Molecular identification method for distant hybridization progeny of Sesame indicum L. and Sesame radiatum

A technology of distant hybridization and molecular identification, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., and can solve the problem of sesame distant hybridization having no affinity, poor reliability, and significant differences, etc. Problems, to achieve the effect of simple and easy pretreatment and test operation steps, stable and reliable data, and short cycle

Active Publication Date: 2013-03-13
HENAN ACAD OF AGRI SCI
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Problems solved by technology

However, due to the incompatibility of sesame distant hybridization, the progeny can hardly self-fertilize. At present, only the immature embryo rescue culture and tissue culture seedling transplanting methods can be used to obtain distant hybrid offspring.
In the identification of the offspring of distant hybrids, due to the limitation of research technology, only identification methods such as appearance and biology are used to judge the authenticity of the F1 generation of sesame distant hybrids
The study found that, unlike the seedlings, the tissue-cultured seedlings of sesame distant hybrids had no obvious differences in appearance, but the differences with their parents were significant.
Therefore, methods such as appearance and biology are used to identify the offspring of distant hybrids, and the reliability of the results is poor, and the identification period is long

Method used

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  • Molecular identification method for distant hybridization progeny of Sesame indicum L. and Sesame radiatum
  • Molecular identification method for distant hybridization progeny of Sesame indicum L. and Sesame radiatum
  • Molecular identification method for distant hybridization progeny of Sesame indicum L. and Sesame radiatum

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Embodiment

[0032] This example is used to cross F 1 Generation identification, the specific operation steps are as follows:

[0033] (1) Culture of distant hybrid parents and offspring: select plump and healthy Sesame indicum L. 11 (Sesame indicum L., 2n=26) and Yezhi 1 (Sesame radiatum, 2n=64) seeds, and plant them in June after sowing In late July, the plants entered the flowering period and were artificially pollinated and hybridized. The immature embryos of the hybrid progeny were cultured by tissue culture method to obtain the distant hybrid progeny F 1 A total of 4 strains ( figure 1 );

[0034] (2) Extract Sesame Yuzhi No. 11, Yezhi No. 1 and the hybrid offspring F 1 Genomic DNA, as a template;

[0035] The extraction of genomic DNA adopts the CTAB mini-extraction method:

[0036] a. Take 2-3 tender leaves of the parent and the F1 offspring of the distant hybrid, add 600 μl of pre-cooled freshly prepared extraction buffer, grind by electroporation, then centrifuge at 5000 rp...

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Abstract

Belonging to the field of biotechnologies, the invention discloses a molecular identification method for a distant hybridization progeny of Sesame indicum L. and Sesame radiatum. The method includes: extracting the genomic DNA of Sesame indicum L., 2n=26, the Sesame radiatum, 2n=64 and the hybridization progeny F1, conducting PCR amplification and gel electrophoresis detection, and determining whether the distant hybridization progeny F1 is true or false according to whether parental specific bands can be amplified simultaneously. The molecular identification method for a sesame distant hybridization progeny established in the invention solves the problems of long distant hybrid identification cycle, low accuracy and the like in the process of sesame distant hybridization breeding study, and fills the vacancy of sesame distant hybridization breeding technology in China. By means of the method, the sesame distant hybridization progeny F1 can be identified efficiently and rapidly, thus laying the material and technology bases for breeding of new disease-resistant sesame varieties.

Description

technical field [0001] The invention relates to a molecular identification method for plant hybrid offspring, in particular to a molecular identification method for distant hybrid offspring of cultivated sesame (2n=26) and wild species (Sesame radiatum, 2n=64), belonging to the field of biotechnology . Background technique [0002] Sesame (Sesame indicum L., 2n=26) is one of the oldest characteristic oil crops in the world. Sesame has been cultivated in my country for more than 3,000 years. Because of its high quality, it occupies an important position in international production and trade. However, due to the poor resistance to disease and water stains of sesame, it is easily affected by environmental conditions, and the stability of yield is not good. In the past two decades, the development of high-yield, stable-yield, disease-resistant and stress-resistant new varieties has always been a key task of sesame genetics and breeding in my country. [0003] Wild sesame spec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 张海洋苗红梅张体德魏利斌李春琚铭王慧丽
Owner HENAN ACAD OF AGRI SCI
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