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cDNA molecular marker of onion fertility restoration and applications thereof

A technology for molecular markers and fertility recovery, applied in recombinant DNA technology, DNA/RNA fragments, microbial measurement/inspection, etc., can solve the problem of increasing farmers' production burden and planting risks, high production costs of onions, and lack of hybrid generation Variety and other issues, to achieve the effect of saving hybridization test cross time, high cost, and long cycle

Inactive Publication Date: 2011-10-12
VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the long breeding cycle of onion varieties, which is 2-4 times that of ordinary vegetable crops such as cabbage and radish, the production is mainly based on imported varieties, and there is a lack of hybrid varieties with independent intellectual property rights. Therefore, it is necessary to spend a lot of foreign exchange from abroad Imported hybrids lead to high production costs of onions, increasing farmers' production burden and planting risks

Method used

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  • cDNA molecular marker of onion fertility restoration and applications thereof
  • cDNA molecular marker of onion fertility restoration and applications thereof
  • cDNA molecular marker of onion fertility restoration and applications thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0075] A cDNA molecular marker for restoring onion fertility, the nucleotide sequence of which is shown in SEQ ID NO.1.

[0076] To identify the specific primers for the above cDNA molecular markers, the nucleotide sequence of the upstream primer F240 is shown in SEQ ID NO.2, and the nucleotide sequence of the downstream primer R240 is shown in SEQ ID NO.3.

[0077] The cDNA molecular marker is obtained by the following steps:

[0078] (1) Detection and analysis of total RNA in samples of different tissue materials

[0079] The total RNA of different samples of onion was extracted by Trizol method, and then the total RNA was treated with DNaseI in order to completely remove the residual DNA contamination. The extracted total RNA was detected by spectrophotometer, where OD 260 / OD 280 The ratio is between 1.8-2.0; non-denaturing agarose gel electrophoresis analysis 28s, 18s bands are clear, no diffusion, no DNA residues (such as figure 1 shown). It is confirmed that the ex...

Embodiment 2

[0092] Application of the cDNA molecular markers and specific primers in Example 1 in the identification of onion nuclear interaction male sterility nuclear restoration materials and fertility maintenance materials The above application, the steps are as follows:

[0093] The Trizol method was used to extract the total RNA of the flower buds of the onion sample, and then the total RNA was reverse-transcribed into cDNA, and after PCR amplification with specific primers (upstream primer F240, downstream primer R240), the product was carried out on 1.5% agarose gel. Electrophoresis, voltage 140v, electrophoresis time 30 minutes, after ethidium bromide staining, compare with ethidium bromide staining DNA molecular markers after electrophoresis on 1.5% agarose gel under the same conditions as above, if it is far from the starting point of the swimming lane If there is a band at the same distance, it contains cDNA molecular markers, which is the sample of onion fertility recovery mat...

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Abstract

The invention relates to a cDNA molecular marker of onion fertility restoration and applications thereof, belonging to the technical field of biological technology. The invention provides a cDNA molecular marker of onion fertility restoration of which the nucleotide sequence is shown in SEQ ID NO.1, a specific primer for identifying the cDNA molecular marker, and applications of the cDNA molecular marker and the specific primer in the identification of the onion cytoplasmic-nuclear interactive male sterile fertility restorer line and maintainer line material, wherein the nucleotide sequence of the forward primer F240 is shown in SEQ ID NO.2 and the nucleotide sequence of the reverse primer R240 is shown in SEQ ID NO.3. The cDNA molecular marker is stable; and the selection is convenient to operate and the cost is saved.

Description

technical field [0001] The invention relates to a cDNA molecular marker for restoring onion fertility and its application, and belongs to the technical field of biotechnology. technical background [0002] Onion (Allium cepa L.), also known as onion, onion, Liliaceae, Allium, biennial plant, has a cultivation history of more than 5,000 years. It is a worldwide vegetable, and at least 175 countries cultivate onions. According to FAO (2009) statistics, the world's onion planting area is 3.7 million hectares, with an output of 72.3 million tons. Among all vegetable crops, its planting area ranks second, and its output ranks third; among them, my country has the world's largest onion production Area and output, the production area exceeds 1 million hectares, and the total output is 21.07 million tons, accounting for about 30% of the world's output. Onion has the reputation of health food. It can not only be eaten fresh, but also used in processed products in large quantities. It...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 吴雄霍雨猛刘冰江缪军杨妍妍张一卉
Owner VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI