CN114410652A - Stripe rust resistant gene QYRsv and swust-1BL tightly linked molecular marker in adult plant stage and application thereof - Google Patents
Stripe rust resistant gene QYRsv and swust-1BL tightly linked molecular marker in adult plant stage and application thereof Download PDFInfo
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Abstract
The invention discloses a stripe rust resistant gene QYRsv.swust-1BL tightly linked molecular marker in an adult plant stage and application thereof, wherein the stripe rust resistant gene is positioned in a segment of 662,307,854-662,550,261 bp of a long arm physical site of a chromosome 1B of an Svevo reference genome of durum wheat; the molecular marker is closely linked with a stripe rust resistant gene QYRsv.swust-1BL and is A012893 and A012894; the nucleotide sequence of the primer of the molecular marker A012893 is shown as SEQ ID NO. 36-SEQ ID NO. 38; the nucleotide sequences of the primers of the molecular marker A012894 are shown as SEQ ID NO. 39-SEQ ID NO. 41. The stripe rust resistant gene QYRsv.swust-1BL positioned by the invention can improve the resistance of wheat varieties to the stripe rust of wheat; the molecular marker closely linked with the disease-resistant gene can be used for evaluating the application of the gene in the stripe rust lasting resistance breeding by marker-assisted selection.
Description
Technical Field
The invention relates to a molecular marker tightly linked with genes, in particular to a strip rust resistant gene QYRsv.swust-1BL tightly linked molecular marker in an adult plant stage and application thereof.
Background
Wheat stripe rust is one of the most serious fungal diseases threatening wheat production. The DNA molecular marker technology is combined to excavate and position the stripe rust resistant gene, so that compared with conventional crossbreeding, the conventional cytogenetic chromosome banding technology and the isozyme technology, the gene positioning research is more convenient and rapid, and the breeding process of disease resistant varieties is accelerated.
With the development of novel molecular markers and the application of high-throughput new-generation sequencing and other technologies in recent years, Single Nucleotide Polymorphism (SNP) has the technical advantages of wide distribution in a genome, high genetic stability, low cost of a single site, co-dominance and the like, and is widely applied to gene positioning, cloning and assisted breeding. Currently developed SNP chips include 9K, 660K, 58K, 55K, 35K, 90K, 820K, which have been published on the web for querying.
KASP (Kompetitive Allele Specific PCR, competitive Allele PCR) belongs to SNP typing detection technology. Compared with the traditional SSR molecular marker and SNP analysis method, the technology greatly simplifies the operation time and steps, not only can detect mutation of a single site, but also can detect deletion of fragments and insertion of large fragments into transgenic materials and the like.
Molecular markers based on PCR (Polymerase Chain Reaction) technology, such as RAPD (Random Amplification Polymorphic DNA), AFLP (Amplified Fragment Length Polymorphism), SSR (Simple Sequence Repeat), STS (Sequence Tagged Site), etc., are commonly used in wheat gene mapping research. However, most of these molecular markers have low flux, are inconvenient to operate when used, have low efficiency, and are very unfavorable for the development of large-scale detection tests. The complexity of wheat genome research hinders the construction of high-density genetic maps of genes.
The stripe rust resistance gene QYRsv.swust-1BL is derived from Svevo of durum wheat at the adult stage, and the markers closely linked on both sides are IWB5732 and IWB4839, respectively, and have an inheritance distance of 0.75cM (721.8Kb) (William et al, 2003; Kolmer, 2015; Zhou et al, 2021). Also contained in this region is a broad spectrum disease resistance gene Yr29/Lr46/Sr58/Pm39, which exhibits resistance to various fungal diseases such as powdery mildew, leaf rust and stem rust. Wherein Yr29/Lr46 shows partial adult-stage rust resistance and is also related to a leaf tip necrosis gene (Ltn 2). In all experimental settings in china and israel, qyrsv. swust-1BL explained phenotypic variation of 11.0% to 34.4%, which is similar to Yr29 (Zhou et al, 2021). Therefore, whether the two genes are the same gene, or allele or closely linked gene cannot be determined, and besides, the two wing-linked markers are not enough to be closely linked with the target gene, and can be effectively applied to molecular marker assisted selective breeding, so that a map is urgently needed to be encrypted, more precise molecular markers are developed to carry out assisted selective breeding research, and whether QYRsv.
