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Molecular marker method for identifying paddy rice Dongxiang wild cytoplasmic male sterility sources

A technology of male sterility and molecular markers, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of long cycle, time-consuming, authenticity and purity identification with certain complexity and other problems to achieve reliable results and good experimental repeatability

Inactive Publication Date: 2016-04-13
江西省农业科学院水稻研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] It can be seen that there are many sources and varieties of sterile cytoplasm of rice three-line hybrid rice, and its authenticity and purity identification has certain complexity
At present, the identification of seed authenticity and purity mostly adopts field planting identification. This method is long-term, labor-intensive and time-consuming, and cannot meet the needs of seed market sales and timely authorization of varieties. Therefore, an identification method that is simple to operate, efficient and convenient is needed.

Method used

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  • Molecular marker method for identifying paddy rice Dongxiang wild cytoplasmic male sterility sources
  • Molecular marker method for identifying paddy rice Dongxiang wild cytoplasmic male sterility sources
  • Molecular marker method for identifying paddy rice Dongxiang wild cytoplasmic male sterility sources

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Embodiment 1

[0021] Example 1 Detection of Cytoplasmic Male Sterile Lines of Rice and Hybrid Combination Cytoplasm

[0022] 1) Test materials

[0023] Table 1 Eight rice materials

[0024]

[0025] 2) Rice total DNA extraction

[0026] Get paddy rice leaf at seedling stage, the extraction of genome total DNA is with reference to the CTAB method of Scott et al. (1988), improves as follows:

[0027] ①Take about 0.1g leaves, put them in a 2.0mL centrifuge tube, add 1 steel ball and 500μL 2×CTAB DNA extract (20gCTAB, 12.1gTris-Base, 7.44gEDTA-Na, 81.9gNaCl, dissolved in 1L distilled water), use tissue The grinder breaks the leaves, bathes in water at 65°C for 30 minutes, and vibrates and mixes several times during the period;

[0028] ②Take out the centrifuge tube from the water bath, add about 500 μL of chloroform (CHCl 3 ), fully shake and mix, and centrifuge at 10000rpm for 5min;

[0029] ③Transfer the supernatant to a new 1.5mL centrifuge tube, add isopropanol 0.7 to 1 times the vo...

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Abstract

The invention discloses a molecular marker method for identifying paddy rice Dongxiang wild cytoplasmic male sterility sources. Molecular marker primers are as shown in SEQ ID No. 1 and SEQ ID No. 2. The method includes: extracting the total DNA of paddy rice materials; using the extracted total DNA as template, and using the primers to perform PCR amplification; performing electrophoresis detection on amplification products, determining that the paddy rice materials contain the Dongxiang wild cytoplasmic male sterility sources if 92bp bands appear, and determining that the paddy rice materials do not contain the Dongxiang wild cytoplasmic male sterility sources if 102bp bands or other bands other than the 92bp bands appear. The primers can simply, fast and accurately identify the Dongxiang wild cytoplasmic male sterility sources and are good in experiment repeatability and reliable in result.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a molecular marker method for identifying the source of rice Higashino-type cytoplasmic male sterility, specifically identifying the cytoplasm source of rice Higashino-type cytoplasmic male sterile lines, Higashino-type hybrid combinations and progeny materials. Background technique [0002] Rice is one of the most important food crops, and more than half of the world's population uses rice as a staple food. With the continuous increase of world population and the continuous decrease of arable land, how to further increase rice yield has always been the primary concern of breeders. The practice of hybrid rice in my country for more than 40 years shows that hybrid rice technology is an effective way to increase rice yield. At present, the annual planting area of ​​hybrid rice in my country is about 15.333 million hm 2 , accounting for 50% of the total rice planting area and 57% of the...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 严松黄仁良沈显华朱珊熊宏亮沈林军魏永清陈莉妹
Owner 江西省农业科学院水稻研究所