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Primers and method for purity identification of hybrid seeds of Chinese pumpkin 'Guangmi NO.1'

A technology of hybrid seed purity and Chinese pumpkin, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of unestablished hybrid seed purity identification methods, and achieve high commercial application value, Simple operation and low cost effect

Active Publication Date: 2013-05-08
INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows for quick detection of pure-hybrids from both females or males that produce them. It also provides technical benefits such as fast response time, accurate results with lower costs compared to current techniques like identifying individual seals by coloring them visually before they mature into edible crops.

Problems solved by technology

Technically speaking, this patented technical problem addressed in this patents relates to testing pure or unclean seals produced through breeding methods that involve removing unwanted parts like stamens after fertilization with spermatozoa collected separately at specific locations within their own fields. Current techniques require multiple steps such as collecting ovules from each plot, isolating them manually, then analyzing these isolated material to determine if any contamination has been introduced into the resulting crops. These procedures take up too much time and cause environmental impact.

Method used

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  • Primers and method for purity identification of hybrid seeds of Chinese pumpkin 'Guangmi NO.1'
  • Primers and method for purity identification of hybrid seeds of Chinese pumpkin 'Guangmi NO.1'
  • Primers and method for purity identification of hybrid seeds of Chinese pumpkin 'Guangmi NO.1'

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Embodiment 1

[0025] Establishment of a method for the detection of seed purity of hybrid pumpkin 'Guangmi No. 1'.

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

[0027] The published pumpkin EST-SSR primers were screened between the parents (the male parent Changqing 2 and the female parent Zaoxuan 1), and the primer VRISSRPMK8 with a co-dominant differential marker band was selected. The sequence is as follows:

[0028] VRISSRPMK8-F: 5'-TAATTAAGGCGTGAAATTGG-3' (SEQ ID NO: 1)

[0029] VRISSRPMK8-R: 5'-GAAAAGAAACATAAACAGGGC-3' (SEQ ID NO: 2)

[0030] The marking band is clear and repeatable. Primer VRISSRPMK8 can produce 193bp maternal specific marker VRISSRPMK8 193 and the 187bp paternal-specific marker VRISSRPMK8 187 .

[0031] 2. Using the above-mentioned specific primers to identify the purity of hybrid pumpkin 'Guangmi No. 1' seeds.

[0032] (1) Extraction of pumpkin DNA

[0033] The experimental materials are the cotyledon DNA of the commercial variety of pumpki...

Embodiment 2

[0058] Adopt the method of embodiment 1 to 100 strains ' Guangmi No. 1 ' that get from the seed purity identification field of Baiyun base, number its single plant, extract single plant DNA and detect (see figure 2 ), the test results are all true hybrids, the seed purity is 100%, which is consistent with the field investigation results, and the accuracy rate is 100%.

Embodiment 3

[0060] Adopt the method of embodiment 1 to 100 strains ' Guangmi No. 1 ' that get from the seed purity appraisal field of Vegetable Institute base, number its single plant, extract single plant DNA and detect (see image 3 ), the results showed that among 100 samples, there were 97 hybrid seeds and 3 female parents, and the seed purity was 97%, which was consistent with the field investigation results.

[0061] The above examples show that the method of the present invention can effectively distinguish the hybrid seeds of 'Guangmi No. 1' from the seeds of its female parent (Zaoxuan No. 1), and quickly and accurately detect the purity of the seeds.

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Abstract

The invention discloses primers and a method for purity identification of hybrid seeds of Chinese pumpkin 'Guangmi NO.1'. The primers are VRISSRPMK8-F:5'-TAATTAAGGCGTGAAATTGG-3' and VRISSRPMK8-R:5'-GAAAAGAAACATAAACAGGGC-3'. The identification method is as follows: 1) extracting a genome DNA (deoxyribonucleic acid) of a pumpkin seedling; 2) carrying out PCR (polymerase chain reaction) amplification by taking the genome DNA of the pumpkin as a template and using the SSR primers VRISSRPMK8; 3) carrying out gel electrophoresis on the amplified product; and 4) analyzing the electrophoresis result, wherein only the single plant simultaneously having parent specific bands is a real hybrid, and the plant lacking of any one band is marked as false hybrid. The primers and the method can be used for effectively distinguishing the hybrid seed of the 'Guangmi NO.1' from the female parent and the male parent, and can identify the purity of the hybrid seeds rapidly and accurately. The method has the advantages of being fast and accurate, low in cost and simple in operation, and the like.

Description

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Claims

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Application Information

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Owner INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI
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