CN107699630B - Molecular marker linked with wheat disease-resistant gene Pm21 and application thereof in breeding - Google Patents
Molecular marker linked with wheat disease-resistant gene Pm21 and application thereof in breeding Download PDFInfo
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Abstract
The invention discloses a molecular marker linked with a wheat disease resistance gene Pm21 and application thereof in breeding. The 25 molecular markers are derived from a wheat-haynaldia villosa tiny fragment translocation line NAU427 containing a Pm21 powdery mildew resistance gene and are tightly linked with a Pm21 gene, and the molecular markers can be used for molecular marker-assisted selection and improve the selection efficiency and accuracy of disease-resistant plants.
Description
Technical Field
The invention belongs to the field of molecular genetic breeding, and relates to a molecular marker linked with a wheat disease-resistant gene Pm21 and application thereof in breeding.
Background
Wheat is an important grain crop of global character, and 35-40% of people all over the world use wheat as staple food. Wheat powdery mildew is one of important diseases worldwide, and can cause yield loss of 13-34% in general onset years and yield loss of more than 50% in severe onset years. The method is the most economic, safe and effective way to cultivate the wheat powdery mildew resistant varieties. In recent years, with the development of molecular marker technology and the construction of high-density molecular genetic maps, molecular marker-assisted breeding plays an increasingly important role in the research of wheat fine trait breeding. The discovery of germplasm resources containing powdery mildew resistance genes and the development of molecular markers linked with disease resistance genes are the basis for the genetic breeding of powdery mildew resistance wheat.
AT present, 85 powdery mildew resistance genes are discovered from 58 gene loci in wheat and its related species, but most of these genes are pedigree-specific resistance and are prone to lose their disease resistance (Wiersma AT, Pulman JA, Brown LK, Cowger C, Olson EL. identification of Pm58from Aeginops tauschii. the applicant Genet,2017,130: 1123-channel 1133). Powdery mildew resistance gene Pm21 from haynaldia villosa shows broad-spectrum high resistance to powdery mildew. Haynaldia villosa (Haynaldia villosa) has 7 pairs of chromosomes (2n ═ 2x ═ 14, VV), each pair of chromosomes comprising two homologous chromosomes, the 7 chromosomes being 1V, 2V, 3V, 4V, 5V, 6V, 7V, respectively. Broad-spectrum high powdery mildew resistance gene Pm21 exists on the 6V short arm of Haynaldia villosa, and can resist more than 120 physiological species of powdery mildew at home and abroad (Huang XQ, Hsam SLK, Zeller FJ. identification of powdery mildew resistance genes in common wheat where. (Triticum aestivum L em.). 9.Cultivars, land roads and weaving lines growth in China wheat where plant Breeding, 1997,116: 233-. A wheat-Haynaldia villosa translocation line T6VS & 6AL (Chen PD, Qi LL, Zhou B, Zhang SZ, Liu DJ.development and molecular cytogenetic analysis of white-Haynaldia villosa 6VS/6AL translocation lines specific resistance to pore tissue milk tissue. TAG,1995, (91):1125 1128) is created by Nanjing agriculture university cytogenetic institute, and the Haynaldia villosa 6VS chromosome arm containing Pm21 gene is substituted for the common wheat 6AL chromosome arm, so that the common wheat is enabled to obtain broad-spectrum resistance to powdery mildew, and more than 30 varieties are bred by the translocation line as a parent for large-area popularization in China. The 6VS chromosome arm is introduced into the common wheat background to introduce the powdery mildew resistance gene Pm21, and other genes on the 6VS chromosome arm are inevitably carried. The Nanjing university of agriculture cytogenetics has created a small fragment translocation line NAU418 carrying the anti-powdery mildew gene Pm21, and developed the molecular markers CINAU273, CINAU274, CINAU275, CINAU15(Chen PD, You CF, Hu Y, Chen SW, Zhou B, Cao AZ, Wang X.radiation-induced translocation with reduced Haynaldia villaldia villosa chromamat at the Pm21locus for a powdered reagent in leather bree patent 2013,31: 477-484-) that can track the translocation fragments. Although the top translocation fragment carried by NAU418 was significantly smaller than the translocation fragment of the full-arm T6VS & 6AL, the translocation fragment was still large and could be detected by the in situ hybridization method. Further creating a Pm21 gene-carrying tiny fragment translocation line and developing a molecular marker linked with the Pm21 gene in the translocation fragment, thereby not only providing new germplasm resources for wheat disease resistance genetic breeding, but also providing a usable molecular marker for molecular marker-assisted selection.
