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Method for detecting three-line hybrid rice honglian-type new restoring gene by using molecule mark sensing

A technology for three-line hybrid rice and gene restoration, which is applied in the field of molecular marker detection, can solve the problems of differences in the relationship between restoration and preservation, and the inability to restore CMS-D2-2, etc., and achieve the effects of convenient operation, increased genetic diversity, and simple equipment

Inactive Publication Date: 2008-07-23
常熟紫金知识产权服务有限公司
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Problems solved by technology

But on the contrary, the latter two cytoplasmic restorer genes cannot restore CMS-D2-2, which shows that there are differences in their restoration relationship (Zhang and McD.Stewart, 2001)

Method used

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  • Method for detecting three-line hybrid rice honglian-type new restoring gene by using molecule mark sensing

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Embodiment Construction

[0061] Using PCR amplification to quickly detect the molecular markers of the rice red lotus type new restorer gene, combined with genetic analysis to screen the red lotus red lotus type new restorer gene as an example, the specific steps are as follows:

[0062] 1. PCR primer design

[0063] The primers were designed based on the co-segregated molecular markers M1 and M2 of the rice red lotus-type cytoplasmic restorer genes Rf5 and Rf6. The sequences are as follows:

[0064] M1 primer:

[0065] Forward primer F1: 5'-ATGACAAATCTGCTCCGATG-3'

[0066] Reverse primer R1: 5'-CTTACTTAGGAAAGACTAC-3'

[0067] M2 primers:

[0068] Forward primer F2: 5'-GGAGATGCTATAGCAGCAGTG-3'

[0069] Reverse primer R2: 5'-ATTGCTCCTTACCACCTTGC-3'

[0070] 2. Extraction of DNA:

[0071] For each rice variety, take a section of rice tender leaves with a length of 3-5cm, a width of 0.5-1cm, and a weight of about 0.1g and 0.2g of powdered quartz sand, put them in a 1.5ml centrifuge tube in turn, gr...

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Abstract

The invention discloses a Honglian type new gene restoring method which utilizes molecule marker to detect the three-line hybrid rice, comprising the steps as follows: first, the design of the specific marker primer; second, the trace extraction of DNA; third, the construction of standard PCR expansion system; forth, the detection analysis of the expanded molecule marker; fifth, the determination of new restoring gene. The Honglian type new gene restoring method is based on the molecule marker of restoring gene co-segregation; the rice germplasm material is screened fast, accurately and reliably in laboratory. The Honglian type new gene restoring method has the advantages of simple method, convenient operation, strong purpose and pertinence for breeding of new restoring line, and increased breeding efficiency of new restoring line.

Description

technical field [0001] The invention relates to the technical field of detection using molecular markers, in particular to a method for using molecular markers to detect a new restoration gene of the three-line hybrid rice red lotus type, which is suitable for the cytoplasm of the red lotus type and the type containing orfH79 gene variation Restoration genetic testing for male sterility. Background technique [0002] The use of molecular markers for variety selection or variety identification is the main content of current plant biotechnology breeding. It has the advantages of accuracy, speed, and no seasonal restrictions, so it is widely used in production. Molecular marker-assisted selection is the product of the combination of modern biotechnology and traditional breeding technology. It transforms the extensive and empirical breeding technology in traditional genetics and breeding into modern fine and directional selection, making breeding more operable. If the DNA sequ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 李绍清谭艳平朱英国
Owner 常熟紫金知识产权服务有限公司
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