InDel molecular markers for assisted selection of early heading genes of rice and application of InDel molecule marker

A molecular marker, rice technology, applied in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of long growth period, affecting popularization and application, and not suitable for late production and planting, etc. achieve the effect of speeding up the cultivation

Active Publication Date: 2020-05-29
GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The indica variety 9311 (Yangdao No. 6) is an excellent backbone parent of hybrid rice. Two super rice combinations Liangyoupeijiu and Y Liangyou 1 have been bred by using it as the male parent. In the double-cropping rice area, due to its long growth period, it is not suitable for late-growing production and planting, which seriously affects its popularization and application, and restricts the increase in grain production.

Method used

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  • InDel molecular markers for assisted selection of early heading genes of rice and application of InDel molecule marker
  • InDel molecular markers for assisted selection of early heading genes of rice and application of InDel molecule marker
  • InDel molecular markers for assisted selection of early heading genes of rice and application of InDel molecule marker

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Experimental program
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Effect test

Embodiment 1

[0028] 1. Identification of heading date of chromosome segment substitution line

[0029] Through the identification of the heading date of 113 chromosomal fragment substitution lines with indica rice 9311 as the genetic background, it was found that a substitution line CL33 headed 15-20 days earlier than the recipient parent 9311. figure 1 shown.

[0030] 2. Genetic analysis of CL33 heading stage genes

[0031] Pass to 9311 / CL33 F 2 524 isolated individual plants were investigated for heading date, and the results were as follows figure 2 As shown, their variation range is 80-109 days, and the segregation ratio of early and late heading is 127:394, which is in line with the segregation ratio of 1:3. Genetic analysis shows that the early heading of CL33 is controlled by a pair of recessive genes, of which late heading is controlled by a pair of recessive genes. Heading is dominant to early heading.

[0032] 3. Whole gene resequencing and BSA analysis based on CL33, 9311 a...

Embodiment 2

[0058] (1) Rice samples

[0059] Early and late heading parents CL33 and 9311, 9311 / CL33 hybrid F 2 In the genetic segregating population, 31 plants showed early heading (80-85 days), 2 plants showed late heading (105-106 days) and 1 plant with a heading period of 94 days.

[0060] (2) DNA sample extraction and detection

[0061] DNA from each rice sample was extracted according to the CTAB method (Murray & Thompson 1980). DNA purity and integrity of each sample were analyzed by agarose gel electrophoresis; 260 / 280 The ratio is between 1.8 and 2.0); Qubit accurately quantifies the sample DNA concentration (sample concentration > 20ng / μL).

[0062] (3) PCR amplification

[0063] PCR amplification was performed according to the method of Panaud et al. (1996) with slight modifications. The 20 μL reaction system includes 0.15 μM primer, 200 μM dNTP, 1× PCR reaction buffer (50 mM KCl, 10 mM Tris-HCl pH8.3, 1.5 mM MgCl 2 , 0.01% gelatin), 50-100ng of DNA template, 1UTaq enzyme...

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Abstract

The invention discloses InDel molecular markers for assisted selection of early heading genes of rice and application of the InDel molecule markers, and belongs to the technical field of molecular markers. A whole-genome resequencing BSA technology is utilized to initially identify a candidate region related to the heading stage of rice, and then the InDel markers which are developed through parental whole-genome resequencing are utilized to carry out marking analysis on 500 early-heading single plants in an F2 segregation population which is constructed through hybridization of 9311/CL33 in the candidate region; and it is revealed that a PSM8-8 marker has coseparation with the genes, and finally the gene qEH8-1 of the early heading stage is finely positioned within a range of about 270 kbbetween InDel-markers PSM8-6 and PSM8-9. Amplification is performed on a rice plant genome through the designed Indel molecular marker PSM8-8, and whether plants have the characteristics of early heading or not is judged and detected through denaturing polyacrylamide gel electrophoresis.

Description

technical field [0001] The invention belongs to the technical field of molecular markers, in particular to an InDel molecular marker for assisting selection of rice early heading genes and its application. Background technique [0002] The heading date (growth period) of rice is one of the important agronomic traits of rice. The length of the heading date of rice directly determines the regional adaptability and seasonal adaptability of rice varieties. It plays an important role in the high and stable yield of rice varieties and is an important goal of rice breeding. . Breeding early-maturing and high-yielding rice varieties has always been the focus of rice breeders. The heading date of a rice variety is mainly determined by its photosensitivity, thermosensitivity and basic vegetative growth; photosensitivity plays a decisive role in controlling the ripening stage for varieties with strong photosensitivity. For the varieties with the same light and temperature response in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 张月雄秦钢黄大辉韦敏益梁海福刘驰马增凤李振经
Owner GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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