The invention content is as follows:
the invention aims to overcome the defects of the prior art and provide a wheat-haynaldia villosa tiny fragment translocation line NAU427 containing a powdery mildew resistance gene Pm21 and a molecular marker which is contained in the translocation line and is closely linked with a Pm21 gene. The NAU427 can be used as a germplasm resource for wheat powdery mildew resistance genetic breeding, and avoids the redundant linkage of a large number of genes on 6 VS; the molecular marker tightly linked with the Pm21 gene in the translocation fragment of the NAU427 can be used for molecular marker-assisted selection, and the breeding efficiency is improved.
The purpose of the invention can be realized by the following technical scheme:
a molecular marker linked with a wheat disease resistance gene Pm21, which is characterized by being selected from any one of CINAU1692, CINAU1693, CINAU1694, CINAU1695, CINAU1696, CINAU1697, CINAU1698, CINAU1699, CINAU1700, CINAU1701, CINAU1702, CINAU1703, CINAU1704, CINAU1705, CINAU1706, CINAU1707, CINAU1708, CINAU1709, CINAU1710, CINAU1711, CINAU1712, CINAU1713, CINAU1714, CINAU1715 and CINAU 1716; the primer sequences are shown in Table 1:
TABLE 1
The sequence of the primer of the molecular marker linked with the disease-resistant gene Pm21 is shown in SEQ ID NO.1-SEQ ID NO. 50.
The primer disclosed by the invention is applied to identification of wheat germplasm resources, lines and varieties containing the Pm21 gene.
The primer disclosed by the invention is applied to wheat breeding for disease resistance.
The minimal fragment disease-resistant translocation line NAU427 containing the wheat powdery mildew resistance gene Pm21 is preserved in China center for type culture collection with the preservation date of 2017.3.17 and the preservation number of CCTCC P201707, and the preservation address is Wuhan university in China. The powdery mildew resistant wheat-haynaldia villosa tiny fragment translocation line NAU427 is from the radiation progeny of the T6 VS.6 AL whole-arm translocation line 92R 137.
The invention relates to application of a tiny fragment disease-resistant translocation line NAU427 containing a wheat powdery mildew resistance gene Pm21 in wheat disease-resistant breeding.
The invention has the beneficial effects that:
(1) the wheat-haynaldia villosa tiny fragment translocation line NAU427 obtained by screening the T6VS & 6AL whole-arm translocation line radiation progeny contains a wheat powdery mildew resistance gene Pm21, has high wheat powdery mildew resistance, can be used as a germplasm resource for wheat powdery mildew resistance genetic breeding, and avoids the unfavorable linkage of a large number of redundant genes of a 6VS whole-arm translocation fragment. (2) The molecular marker linked with the disease-resistant gene Pm21 developed by the invention can be used for molecular marker-assisted selective breeding, and the disease-resistant breeding efficiency of wheat is greatly improved. (3) 24 of the 25 molecular markers linked with the Pm21 are designed aiming at specific insertion deletion sites in a haynaldia villosa genome sequence, can specifically amplify specific bands on haynaldia villosa chromatin gradually permeating into the NAU427 and can be quickly detected by 1% agarose gel electrophoresis, wherein the CINAU1704 is a marker developed based on the size difference of gene introns, and an amplification product after PCR amplification reaction is detected by using non-denaturing polyacrylamide gel (acrylamide: methylene acrylamide: 39:1), and the electrophoresis detection is convenient for quick identification of molecular marker assisted selection.
