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Method for rapidly breeding sugarcane stem tip by tissue culture and virus removal

A stem tip and sugar cane technology, applied in the field of plant detoxification and rapid propagation, can solve the problems of waste of manpower and material resources, easy variation, and low survival rate of shoot tip meristem, and achieve increased yield and quality, normal shape, and resolved The effect of germplasm degradation

Inactive Publication Date: 2013-06-05
广州甘蔗糖业研究所湛江甘蔗研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the problem that above-mentioned approach exists is: ① callus approach: although a large amount of tissue culture plantlets can be obtained through callus in a short period of time, it has the weakness of easy variation; The following shoot apical meristems need to be operated under a microscope, which is difficult, and the survival rate of such tiny shoot apical meristems is very low, and there is a waste of manpower and material resources to a certain extent

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] (Take the tip of the top bud of sugarcane as the explant)

[0037] (1) Selection and sterilization of explants: During April to May, the terminal buds of sugarcane were taken, and the older bracts were peeled off, treated with hot water at 52°C for 30 minutes, and then soaked in 75% alcohol 0.5~1.0min, then treated with 0.1% mercuric chloride for 4min, rinsed with sterile water 3~5 times;

[0038] (2) Basic medium: use MS medium, sucrose 30g / L, pH value 5.8~6.2;

[0039] (3) Initiate culture: Remove the leaf sheath carefully from the terminal buds treated in step (1) under sterile conditions, pick out 1~2mm shoot tips, and inoculate them in MS+IBA0.02mg / L+activated carbon 1.0g / L On the filter paper bridge culture medium, induce germination;

[0040] (4) Differentiation culture: transfer the buds induced in step (3) to the differentiation medium to induce cluster buds. The differentiation medium is: MS medium, 6-BA0.5mg / L and IBA0.02mg / L;

[0041] (5) Cultivation of s...

Embodiment 2

[0046] (Take the tip of the sugarcane axillary bud as the explant)

[0047](1) Selection and sterilization of explants: From July to October, cut sugarcane stems according to the double buds → pretreat with 75% alcohol and bromogeramine → treat with hot water at 52°C for 30 minutes → place Germinate at high temperature in an artificial climate box (38°C, cultivate for 1 week), or place in a greenhouse for 1 week in fine river sand, take the germinated axillary buds, peel off the older bracts, soak in 75% alcohol for 0.5~1.0min , and then treated with 0.1% mercuric chloride for 3 minutes, rinsed with sterile water 3 to 5 times;

[0048] (2) Basic medium: use MS medium, sucrose 30g / L, pH value 5.8~6.2;

[0049] (3) Initiate culture: Carefully remove the leaf sheaths from the axillary buds treated in step (1) under sterile conditions, pick out 1~2mm shoot tips, and inoculate them on the filter paper of MS+IBA0.02mg / L+activated carbon 1.0g / L on the bridge culture medium to induc...

Embodiment 3

[0056] Test case (virus detection)

[0057] Control materials: sugarcane diseased leaves, cane stems, and cane roots collected from the field were used as materials.

[0058] Processing material: take the tissue culture seedling obtained in embodiment 1 and embodiment 2 as material.

[0059] The test results are shown in the table below:

[0060] deal with test results control Positive Virus-free tissue culture seedlings Negative

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Abstract

The invention relates to a method for rapidly breeding sugarcane stem tip by tissue culture and virus removal, belonging to the technical field of rapid breeding of plant by virus removal. The method comprises the following steps: carrying out disinfection pretreatment on a terminal bud of sugarcane and a cut double-bud stem section, with the terminal bud of the sugarcane and a stem tip tissue ofan axillary bud as explants, and carrying out stem tip initial culture, sprout differentiation culture, crowd sprout strong stock culture, plant root culture and virus checking by adopting a filter paper bridge culture way, thus massive virus-removed tissue culture seedlings are obtained. The invention has the advantages: the stem tip with the length of 1-2mm is taken as the explant, thus the processing operation is easy; the filter paper bridge culture way is adopted during the initial culture, thus the initiating speed is high and the perennial root dwarfing virus removing effect is thorough; and the stem tip is induced and germinated and is rapidly initiated and differentiates massive crowd sprouts, the month breeding coefficient reaches 5-8, and normomorph is realized in a long-term breeding process, thus the method is completely applicable to factory production on a large scale.

Description

technical field [0001] The invention relates to a method for detoxification and rapid propagation of sugarcane stem tips through tissue culture, in particular to a method for detoxification and rapid propagation of sugarcane stem tips through tissue culture, and belongs to the technical field of plant detoxification culture and rapid propagation. Background technique [0002] Sugarcane ratoon stunting disease (RSD) was first discovered in the sugarcane variety Q28 in Queensland, Australia from 1944 to 1945. Now it occurs widely in the main sugarcane producing areas in China and around the world, leading to the decline of species and seriously affecting The quality greatly shortens the economic service life of fine varieties. [0003] Sugarcane ratoon dwarf disease is caused by (Clavibacter xyli subsp.Xyli, Lxx), and the average infection rate of RSD cane plants is about 50%, which usually results in a 12-37% reduction in sugarcane yield, up to 60% in drought conditions, and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 谭嘉娜陈月桂李奇伟杨俊贤吴文龙潘方胤
Owner 广州甘蔗糖业研究所湛江甘蔗研究中心
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