Disclosure of Invention
The invention aims to provide a stripe rust resistant gene QYRsv.swust-1BL tightly-linked molecular marker in an adult plant stage and application thereof, wherein the marker is tightly linked with the stripe rust resistant gene QYRsv.swust-1BL, and the marker can be used for assisting in selecting and evaluating the application of the gene in stripe rust lasting resistance breeding.
In order to achieve the purpose, the invention provides a stripe rust resistant gene QYRsv.swust-1BL in an adult stage, wherein the stripe rust resistant gene QYRsv.swust-1BL in the adult stage is positioned in a physical segment of a long arm of a chromosome 1B of an Svevo reference genome of durum wheat, and a nucleotide sequence of the stripe rust resistant gene QYRsv.swust-1BL in the adult stage is a sequence of 662,307,854-662,550,261 bp of the Svevo reference genome of durum wheat.
The invention also aims to provide a primer of a molecular marker tightly linked with the stripe rust resistant gene QYRsv.swust-1BL in an adult stage, wherein the molecular marker is a molecular marker tightly linked with two wings of the stripe rust resistant gene QYRsv.swust-1BL and is A012893 and A012894; the nucleotide sequence of the primer of the molecular marker A012893 is shown as SEQ ID NO. 36-SEQ ID NO. 38; the nucleotide sequences of the primers of the molecular marker A012894 are shown as SEQ ID NO. 39-SEQ ID NO. 41.
The invention also aims to provide a kit for identifying the stripe rust resistant gene QYRsv.switch-1 BL in the adult plant stage, wherein the primer in the kit is the primer of the molecular marker closely linked with the stripe rust resistant gene QYRsv.switch-1 BL.
The invention also aims to provide application of the primer of the molecular marker tightly linked with the stripe rust resistant gene QYRsv.swust-1BL in the adult stage in molecular marker assisted breeding or stripe rust resistant gene QYRsv.swust-1BL research.
The stripe rust resistant gene QYRsv. swust-1BL tightly linked molecular marker in the adult plant period and the application thereof have the following advantages:
the invention identifies a QTL of adult disease resistance (APR) in the Svevo 1BL of durum wheat, does not share common candidate genes with other disease resistance genes on the chromosome, and is a new adult disease resistance locus. By constructing MM37/Svevo F2And Svevo/Zavitan RIL population, using 90K and 660K SNP chip scanning to develop 15 KASP markers, comparing the Chinese spring reference genome with the durum wheat Svevo reference genome, and mapping the initial positioning interval of QYRsvThe 7kb was shortened to 242kb (3.4 cM). Among them, the molecular markers A012893 and A012894 closely linked to the disease-resistant gene can be used in auxiliary selective breeding.
