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Xinhui mandarin orange virus-free test tube micro-grafting method

A micro-grafting and test-tube seedling technology, applied in the field of citrus seedling breeding and reproduction, can solve the problems of the receptor system species specificity, low regeneration efficiency, low survival rate, genetic transformation limitation of Xinhui Dahong citrus, etc., and achieves excellent color and quality. The effect of improving peel development and quality, and improving resistance to harsh environments

Inactive Publication Date: 2016-10-12
JIANGMEN XINHUI FORESTRY SCI RES INST
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AI Technical Summary

Problems solved by technology

[0005] The genetic transformation of citrus is based on the development of the receptor system. Having a good tissue culture regeneration system is often the key to success. The variety specificity and low regeneration efficiency of the receptor system have greatly hindered the genetic transformation of Xinhui Dahonggan. limit
Traditional citrus grafting is mostly detoxified by heat treatment, and then grafted at the tip of the stem in the field, cut open and exposed to the air. Bacteria or viruses are easy to infect or latent through the cut, and the survival rate is low, especially in the humid and rainy climate in the south. prone to the above problems

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The method for micro-grafting of Xinhui Dahonggan detoxified test-tube seedlings mainly includes the following steps:

[0029] Step 1. Acquisition of cluster buds of detoxified Xinhui big red mandarin orange:

[0030] Cut the sterilized and sterilized mother branch of Xinhui Dahongkan on the ultra-clean workbench to cut the shoot tip 2mm, transfer it to the improved MT medium for pre-cultivation, and pre-cultivate it in the dark at 4°C for 3 days, and the shoot tip tissue Transfer to the protection medium to freeze and adapt for 1 hour, then transfer it to a cryotube and quickly put it into liquid nitrogen to rapidly cool down the shoot tip tissue to -196°C for 1 hour, then quickly transfer it to 45°C water for thawing and recovery, and then Inoculate into the subculture medium, cultivate in the dark at 25°C for one week, and then perform conventional light culture to obtain clustered buds of the virus-free Xinhui big red mandarin orange; strict control of the above cul...

Embodiment 2

[0042] The method for micro-grafting of Xinhui Dahonggan detoxified test-tube seedlings mainly includes the following steps:

[0043] Step 1. Acquisition of cluster buds of detoxified Xinhui big red mandarin orange:

[0044] Cut the sterilized and sterilized mother branch of Xinhui Dahong tangerine on the ultra-clean workbench to cut the shoot tip 1mm, transfer it to the improved MT medium for pre-cultivation, and pre-cultivate it in the dark at 4°C for 5 days, and the shoot tip tissue Transfer to the protection medium to freeze and adapt for 1 hour, then transfer it to a cryotube and quickly put it into liquid nitrogen to rapidly cool down the shoot tip tissue to -196°C for 1 hour, then quickly transfer it to 45°C water for thawing and recovery, and then Inoculate into the subculture medium, cultivate in the dark at 25°C for one week, and then culture in the conventional light to obtain clustered buds of the virus-free Xinhui Dahonggan;

[0045] The improved MT medium compri...

Embodiment 3

[0056] The method for micro-grafting of Xinhui Dahonggan detoxified test-tube seedlings mainly includes the following steps:

[0057] Step 1. Acquisition of cluster buds of detoxified Xinhui big red mandarin orange:

[0058] Cut the sterilized and sterilized mother branch of Xinhui Dahongkan on the ultra-clean workbench to cut the shoot tip 2mm, transfer it to the improved MT medium for pre-cultivation, and pre-cultivate it in the dark at 4°C for 5 days, and the shoot tip tissue Transfer to the protection medium to freeze and adapt for 1 hour, then transfer it to a cryotube and quickly put it into liquid nitrogen to rapidly cool down the shoot tip tissue to -196°C for 1 hour, then quickly transfer it to 45°C water for thawing and recovery, and then Inoculate into the subculture medium, cultivate in the dark at 25°C for one week, and then culture in the conventional light to obtain clustered buds of the virus-free Xinhui Dahonggan;

[0059] The improved MT medium comprises the...

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Abstract

The invention relates to the technical field of Xinhui big red mandarin seedling breeding and propagation, in particular to a method for micro-grafting of Xinhui big red mandarin detoxified test-tube seedlings. The micro-grafting method mainly includes the following steps: take the 1-2mm stem tip of the target plant, adopt the improved MT medium for pre-cultivation, drop into liquid nitrogen for detoxification, restore inoculation to the subculture medium for cultivation; rootstock seeds are sterilized, sterilized , germinated; the clustered buds developed by the axillary buds of Xinhui big red mandarin orange were grafted onto the rootstock, and then the seedlings were cultured with liquid in test tubes. This culture method uses the improved MT medium for pre-cultivation. After the improvement, the subculture value-added coefficient is greater than 4, basically no vitrification phenomenon occurs, and the variation is greatly reduced; this culture method makes the root system of the plant more developed, strong in resistance to harsh environments, and easier to adapt In the humid and rainy climate in the south, the survival rate is also greatly improved.

Description

technical field [0001] The invention relates to the technical field of citrus seedling cultivation and propagation, in particular to a method for micro-grafting of Xinhui citrus tangerine detoxified test-tube seedlings. Background technique [0002] Xinhui tangerine peel is the dried peel of Xinhui tangerine. Because of its unique quality, it has been well-known as early as before the Ming and Qing Dynasties, and was listed as a "tribute" and sold at home and abroad. [0003] Tangerine peel is better as long as it is stored, so it is called "tangerine peel". Tangerine peel is best produced in Guangdong. In the historical trade, it is specially called "Guang Chenpi" to distinguish it from other provinces. [0004] The peel of Xinhui big red tangerine is the famous specialty of Xinhui—Chen peel. Because it has high medicinal value and is a traditional spice and condiment, it has always enjoyed a high reputation. As early as the Song Dynasty, it has become one of the "Guang...

Claims

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Application Information

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IPC IPC(8): A01G1/06A01G31/00A01H4/00
CPCA01G2/30A01G31/00A01H4/001A01H4/005
Inventor 王立黄宏健谭沛涛胡杨陈小玲
Owner JIANGMEN XINHUI FORESTRY SCI RES INST
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