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Primer and method for identifying purity of cucumber 'Yuexiu No.3' hybrid seed

A technology for hybrid seed purity and cucumber, applied in biochemical equipment and methods, microbe determination/inspection, DNA/RNA fragments, etc., can solve problems such as false hybrids, decreased seed genetic purity, production loss, etc., to achieve low cost, High commercial application value, simple operation effect

Active Publication Date: 2015-06-17
INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Limited parental resources and the continuous emergence of hybrid varieties have made the genetic differences between varieties, especially hybrid seeds, smaller and smaller, and the authenticity and purity of vegetable seeds have become more and more difficult to identify. In addition, cucumber is a self-pollinating crop. , Artificial detasseling is not timely, false hybrids often appear, resulting in a decline in the genetic purity of the seeds, causing huge losses to production

Method used

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  • Primer and method for identifying purity of cucumber 'Yuexiu No.3' hybrid seed
  • Primer and method for identifying purity of cucumber 'Yuexiu No.3' hybrid seed
  • Primer and method for identifying purity of cucumber 'Yuexiu No.3' hybrid seed

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Establishment of a method for testing the purity of hybrid cucumber 'Yuexiu 3' seeds.

[0033] 1. Screen the SSR primers for purity identification.

[0034] Screen between the parents of the published cucumber SSR primers, and select two sets of primers for co-dominant differential marker bands, wherein,

[0035] The sequence of primer set 1 is as follows:

[0036] 3B-10-F: 5'-CATCACGACCCTCTCCATCT-3' (SEQ ID NO: 1);

[0037] 3B-10-R: 5'-GGGTTTGATAGTGGAGATTATTCA-3' (SEQ ID NO: 2);

[0038] The sequence of primer set 2 is:

[0039] 2B-41-F: 5'-CACCGAGAACGAAAGAAGGA-3' (SEQ ID NO: 3);

[0040] 2B-41-R: 5'-TCTAAGGTGGGAGGGAAACC-3' (SEQ ID NO: 4).

[0041] The marker bands of the two sets of primers are clear and reproducible, and the primers can produce maternal-specific markers and paternal-specific markers.

[0042] 2. Using the above-mentioned specific primers to identify the purity of hybrid cucumber 'Yuexiu 3' seeds.

[0043] (1) Extraction of Cucumber...

Embodiment 2

[0068] Embodiment 2 specificity experiment

[0069] Take the seeds of the same type of cucumber varieties on the market: Zhongnong 108, Yuanfeng, Jinlv 5, Zhongnong 8, Jinyan 4, Yuexiu 3, female parent A-2, male parent C-8, and extract them respectively DNA was amplified by PCR with primer set 1 and primer set 2, respectively. The results showed that only Yuexiu 3 had two specific bands, and other varieties had only one band or no band. image 3 is the electropherogram of the amplification product of primer set 1, Figure 4 Electropherogram of the amplification product of primer set 2.

Embodiment 3

[0070] Example 3 field application

[0071] Using the method of Example 1, 42 strains of 'Yuexiu No. 3' taken from the seed purity identification field of Baiyun base were numbered, and the DNA of the individual strains was extracted for detection. The results showed that the detection results of the two sets of primers were consistent, and the seed purity was 97.6%, which was consistent with the field investigation results, and the accuracy rate was 100%. Figure 5 is the electropherogram of the amplification product of primer set 1, Figure 6 Electropherogram of the amplification product of primer set 2.

[0072] The above examples show that the method of the present invention can effectively distinguish the 'Yuexiu No. 3' hybrid seeds from their parent seeds or seeds of other cucumber varieties, and quickly and accurately detect the purity of the seeds. And choose any one of the primers can be accurately identified.

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Abstract

The invention discloses a primer and a method for identifying purity of a cucumber 'Yuexiu No.3' hybrid seed. The method comprises the following steps of: (1) extracting deoxyribonucleic acid (DNA) of cucumber genome; (2) carrying out polymerase chain reaction (PCR) augmentation by using a screened specific primer; (3) carrying out gel electrophoresis on a PCR augmentation product; and (4) judging the purity of the seed according to the relative position on a gel band. By adopting the method disclosed by the invention, the 'Yuexiu No.3' hybrid seed can be distinguished from a female parent seed and a male parent seed; the purity of the hybrid seed can be rapidly detected. The method has the advantages of being rapid, accurate, low in cost, simple to operate and the like, and has high commercial application value; a traditional method for identifying the purity of the hybrid seed can be replaced.

Description

technical field [0001] The invention belongs to the field of molecular detection, and in particular relates to a primer and a method for identifying the purity of cucumber 'Yuexiu No. 3' hybrid seeds. Background technique [0002] Seed quality is an important element in agricultural production, and its quality directly affects the quality and yield of agricultural products. The traditional identification of purity and authenticity of varieties is based on morphological markers. This identification method has the advantages of stability and reliability from the perspective of agricultural production, but from the perspective of genetics, the identification of purity and authenticity of varieties is essentially a For the identification of quality genotype, only by identifying the DNA molecule itself can the genotype of the variety be accurately and reliably identified. DNA molecular marker technology is a new type of identification method developed with the development of mol...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 林毓娥王瑞吴廷全梁肇均黄河勋田永红
Owner INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI
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