Drawings
FIG. 1 is a high-density genetic linkage map of the adult-plant disease-resistant gene QYRsv.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Experimental example 1 first round of KASP marker development of hard wheat stripe rust resistance gene QYRsv. switch-1 BL
(1) Determination of hybridization combinations to construct fine mapping populations
The test material is wild two-grain wheat Zavidian, national examined variety Mian 37(MM 37) and donor material durum wheat Svevo (T.turgidum ssp.durum) of stripe rust resistant gene QYRsv.swust-1 BL. High-susceptible varieties of Mingxian 169(MX 169) and Jinmai 47(JM 47) are used as susceptible controls. Wild type gemfibrozil (t.turgidum ssp. dicoccuides) belongs to the wild kindred species, which is an important source of genetic diversity in hexaploid wheat (mujing mei, 2019). When resistance genes are difficult to find in common wheat varieties, resistance genes of wild kindred species can be explored. Many of the disease-resistant genes found in the kindred or wild relatives of triticum are such as Yr24/Yr26, Yr53, Yr56, Yr64, Yr65 from t.turgidum and Yr15, Yr35 and Yr46 from t.dicoccoides (mu. jingmei, 2019). Currently, zavidian and svivo genome sequences have been released, which provides convenience for breeding work such as studying wheat genetic diversity, identifying and mining more disease-resistant genes, and searching for yield-related loci (Vendramin et al, 2019). MM37 is bred by cooperation of the institute of agricultural science of Sichuan province and the institute of crop science of Chinese academy of agricultural sciences, is widely planted in Sichuan, and is a bone for breeding Sichuan wheatDry parents, containing 6VS/6AL translocating chromosomes, are highly resistant to powdery mildew (Duhaimei et AL, 2019). In recent years, the increasing frequency of CYR34 in the whole country results in the loss of disease resistance of a batch of wheat varieties containing Yr24/Yr26, such as materials of Guinong line, 92R line and Moro (yellow and bright, etc., 2019). Ren et al (2015) identified the disease resistance gene YrMY37 in MM37 to exhibit resistance to the current epidemic race of chinese stripe rust. MM37 can be used in this test for breeding of polymeric varieties. 137 recombinant inbred families (RlLs) obtained by continuously inbreeding the wheat variety Svevo/Zavidian are used for positioning QYRsv. swust-1BL, and 12F are obtained after hybridization of the wheat variety MM37 (Mianmai 37)/Svevo1Inbreeding of the lines to give 318F which can be used to propagate progeny2Plants and F2:3Family is used for fine positioning.
(2) Conventional breeding and field management
In 2019 and 2020, 137 RILs of Zavidian/Svevo hybrid were planted at two test points of Shaanxi Yangling (YL, 34.292N,108.077E) and Sichuan Mianyang (MY,31.682N,104.663E), 318 MM37/Svevo hybrid F2And F2:3The family is planted in Sichuan Mianyang to identify the disease resistance. Each family is repeated twice, the line length is 1m, the line spacing is 25cm, and each line contains 25-30 seeds; MM37/Svevo hybrid F210 grains were planted in each row. Susceptible varieties JM47 and MX169 were planted every 20 rows to increase field morbidity. The seeding time of Yangling is 10 months (No. 6-10) every year, and the seeding time of Mianyang is 11 months (No. 12-15) every year.
Inoculating mixed bacteria (wheat stripe rust bacteria CYR32, CYR33 and CYR34 in China) when the flagleaf is extracted at the bottom of 3 months in Yangling; the Szechuan sheep Yang is used as a fungus source base for the overwintering of the stripe rust fungus, and naturally attacks the disease in the field at the test point.
Disease resistance was investigated by recording the response type (IT) and severity (DS) when the severity of the rows MX169 and JM47 reached 60% and 90%, respectively. IT is recorded according to 0-9 classification standards, and DS is divided into (%) 0,5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 and 100 according to the incidence area of the whole leaf. The parent and the recombinant inbred line were investigated for IT and DS in two separate runs.
(3) Genomic DNA extraction
137 RILs and 318F are collected2、3F2:3Extracting genome DNA from seedling stage leaves of the pedigree and the three parents by adopting a 1 xCTAB method.