Drawings
FIG. 1 shows the identification of resistance of the extremely small fragment translocation line NAU427 and the susceptible control Chinese spring in seedling stage (A) and adult stage (B);
FIG. 2 is a photograph of in situ hybridization (A) and a photograph of molecular marker identification (B) of the very small fragment translocation line NAU427 and its control NAU 418;
1: marker; 2: CS (disease resistance gene without Pm 21); 3: VV (haynaldia villosa, containing disease resistance gene Pm 21); 4: ABV (synthetic wheat, containing Pm 21); 5: t6 VS.6 AL (containing Pm21 disease-resistant gene); 6: NAU418 (small fragment translocation line containing Pm21 disease resistance gene); 7: NAU427
FIG. 3 shows the disease resistance identification and the molecular marker CINAU1704 identification of the backcross progeny plants of the disease-resistant material NAU427 and the powdery mildew-susceptible varieties Yangmai 158 and Shi 4185;
y158: yangmai 158; y158(R)/SI +: the progeny of backcrossing NAU427 with Y158 segregate the disease-resistant material in the population. Stone 4185(R)/SI +: the progeny of backcross of NAU427 with stone 4185 segregates the disease-resistant material in the population.
FIG. 425 shows the amplification results of the linked markers in various materials.
1: marker; 2: CS (disease resistance gene without Pm 21); 3: VV (haynaldia villosa, containing disease resistance gene Pm 21); 4: ABV (synthetic wheat, containing Pm 21); 5: t6 VS.6 AL (containing Pm21 disease-resistant gene); 6: NAU418 (small fragment translocation line containing Pm21 disease resistance gene); 7: NAU427
Biological sample preservation information
The wheat seed NAU427 is classified and named as Triticum aestivum NAU427 and is preserved in China center for type culture collection with the preservation date of 2017, 3 and 17 months and the preservation number of CCTCC NO: P201707 and the preservation address of Wuhan university in China.
Detailed Description
Example 1 creation of wheat-Haynaldia villosa minimal fragment translocation line NAU427 carrying Pm21 Gene
(1) 1600 doses for farm yard of Jiangsu province60CO gamma-ray irradiation of ears of T6 VS.6 AL entire arm translocation line 92R137 2-3 days before flowering, emasculation of the irradiated ears on the same day, pollination of the irradiated ears with Chinese spring normal pollen 2-3 days later, obtaining structural variant M1(Chen PD, You CF, Hu Y, Chen SW, Zhou B, Cao AZ, Wang X (2013) Radiation-induced translocation with reduced Haynaldia villosa chromium mat at the Pm21 location for powder mill restriction in the woheat. mol Breed 31:477 484:).
(2) Planting M1 seed in field, inoculating and identifying powdery mildew in seedling and adult stage, screening high wheat powdery mildew resistant plant, and harvesting seed in mature stage.
(3) Cutting the root tip of the seed harvested from the disease-resistant plant, carrying out in-situ hybridization experiment by using the haynaldia villosa genome DNA as a probe, and detecting the size of the chromosome introgression segment of the haynaldia villosa in the disease-resistant plant. A single plant is screened from a disease-resistant population, a hybridization signal of a haynaldia villosa genome cannot be detected on a chromosome of a root tip cell of the material, a tiny fragment translocation introgression event is presumed to occur, and the introgression fragment carries a Pm21 gene.
(4) The material is subjected to PCR amplification by using 6VS specific molecular markers CINAU273, CINAU274, CINAU275 and CINAU15 developed in NAU418, wherein the specific molecular marker on 6VS can be amplified by using the marker CINAU273, and the chromosome segment carrying the Pm21 gene is further clarified to be introgressed into the disease-resistant material.
(5) The obtained disease-resistant and small-fragment translocation introgression structural variant is backcrossed with Chinese spring for multiple times, and each backcross generation is subjected to disease resistance identification and molecular marker CINAU273 amplification analysis, so that a stable material with high resistance to wheat powdery mildew in the whole growth period is obtained and is named as NAU 427. The material is preserved in China center for type culture Collection with the preservation date of 2017.3.17 and the preservation number of CCTCC NO: P201707.
