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Method for identifying authenticity of Indian pumpkin*Chinese pumpkin hybrid rootstock varieties

A technology of Chinese pumpkin and Indian pumpkin, applied in biochemical equipment and methods, determination/inspection of microorganisms, etc., can solve the problems of ineffective identification of authenticity, time-consuming, poor reliability, etc., and achieve accurate and reliable identification results. Environmental impact, easy-to-use effects

Active Publication Date: 2018-03-16
NINGBO ACAD OF AGRI SCI
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Problems solved by technology

These methods are time-consuming, easily affected by the environment, and have poor reliability. Faced with a variety of India-China hybrid pumpkin rootstock materials with similar appearance and phenotype, it is difficult to identify the authenticity.
[0005] The new rootstock varieties 'Yongzhen No. 7', 'Yongzhen No. 8' and 'Yongzhen No. 10' have only been identified by field morphological purity so far, and there is still a lack of convenient and effective methods for authenticity identification of varieties, which is far from enough for variety protection. not enough

Method used

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  • Method for identifying authenticity of Indian pumpkin*Chinese pumpkin hybrid rootstock varieties
  • Method for identifying authenticity of Indian pumpkin*Chinese pumpkin hybrid rootstock varieties
  • Method for identifying authenticity of Indian pumpkin*Chinese pumpkin hybrid rootstock varieties

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Embodiment Construction

[0011] 1. Sampling: sow 'Yongzhen No. 7', 'Yongzhen No. 8' and 'Yongzhen No. 10' and their controls, and when the plants grow to 5-6 true leaves, collect 1g of young leaves and store them at -80°C .

[0012] 2. DNA extraction: Refer to the CTAB method (see: Murray HG, Thompson WF. Rapid isolation of higher weight DNA. Nucleic Acids Res, 1980, 8: 4321), extract the total genomic DNA from fresh leaves.

[0013] 3. PCR amplification: 15 μl of mixed sample system (Fazio G, Staub JE, Stevens MR. Genetic mapping and QTL analysis of horticultural traits in cucumber (Cucumis sativus L.) using recombinant inbred lines. Theor Appl Genet, 2003, 107: 864- 874), including 1.5μl dNTP (1.25mM), 1.5ul 10×buffer, 1.5ul MgCl 2 (25mM), 2.0ul primers (5mM; add 1.0ul to the two SSR primers respectively), 5ul DNA (10ng / ul), 0.05ul Taq (1U), 3.45ul ultrapure water. The PCR amplification program was: pre-denaturation at 94°C for 1 min; denaturation at 93°C for 1 min; annealing at 40°C for 1 min; ex...

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Abstract

The present invention relates to a method for identifying the authenticity of Cucurbita maxima*Cucurbita moschata hybrid rootstock varieties, and belongs to the field of biology. In the prior art, identification of the variety authenticity is increasingly important for the maintaining of the interests of producers and breeders; the Indian pumpkin*Chinese pumpkin hybrid rootstock varieties are more, and are similar in appearance; and the application of the morphological indexes to identify the authenticity has disadvantages of long time consuming and poor reliability. In order to solve the problems in the prior art, the invention provides the method for identifying the authenticity of the variety by using molecular markers. The technical scheme comprises that three rootstock varieties and control materials are subjected to PCT amplification with 5 pairs of SSR primers (SEQ ID NO.1-10), the generated 9 difference sites are assigned with 0 or 1, and a variety fingerprint for variety authenticity identification is constructed. According to the present invention, the Cucurbita maxima*Cucurbita moschata hybrid rootstock varieties such as Yongzhen 7, Yongzhen 8 and Yongzhen 10 are accurately and effectively identified and protected through the sensitive identification method at the low time cost and the low labor cost.

Description

1. Technical field [0001] The invention belongs to the field of biology, and in particular relates to a method for identifying the authenticity of rootstock varieties 'Yongzhen No. 7', 'Yongzhen No. 8' and 'Yongzhen No. 10' of Indian pumpkin x Chinese pumpkin hybrids of the Yongzhen series. 2. Background technology [0002] Continuous cropping obstacles caused by watermelon wilt are common in melon growing areas and are a major problem in melon production. According to statistics, the incidence rate of watermelon wilt can reach 10-30% every year, serious plots or even production failure, resulting in huge economic losses. At present, due to the limitation of land resources and the stubbornness of the pathogenic bacteria of Fusarium wilt, crop rotation and chemical control are not effective, and there are problems such as environmental pollution and food safety. Therefore, grafting is an effective means to overcome continuous cropping obstacles and Fusarium wilt in watermelon...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895
CPCC12Q1/6895C12Q2600/156
Inventor 宋慧张香琴王迎儿黄芸萍
Owner NINGBO ACAD OF AGRI SCI
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