(4) Development design and detection of KASP molecular marker
Known disease-resistant QTL QYRsv.swst-1 BL positioned on Svevo 1BL has 90K SNP markers IWB8812 and IWB5732(Zhou et al, 2021) with two wings closely linked, The two wing sequences of The two SNP markers are aligned to Wheat Chinese spring reference Genome version 1.0 (The International Wheat Genome Consortium, IWGSC) and Svevo reference Genome to obtain The physical positions thereof, and The interval for primarily positioning QYRsv.sw-1 BL is 827kb (physical positions: 670.171-670.998 Mb). The APR disease-resistant gene QYR. ucw-1BL/YR29(Cobo et al, 2019) is also included in this interval. In order to further confirm whether QYr.ucw-1BL/Yr29 and QYRsv.swust-1BL are at the same position, 26 660K SNP chips were selected in this interval and developed as KASP markers, and 15 chromosome-specific KASP markers were obtained (see Table 1).
For the designed KASP tag sequences (see table 1), FAM or VIC fluorescent linker sequences were added to the 5' ends of the two forward primers.
FAM linker sequence (SEQ ID NO.1) is:
5’-GAAGGTGACCAAGTTCATGCT-3’；
the VIC linker sequence (SEQ ID NO.2) is:
5’-GAAGGTCGGAGTCAACGGATT-3’。
the synthesized primers were dissolved and prepared as primers mix (12 mM for both forward primers and 30mM for the reverse primer).
The KASP test was performed according to the procedures provided by LGC Genomics, after the reaction was completed, the fluorescence data at the end was read in a microplate reader, and then the data was introduced into Klustercaller software for genotyping. If typing is not apparent, it is generally necessary to add 3 additional cycles to re-type, but the total number of cycles does not exceed 48.
The synthesized KASP marker is marked by using parent Svevo and Zavitan, 23 disease-resistant homozygous RIL families and 22The homozygous RIL pedigree for the disease was checked for polymorphism in the markers and it was found that 9 of the 15 KASP markers developed exhibited polymorphism in the population (see table 2). 137 RILs families were screened with these 9 markers and 4 key recombinants were found to be families RIL-28, RIL-109, RIL-7 and RIL-71, respectively (see Table 2). The markers closely linked with the target gene are A009271 and A007972, the positioning interval is arranged at one side of the markers, and the corresponding physical position in the Svevo reference genome of the durum wheat is 661,593,905-661,885,275 bp by comparison; the physical position corresponding to the version 1.0 of Chinese spring is 670,382,321-670,783,618 bp, and the positioning interval is 401.297kb (see Table 3). Therefore, QYRsv. switch-1 BL and QYR.ucw-1BL/YR29 are not in the same region, and QYR.ucw-1BL/YR29 is located in the physical site fragment 669.902-670.234 Mb. Labeled with 9 KASP's at MM37, Svevo and 87F2In the population, only the A009289 marker was found to be polymorphic by typing, and the markers that exhibited the polymorphism were then found to be at 318F2The key recombinant cross-over strains screened by the detection in the generation plants are shown in Table 5.
15 KASP marker sequences developed in Table 1
Table 2 shows the gene locations of the 9 KASP markers and the genotypes and field morbidity in the RIL population
Note: a: 9 KASP markers developed in the first round; b: marking a physical location in the durum wheat svivo reference genome; c: the number of RILs with the same genotype as the parent.
TABLE 3 SNP positions and sequences of the developed 15 KASP markers
Note: bold indicates the base corresponding to Svevo.
Experimental example 2 second round of KASP marker development of hard wheat stripe rust resistance gene QYRsv. shock-1 BL
The wheat 660K chip was used to genotype the SNPs of the parents Svevo, Zavitan and MM 37. According to the scanning results of 630,517 660K chips, 66 SNPs with polymorphism are counted, and are compared to an Svevo reference genome, and the 66 SNPs are found to be distributed in an interval 661,593, 990-663, 334,719bp on QYRsv. Comparing to Chinese spring, wherein 57 polymorphic SNPs exist in the interval of 670,783,568-672,779,761 bp; there were 42 overlapping differential SNPs between the two reference genomes.
