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SSR primers and method for purity identification of luffa hybrid seeds

A technology for the purity and identification method of hybrid seeds, which is applied in the field of identified SSR primers and the identification of the purity of loofah hybrid seeds, can solve the problems that have not been seen in identifying the purity of loofah hybrid seeds, and achieve high commercial application value, strong specificity, and convenient operation Effect

Active Publication Date: 2016-08-03
CROP RES INST OF FUJIAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there has been no report on the use of SSR molecular marker technology to identify the purity of loofah hybrid seeds

Method used

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  • SSR primers and method for purity identification of luffa hybrid seeds
  • SSR primers and method for purity identification of luffa hybrid seeds
  • SSR primers and method for purity identification of luffa hybrid seeds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The establishment of embodiment 1 hybrid luffa seed purity detection method

[0026] 1. Screen the SSR primers for purity identification

[0027] Several pairs of primers designed according to the sequencing results of the loofah transcriptome were screened between parents and offspring, and the primer SGSSR4 with a co-dominant differential marker band was selected. The marker band pattern is clear and reproducible. The sequence is as follows:

[0028] SGSSR4-F:5'-AGGGGTTAGTCGACCGATTT-3' (SEQ ID NO.1);

[0029] SGSSR4-R: 5'-TGCTTGCCACTTGAAGAAAC-3' (SEQ ID NO. 2).

[0030] Using loofah genomic DNA as a template, PCR amplification was performed using the above-mentioned SSR primer SGSSR4, and the amplified product was subjected to gel electrophoresis. The electrophoresis results showed that the SSR primer SGSSR4 could produce a 151bp paternal-specific band and a 135bp maternal-specific band Bands.

[0031] 2. Using the above-mentioned specific primers to identify the p...

Embodiment 2

[0047] Adopt the method of embodiment 1 to 12 strains of loofahs that are taken from the seed purity identification field of Dongzhang base, number its individual plant, extract individual plant DNA and detect, see figure 2 , which is the electrophoretic analysis pattern of 12 individual plants and their parental amplification products in Example 2, wherein, M: Marker; P1: male parent; P2: female parent; 10: pseudohybrid; 1-9,11- 12: Sampling loofah hybrids. Test results: 11 hybrid seeds and 1 male parent (namely pseudo-hybrid), the seed purity is 91.67%, which is consistent with the field test results.

Embodiment 3

[0049] The method of embodiment 1 is adopted to identify 14 strains of luffa from the seed purity identification field of the vegetable center base, and its single plant numbering is carried out, and the single plant DNA is extracted for detection, see image 3 , which is the electrophoretic analysis pattern of the 14 individual plants and their parental amplification products in Example 3. Among them, M: Marker; P1: male parent; P2: female parent; 1-14: sampling loofah hybrids. Test results: All are real hybrids, the seed purity is 100%, which is consistent with the field test results.

[0050] The above examples show that the method of the present invention can effectively distinguish luffa hybrid seeds from their parent seeds, and quickly and accurately detect the purity of the seeds.

[0051] In a word, the advantages of the present invention are as follows: the present invention has the following advantages:

[0052] (1) The SSR primer SGSSR4 in the present invention ca...

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Abstract

The invention provides SSR primers and method for purity identification of luffa hybrid seeds. The primers comprise an upstream primer SGSSR4-F of a primer SGSSR4 and a downstream primer SGSSR4-R of the primer SGSSR4, wherein the nucleotide sequence of the upstream primer SGSSR4-F is as shown in SEQ ID NO. 1, and the nucleotide sequence of the downstream primer SGSSR4-R is as shown in SEQ ID NO.2. The method includes the following steps that a luffa parent and a hybrid progeny genome DNA are extracted; the hybrid progeny genome DNA serves as a template, and the primer SGSSR 4 is utilized to conduct PCR amplification; an amplification product is subjected to gel electrophoresis; an electrophoresis result is analyzed, only a single plant with parent specificity bands at the same time becomes a real hybrid seed, the plant in lack of any band is marked as a false hybrid seed, and seed purity is calculated; the primer SGSSR4 can generate 151bp male parent specificity bands and 135bp female parent specificity bands. The primers can generate male parent specificity marks and female parent specificity marks, and specificity is high; by means of the SSR primers, purity of the luffa hybrid seeds can be identified rapidly, conveniently, accurately and effectively.

Description

【Technical field】 [0001] The invention belongs to the technical field of agricultural vegetable breeding and application, and in particular relates to an SSR primer for identifying the purity of luffa hybrid seeds and a method for identifying the purity of luffa hybrid seeds. 【Background technique】 [0002] The test of Distinctness, Uniformity and Stability of new plant varieties (DSU test for short) is the most important work content in the substantive examination of the new variety rights granted by the Plant New Variety Protection Office of the Ministry of Agriculture. . Seed purity is one of the most important quality indicators for seed quality inspection. Due to the problem of seed purity, the incidents of sharply reduced production of farmers have occurred from time to time, which has seriously affected the increase in agricultural production and income and the stability of rural society. The traditional identification method of seed purity is based on the field obs...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 朱海生温庆放李祖亮刘建汀陈敏氡王彬张前荣
Owner CROP RES INST OF FUJIAN ACAD OF AGRI SCI
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