SSR markers linked with resistance genes Bs_t of physiological races No. 0 and No. 1 of tobacco black shank and applications thereof

A technology of tobacco black shank and physiological race, applied in the field of SSR marker, can solve the problems of chain burden, medium resistance, difficulty in maintaining the quality of flue-cured tobacco, etc., and achieve the effect of low cost and fast cost

Active Publication Date: 2018-01-23
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above-mentioned tobacco germplasms with resistance to black shank No. 0 and No. 1 all have the following disadvantages: 1) It is a quantitative trait with moderate resistance and it is not easy to further use the resistance QTL for breeding
2) There is a linkage burden between the QTL controlling black shank resistance and unfavorable traits. For cigar tobacco Florida301 and Beinhart1000-1, when the resistance QTL is transferred to flue-cured tobacco, there will be some cigar genome in flue-cured tobacco. Fragments make it difficult to maintain the excellent quality of flue-cured tobacco itself

Method used

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  • SSR markers linked with resistance genes Bs_t of physiological races No. 0 and No. 1 of tobacco black shank and applications thereof
  • SSR markers linked with resistance genes Bs_t of physiological races No. 0 and No. 1 of tobacco black shank and applications thereof
  • SSR markers linked with resistance genes Bs_t of physiological races No. 0 and No. 1 of tobacco black shank and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Screening of SSR markers linked to Bs_t genes in tobacco race 0 and race 1 for resistance to black shank by segregating population analysis (BSA)

[0035] 1. Experimental materials

[0036] Flue-cured tobacco Honghua Dajinyuan, which is good in quality but susceptible to tobacco black shank physiological race 0 and 1, was used as the female parent, and the flue-cured tobacco breeding material GH14-62 (its The resistance of race 0 and race 1 to black shank disease is controlled by the Bs_t gene (nucleic acid fragment), and Nicotiana japonica ( N. rustica ) was formed by transferring the genome fragments of races 0 and 1 resistant to black shank into flue-cured tobacco safflower Dajinyuan) as the male parent, crossed and selfed to obtain F1 and F2 populations respectively.

[0037] 2. Identification of resistance to black shank race 0 and 1 in parents and F2 segregation populations

[0038]The test materials were transplanted to the field after they became seedlings, w...

Embodiment 2

[0047] Map Distance of Linkage Markers and Its Validation in Individual Plants of F2 Population

[0048] 1. Data Analysis

[0049] First, the tobacco genomic DNA was extracted according to the method described in Example 1, and the F2 population was cultured and multiplied for the identification of resistance to race 0 and race 1 of blackleg disease and the analysis of SSR markers. Secondly, 300 individual plants in the F2 population were genotyped using markers TM51207 and TM55805 screened by BSA method. Finally, carry out data statistics on the band type of each individual plant. The resistant homozygous band is marked as "B", the resistant heterozygous band is marked as "H", the susceptible band is marked as "A", and the band Unclear or no amplified bands were recorded as "U".

[0050] 2. Calculation of genetic distance of linkage markers

[0051] Using JoinMap 4.0 software combined with the identification data of black shank race 0 and race 1 resistance of individual F2...

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Abstract

The invention discloses SSR markers linked with resistance genes Bs_t of physiological races No. 0 and No. 1 of tobacco black shank and applications thereof. The SSR markers linked with resistance genes Bs_t of physiological races No. 0 and No. 1 of tobacco black shank are numbered as TM51207 and TM55805, and nucleotide sequences of amplified products of the SSR markers are shown as SEQ ID No. 1,SEQ ID No. 2, SEQ ID No. 3 and SEQ ID No. 4 respectively. The applications refer to applications of the SSR markers linked with resistance genes Bs_t of physiological races No. 0 and No. 1 of tobaccoblack shank in detecting whether resistance genes Bs_t of physiological races No. 0 and No. 1 of tobacco black shank exist in tobacco genomic DNA. The SSR markers disclosed by the invention have the characteristics of being stable, reliable, simple and convenient, rapid and low in cost, and therefore, the molecular markers can be applied as a molecular marker-assisted selection of Bs_t genes in breeding for disease resistance of tobacco black shank (physiological races No. 0 and No. 1).

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an SSR marker linked with tobacco black shank disease No. 0 and No. 1 physiological race resistance gene Bs_t and its application. Background technique [0002] Tobacco black shank is caused by Phytophthora nicotianae ( Phytophtora parasitica var. nicotianae Tucker), which can infect all cultivated tobacco, is one of the most devastating diseases in tobacco production. It was first reported in China in 1950 in the Huanghuai tobacco area (Shang Zhiqiang. Pathogen of tobacco black shank , Occurrence law and research progress of comprehensive control. China Agricultural Science and Technology Herald, 2007, 9(2):73-76.). There are 4 identified physiological races of tobacco black shank: No. 0 and No. 1 physiological races were discovered in North Carolina, USA in 1962 (Apple J L. Physiological specialization within Phytophthora nicotianae var. nicotianae .Phytopatholog...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 童治军肖炳光曾建敏陈学军方敦煌吴兴富李永平
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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