Method for quickly testing purity of hybrid watermelon seed Quankanglvba

A technology of hybrids and hybrids, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of poor accuracy, long time and high cost, and achieve the effect of high accuracy and simple operation.

Active Publication Date: 2015-04-01
WIN ALL HI TECH SEED CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The traditional identification of the purity of watermelon hybrid seeds is still based on the identification of field planting. The identification by comparison of the appearance and shape of adult plants gen

Method used

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  • Method for quickly testing purity of hybrid watermelon seed Quankanglvba
  • Method for quickly testing purity of hybrid watermelon seed Quankanglvba
  • Method for quickly testing purity of hybrid watermelon seed Quankanglvba

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Experimental program
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Effect test

Embodiment 1

[0044] Three samples of Quankang Luba (hybrid F1) with different purity (the purity of sample 1, sample 2 and sample 3 were 99.5%, 97.3% and 90% respectively), all seeds were purchased from Anhui Quanyin Hi-Tech Seed Co., Ltd. provided by the company.

[0045] 1) Rapid extraction of sample DNA: Cut out the 1cm sprouts of 3 sample seeds 3-4 days after germination, put them into a 2ml centrifuge tube, put 2 small steel balls into the centrifuge tube at the same time, add 100μl, 0.4M For NaOH solution, crush the sample with a tissue masher, boil in boiling water for 1 min, add 100 μl of 100 mM Tris-HCl (pH 8.0) buffer, mix well, and store at 4 °C for later use.

[0046] 2) PCR amplification

[0047] ①Reaction system

[0048] The reaction system consists of sample genomic DNA, containing Mg 2+ buffer, dNTPs, primers, Taq enzyme and ddH 2 O composition. Its volume ratio is: 2:1.5:1:2:0.2:3.3.

[0049] During the preparation of the PCR reaction mixture, always keep it on ice. A...

Embodiment 2

[0070] The present invention utilizes primers QYXG3 and QYXG7 to test 48 samples of the hybrid watermelon variety Quankang Luba in 2014 respectively, perform PCR amplification and polyacrylamide gel electrophoresis separation on them, calculate the purity of the tested seeds, and conduct detection.

[0071] A described method for quickly testing the purity of hybrid watermelon seeds Quan Kang Luba comprises the following steps:

[0072] 1) Cut the 1cm sprouts of the watermelon hybrid variety Quankang Luba 3-4 days after germination, put them into a 2ml centrifuge tube, add 0.4M NaOH solution, crush the samples with a tissue smasher, and bathe in boiling water for about 1min. Add Tris-HCl buffer solution with a pH of 8.0, mix thoroughly, and the obtained DNA extract of the watermelon hybrid variety Quankang Luba sample is the genomic DNA of the watermelon hybrid variety Quankang Luba to be tested, and store it at 4°C for future use;

[0073] The volume ratio of NaOH solution t...

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Abstract

The invention discloses a method for quickly testing the purity of a hybrid watermelon seed Quankanglvba, and particularly relates to a method for quickly testing the purity of the hybrid watermelon seed Quankanglvba by using an SSR molecular marker, which comprises the following steps: by using an SSR primer QYXG3 or QYXG7, performing amplification and polyacrylamide gel electrophoresis separation on genome DNAs of the watermelon hybrid variety Quankanglvba and parents thereof, thus finding out a key band indicating the difference between a first filial generation and the parents; and calculating the purity of the tested seed to test the Quankanglvba sample, wherein the purity test result is highly consistent with a purity control result identified in field planting. The test method disclosed by the invention can finish seed purity identification within 5 hours, and has the characteristics of being simple and easy to implement, accurate, quick, low in cost and convenient to popularize and the like. The method is applicable to the authentication and variety purity test of the Quankanglvba watermelon hybrid and parents thereof.

Description

technical field [0001] The invention belongs to the technical field of seed quality inspection, and relates to a method for inspecting the purity of hybrid watermelon seeds, in particular to a method for quickly inspecting the purity of hybrid watermelon seeds. Background technique [0002] Quankang Luba is a new high-yield and high-quality hybrid watermelon variety (Jin Shen Xi Melon 2011006, Gan Ren Gua 2011005, Wan Pin Jian Deng Zi No. 1001007) bred by Anhui Quan Yin High-Tech Seed Industry Co., Ltd., which has excellent field agronomic characteristics. During the hybrid seed production process of this variety, the female parent needs to be detasselled manually, and the male parent pollen is given to complete the hybridization process. Incomplete or missing emasculation of the female parent will greatly affect the purity of the seed production. Therefore, it is very important to identify the purity of hybrid watermelon seeds. [0003] The traditional identification of th...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/156
Inventor 王慧张从合陈金节严志周桂香黄艳玲钱奕道杨坤方玉王林
Owner WIN ALL HI TECH SEED CO LTD
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