Example 2 analysis of disease resistance Effect of wheat-Haynaldia villosa very Small fragment translocation line NAU427
(1) In order to analyze the disease-resistant effect of the translocation segment in the wheat-haynaldia villosa tiny segment translocation line NAU427, the NAU427 is hybridized with powdery mildew promotion varieties stone 4185 and Yangmai 158 in different ecological regions to obtain F1, then multiple backcrosses are respectively carried out by taking the promotion varieties stone 4185 and Yangmai 158 as recurrent parents, each generation is subjected to disease resistance identification and molecular marker CINAU1704 identification, and the disease-resistant plants are subjected to selfing in the backcross BC4 generation to obtain seeds BC4F 1.
(2) Planting the selfed seeds in a Jianpu farm plastic greenhouse of Nanjing university of agriculture, wherein the row length is 1.2m, the row spacing is 20cm, 10 seeds are sowed in each row, performing powdery mildew resistance identification on a single plant in the seedling stage and the adult stage, and performing disease resistance identification for at least 3 times in the late 3 months and the early 4 months.
(3) Extracting DNA of disease-resistant plants and disease-susceptible plants, and analyzing whether translocation segments exist by using a molecular marker CINAU1704, wherein the plants capable of amplifying the 6VS specific bands are all disease-resistant plants, and the plants incapable of amplifying the 6VS specific bands are all disease-susceptible plants.
The above results show that: NAU427 carries a haynaldia villosa 6VS translocation chromosomal fragment, which carries the Pm21 gene. The disease resistance effect of the tiny translocation fragment carried by NAU427 can be stably inherited.
Example 3 development of molecular markers linked to Pm21 in NAU427 of wheat-Haynaldia villosa very small fragment translocation line (1) development of molecular markers linked to Pm21 in NAU427
1.1 determination of collinearity region of translocation fragment of Haynaldia villosa introgressed with NAU427 with brachypodium and rice
Both NAU418 and NAU427 contain Pm21 gene, and the in situ hybridization result shows that the translocation fragment of NAU418 is larger than that of NAU 427. To develop molecular markers linked to the Pm21 gene in the NAU427 translocation fragment, EST sequences of 4 markers CINAU273, CINAU274, CINAU275, CINAU15 amplified to give 6 VS-specific bands were first extracted. The 4 EST sequences are respectively aligned with genome sequences of Brachypodium distachyon (http:// plants. ensembl. org/Brachypodium _ distachyon/Info/Index) and rice (http:// plants. ensembl. org/Oryza _ sativa/Info/Index), and the alignment result shows that the 4 EST sequences are consistent in the arrangement sequence of the third chromosome of the Brachypodium and the 2 nd chromosome of the rice, which indicates that the genomic sequences of the dasypyrum villosum, the Brachypodium and the rice have better collinearity relation in the interval. Further extracting gene sequences of genes BRADI3G03270 to BRADI3G04940 on the third chromosome of brachypodium, respectively searching the dasypodium villosum genome sequence by the extracted brachypodium gene sequence, and obtaining and extracting the sequences of the dasypodium villosum homologous genes.
1.2 determination of specific insertion/deletion sites in Haynaldia villosa Gene sequences
The extracted sequence of the haynaldia villosa homologous gene is respectively compared with the sequences (https:// urgi. versals. inra. fr/blast /) of the corresponding homologous genes on the chromosomes of Chinese spring 6A, 6B and 6D for multi-sequence comparison analysis, if a specific insertion/deletion section exists in a certain haynaldia villosa gene compared with the homologous gene of 6A, 6B and 6D, and the insertion/deletion section has no difference in the sequences of Chinese spring 6A, 6B and 6D, an amplification primer aiming at the insertion/deletion site is designed according to the haynaldia villosa sequence. Primer utilizationhttp://primer3.ut.eeOn-line design, synthesized by Shanghai Yingjun Biotechnology Ltd. A series of cytological materials are used for verification, whether the designed primer can amplify a specific strip of the haynaldia villosa is detected, if the specific strip of the haynaldia villosa can be amplified in NAU427, a molecular marker linked with Pm21 is successfully developed.
