Rapid identification method of genetic purity of muskmelon hybrid seeds based on PCR

Abstract

The invention discloses a rapid identification method of genetic purity of muskmelon hybrid seeds based on PCR, characterized by respectively utilizing screened SRAP primer combination NAUSRem6 / NAUSRfc8 and / or RAPD primer NAURP401, using Haimi No.2 muskmelon parent and F1 individual genome DNA as a template to conduct PCR amplification, and using non-denaturing polyacrylamide gel electrophoresis to detect PCR amplification products; wherein only the individual plant with parent special bands can be really hybrid, and the individual plant without any parent band is false hybrid. The method provided by the invention overcomes the disadvantages of complicated operation, low accuracy and the like of conventional field purity detection, has the advantages of rapidness, simpleness, low cost, stability and reliability, immunity to the growth stages and environment, and can be one of the important detection methods of genetic purity identification of muskmelon hybrid seeds.

Description

Technical field [0001] The invention belongs to the field of biotechnology and relates to a method for quickly identifying the genetic purity of melon hybrid seeds based on PCR. Background technique [0002] Cucumis melo L. is an annual vegetable in the Cucurbitaceae family. Melon hybrids have strong heterosis in terms of yield, quality and disease resistance. At present, the demand for melon hybrids on the market is increasing year by year. Most melons are monoecious plants (same plants for both sexes) or monoecious plants (edited by Zhengzhou Institute of Fruit Science, Chinese Academy of Agricultural Sciences. China Watermelon and Melon. Beijing: China Agriculture Press, 2000.2). During the seeding process, due to the lack of timely emasculation of the female parent, inadequate or incomplete isolation, etc., muskmelon hybrids are mixed with a small number of female parent selfed or'false hybrids' caused by pollen from other unknown sources. In addition, due to the high degre...

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Problems solved by technology

[0003] At present, the commonly used methods for the identification of seed genetic purity are field morphological identification and isoenzyme identification, but the field identification method is time-consuming, laborious, and costly, which cannot meet the requirements of the market, and is easily affected by seasonal restrictions and environmental factors, with low accuracy.

Sometimes because the morphological difference between the false hybrid and the true hybrid is not obvious, it is difficult to distinguish between the seedling stage and the adult stage

Isozyme analysis is easily affected by the environment and the location of the material, especially for some hybrid varieties with small genetic differences between parents, it is difficult to perform purity detection and identification, and the accuracy is not high (Liu Liwang et al., Molecular marker technology in the identification and identification of vegetable crop varieties The role of purity detection. Molecular Plant Breeding, 2004,2(4):563~568)

In recent years, with the continuous development of modern biotechnology, using SSR (Simple Sequence Repeats, simple sequence repeats), ISSR (Inter-Simple Sequence Repeats, simple sequence repeats inter-sequence), RAPD (Randomly Amplified Polymorphic DNA, random amplification multiple There have been related reports on the identification of genetic purity of hybrids by molecular marker techniques such as morphological DNA), but most of them still use a certain molecular marker alone for analysis, and the accuracy is low (Pallavi, H.M., et al. Identification of SSR markers for hybridity and seed genetic purity testing in sunflower (Helianthus annuus L.).Helia,2011(34):59-66)

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