CN104365471A - Breeding and identifying method of soft and powdery mildew resistant triticum aestivum-Dasypyrum villosum translocation line - Google Patents
Breeding and identifying method of soft and powdery mildew resistant triticum aestivum-Dasypyrum villosum translocation line Download PDFInfo
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Abstract
The invention discloses a breeding and identifying method of a soft and powdery mildew resistant triticum aestivum-Dasypyrum villosum translocation line. The method comprises the steps of hybridizing and backcrossing a triticum aestivum-Dasypyrum villosum 5V alien addition line DA5V and a Chinese spring ph1b1b mutant, identifying individual plants relevant to 5VS and ph1b1bph1b1b from BC1F1 by use of a molecular marker, identifying individual selfed seeds by use of GISH (genomic in situ hybridization) technology, and analyzing individual plants containing translocated chromosomes by use of C-zoning, GISH and molecular markers to prove that the translocated chromosome is T5VS.5AL. A PCR (polymerase chain reaction) primer is designed by use of an EST (Expressed Sequence Tag) located on the homologous chromosome group of the fifth part of triticum, a co-dominant marker capable of identifying the translocated chromosome specially can be screened, and homozygosis and heterozygosis translocations can be distinguished. The breeding utilization value of the translocation line can be disclosed through agronomic traits, kernel hardness analysis and powdery mildew resistance identification.
Description
Technical field
Patent of the present invention discloses soft a, seed selection of mildew-resistance Triticum aestivum-Haynaldia villosa T5VS5AL translocation line NAU421, authentication method, belongs to field of plant variety breeding technology.
Background technology
Haynaldia villosa (Dasypyrum villosum (L.) Schur) (2n=2x=14, VV) be the common wheat dliploid affinity species originating in Mediterranean northeast and Southwest Asia, the proterties that has fine qualities and anti-multiple wheat diseases, that wheat breed is improved three grades of important genetic resourceses (seen reference document: Agnieszka Gradzielewska.The genus Dasypyrum-part 2.Dasypyrum villosum-a wild species used in wheat improvement.Euphytica, 2006, 152:441-454).Agricultural University Of Nanjing's cell utilize from Britain Camb botanical garden introduce haynaldia villosa 91C43 be donor, (see reference a set of Triticum aestivum-Haynaldia villosa alien addition line of seed selection document: Zhang W, Zhang RQ, Feng YG, Bie TD, Chen PD.Distribution ofhighly repeated DNA sequences in Haynaldia villosa and its application in the identification of alienchromatin.Chin Sci Bull, 2013,58:890 – 897).
Grain hardness is the major genetic factors determining wheat quality, and the grain hardness gene Pina/Pinb controlling common wheat is positioned at 5DS end, and wheat 5AS, 5BS lack grain hardness gene.In addition, also there is the gene controlling grain hardness near isogenic wheat line, present patent application person has confirmed that the soft gene location of haynaldia villosa seed is in the end of 5VS, and genotype is Dina-D1a/Dinb-D1a.(see reference document: Zhang Ruiqi, Wang Xiue, Chen Peidu.Molecular andcytogenetic characterization of a small alien-segment translocation line carrying the softness genes ofHaynaldia villosa.Genome, 2012,55:639-646).If the soft gene transfer of being carried by 5VS is on common wheat 5A or 5B chromosome, then common wheat just has two sites to control Germplasms quality simultaneously, thus forms the new germ plasm of ultra-soft endosperm structure.
Powdery mildew is the Major Diseases of wheat, powdery mildew resistance gene Pm 21 from haynaldia villosa is the main anti-source of the current breeding for disease resistance of China, the kind that existing dozens of contains Pm21 gene utilizes in production that (see reference document: Li, G P, Chen P D, Zhang H Z, Wang X E, He Z H, Zhang Y, Zhao H, Huang, H Y, Zhou X C.Effects of the 6VS.6ALtranslocation on agronomic traits and dough properties of wheat.Euphytica, 2007,155,305-313).Along with constantly extensively utilizing of this gene, its risk overcome by new pathogen biological strain, also in increasing, therefore, excavates new disease-resistant gene very necessary further from haynaldia villosa.In addition, the practice of nearly mildew-resistance breeding decades of China shows, breeding work is always in the change of catching up with pathogen biological strain passively, because biological strain constantly changes, cause new varieties resistance after spread 3-5 just obviously to be lost, the kind even had also is not authorized resistance and is just lost.Therefore, Plant resistance gene is utilized may to be the optimal selection realizing kind durable resistance future.Research has confirmed haynaldia villosa 5VS to carry a powdery mildew Plant resistance gene, and therefore, soft, the disease-resistant gene carried by haynaldia villosa 5VS imports common wheat, very necessary for cultivation high-quality, disease-resistant weak muscle kind.