1.3 development of molecular marker linked with Pm21 in NAU427
(1) Extraction of DNA: haynaldia villosa (2 n-2 x-14, VV) containing the Pm21 gene, synthetic wheat hard haynaldia villosa (AABBVV), wheat-haynaldia villosa whole-arm translocation line T6 VS.6 AL, apical translocation line NAU418, very small fragment translocation material NAU427, and triticum aestivum chinese (AABBDD) not containing the Pm21 gene were extracted by the CTAB method.
(2) PCR procedure: and performing PCR amplification by taking the extracted genomic DNA of each cytological material as a template, performing statistical analysis on polymorphism, and developing a molecular marker tightly linked with Pm 21. . The PCR reagent in the PCR amplification reaction system comprises: 1. mu.L of DNA template (20-100ng), 1.0. mu.L of 10 XPCR buffer, 0.8. mu.L of dNTP, 0.2. mu.L of each of the primers, 0.1. mu.L of Taq DNA polymerase, and 6.7. mu.L of ddH 2O. The PCR procedure was: pre-denaturation at 94 ℃ for 3 min; then denaturation at 94 ℃ for 30 seconds, renaturation at 57 ℃ for 45 seconds, extension at 72 ℃ for 1 minute, 35 cycles; final extension at 72 ℃ for 10 min; storing at 10 deg.C. After the PCR amplification reaction, the amplified product except the marker CINAU1704 was electrophoretically detected on a non-denaturing polyacrylamide gel (acrylamide: methylene acrylamide: 39:1), and the remaining markers were electrophoretically detected on a 1% agarose gel.
(3) Determination of molecular markers linked to Pm21 in NAU 427: when a certain pair of primers amplifies a consistent band in the haynaldia villosa, T6 VS.6 AL, NAU418 and NAU427, but the band cannot be amplified in Chinese spring, the corresponding marker of the primer is a molecular marker linked with Pm21 in the NAU 427. According to the standard, the invention develops 25 molecular markers linked with Pm21 in NAU 427: CINAU1692, CINAU1693, CINAU1694, CINAU1695, CINAU1696, CINAU1697, CINAU1698, CINAU1699, CINAU1700, CINAU1701, CINAU1702, CINAU1703, CINAU1704, CINAU1705, CINAU1706, CINAU1707, CINAU1708, CINAU1709, CINAU1710, CINAU1711, CINAU1712, CINAU1713, CINAU1714, CINAU1715, CINAU 1716. The primer sequence information is shown in table one.
Example 4 application of molecular marker linked with Pm21 in marker-assisted selection
And (3) carrying out PCR amplification on the genome DNA of the extremely-small fragment translocation line NAU427 or the progeny material derived from the extremely-small fragment translocation line NAU427 or the parent by using a primer of a molecular marker linked with Pm21, carrying out gel electrophoresis separation on the amplification product, and if the specific target band of the haynaldia villosa corresponding to the molecular marker can be amplified, indicating that the wheat variety to be identified contains the gene Pm21, otherwise, not containing the gene. Through the development and the utilization of molecular markers linked with Pm21, the selection efficiency of wheat breeding for disease resistance can be obviously improved.
