SSR primer SSR81 used for identifying purity of hybrid seeds of Xiaguan No.1 zucchinis and identification method

A technology for hybrid seed purity and identification method, which is applied in the field of hybrid seed purity identification, can solve the problems of hybrid seed impurity, untimely emasculation, biological mixing, etc., and achieve the effects of short identification period, simple operation and low cost

Active Publication Date: 2016-05-04
INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reasons for the impurity of hybrid seeds may be: the female parent is not castrated in time or completely in the field during seed production, resulting in biological mixing caused by the female parent's self-fertilization; the field is not well isolated and the pollen of other varieties is mixed. Biological mixing; mechanical mixing caused by male parents not being pulled out in time before harvest or mixed with other varieties during drying and processing after harvest

Method used

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  • SSR primer SSR81 used for identifying purity of hybrid seeds of Xiaguan No.1 zucchinis and identification method
  • SSR primer SSR81 used for identifying purity of hybrid seeds of Xiaguan No.1 zucchinis and identification method
  • SSR primer SSR81 used for identifying purity of hybrid seeds of Xiaguan No.1 zucchinis and identification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The screening process of the SSR primer SSR81 for the purity identification of Xiaguan No. 1 Zucchini hybrid seeds provided in this example is as follows:

[0036] Several pairs of primers designed according to the sequencing results of the wax gourd transcriptome were screened between the parents (Xiaguan No. The primer SSR81 with the band, the marker band pattern is clear and reproducible, and the sequence is as follows:

[0037] SSR81-F: 5'-GACAGAATTGGGGGTTTGTG-3' (SEQ ID NO.1);

[0038] SSR81-R: 5'-TCAACAGCAGTTGGTGGAAG-3' (SEQ ID NO. 2).

[0039] Using zucchini genomic DNA as a template, use the above SSR primer SSR81 for PCR amplification—PAGE for polyacrylamide gel electrophoresis of the amplified product—recover DNA from specific fragments (polyacrylamide non-denaturing gel)—PCR amplification—agarose Gel recovery—after TA clone sequencing, the analysis results showed that the primer SSR81 can produce a 181bp maternal specific marker SSR81 181 and the 188bp pat...

Embodiment 2

[0041] The method for identifying the purity of Xiaguan No. 1 Zucchini hybrid seeds provided in this example comprises the following steps:

[0042] (1) Genomic DNA extracted from the cotyledons of Chickpea

[0043] The experimental materials are "Xiaguan No. 1" Chinese zucchini hybrid experimental species and its male parent (Xiaguan No. 1 male parent) and female parent (Xiaguan No. 1 female parent) cotyledons. The steps for extracting cotyledon DNA are as follows:

[0044] ①Put a piece of cotyledon into a 2mL centrifuge tube, add liquid nitrogen and grind it with a grinding pestle, quickly add 800 μL 2% CTAB extract when the liquid nitrogen is quickly evaporated to dryness, mix well and place in a 65°C water bath for 45 minutes (shake well once every 10 minutes) );

[0045] ② After standing at room temperature, add 800 μL of chloroform:isoamyl alcohol (24:1), mix gently, let stand for 2 minutes, centrifuge at 12000 rpm for 15 minutes, transfer the supernatant (about 510 μL) t...

Embodiment 3

[0063] The method of Example 2 is used to plant and sample the Xiaguan No. 1 Chinese gourd hybrid that needs to be identified for its purity. There are 294 samples in total, and the individual plants are numbered sequentially, and the DNA of the individual plants is extracted for detection (see Figure 2-9 ), test results: 213 hybrid seeds, 81 female parents, seed purity of 72.45%, consistent with field test results.

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Abstract

The invention discloses an SSR primer SSR81 used for identifying the purity of hybrid seeds of Xiaguan No.1 zucchinis. The primer SSR81 comprises an upstream primer SSR81-F and a downstream primer SSR81-R, wherein the nucleotide sequence of the upstream primer SSR81-F is shown as SEQ ID NO.1; the nucleotide sequence of the downstream primer SSR81-R is shown as SEQ ID NO.2. The invention further discloses an identification method for the purity of the hybrid seeds of the Xiaguan No.1 zucchinis. The primer disclosed by the invention can generate a female parent specific marker and a male parent specific marker, and has good repeatability and high specificity. By utilizing the SSR primer, the method can be used for accurately, rapidly, conveniently and effectively identifying the purity of the hybrid seeds of the Xiaguan No.1 zucchinis.

Description

technical field [0001] The invention belongs to the technical field of purity identification of hybrid seeds, and in particular relates to an SSR primer SSR81 and an identification method for the identification of the purity of Xiaguan No. 1 zucchini hybrid seeds. Background technique [0002] Seeds are important means of agricultural production, and the quality of seeds directly affects the effect of agricultural production. Seed purity is an important index to measure seed quality. Establishing a reliable and convenient method for detecting seed purity is of great significance for accurately and quickly evaluating seed quality and ensuring the interests of seed producers, sellers and users. [0003] "Xiaguan No. 1" zucchini is a first-generation hybrid zucchini variety bred by the Vegetable Research Institute of Guangdong Academy of Agricultural Sciences. During the production process of its hybrids, the parental inbred lines are planted in rows according to a certain rat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 何晓明彭庆务马金露刘文睿谢大森江彪罗少波林毓娥郭汉权刘洪彪王瑞孙保娟梁肇均
Owner INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI
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