Patent of the present invention is hybridized by China spring ph1b1b mutant and haynaldia villosa 5V alien addition line DA5V, is backcrossed, utilize in offspring and follow the trail of the qualification of 5VS specific molecular marker, genomic in situ hybridization (GISH) and Chromosome C banding, select the Triticum aestivum-Haynaldia villosa T5VS5AL translocation line NAU421 of 1 China spring background.The codominant marker filtered out is utilized to identify from the F raising wheat 15 × NAU421 in conjunction with GISH
2colony and F
2:3the hereditary effect of pedigree analysis T5VS5AL translocation chromosome is worth with the breeding utilization of clear and definite this translocation line.
Summary of the invention
The object of the invention is to disclose the selection of soft, a mildew-resistance Triticum aestivum-Haynaldia villosa T5VS5AL translocation line NAU421 and identify the codominant marker Xcinau5VS-1 of this transposition, and evaluating the effect of this translocation line to grain endosperm quality and powder mildew resistance.
The selection of a kind of soft, mildew-resistance Triticum aestivum-Haynaldia villosa T5VS5AL translocation line NAU421, China spring mutant CSph1bph1b and Triticum aestivum-Haynaldia villosa 5V chromosome alien addition line DA5V is hybridized, then carries out with China spring mutant CSph1bph1b the acquisition BC that backcrosses
1; Plantation BC
1seed, utilizes the primer pair DNA in seedling stage of molecular labeling CINAU41 to identify, through electrophoresis detection, has the individual plant of 745bp specific band to show containing 5V chromosome; Carry out the screening of ph1b recessive homozygote to the primer containing 5V chromosomal individual plant recycling ph1b gene specific molecular marker ABC302.3, through electrophoresis detection, the individual plant lacking 920bp specific band is shown to be ph1bph1b and isozygotys individual plant; Will containing exogenous chromosome 5V and the homozygotic plant of ph1bph1b carry out pollen mother cells chromosome pairing in mid-term I observe, statistics exogenous chromosome and chromosome of wheat pair frequency, by the plant bagging selfing containing Homoeologous chromosome pairing, obtain BC
1f
1, by BC
1f
1seed germinates in culture dish, and a point individual plant clip tip of a root carries out fluorescence in situ hybridization qualification, and point individual plant extracts DNA, utilizes 5VS specific molecular marker CINAU41, carries out PCR qualification, obtains BC to identifying the translocation line individual plant bagging selfing relating to 5VS
1f
2seed; By BC
1f
2seed is planted in green house and is extracted BC
1f
2individual plant DNA, the primer pair individual plant DNA again applying CINAU41 carries out pcr amplification, through electrophoresis detection, the individual plant of 745bp specific band is had to show containing 5VS chromosome arm, and then utilize the individual plant DNA that the primer pair of 5AS specific molecular marker Xgwm443 contains 5VS chromosome arm to carry out pcr amplification, through electrophoresis detection, the individual plant of the 245bp specific band that can not increase shows to lack 5AS chromosome, is the T5VS5AL translocation line isozygotied.
Described method also comprises the T5VS5AL translocation line individual plant bagging selfing to isozygotying, and obtains BC
1f
2:3; By BC
1f
2:3seed germinates in culture dish, and a point individual plant clip tip of a root carries out cytological Identification successively, determines the identity of translocation chromosome further.
The primer of described molecular labeling CINAU41 is CINAU41F:SEQ ID NO.1, CINAU41R:SEQ ID NO.2; The primer of described molecular labeling ABC302.3 is ABC302.3F:SEQ ID NO.3, ABC302.3R:SEQ ID NO.4; The primer of described 5AS specific molecular marker Xgwm443 is: Xgwm443F:SEQ ID NO.5, Xgwm443R:SEQ ID NO.6.