Primer sequences of molecular markers linked to Pm21 in Table-NAU 427
Sequence listing
<110> Nanjing university of agriculture
Nanjing Xinmaixiu Biotech Co., Ltd
<120> molecular marker linked with wheat disease resistance gene Pm21 and application thereof in breeding
<160> 50
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<212> DNA
<213> Artificial Sequence (Artificial Sequence)
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<212> DNA
<213> Artificial Sequence (Artificial Sequence)
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agctacagaa acacggtaac actgcta 27
<210> 3
<211> 31
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
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gtggtgattt tagtttcatg gtgatagtat g 31
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<212> DNA
<213> Artificial Sequence (Artificial Sequence)
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agtctacctc acccgttgga cttg 24
<210> 9
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<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 9
ctctgacatg atatcatgag caattaactc 30
<210> 10
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 10
ggtgatcttc attgcctcca 20
<210> 11
<211> 26
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 11
acttcatgcg ccaatttgga ggatgt 26
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<212> DNA
<213> Artificial Sequence (Artificial Sequence)
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caaggccgcc gaatcctaca at 22
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<212> DNA
<213> Artificial Sequence (Artificial Sequence)
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ctgtacgcgc gagacactgg a 21
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<213> Artificial Sequence (Artificial Sequence)
<400> 14
actgttgaat ttgttccact taatacgaga gt 32
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<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 15
tcttctgacc cgtgcaaatc cagaa 25
<210> 16
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 16
tccagtgtct cgcgcgtaca g 21
<210> 17
<211> 23
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<400> 17
aaattcctac ttttgcgatg tca 23
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<400> 18
aaatcttgcc atgagttgta gggaaaata 29
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<213> Artificial Sequence (Artificial Sequence)
<400> 19
aaatcttgcc atgagttgta gggaaaata 29
<210> 20
<211> 29
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 20
caagaggttt ataatatagt gtctgtaga 29
<210> 21
<211> 23
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 21
aagctgatgc ctggtcactc tca 23
<210> 22
<211> 26
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 22
ggctctactt tgtctacaga aaagcc 26
<210> 23
<211> 29
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 23
ggttataggc tgaaatacgt atttcatag 29
<210> 24
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 24
tcatgtattg acgaacggat gt 22
<210> 25
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 25
tctcctattc tgcgtcttcc a 21
<210> 26
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 26
gccagtgaag gcttgatgta 20
<210> 27
<211> 28
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 27
tgttttcttg cagtcgacca tgacaatc 28
<210> 28
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 28
tgttgacatc gcaaaagtag g 21
<210> 29
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 29
gcatccgttc gacgttggct tc 22
<210> 30
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 30
ctctggttgg agggggaggt 20
<210> 31
<211> 27
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 31
gtagcattat agtccgctga attaacc 27
<210> 32
<211> 25
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 32
ccatggcgca tagcataacc attgg 25
<210> 33
<211> 29
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 33
tgttcagagt tcagacagta agcaatgaa 29
<210> 34
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 34
ctgattatgc gattgcctgt 20
<210> 35
<211> 30
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 35
gtttattttt tagttggacc aagttggacc 30
<210> 36
<211> 29
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 36
ttcattgctt actgtctgaa ctctgaaca 29
<210> 37
<211> 28
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 37
catcgttaac caaatcacat cgatctgc 28
<210> 38
<211> 24
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 38
caaggtccaa ttctgctttg ctcc 24
<210> 39
<211> 26
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 39
gatggtcttc ttctttagcg caaatg 26
<210> 40
<211> 31
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 40
tagagataaa tattgtattc cctgagtccc a 31
<210> 41
<211> 28
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 41
gctgaaatga ttgtctgtac actagggt 28
<210> 42
<211> 26
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 42
gtgaactttc tcgagagcac agaaga 26
<210> 43
<211> 27
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 43
gagcgtttag atcactaaag tagattc 27
<210> 44
<211> 29
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 44
gggtttatta tgatatatac tggctatag 29
<210> 45
<211> 26
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 45
tagtgcatcc atacttcaac ttagtc 26
<210> 46
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 46
tcaggatcaa tgtagcagaa gg 22
<210> 47
<211> 28
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 47
gacggaaagg accattttag tgattagc 28
<210> 48
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 48
agatgggagc cataggatca 20
<210> 49
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 49
caacatgttc cggaccctct g 21
<210> 50
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 50
agcgcatctt gaatccttga 20
Claims (5)
1. A molecular marker linked with a wheat disease resistance gene Pm21, which is characterized by being selected from any one of CINAU1692, CINAU1693, CINAU1694, CINAU1695, CINAU1696, CINAU1698, CINAU1699, CINAU1700, CINAU1701, CINAU1702, CINAU1703, CINAU1704, CINAU1705, CINAU1706, CINAU1707, CINAU1708, CINAU1709, CINAU1710, CINAU1711, CINAU1712, CINAU1713, CINAU1714, CINAU1715 or CINAU 1716; the primer sequences are shown in Table 1:
TABLE 1
2. The primer of the molecular marker linked with the disease-resistant gene Pm21 in claim 1, wherein the sequence is shown in SEQ ID NO.1-SEQ ID NO.10 and SEQ ID NO.13-SEQ ID NO. 50.