Based on two alignments of SNPs overlapping in a target interval, the flanking sequences are aligned to a wheat reference genome to obtain physical position information, 4 specific KASP markers with polymorphism between Svevo and Zavidian are successfully developed in the target interval of 670,783,568-672,779,761 bp and are respectively A012891, A012893, A012894 and A012896 (Table 4), and 6 KASP markers (A012891, A012892, A012893, A012894, A012896 and A012897) (Table 5) have polymorphism between Svevo and MM37 and can be used as auxiliary selection breeding.
Experimental example 3 screening of recombinant crossover strain of hard wheat stripe rust resistance gene QYRsv. swust-1BL and construction of high-density genetic map
A total of 13 KASP markers were polymorphic between the parent and RIL populations and could be successfully typed (9 in the first round and 4 in the second round) through two rounds of KASP marker development, and these markers were used to screen for Svevo/Zavidian RILs, yielding 32 recombinant crossovers in total, with 11 recombination events and markers A007972, A012891, A012893, A012894, A012896 closely linked, and 6 recombination events occurring between markers A007972 and A012896 (see Table 4). The markers of close linkage at two ends of QYRsv.swst-1 BL are A007972 and A012896, and the target interval is shortened to 1550.246kb (physical distance: 670,783,618-672,333,864 bp) when the gene group of Chinese spring wheat is compared with the 1.0 version; the target interval was shortened to 1295.343kb (physical distance: 661,593,905-662,889,248 bp) by alignment to the Svevo reference genome (see tables 3 and 4).
TABLE 4 polymorphisms of 13 KASP markers between parents and RIL populations developed for first and second rounds of KASP markers
Note: and b, superscript a: 9 markers developed in the first round; and b, superscript b: 4 markers developed in the second round; and c, superscript c: the KASP marker corresponds to the physical location in the durum wheat svivo reference genome.
A total of 7 KASP markers can be successfully developed in MM37/Svevo through two rounds of F development2Group typing (Table 5) was carried out for A009289 screened in the first round of development and 6 markers screened in the second round of development (A012891, A012892, A012893, A012894, A012896 and A012897). Screening of 318F with these 7 markers2Plants, 37 recombinant cross-over strains were obtained, of which 6 key recombinants were associated with qyrsv. 5 recombination events occurred between a012893 and a012894, 4 recombinants occurred between markers a012892 and a012893, and 3 recombinants occurred between a0128924 and a 012896. There were two heterozygous markers (a012893 and a012894) with a significantly different disease-resistant phenotype among 8 plants. Based on the 6 key recombinants obtained by screening, qyrsv. switch-1 BL was finally locked between a012893 and a012894 (3.4cM) (see table 5 and fig. 1). The 1.0 version region of the Svevo reference genome of durum wheat is approximately 242kb by alignment, and the genetic map construction is carried out according to the JoinMap 4.0 software.
TABLE 57 KASP markers developed for the first and second rounds of KASP markers in the parent and F2Polymorphism between populations
Note: and b, superscript a: one marker developed in the first round and 6 markers developed in the second round; and b, superscript b: phenotype of strain F2:3 (R: homozygous resistant strain, Seg: segregating strain, S: homozygous susceptible strain); and c, superscript c: the KASP marker corresponds to the physical location in the durum wheat svivo reference genome (refseqrel.1.0); and (4) superscript d: a-disease resistant parent (svivo); b-susceptible parent (MM 37); h-heterozygosis; and (4) superscript e: the number of F2 plants having the same genotype as the parent.
SNP analysis (containment) and Clustering (Clustering) were performed by Genome Studio polymeric Clustering v1.0 software from Illumina and GTC software from Affymetrix, respectively. Eliminating SNP with no polymorphism and low quality, wherein deletion value is more than 20%, grouping is not obvious, allelic frequency is less than 5%, and the like. After the differential SNP in the parent and the group is extracted by using Excel, local blast is carried out on the two wing sequences of the differential SNP and the genome according to the genome information of the spring version 1.0 of China and the genome information of the durum wheat svivo, and the reference physical position information of the SNP is obtained. And (4) screening SNP closely linked with the target gene. By chi-square test (chi)2) Significance of SNP site-to-phenotype differences.
