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Watermelon efficient genetic transformation method and transgenic plant identification method

A genetic transformation method and genetic transformation technology are applied in the fields of efficient genetic transformation of watermelon and identification of transgenic plants, which can solve the problems of lack of germplasm resources and narrow genetic foundation of watermelon, and achieve the effect of high transformation frequency.

Inactive Publication Date: 2019-01-04
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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Problems solved by technology

The genetic basis of watermelon is narrow, the germplasm resources are lacking, and many important traits cannot be improved through conventional breeding methods

Method used

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  • Watermelon efficient genetic transformation method and transgenic plant identification method
  • Watermelon efficient genetic transformation method and transgenic plant identification method
  • Watermelon efficient genetic transformation method and transgenic plant identification method

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Experimental program
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Effect test

Embodiment 1

[0073] Example 1. Agrobacterium-mediated genetic transformation of watermelon

[0074] The watermelons to be transferred are: cultivated watermelon variety 97103 and wild watermelon variety PI296341. The experiment was repeated three times.

[0075] 1. Preparation of explants

[0076] Select the plump, normal-looking seeds of cultivated watermelon variety 97103 and wild watermelon variety PI296341, carefully remove the seed husks, do not damage the cotyledons and growth points, sterilize, plant and grow in the MS solid medium at 28°C in the dark for 3 days about. Select plants in the period when the radicle is elongated but the two cotyledons have not yet separated, cut the middle section of the cotyledon, remove the tip and the part near the petiole, and cut it into small pieces of 1.5mm×1.5mm as explants.

[0077] 2. Preparation of infection solution

[0078] Recombinant expression vector preparation: the target gene 1 (the DNA fragment shown in sequence 3 in the sequence list) was...

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Abstract

The invention discloses a watermelon efficient genetic transformation method and a transgenic plant identification method. The invention discloses the watermelon genetic transformation method. The watermelon genetic transformation method comprises the following steps: (1) using an infection solution containing recombination agrobacterium to infect watermelon explants, co-culturing the watermelon explants after infection on a co-culture culture medium to obtain explants after co-culture, wherein the recombination agrobacterium contains a recombinant expression vector carried with target genes and BASTA genes; (2) culturing the explants after co-culture on a screening culture medium to obtain a plant tissue containing positive buds; (3) culturing the plant tissue containing the positive budson a bud extension culture medium to obtain seedlings with watermelon ground parts; (4) culturing the seedlings with watermelon ground parts on a rooting culture medium to obtain watermelon plants; and (5) screening the plants containing the target genes from the watermelon plants to obtain the watermelon plants after genetic transformation. By using the method disclosed by the invention, the high conversion frequency and high repeatability conversion for watermelon can be realized.

Description

Technical field [0001] The invention relates to a method for efficient genetic transformation and transgenic plant identification of watermelon in the field of plant genetics and genetic engineering. Background technique [0002] Watermelon (Citrullus lanatus (Thunb.) Mansfeld.et Nakai) is an annual herbaceous vine in the Cucurbitaceae family. It is an important horticultural crop. Watermelon has a narrow genetic basis and lack of germplasm resources. Many important traits cannot be improved by conventional breeding methods. Genetic engineering technology has become an effective supplement to conventional breeding. With the continuous development of biotechnology, human manipulation of genetic material using transgenic technology and gene editing technology has begun to be widely used in various plants, providing a new method for creating new germplasm. Summary of the invention [0003] The technical problem to be solved by the present invention is how to carry out genetic trans...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H4/00A01H5/00A01H6/34
CPCA01H4/008C12N15/8205
Inventor 许勇张洁田守蔚宗梅张海英宫国义刘凡郭绍贵任毅李茂营
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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