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Marker and method for identifying cytoplasmic sterility homozygous restorer line of Gossypium harknessii

A technology for Harknessy cotton and homozygous restorer lines, which is applied in the field of identification of markers for homozygous restorer lines of Harknessy cotton cytoplasmic sterility, can solve the problems of poor accuracy, large consumption, long time required, and the like, Achieve the effect of high accuracy, fast speed and improved accuracy

Active Publication Date: 2013-12-25
COTTON RES INST SHANXI ACAD OF AGRI SCI
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AI Technical Summary

Problems solved by technology

[0004] In order to solve the problems of long time required for field test crossing and identification of the cytoplasmic sterile restorer line of Harknessy cotton, large consumption and poor accuracy, the present invention provides a method for identifying the cytoplasmic sterile of Harknessy cotton Primers and methods for homozygous restorer lines, so as to speed up the process of restorer line selection and improvement, while ensuring the homozygosity of the restorer lines

Method used

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  • Marker and method for identifying cytoplasmic sterility homozygous restorer line of Gossypium harknessii
  • Marker and method for identifying cytoplasmic sterility homozygous restorer line of Gossypium harknessii

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preparation example Construction

[0022] Material preparation and method:

[0023] (1) (19R × Jinmian 50) BC 3 F 2 : The cytoplasmic sterile restorer line of Harknessy cotton was used as the female parent, and Jinmian 50 was used as the male parent (recurrent parent). After crossing with the restorer line for one generation and backcrossing for three generations, BC 3 F 1 Fertile plants in segregating populations are selfed to obtain BC 3 F 2 separate groups.

[0024] (19R×GK44)F 2 : Taking the restorer line of Harknessey cotton cytoplasmic sterility as the female parent and GK44 as the male parent, after crossing with the restorer line for 2 generations, (19R×GK44)F was obtained 2 .

[0025] (2) Pollen fertility survey: The fertility survey method refers to the method of Justus (Justus N, Leinweber CL. A heritable partially male sterile character in cotton. J Hered (1960) 51 (4): 191-192), using hand twist Whether pollen grains appear in broken anthers is the criterion for distinguishing fertile and s...

Embodiment 1

[0028] In the summer of 2010, it was planted on the farm of the Cotton Research Institute of Shanxi Academy of Agricultural Sciences (19R × Jinmian 50) BC 3 F 2 Divide colonies and plant 8 rows of 23-25 ​​plants each. Since the female parent is derived from a restorer line with cytoplasmic sterility, fertility segregation occurs in the progeny. There are 196 isolates in this segregated population. First, the anthers were broken to identify the fertility of a single plant. Among them, 125 were fertile plants, and among the fertile plants , a total of 48 individual strains amplified by the SSR marker NAU5121 primer with an amplified band at 170 bp and without an amplified band at 190 bp were selected.

[0029] In the winter of 2010, field test-crossing and identification of the above-mentioned individual plants with 170 bp amplification band and no 190 bp amplification band were carried out in Hainan. The results showed that 47 of them were confirmed to be homozygous restorer p...

Embodiment 2

[0031] Planted on the farm of the Cotton Research Institute of Shanxi Academy of Agricultural Sciences in the summer of 2010 (19R×GK44)F 2 Divide colonies and plant 20 rows of 23-25 ​​plants each. There were 483 plants in this isolated population, and DNA was extracted from each individual plant of the isolated population before flowering. In this isolated population, a total of 349 individual strains with two amplified bands at 1200 bp and 1045 bp amplified by the marker PF1 primer, 126 strains with only 1045 bp band type, and 126 strains with no amplified band type 8 strains, see PCR amplification results figure 1 ; 349 strains were identified by SSR marker NAU5121 primer in the second round, and there were 117 individual strains with 170 bp amplification band and no 190 bp amplification band. The PCR amplification results are shown in figure 2 , 117 individual plants were test-crossed and combined with sterile lines.

[0032] In the winter of 2010 in Hainan, the above-...

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Abstract

The invention relates to the technical field of molecular breeding of cotton, in particular relates to a marker and a method for identifying a cytoplasmic sterility homozygous restorer line of Gossypium harknessii. Compared with the prior art, the marker and the method have the beneficial effects of high speed and high accuracy as well as practicability and simplicity, wherein when a primer provided by the invention is used for identifying the molecular marker, the homozygosity of the restorer line (or strain) can be quickly identified, and the workload can be reduced by more than 70% compared with that of traditional identification methods; the primer has the advantages of simplicity, stability, reliability, high repeatability and the like and can be used for quickly identifying the homozygosity of restore genes rapidly, the genetic condition of the restore genes can be truly identified in the level of DNA, and thus the selection accuracy is improved to be above 97% at least; all procedures of the identification method provided by the invention are easy to operate, and thus the method has good scientific research and commercial application prospects.

Description

technical field [0001] The invention relates to the technical field of cotton molecular breeding, in particular to a marker and a method for identifying a homozygous restorer line of cytoplasmic sterile Hackney West cotton. Background technique [0002] Hybrid cotton is an important way to obtain high yield of cotton in my country. Artificial hybrid cotton has been widely used in the cotton areas of the Yangtze River Basin and the Yellow River Basin. In recent years, with the increase of labor costs, the cost of manual castration hybrid seed production remains high, and the promotion of hybrid cotton has been affected. The use of cotton cytoplasmic male sterile three-line matching seed production has the advantages of labor saving, high seed purity, and low cost. In the case of the declining cotton planting area in my country, the use of cotton cytoplasmic male sterile three-line matching seed production is an inevitable trend in the development of cotton hybrid productio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 李朋波曹美莲吴翠翠杨六六曹彩荣刘惠民王长彪
Owner COTTON RES INST SHANXI ACAD OF AGRI SCI
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