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Identifying and utilizing method of rice wide-compatibility recessive male nuclear sterile line

A technique of recessive sterility and wide-breeding, which is applied in the fields of botany equipment and methods, biochemical equipment and methods, and microbial measurement/inspection. advanced questions

Inactive Publication Date: 2016-10-05
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the semi-sterile characteristics of indica-japonica hybrids, indica-japonica heterosis encounters great difficulties in practical application, including a series of problems such as high plant height, poor flowering period, and low seed-setting rate. The seed setting rate of hybrid F1 generation is low, and the seed setting rate of most hybrids is between 5% and 40%.

Method used

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  • Identifying and utilizing method of rice wide-compatibility recessive male nuclear sterile line
  • Identifying and utilizing method of rice wide-compatibility recessive male nuclear sterile line
  • Identifying and utilizing method of rice wide-compatibility recessive male nuclear sterile line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Constructing a recombinant inbred line and obtaining a sterile line.

[0036] according to figure 1 In the technical route shown, F 1 , using F 1 F 2 , from F 2 Randomly select 200 seeds in the middle, self-cross to get 200 families, choose 1 seed for each family to continue self-crossing, and get F 3 , also in F 3 In each family, one seed is self-crossed, and self-crossed to F 6 , 177 recombinant inbred lines (see figure 1 ), in which the 127th recombinant inbred line had a completely sterile mutant individual plant and a normal fertile individual plant, and their segregation ratio was 1:3; the sterile individual plant showed abnormal stamen development, and some stamens became pistil-like structure, or completely degenerated stamens. The genotype composition of RI127 was identified with 150 pairs of markers on the genome, and it was found that 55% of its genotype came from indica rice and 45% from japonica rice. At the same time, it was also foun...

Embodiment 2

[0077] Example 2: Map-based cloning of the male sterility gene.

[0078] Specific steps are as follows:

[0079] (1) Mapping population: cross the CMS line RI127S of mutant RI127 with the indica rice variety "MH63", and obtain F 2 The segregated population was used to map the male sterility gene. The investigation of field traits shows that the segregation ratio of sterile single plant: fertile single plant is 1:3, and the sterile single plant shows abnormal development of stamens, some stamens become a structure similar to pistils, or stamens are completely degenerated, such as Figure 4 shown.

[0080] (2) Total DNA extraction of rice: DNA extraction method refers to CTAB method (see Zhang et al., genetic diversity and differentiation of indica and japonica rice detected by RFLP analysis, 1992, TheorAppl Genet, 83, 495-499).

[0081] (3) Bulked segregant analysis (bulked segregant analysis) to locate target genes: Randomly select 120 (10 on each chromosome) SSR markers on...

Embodiment 3

[0092] Example 3 Obtaining Insertion Sequence and Verification of Genotype Identification by Gene Internal Markers

[0093] (1) Obtaining the insertion sequence

[0094] Using the total DNA of a single sterile rice plant RI127S as a template, TAIL-PCR amplification was performed, sequenced, and the insertion sequence was isolated. The specific method of TAIL-PCR is as follows: firstly, primary PCR is used, the total volume of the system is 20 μl: total DNA 40ng, 10×PCR buffer 2μl, 2mM dNTP 1.5μl, 25mMMg 2+ 1.5 μl, 10 μM specific primer (specific primer) 10.3 μl, 100 μM AD primer AD2a (primer) 0.2 μl, rTaq enzyme 0.1 μl, add double distilled water to 20 μl.

[0095] Reaction program: 94°C 5min; (94°C 30sec; 62°C 1min; 72°C 2.5min) × 5cycles; 94°C 30sec; 25°C 2min; 72°C (32% ramp), 2.5min; (94°C 20sec; 65°C 1min; 72℃2.5min; 94℃20sec; 65℃1min; 72℃2.5min; 94℃20sec; 45℃1min; 72℃2.5min)×15cycles; 72℃7min; 25℃10min. Then Secondary PCR, the total volume of the system is 20μl: the l...

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Abstract

The invention belongs to the field of molecular breeding of crops, and relates to an identifying and utilizing method of a rice wide-compatibility recessive male nuclear sterile line. Nonglutinous rice 'Teqing' and the wide-compatibility variety '02428' are adopted for constructing a recombinant inbred line (RIL), a selfing line with the fertility feature is selected, a novel sterile line RI127S is cultivated and screened, and the sterile line can be used as an intermediate material for rice rotational breeding. The identifying and utilizing method has the advantages that (1) the sterility of the male sterile line is controlled by recessive genes, and is not influenced by length of illumination and temperature under the normal growth condition, and the male sterile line is an ideal recurrent selection intermediate material for rice; (2) the sterile line RI127S contains wide-compatibility genes, the genome of the sterile line RI127S is a mixture of nonglutinous rice and japonica rice varieties, after hybridization with nonglutinous rice and japonica rice, the hybridization seeds are fertile, the F1 fruitage rate of nonglutinous rice is 85%, and the F1 fruitage rate of japonica rice is 80%; (3) the inbred line RI127 has the high yield property intrinsically, and can be taken as a parent of the good stock, and when the RI127 is taken as the sterile line, the process of artificial emasculation for hybridization can be saved.

Description

technical field [0001] The invention relates to the technical field of rice breeding, in particular to the utilization of a wide-compatibility recessive male sterile line in rice breeding. Background technique [0002] Fertility is an important trait of rice and is of great significance in rice production. The research on male sterility in rice was first conducted in the 1920s. Since the 1960s, many different genetic types of male sterile rice materials have been discovered, most of which are recessive genital sterility, such as photothermosensitive male sterility. In the 1970s and 1980s, the location of the rice male sterility gene was mainly based on the analysis of rice trisomies and trait markers to determine the chromosome where the gene is located, such as the recessive genic male sterile genes ms-1, ms-7, ms- 8. ms-9, ms-10, and ms-14 (Khush G S et al., 1991), now mainly use molecular markers to construct genetic linkage maps to locate genes, such as photosensitive s...

Claims

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Application Information

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IPC IPC(8): C12Q1/68A01H1/02A01H1/04
Inventor 邢永忠毛东海张莉
Owner HUAZHONG AGRI UNIV
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