Method using protoplast asymmetric fusion technology to obtain hybrid grape plants

A protoplast and asymmetric technology, which is applied in botany equipment and methods, biochemical equipment and methods, fusion cells, etc., can solve the problem of few successful examples of grape protoplast separation and purification, and solve the problems of cultivation and fusion difficulties, consumption The effect of short time and low NH4+ concentration

Active Publication Date: 2016-06-08
ZHEJIANG WANLI UNIV
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There are very few successful examples of the isolation,

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  • Method using protoplast asymmetric fusion technology to obtain hybrid grape plants
  • Method using protoplast asymmetric fusion technology to obtain hybrid grape plants
  • Method using protoplast asymmetric fusion technology to obtain hybrid grape plants

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[0036] The present invention will be further described in detail below in conjunction with the embodiments. It should be noted that the following specific descriptions are all illustrative and are intended to provide further description of the present invention. Unless otherwise specified, all scientific and technical terms used in the present invention have the same meanings as commonly understood by those skilled in the art to which the present invention belongs.

[0037] In this embodiment, the method for obtaining grape hybrid plants by using protoplast asymmetric fusion technology includes the following steps:

[0038] 1. Separation and purification of grape protoplasts

[0039] The separation of protoplasts is an important part of the success of protoplast culture. The enzymatic hydrolysis of protoplasts is not only affected by the separation material, but also related to the type and concentration of enzymes, enzymatic hydrolysis time, temperature, pH, osmotic pressure and ot...

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Abstract

The invention provides a method using protoplast asymmetric fusion technology to obtain hybrid grape plants. The method includes: taking fresh embryonic callus, and adding enzyme liquid for enzymolysis; respectively performing IOA passivation and UV passivation on protoplast to obtain receptor protoplast and donor protoplast, mixing the receptor protoplast and the donor protoplast according to the ratio of 1:1, sequentially adding PEG fusion induction liquid and a high-Ca2+ high-pH solution to obtain fused hybrid cells; using a KM8P solid culture medium to culture the fused hybrid cells in an embedded manner; sequentially transferring the particles of the embryonic callus to a differential culture medium and a rooting culture medium for culturing; when tissue cultured seedlings grow to 5-7cm, planting into a seedling exercising room for seedling exercising so as to obtain somatic cell hybrid plants. The method has the advantages that the grape embryonic callus is used as the material, the protoplast asymmetric fusion technology is used to successfully obtain the hybrid grape plants, and a foundation is laid on the culture of new grape varieties, grape resistance enhancing and grape quality increasing.

Description

technical field [0001] The invention belongs to the technical field of somatic cell hybridization of grapes, and in particular relates to a method for obtaining grape hybrid plants by utilizing the protoplast asymmetric fusion technology. Background technique [0002] Sexual hybridization of plants is limited to some close relatives between cultivated species and wild species or within species. Some plants have low or no sexual reproductive ability, so it is difficult to improve and cultivate new varieties through sexual hybridization. , severely limited the progress of breeding work. The methods of conventional grape breeding in southern my country include cross breeding, introduction, seed selection, mutation breeding and bud selection. The plant somatic cell hybridization technology can widely recombine the excellent genetic traits of the plant kingdom through the fusion of heterologous protoplasts, overcome the incompatibility of distant sexual hybridization, shorten th...

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Application Information

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IPC IPC(8): C12N15/05A01H4/00A01H5/00
CPCA01H4/008A01H5/00C12N5/14C12N2510/00
Inventor 吴月燕饶慧云钱萍仙刘蓉卢丹
Owner ZHEJIANG WANLI UNIV
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