QTL mapping uses complete complex interval mapping (ICIM) function, marker clustering and ordering. The genetic distance calculation uses the Kosambi function. The proportion of each population separated was determined by Chi-square analysis (chi-square)2) The significance of the actual value to the expected value is determined. The LOD (the likelihood odds) threshold was set to 3.0, obtained by software alignment check calculation, and the alignment number (permatation) was set to 1000. Data from DS and IT were used for QTL analysis including averages of points throughout the year.
Experimental example 4 prediction of candidate Gene of hard wheat stripe rust resistance Gene QYRsv. switch-1 BL
To F2The population was screened for recombinant cross-over, QYRsv. swust-1BL mapped to 3.4cMInterval, the close linkage of the wings is labeled a012893 and a012894 (see fig. 1). The mapping interval of the Svevo reference genome is 242 kb. And performing annotation analysis according to the chip difference SNP sequence falling in the interval, the annotation of the version 2.0 of the Chinese spring wheat reference genome and the Svevo genome. Within this interval 4 annotated genes were included (see table 6). There are 4 high-confidence genes and disease resistance related, including: nucleotide binding site-Leucine-rich repeat-coiled-coil (Nucleotide-binding-site-Leucine-rich repeat-coiled-coil, NBS-LRR-CC) (TRITD1Bv1G221180.1), Ral GTPase activator protein subunit alpha-1(TRITD1Bv1G221200.1), DNAJ heat shock N-terminal domain protein (TRITD1Bv1G221230.1), S-adenosylmethionine: tRNA ribosyltransferase-isomerase (TRITD1Bv1G221240.1) (Table 6). These were preliminarily presumed to be candidate genes for QYRsv.
TABLE 6 candidate genes for QYRsv
While the present invention has been described in detail with reference to the preferred embodiments, it should be understood that the above description should not be taken as limiting the invention. Various modifications and alterations to this invention will become apparent to those skilled in the art upon reading the foregoing description. Accordingly, the scope of the invention should be determined from the following claims.
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<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 23
gtgttcgttg tcttgtaaga ctctaagtt 29
<210> 24
<211> 44
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 24
gaaggtgacc aagttcatgc tcgtgctttg cgttcaccat atga 44
<210> 25
<211> 44
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 25
gaaggtcgga gtcaacggat tcgtgctttg cgttcaccat atgc 44
<210> 26
<211> 25
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 26
gcatggccag gaagaactgt gaaat 25
<210> 27
<211> 42
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 27
gaaggtgacc aagttcatgc tagaaatcat tgcggtagcc ga 42
<210> 28
<211> 44
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 28
gaaggtcgga gtcaacggat tctagaaatc attgcggtag ccgg 44
<210> 29
<211> 25
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 29
gcagtactcc tagcgtaact ggttt 25
<210> 30
<211> 26
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 30
ataacctaag ctgcagcata acagta 26
<210> 31
<211> 23
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 31
acctaagctg cagcataaca gtc 23
<210> 32
<211> 26
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 32
cagtaagtac tacatgctct gccctt 26
<210> 33
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 33
cattggggtg ggtcaaggca t 21
<210> 34
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 34
attggggtgg gtcaaggcag 20
<210> 35
<211> 25
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 35
ctatgaggga aggggaacca ctaaa 25
<210> 36
<211> 23
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 36
caggcacatg cttagggatt gag 23
<210> 37
<211> 23
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 37
caggcacatg cttagggatt gac 23
<210> 38
