Detoxification method for excised stem segment of grape

A grape and in vitro technology, which is applied in the field of detoxification of grape isolated stems, can solve the problems of high detoxification rate, complicated procedures, and long time required for detoxification, and achieve strong temperature adaptability and prevent water loss Effect

Inactive Publication Date: 2011-11-09
CHANGLI INST OF POMOLOGY HEBEI ACADEMY OF AGRI & FORESTRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 condit...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Select subculture 60d, the test-tube plantlet (the poisoning rate is 100%) of the vigorous grape variety Kyoho, under aseptic condition, cut the semi-lignified single-bud stem section and inoculate it in the improved B5+0.2mg / L IAA +20mg / L white granulated sugar+5.0mg / L agar, in medium of pH5.8, placed in 27℃, 2000lx culture room for 7 days, exposed the small white root, put it in a foam box, covered it, and put it in the incubator . After 10 days, the yellowed young shoots grow to the mouth of a 100ml triangular flask (about 8-9cm), and the micro shoot tips of 0.5mm are stripped under aseptic conditions and cultured in improved B5+0.2mg / L IAA+20mg / L white granulated sugar+ 5.0mg / L agar, pH5.8 culture medium, cultured in 27℃, 2000lx culture room.

[0035] After the test-tube plantlets were formed at the shoot tip, the virus was detected by the enzyme-linked immunosorbent double-antibody sandwich method. As a result, the virus-free rate of grape fan leaf virus and leafr...

Embodiment 2

[0037] Select subculture 57d, the test-tube plantlet of the grape variety Red Globe that grows vigorously, under aseptic condition, cut the single-bud stem section and inoculate on improved B5+0.5mg / L IAA+30mg / L white granulated sugar+8mg / L agar, In the culture medium of pH5.8, place it in a culture room at 25°C and 2500 lx for 9 days. After exposing the small white root, put it in a foam box, cover it, and put it in the culture box. After 12 days, the yellowed young shoots are 6-8cm long, and the micro shoot tips of 0.4-0.7mm are stripped under sterile conditions and cultured in the improved B5+0.24mg / L IAA+25mg / L white sugar+7.0mg / L agar, pH5 .8 culture medium, as 25 ℃, 2500lx culture room culture.

[0038] After the test-tube plantlets were formed at the shoot tips, virus detection was carried out. As a result, the virus-free rate of grape fan leaf virus and leafroll virus was 78.8%.

Embodiment 3

[0040]Select subculture 60d, the test-tube seedling of vigorously growing grape rootstock 5BB (toxic rate is 100%), under aseptic condition, cut single-bud stem segment and inoculate in improved B5+0.2mg / LIAA+20mg / L white Sugar + 5.0 mg / L agar, pH 5.8 culture medium, placed in 27°C, 2000 lx culture room for 7 days, after exposing the small white root, put it in a foam box, cover it, and put it in the incubator. After 15 days, the yellowed young shoots grow to 8-9cm, and the micro shoot tips of 0.5mm are stripped under aseptic conditions and cultured in improved B5+0.2mg / L IAA+20mg / L white granulated sugar+5.0mg / L agar, pH5. 8 medium, as for 27 ℃, 2000lx culture room culture.

[0041] After the test-tube seedlings were formed at the shoot tips, the virus-free rate of grape fan leaf virus and leafroll virus was 92.8%.

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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...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 赵艳华吴雅琴吴永杰程和禾李玉生
Owner CHANGLI INST OF POMOLOGY HEBEI ACADEMY OF AGRI & FORESTRY SCI
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