Described cytological Identification comprises C-and divides band and hybridization histochemistry: utilize fluorescence in situ hybridization to CS ph1b BC
1individual plant carries out pollen mother cell observation in mid-term I, green is haynaldia villosa 5V chromosome, if existing green has danger signal again in the bivalent that danger signal is chromosome of wheat to be formed, show that there occurs homologous chromosome between haynaldia villosa 5V chromosome and chromosome of wheat matches; Haynaldia villosa 5VS chromosome arm has strong end C-to divide band band line feature, and wheat 5AL chromosome arm has strong middle C-to divide band band line feature, according to these features, to BC
1f
2:3the translocation line material that generation obtains carries out C-successively and divides band to detect and fluorescence in situ hybridization detection, if the galianconism of translocation chromosome is green and has strong end C-to divide band band line feature, long-armed have strong middle C-to divide band to be with line feature for danger signal, shows that this transposition is T5VS5AL Chromosome translocation.
For following the trail of 1 codominant marker Xcinau5VS-1 of Triticum aestivum-Haynaldia villosa T5VS5AL translocation line NAU421 of the present invention, this molecular labeling primer is XCINAU5VS-1F:SEQ ID NO.7, XCINAU5VS-1R:SEQ ID NO.8; Utilize this molecular marker primer pair T5VS5AL Chromosome translocation to carry out pcr amplification, the specific band of 5VS 700 bp can be amplified, and lack the specific band of wheat 5AS 750 bp.
The described application of molecular labeling Xcinau5VS-1 in molecular marker assisted selection breeding.
Beneficial effect:
1, common wheat China spring-haynaldia villosa T5VS5AL translocation line NAU421 is compensatory whole arm translocation system, and economical character is excellent and carry a dominant wheat powdery mildew strain-forming period resistance gene.
2, common wheat China spring-haynaldia villosa T5VS5AL translocation line NAU421 carries the gene loci that two control grain hardness simultaneously, and lay respectively on 5VS and 5DS, this translocation line can significantly reduce grain hardness value, has ultra-soft endosperm structure.
3,1 codominant marker Xcinau5VS-1 disclosed by the invention can identify this translocation chromosome effectively, and can distinguish and isozygoty or heterozygosis transposition, for molecular marker assisted selection breeding.
Accompanying drawing explanation
Fig. 1 is the root-tip cells Metaphase Chromosomes genomic in situ hybridization (2n=42) (a) of T5VS5AL translocation line NAU421, pollen mother cells I in mid-term chromogene group in situ hybridization (21 II) (b), the chromosomal C-of common wheat China spring 5A divides band (c), the chromosomal C-of haynaldia villosa 5V divides band (d), the C-of T5VS5AL translocation chromosome divides band (e) and genomic in situ hybridization (f) .a, that send green florescent signal in b and c is haynaldia villosa translocation chromosome fragment 5VS, what send redness or blue-fluorescence signal is chromosome of wheat.
Fig. 2 is codominant marker Xcinau5VS-1 at the amplification of T5VS5AL translocation line NAU421, parent and China spring the 5th group of nulli-tetrasomes systems and chromosome mapping. is from left to right followed successively by, swimming lane 1:DNA Marker, DL2000; 2: China spring; 3: haynaldia villosa; 4:T5VS5AL translocation line NAU421-1; 5:T5VS5AL translocation line NAU421-2; 6:T5VS5DL translocation line NAU415; 7: in durum wheat, draw 1286; 8: China spring 5A nulli-tetrasomes (N5AT5D); 9: China spring 5B nulli-tetrasomes (N5BT5D); 10: China spring 5D nulli-tetrasomes (N5DT5A).
Fig. 3 is the fringe type of China spring and T5VS5AL translocation line NAU421, and a left side is China spring fringe type, and the right side is NAU421 fringe type.
Fig. 4 is the qualification of Triticum aestivum-Haynaldia villosa T5VS5AL translocation line NAU421 Adult plant powder mildew resistance, is from left to right followed successively by China spring, T5VS5AL (NAU421), (NAU421/ raises wheat 15) F1, flowering stage of raising wheat 15 fall two leaf blades.
Fig. 5 is the qualification of Triticum aestivum-Haynaldia villosa T5VS5AL translocation line NAU421 resistance to powdery mildew in seedling stage, is from left to right followed successively by the two leaf stage plant of China spring, T5VS5AL (NAU421), disease-resistant contrast 09P190 (Pm21 gene).