<211> 29
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 38
gaacagcgca tttccagaat ttcctaatt 29
<210> 39
<211> 25
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 39
aagaagttca aggcatgggc agata 25
<210> 40
<211> 23
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 40
gaagttcaag gcatgggcag atg 23
<210> 41
<211> 25
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 41
ctacttcggg aagtacttgt cccaa 25
<210> 42
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 42
gtacgtccac tcgctcaagg a 21
<210> 43
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 43
gtacgtccac tcgctcaagg t 21
<210> 44
<211> 25
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 44
cgttatcttt ggtgaccgca ggata 25
<210> 45
<211> 24
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 45
atctttggtg accgcaggat agtt 24
<210> 46
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 46
ctttggtgac cgcaggatag tc 22
<210> 47
<211> 23
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 47
ctcaaggtga agcccatccg cat 23
<210> 48
<211> 71
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 48
gtgttcgttg tcttgtaaga ctctaagttt gtgtaggctg cctgttgtgg gctttattca 60
gttaaagccg g 71
<210> 49
<211> 71
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 49
atcactgtgg cctcgtgctt tgcgttcacc atatgaattt cacagttctt cctggccatg 60
ctatgccaga t 71
<210> 50
<211> 71
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 50
tagaataatg caccgcagcc attcgactta gatcccacgg ctaaaaaatc gagaatttca 60
ttgtgttttt c 71
<210> 51
<211> 71
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 51
tactagcaag agatcaaaac atacacgcaa cgaaattatt aaaacaaaca aacacgatat 60
atagatacag c 71
<210> 52
<211> 71
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 52
agtggagatc atatgcttcc atctgaaaaa taatatggtg tgctggagag acgacctaca 60
aacatgtaaa g 71
<210> 53
<211> 71
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 53
aaaagcaaat caatatagca tgttaaacaa aaaacgaacc tgtgccaaac cagaaatagc 60
caaattgggc t 71
<210> 54
<211> 71
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 54
ataaccagaa gaacaaaggc gtgatggcgt acacttacga caccctcagc tttggagatg 60
ggcggaacct c 71
<210> 55
<211> 71
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 55
acctgcagta ctcctagcgt aactggttta gctttacggc taccgcaatg atttctagcg 60
ttccattgtg t 71
<210> 56
<211> 72
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 56
accccttggc cttgtccaac aggcccgcca tggccgtccc ttgctctctt cccgctcctt 60
cccgcctctg at 72
<210> 57
<211> 71
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 57
acttctgtgg ttatagatag ctaaatctat attgtgactg ttatgctgca gcttaggtta 60
tgatctgttg t 71
<210> 58
<211> 71
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 58
acttctatag aatatcattg gggtgggtca aggcattgcc tgcagaatat tttagtggtt 60
ccccttccct c 71
<210> 59
<211> 71
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 59
cctaatttca ttgtttgtga acagtgtctt ccccagtcaa tccctaagca tgtgcctgcg 60
ccgggaaacc t 71
<210> 60
<211> 71
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 60
aatctttggc tgagaagttc aaggcatggg cagatgacac ctccaggggg tttaccttct 60
attcgatcgg g 71
<210> 61
<211> 71
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 61
cgtcgaccgc atcgcgtacg tccactcgct caaggtgaag cccatccgca tccccaacta 60
tcctgcggtc a 71
<210> 62
<211> 71
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 62
tccactcgct caaggtgaag cccatccgca tccccaacta tcctgcggtc accaaagata 60
acgccttgat c 71
Claims (4)
1. A stripe rust resistant gene QYRsv.swust-1BL in an adult stage is characterized in that the stripe rust resistant gene QYRsv.swust-1BL in the adult stage is positioned in a physical segment of a long arm of a chromosome 1B of an Svevo reference genome of durum wheat, and a nucleotide sequence of the stripe rust resistant gene QYRsv.swust-1BL in the adult stage is a sequence of 662,307,854-662,550,261 bp of the Svevo reference genome of durum wheat.
