Detoxification method for excised stem segment of grape

Abstract

The invention relates to a detoxification method for an excised stem segment of grape. The method comprises the following steps: 1) selecting a grape variety and a test-tube plantlet related to parental stock, wherein the grape variety is subjected to a secondary culture until a stem is semi lignified; under a sterile condition, clipping a single bud stem segment and inoculating in a culture media, followed by placing in a 2000-3000 lx culture room having a temperature of 22-28 DEG C until a white rootlet is exposed to obtain a excised stem segment of the grape; 2) placing the excised stem segment of the grape in a incubator having a temperature of 35-40 DEG C, followed by carrying out a dark treatment while carrying out a heat treatment; 3) observing a growth condition of the lateral bud after heat treating for 10 days, visual peeling 0.2-0.8 mm of the micro stem apex under a sterile condition when a length of the yellow young shoot is 6-10 cm, then placing the micro stem apex in the culture media to obtain the propagation material. According to the present invention, illumination and frequent temperature change process do not require, such that the method has characteristics of simple operation, time saving, electricity saving, labor saving, high efficiency and low cost. In addition, the method has not strict requirements on research environments and research conditions and facilities, such that the method can be applicable for the research and teaching unit with the tissue culture condition.

Description

technical field [0001] The invention relates to the field of fruit tree biotechnology, in particular to a detoxification technology for isolated grape stems. Background technique [0002] At present, there are mainly 9 kinds of grape virus diseases reported in China: fan leaf disease, leaf curl disease, stem pox disease, cork disease, spot disease, vein necrosis disease, ear protrusion disease, stellate mosaic disease and leaf shrunken disease. Among them, fan leaf disease and leaf roll disease are the most widely distributed and most harmful viral diseases in grape production in my country. According to the survey, at present in my country's viticulture areas, more than 85% of the vineyards are generally affected, which has brought great losses to grape production. Guo Deyin and others used the indirect ELISA method to randomly detect 35 grape varieties, and the virus rate with fan leaves was as high as 74.3%. He Shuitao et al. conducted PAS-ELISA detection on 78 grapes o...

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Problems solved by technology

[0005] According to current reports, although the detoxification of grape planting materials has made good progress, there are still many problems, mainly: (1) the test tube seedling materials used for detoxification are not heat-resistant, and the plants die in the detoxification process , the number of micro-shoot tips obtained is small; (2) the temperature needs to be raised gradually or changed day and night, and the humidity and light should be controlled at the same time, which has strict requirements on the research environment and conditions and facilities, and poor versatility; (3) the procedures are complicated and require detoxification. Therefore, the period of cultivating grape virus-free seedlings is relatively long; (4) different media are required from the preparation of test tube materials, rooting to shoot tip culture; (4) although some articles published by the predecessors have obtained more High detoxification rate, but it has not been compared with the poisonous situation of the material before detoxification, so its higher detoxification rate needs further experiments