Biological deposits
NAU421, Classification And Nomenclature is common wheat Triticumaestivum, Seed storage is in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), preservation date is 2014-9-12, preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, and deposit number is CGMCC NO.9598.
Embodiment
(1) control homologous chromosome pairing systematic mutation body is utilized, induction haynaldia villosa 5V chromosome and common wheat 5A chromosome translocation
By China spring mutant CSph1bph1b, (see reference document: Gill K S and Gill B S.A DNA fragment mappedwith in the submicroscopic deletion of Ph1, a chromosome pairing regulator gene in polyploid wheat.Genetics.1991, 129:251-259. Wang Xin hopes, Lai Jingru, Chen Liang letter, Liu Guangtian. the Molecular Identification of Chinese Spring ph 1 b Mutant. Scientia Agricultura Sinica, 1998, 31 (5): 31-34) (see reference document: Zhang Wei with Triticum aestivum-Haynaldia villosa 5V alien addition line DA5V, Zhang Ruiqi, Feng Yigao, Bie Tongde, Chen Peidu.Distribution of highly repeated DNAsequences in Haynaldia villosa and its application in the identification of alien chromatin.Chinesescience bulletin, 2013, 58:890 – 897.) hybridize, carry out backcrossing with CSph1bph1b again and obtain BC1, by BC
1seed is planted in green house, utilize molecular labeling CINAU41 (see reference document: Cao Yaping, Cao Aizhong, Wang Xiue, Chen Peidu. based on EST-PCR H. villosa chromosome specific molecular marker screening and application.Acta Agronomica Sinica, 2009,35 (1), 1-10.) (primer sequence: F:CTCCAAGAGCCCAGATAG (SEQ ID NO.1)); R:AGGGTGGGGAGAAAGTTA (SEQ ID NO.2)) NDA in seedling stage is identified to, through electrophoresis detection, have the individual plant of 745bp specific band to show containing 5V chromosome.To containing 5V chromosomal individual plant recycling ph1b gene specific molecular marker ABC302.3, (see reference document: Wang Xinwang, Lai Jingru, Chen Liang letter, Liu Guangtian. the Molecular Identification of Chinese Spring ph 1 b Mutant. Scientia Agricultura Sinica, 1998,31 (5): 31-34) (primer sequence: F:ATAAAGGAGAAGATTGAGTC (SEQ IDNO.3); R:A TAAGAAACAGGAACAGAG (SEQ ID NO.4)) carry out the screening of ph1b recessive homozygote, through electrophoresis detection, the individual plant lacking 920bp specific band is shown to be ph1bph1b and isozygotys individual plant.Will containing exogenous chromosome 5V and the homozygotic plant of ph1bph1b carry out pollen mother cells chromosome pairing in mid-term I observe, observe the pollen mother cell of 380 plant altogether, what wherein 5V chromosome and wheat Part V homology group chromosome occurred to match only has 3 plant, and pair frequency is only about 0.8%.By the plant bagging selfing containing Homoeologous chromosome pairing, obtain BC
1f
1.By BC
1f
1seed germinates in culture dish, divide the individual plant clip tip of a root to be stored in 4 DEG C of refrigerators and carry out cytological Identification, fluorescence in situ hybridization technique is utilized to identify 500 seeds altogether, what wherein relate to the transposition of 5VS or 5VL chromosome arm has 15 strains, then by this 15 strain seedling replanting to green house, individual plant extract DNA, application 5VS specific molecular marker CINAU41, carry out PCR qualification, have 5 strain DNA cloning to go out the 5VS specific band of 745bp, show that these 5 individual plants relate to the translocation line individual plant of 5VS; Respectively by this 5 individual plant baggings selfing, obtain BC
1f
2seed; Each individual plant gets 50 BC
1f
2seed plants green house, get blade seedling stage and extract individual plant DNA, utilize molecular labeling CINAU41 to carry out PCR qualification simultaneously, through electrophoresis detection, there is the individual plant of 745bp specific band to show containing 5VS chromosome arm, and then utilize 5AS specific molecular marker Xgwm443 primer (primer sequence: F:GGGTCTTCATCCGGAACTCT (SEQ ID NO.5); R:CCATGATTTATAAATTCCACC (SEQ ID NO.6)) pcr amplification is carried out to the individual plant DNA containing 5VS chromosome arm, through electrophoresis detection, the individual plant of 245bp specific band of can not increasing shows to lack 5AS chromosome arm, is the T5VS5AL translocation line isozygotied; BC is obtained to the translocation line individual plant bagging selfing of isozygotying
1f
2:3; By BC
1f
2:3seed germinates in culture dish, and a point individual plant clip tip of a root carries out cytological Identification.Haynaldia villosa 5VS chromosome arm has strong end C-to divide band band line characteristic, and wheat 5AL chromosome arm has strong middle C-to divide band band line characteristic, according to these features, to BC
1f
2:3generation obtain translocation line material carry out successively Chromosome C banding detect and fluorescence in situ hybridization detection (green is haynaldia villosa 5VS chromosome arm, danger signal is chromosome of wheat), if the galianconism of translocation chromosome is green and has strong end C-to divide band band line feature, long-armed have strong middle C-to divide band to be with line feature for danger signal, determines that this transposition is T5VS5AL translocation line further.