2. A primer of a molecular marker tightly linked with a stripe rust resistant gene QYRsv.swust-1BL in an adult plant stage is characterized in that the molecular marker tightly linked with two wings of the stripe rust resistant gene QYRsv.swust-1BL is A012893 and A012894; the nucleotide sequence of the molecular marker A012893 primer is shown in SEQ ID NO. 36-SEQ ID NO. 38; the nucleotide sequence of the molecular marker A012894 primer is shown in SEQ ID NO. 39-SEQ ID NO. 41.
3. A kit for identifying a stripe rust resistant gene QYRsv.switch-1 BL in an adult plant stage is characterized in that a primer in the kit is a primer of a molecular marker closely linked with the stripe rust resistant gene QYRsv.switch-1 BL as described in claim 2.
4. The application of the primer of the molecular marker tightly linked with the stripe rust resistant gene QYRsv.switch-1 BL in the adult stage as claimed in claim 2 in molecular marker assisted breeding or stripe rust resistant gene QYRsv.switch-1 BL research.
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Citations (6)
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|---|---|---|---|---|
| CN109321671A (en) * | 2018-10-10 | 2019-02-12 | 四川农业大学 | Wheat stripe rust resisting ospc gene QYr.sicau-1B-1 SSR molecular marker and application |
| CN109706263A (en) * | 2019-02-22 | 2019-05-03 | 四川农业大学 | Chain SNP marker and application with wheat stripe rust resisting ospc gene QYr.sicau-1B-1 |
| CN109797240A (en) * | 2019-03-20 | 2019-05-24 | 西北农林科技大学 | A kind of molecular labeling, primer and its application with wheat stripe rust resisting ospc gene Yr26 close linkage |
| CN110607388A (en) * | 2019-09-29 | 2019-12-24 | 四川农业大学 | Wheat stripe rust resistant gene related SNP molecular marker in adult stage, primer and application thereof |
| CN112430606A (en) * | 2020-12-11 | 2021-03-02 | 四川农业大学 | Wheat stripe rust resistant gene yrZ15-1949 and molecular marker and application thereof |
| CN112481275A (en) * | 2020-12-11 | 2021-03-12 | 四川农业大学 | Wheat stripe rust resistant gene yrZ15-1370 and molecular marker and application thereof |
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| CN109321671A (en) * | 2018-10-10 | 2019-02-12 | 四川农业大学 | Wheat stripe rust resisting ospc gene QYr.sicau-1B-1 SSR molecular marker and application |
| CN109706263A (en) * | 2019-02-22 | 2019-05-03 | 四川农业大学 | Chain SNP marker and application with wheat stripe rust resisting ospc gene QYr.sicau-1B-1 |
| CN109797240A (en) * | 2019-03-20 | 2019-05-24 | 西北农林科技大学 | A kind of molecular labeling, primer and its application with wheat stripe rust resisting ospc gene Yr26 close linkage |
| CN110607388A (en) * | 2019-09-29 | 2019-12-24 | 四川农业大学 | Wheat stripe rust resistant gene related SNP molecular marker in adult stage, primer and application thereof |
| CN112430606A (en) * | 2020-12-11 | 2021-03-02 | 四川农业大学 | Wheat stripe rust resistant gene yrZ15-1949 and molecular marker and application thereof |
| CN112481275A (en) * | 2020-12-11 | 2021-03-12 | 四川农业大学 | Wheat stripe rust resistant gene yrZ15-1370 and molecular marker and application thereof |
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| XINLI ZHOU等: "Genome-Wide Mapping of Loci for Adult-Plant Resistance to Stripe Rust in Durum Wheat Svevo Using the 90K SNP Array", 《PLANT DIS》, vol. 105, no. 4 * |
| XINLI ZHOU等: "Genome-Wide QTL Mapping for Stripe Rust Resistance in Winter Wheat Pindong 34 Using a 90K SNP Array", 《FRONT PLANT SCI》 * |
| 钟晓: "小麦品冬34的重要性状QTL分析及Yr62基因的应用研究", 《中国优秀硕士学位论文全文数据库》 * |
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| CN114410652B (en) | 2023-07-21 |
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