3. The primer of claim 2 is applied to identification of wheat germplasm resources, lines and varieties containing the Pm21 gene.
4. The use of the primers of claim 2 in wheat breeding for powdery mildew resistance.
5. The application of the tiny fragment disease-resistant translocation line NAU427 containing the wheat powdery mildew resistance gene Pm21 in wheat powdery mildew resistance breeding is characterized in that the NAU427 is preserved in China center for type culture Collection with the preservation date of 2017.3.17 and the preservation number of CCTCC P201707, and the preservation address is Wuhan university in China.
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Citations (3)
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|---|---|---|---|---|
| CN105969778A (en) * | 2016-05-10 | 2016-09-28 | 中国农业大学 | Haynaldia villosa gene NAM-V1 and molecular marker and application thereof |
| CN108342401A (en) * | 2017-01-22 | 2018-07-31 | 南京农业大学 | One kinase genes LeRLK1-V and its expression vector and application |
| CN109136232A (en) * | 2017-06-19 | 2019-01-04 | 江苏大学 | Haynaldia villosa mildew-resistance gene DvRGA-1, DvRGA-2 and its application |
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| CN1328389C (en) * | 2004-12-30 | 2007-07-25 | 南京农业大学 | Tagged primer chained to Pm21 gene of anti powdery mildew of wheat and application |
| CN101760550A (en) * | 2008-12-19 | 2010-06-30 | 曹淑兰 | Research on location of molecular markers of powdery mildew resistance genes |
| CN102925544A (en) * | 2012-07-03 | 2013-02-13 | 江苏里下河地区农业科学研究所 | Molecular marker for specifically tracking dasypyrumvillosum 6VS chromosome and use method thereof |
| CN104278028B (en) * | 2014-09-16 | 2016-10-19 | 天津师范大学 | It is positioned at haynaldia villosa 6VS DNA and penetrates into wheat anti-powdery mildew NIL sequence and application |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105969778A (en) * | 2016-05-10 | 2016-09-28 | 中国农业大学 | Haynaldia villosa gene NAM-V1 and molecular marker and application thereof |
| CN108342401A (en) * | 2017-01-22 | 2018-07-31 | 南京农业大学 | One kinase genes LeRLK1-V and its expression vector and application |
| CN109136232A (en) * | 2017-06-19 | 2019-01-04 | 江苏大学 | Haynaldia villosa mildew-resistance gene DvRGA-1, DvRGA-2 and its application |
Non-Patent Citations (3)
| Title |
|---|
| Map-based cloning of the gene Pm21 that confers broad spectrum resistance to wheat powdery;Huagang He et al.;《bioRxiv preprint doi: https://doi.org/10.1101/177857.》;20170818;第1-26页 * |
| Pm21 from Haynaldia villosa Encodes a CC-NBS-LRR Protein Conferring Powdery Mildew Resistance in Wheat;Liping Xing et al.;《Mol Plant》;20180320;第874-878页 * |
| Pm21, Encoding a Typical CC-NBS-LRR Protein, Confers Broad-Spectrum Resistance to Wheat Powdery Mildew Disease;Huagang He et al.;《Mol Plant》;20180320;第879-882页 * |
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