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Tobacco cytoplasmic male sterile line tab1-CMS specific molecular marker and application thereof

A technology of male sterile lines and molecular markers, applied in the field of plant genetic engineering, can solve problems such as no relevant technical reports, and achieve the effect of enriching the source of tobacco sterile cytoplasm

Pending Publication Date: 2021-07-23
TOBACCO RES INST CHIN AGRI SCI ACAD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, for the identification of natural mutants, there are no related technical reports

Method used

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  • Tobacco cytoplasmic male sterile line tab1-CMS specific molecular marker and application thereof
  • Tobacco cytoplasmic male sterile line tab1-CMS specific molecular marker and application thereof
  • Tobacco cytoplasmic male sterile line tab1-CMS specific molecular marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] A total of 8 varieties of fertile tobacco were selected from sua-CMS and tab1-CMS sterile lines, and total leaf DNA was extracted with a kit (TIANGEN Biotech, China). The types and characteristics of the 8 varieties are shown in Table 1.

[0051] Table 1 Material types and characteristics identified by tobacco tab1-CMS sterility-specific markers

[0052]

[0053] According to the tobacco chloroplast genome sequence (GenBank No: Z00044), two pairs of specific primers were designed (S1: 5'-GAGTGGTAAGGCAGAGGAC-3' and 5'-AAATCAATGAAGGAAAAGC-3'; S2: 5'-TATCGCTTTTCCTTCATTG-3' and 5'- ATTTTCTGCTAGATCCCGT-3') amplifies the chloroplast trnC-trnD intergenic region. The PCR program was as follows: 94°C pre-denaturation for 5 min, PCR amplification for 30 cycles, each cycle of 94°C denaturation for 30 s, 51°C annealing for 30 s, 72°C extension for 45 s, and finally 72°C extension for 7 min. The amplified PCR products were detected on 1% agarose gel.

[0054] DNA fragments wer...

Embodiment 2

[0060] A method for identifying or assisting in identifying tobacco cytoplasmic male sterile line tab1-CMS varieties:

[0061] Use specific primer pair S3: forward primer 5'-AGGGGGCAGCAACTACAAT-3'; reverse primer 5'-CGGAAAAAATGATAAATAAAT-3' to amplify the specific marker of tab1-CMS: located at the s1738 site in the chloroplast trnC-trnD intergenic region The SNP site, the SNP site is GGGGGCAA;

[0062] The InDel site located at the s2321 site in the chloroplast trnC-trnD intergenic region has a base deletion. If the obtained result is the same as the above-mentioned locus, then the tobacco plant is a cytoplasmic male sterile line tab1-CMS variety.

Embodiment 3

[0064] A method for identifying or assisting in identifying tobacco cytoplasmic male sterile line tab1-CMS varieties:

[0065] Utilize the S3 primer pair: forward primer 5'-AGGGGGCAGCAACTACAAT-3' and reverse primer 5'-CGGAAAAAATGATAAATAAAT-3' to carry out PCR amplification of the tobacco to be tested, carry out agarose gel electrophoresis to the PCR amplified product, if amplified If there is a specific band with a size of 588bp, the tobacco plant is a cytoplasmic male sterile line tab1-CMS variety.

[0066] The total genomic DNA of ms Zhongyan 100, Zhongyan 201, NC55, SK01, YL-1, YL-6, Zhongyan 100, and K326 were extracted respectively; eight kinds of tobacco materials were amplified by PCR with S3 (S3:5'-AGGGGGCAGCAACTACAAT -3' and 5'-CGGAAAAAATGATAAATAAAT-3'); The amplified PCR product is subjected to agarose gel electrophoresis, such as figure 1 As shown, a 588bp size-specific band can only be amplified in tab1-CMS. Therefore, the method can be used to identify or assist...

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Abstract

The invention belongs to the field of plant genetic engineering, and relates to a tobacco cytoplasmic male sterile line tab1-CMS specific molecular marker and application thereof. According to the invention, a specific molecular marker of the tobacco tab1-CMS variety is obtained through screening; and the obtained specific molecular marker can be used for rapidly and accurately identifying the tobacco tab1-CMS variety. The specific molecular marker lays a foundation for enriching tobacco sterile cytoplasm sources and using tobacco excellent sterile cytoplasm in breeding and production.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and relates to a tobacco cytoplasmic male sterile line tab1-CMS specific molecular marker and application thereof. Background technique [0002] The methods for identifying the types of cytoplasmic male sterile lines include morphology, cytology, and molecular markers, among which molecular markers are the main method for identifying the types of cytoplasmic male sterile lines, including mitochondrial specific genes, mitochondrial / chloroplast SSR, SNP, etc. Chloroplast is an organelle related to CMS. There are a large number of SNPs, InDels, translocations, duplications, point mutations, and genomic structural variations in the intergenic and intronic regions of the chloroplast genome, with higher nucleotides than the nuclear genome Replacement rate, which can be used to distinguish cytoplasmic male sterile lines. Chloroplast trnC-trnD is about 3000bp, including four genes trnC, petN, ps...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2600/13C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 李凤霞刘艳芳刘志文杨爱国孙玉合
Owner TOBACCO RES INST CHIN AGRI SCI ACAD
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