on the non-denaturing polyacrylamide gel of acrylamide and methene acrylamide mass ratio 39:1, detects with argentation again;
(2) two pairs of labeled primers are identified
QYm.nau-5A.1The result be:
Use mark
CINAU152Primer carry out pcr amplification, the plant that can amplify about 1100bp specific band is for containing
QYm.nau-5A.1Plant;
Perhaps use mark
CINAU153Primer carry out pcr amplification, the plant that can amplify about 900bp specific band is for containing
QYm.nau-5A.1Plant.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 201110194389 CN102260669B (en) | 2011-07-12 | 2011-07-12 | Molecular marking method for anti-wheat yellow mosaic virus active quantitative trait locus (QTL) in Xifeng wheat |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110194389 CN102260669B (en) | 2011-07-12 | 2011-07-12 | Molecular marking method for anti-wheat yellow mosaic virus active quantitative trait locus (QTL) in Xifeng wheat |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102604972A (en) * | 2011-12-15 | 2012-07-25 | 浙江省农业科学院 | Replicase gene segment for efficiently exciting wheat yellow to resist mosaic virus and application |
| CN102807984A (en) * | 2012-07-17 | 2012-12-05 | 南京农业大学 | Molecular marker for wheat yellow mosaic bymovirus resistant major-effect quantitative trait locus (QTL) of Yining wheat and application thereof |
| CN108642204A (en) * | 2018-04-24 | 2018-10-12 | 南京农业大学 | The combination of SNP marker primer and its application of one anti-wheat yellow mosaic QTL QYm.nau-5A.1 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN100519763C (en) * | 2007-01-11 | 2009-07-29 | 江苏省农业科学院 | Molecule tag close linked to resistance QTL of wheat sharp eyespot, and application |
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| CN100519763C (en) * | 2007-01-11 | 2009-07-29 | 江苏省农业科学院 | Molecule tag close linked to resistance QTL of wheat sharp eyespot, and application |
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| 《江苏省遗传学会第八届会员代表大会暨学术研讨会论文集》 20101231 朱晓彪 等 普通小麦品种西风小麦黄花叶病抗性相关分析及主效抗性 59-60 1-3 , * |
| 《遗传学报》 20050731 张清平 等 小麦品种梭条花叶病抗性基因遗传 733-737 1-3 第32卷, 第7期 * |
| 《麦类作物学报》 20081231 颜伟 等 小麦抗梭条花叶病的分子标记及QTL 定位 900-904 1-3 第28卷, 第5期 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102604972A (en) * | 2011-12-15 | 2012-07-25 | 浙江省农业科学院 | Replicase gene segment for efficiently exciting wheat yellow to resist mosaic virus and application |
| CN102807984A (en) * | 2012-07-17 | 2012-12-05 | 南京农业大学 | Molecular marker for wheat yellow mosaic bymovirus resistant major-effect quantitative trait locus (QTL) of Yining wheat and application thereof |
| CN102807984B (en) * | 2012-07-17 | 2013-07-17 | 南京农业大学 | Molecular marker for wheat yellow mosaic bymovirus resistant major-effect quantitative trait locus (QTL) of Yining wheat and application thereof |
| CN108642204A (en) * | 2018-04-24 | 2018-10-12 | 南京农业大学 | The combination of SNP marker primer and its application of one anti-wheat yellow mosaic QTL QYm.nau-5A.1 |
| CN108642204B (en) * | 2018-04-24 | 2021-11-23 | 南京农业大学 | SNP (Single nucleotide polymorphism) marker primer combination for resisting wheat yellow mosaic disease QTL QYm. nau-5A.1 and application thereof |
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