(2) according to being positioned common wheat Part V homology group est sequence design primer, the codominant marker of Screening and Identification T5VS5AL translocation chromosome
The Unigene ESTs sequence (http://www.ncbi.nlm.nih.gov/unigene) of wheat is utilized to design PCR primer, therefrom filter out 1 codominant marker Xcinau5VS-1, (XCINAU5VS-1F:GTTTATCAGGCGGTGCCATA (SEQID NO.7); XCINAU5VS-1R:GGACTTCTTGCTCCCCTTTC (SEQ ID NO.8)) T5VS5AL translocation line NAU421 can amplify the specific band of 5VS 700bp, and lack the specific band (table 1, Fig. 2) of wheat 5AS 750bp.
(3) grain hardness of Triticum aestivum-Haynaldia villosa T5VS5AL translocation line NAU421 and Analysis of agronomic characters
In order to understand the effect of this translocation chromosome to Other Main Agronomic Characters, disclose the value of this translocation line, this patent is annual in experimental farm, Agricultural University Of Nanjing Jiangpu in 2013-2014, to China spring, grain hardness and the Other Main Agronomic Characters of two strain NAU421-1 and NAU421-2 of T5VS5AL translocation line compare discovery, translocation chromosome has no significant effect plant height, spike length, but has positive result to spikelet number, grain number per spike and thousand kernel weight; And the grain hardness value of translocation line is significantly lower than background parent China spring, the translocation line showing to carry two soft gene locis has the endosperm structure (table 2) of ultra-soft.
(4) Triticum aestivum-Haynaldia villosa T5VS5AL translocation line NAU421 resistance to powdery mildew effect analysis
For understanding T5VS5AL translocation line to the resistance of powdery mildew, utilizing the powdery mildew of In Nanjing to mix biological strain respectively at 2013,2014 to the NAU421 planted in Jiangpu booth, raising wheat 15, raise wheat 15/NAU421 F
1and raise wheat 15/NAU421F
2inoculated identification.Respectively in heading stage, pustulation period investigation powder mildew resistance.All investigation materials are divided into two types, i.e. high resistance plant (without scab) and disease plant (having scab).Result shows all high mildew-resistances of individual plant containing translocation chromosome, without all high sense powdery mildew (Fig. 4, table 3) of external source individual plant; Meanwhile, seedling stage assay utilizes hybrid bacterial strain to carry out inoculated identification, China spring and NAU421 all susceptible (Fig. 5) to the experiment material China spring of 2 leaf phases, NAU421 and disease-resistant control material 09P190 in greenhouse.Confirm that T5VS5AL translocation line carries a powdery mildew Plant resistance gene, name as Pm5VS.
The codominant marker of Triticum aestivum-Haynaldia villosa T5VS5AL translocation line NAU421 identified by table 1
Table 2 Triticum aestivum-Haynaldia villosa T5VS5AL translocation line NAU421 compares with its background parent China spring grain hardness and economical character
Note: in the same genetic background of different letter representation, translocation line offspring and parent's difference reach 5% significance level
Wheat 15/NAU421 F raised by table 3
2colony's powdery mildew strain qualification
Claims (6)
1. the selection of soft, a mildew-resistance Triticum aestivum-Haynaldia villosa T5VS5AL translocation line NAU421, it is characterized in that China spring mutant CSph1bph1b and Triticum aestivum-Haynaldia villosa 5V chromosome alien addition line DA5V to hybridize, then carry out with China spring mutant CSph1bph1b the acquisition BC that backcrosses
1; Plantation BC
1seed, utilizes the primer pair DNA in seedling stage of molecular labeling CINAU41 to identify, identifies containing the chromosomal individual plant of 5V; The screening of ph1b recessive homozygote is carried out to the primer containing 5V chromosomal individual plant recycling ph1b gene specific molecular marker ABC302.3, obtain containing exogenous chromosome 5V and ph1bph1b isozygotys individual plant; Will containing exogenous chromosome 5V and the homozygotic plant of ph1bph1b carry out pollen mother cells chromosome pairing in mid-term I observe, statistics exogenous chromosome and chromosome of wheat pair frequency, by the plant bagging selfing containing Homoeologous chromosome pairing, obtain BC
1f
1, by BC
1f
1seed germinates in culture dish, and a point individual plant clip tip of a root carries out fluorescence in situ hybridization qualification, and point individual plant extracts DNA, utilizes 5VS specific molecular marker CINAU41, carries out PCR qualification, obtains BC to identifying the translocation line individual plant bagging selfing relating to 5VS
1f
2seed; By BC
1f
2seed is planted in green house and is extracted BC
1f
2individual plant DNA, the primer pair individual plant DNA again applying CINAU41 carries out pcr amplification, through electrophoresis detection, the individual plant of 745bp specific band is had to show containing 5VS chromosome arm, and then utilize the individual plant DNA that the primer pair of 5AS specific molecular marker Xgwm443 contains 5VS chromosome arm to carry out pcr amplification, through electrophoresis detection, the individual plant of the 245bp specific band that can not increase shows to lack 5AS chromosome, is the T5VS5AL translocation line isozygotied.
2. selection according to claim 1, is characterized in that described method also comprises the T5VS5AL translocation line individual plant bagging selfing to isozygotying, and obtains BC
1f
2:3; By BC
1f
2:3seed germinates in culture dish, and a point individual plant clip tip of a root carries out cytological Identification successively, determines the identity of translocation chromosome further.
3. selection according to claim 1, is characterized in that the primer of described molecular labeling CINAU41 is CINAU41F:SEQ ID NO.1, CINAU41R:SEQ ID NO.2; The primer of described molecular labeling ABC302.3 is ABC302.3F:SEQ ID NO.3, ABC302.3R:SEQ ID NO.4; The primer of described 5AS specific molecular marker Xgwm443 is: Xgwm443F:SEQ ID NO.5, Xgwm443R:SEQ ID NO.6.
4. selection according to claim 1, is characterized in that described cytological Identification comprises C-and divides band and hybridization histochemistry: utilize fluorescence in situ hybridization to CS ph1b BC
1individual plant carries out pollen mother cell observation in mid-term I, green is haynaldia villosa 5V chromosome, if existing green has danger signal again in the bivalent that danger signal is chromosome of wheat to be formed, show that there occurs homologous chromosome between haynaldia villosa 5V chromosome and chromosome of wheat matches; Haynaldia villosa 5VS chromosome arm has strong end C-to divide band band line feature, and wheat 5AL chromosome arm has strong middle C-to divide band band line feature, according to these features, to BC
1f
2:3the translocation line material that generation obtains carries out C-successively and divides band to detect and fluorescence in situ hybridization detection, if the galianconism of translocation chromosome is green and has strong end C-to divide band band line feature, long-armed have strong middle C-to divide band to be with line feature for danger signal, shows that this transposition is T5VS5AL Chromosome translocation.
5. for following the trail of 1 codominant marker Xcinau5VS-1 of Triticum aestivum-Haynaldia villosa T5VS5AL translocation line NAU421 described in claim 1, it is characterized in that this molecular labeling primer is XCINAU5VS-1F:SEQ ID NO.7, XCINAU5VS-1R:SEQ ID NO.8; Utilize this molecular marker primer pair T5VS5AL Chromosome translocation to carry out pcr amplification, the specific band of 5VS 700bp can be amplified, and lack the specific band of wheat 5AS 750bp.
6. the application of molecular labeling Xcinau5VS-1 according to claim 5 in molecular marker assisted selection breeding.
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| CN105145339A (en) * | 2015-09-07 | 2015-12-16 | 江苏丘陵地区镇江农业科学研究所 | Creation method for dasypyrum villosum-based high-quality powdery mildew-resistant breeding element |
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