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Molecular marker for identifying mitochondrion gene related to CMS (cytoplasmic male sterility) in green Chinese onion

A technology of mitochondrial genes and molecular markers, which is applied in the direction of introducing foreign genetic material by using vectors, cells modified by introducing foreign genetic material, DNA/RNA fragments, etc. Type detection and other issues, to achieve the effect of convenient identification, high reproducibility and wide application range

Inactive Publication Date: 2014-08-13
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Application Information

AI Technical Summary

Problems solved by technology

There is only one SCAR molecular marker for identifying the cytoplasmic type of scallions that has been reported, that is, SCS13 above, but the variety of scallions detected by this marker is limited, and it cannot be widely used in the detection of cytoplasmic types of multiple varieties of scallions

Method used

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  • Molecular marker for identifying mitochondrion gene related to CMS (cytoplasmic male sterility) in green Chinese onion
  • Molecular marker for identifying mitochondrion gene related to CMS (cytoplasmic male sterility) in green Chinese onion
  • Molecular marker for identifying mitochondrion gene related to CMS (cytoplasmic male sterility) in green Chinese onion

Examples

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Embodiment 1

[0040] This example is a molecular marker for identifying mitochondrial genes related to cytoplasmic male sterility (CMS) in scallions and a preparation method thereof.

[0041] A molecular marker for identifying the mitochondrial gene related to CMS in scallions, the nucleotide sequence of the molecular marker of the mitochondrial gene related to CMS is shown in SEQ No: 1 in the sequence listing.

[0042] The nucleotide sequences of the primers used to amplify the molecular markers of mitochondrial genes related to CMS are shown in SEQ No: 2 and SEQ No: 3 in the sequence listing.

[0043] A recombinant expression vector, a transgenic cell line, and a host bacterium all contain the molecular marker of the mitochondrial gene related to CMS.

[0044] A preparation method for identifying molecular markers of mitochondrial genes related to CMS in scallions, the nucleotide sequence of the molecular markers of mitochondrial genes related to CMS is shown in SEQ No: 1; the steps of th...

Embodiment 2

[0091] This example is the application of the molecular markers of mitochondrial genes related to CMS in identifying the cytoplasmic types of scallions.

[0092] In order to detect the reliability of the molecular markers of the mitochondrial genes associated with CMS, 4 pairs of sterile lines and maintainer lines (respectively selected from Zhangqiu scallions, whip scallions, Wuyeqi, Dongling Bai) and 4 hybrid varieties introduced from Japan (Tianguang Ichiba, Yamamoto Wanshou F1, Shoufeng F1, Guangsaber F1) were used as the scallions to be tested.

[0093] The steps of the detection method include:

[0094] A, get the blade of fresh scallion, adopt CTAB method to carry out the extraction of scallion DNA, obtain the total DNA of the detected scallion;

[0095] The steps of extracting and processing the total DNA of scallions are:

[0096] a. Take 0.05-0.2g of leaves from the scallions of the above varieties, preferably 0.1g, put them into a mortar, add 1ml CTAB buffer solut...

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Abstract

The invention provides a molecular marker for identifying a mitochondrion gene related to CMS (cytoplasmic male sterility) in a green Chinese onion, a preparation method and an application method thereof. The nucleotide sequence of the molecular marker of the mitochondrion gene related to CMS is shown as SEQ No:1 in a sequence table; and the nucleotide sequences of primers for amplifying the molecular marker are shown as SEQ No:2 and SEQ No:3 in the sequence table. According to the invention, the AFLP (amplified fragment length polymorphism) molecular marker of the mitochondrion gene related to CMS in a green Chinese onion is converted into the SCAR (sequence characterized amplified region) molecular marker, so that a large number of samples can be quickly detected, the result is good in stability, the reproducibility is high, and the operation is facilitated. The SCAR marker in the invention can detect all varieties in the experiment, namely 8 sterile line varieties, 8 maintainer line varieties and 4 hybrid varieties; and the marker has a wide application range, can conveniently identify the cytoplasm type of the green Chinese onion and assists the development of molecular breeding.

Description

technical field [0001] The invention belongs to the technical field of molecular markers, and specifically relates to a molecular marker for identifying mitochondrial genes related to cytoplasmic male sterility (CMS) in scallions, and a preparation method and application method of the molecular marker. Background technique [0002] Cytoplasmic male sterility (CMS) is maternally inherited and cannot produce fertile pollen. Cytoplasmic male sterility has been found in more than 300 kinds of plants and is widely used in hybrid seed production. So far, most of the genes related to CMS reported are located in the mitochondrial genome, and these genes are usually rearranged, including translocation, insertion and deletion. [0003] Green onion (Allium fistulosum L.) is one of the important vegetable crops widely cultivated in East Asia. Zhang Qipei et al first reported the male sterility of scallions in China (Zhang Q P, Wei YY, Zhang Q. 1987. Male sterility of welsh onion (Alli...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21
Inventor 王永勤王婵李洪有
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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