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Method for constructing fructus alpiniae oxyphyllae tissue culture regeneration system

A technology of tissue culture and construction method, which is applied in the field of tissue culture regeneration system of Yizhiren, can solve the problems of browning of explants, low survival rate of transplanted field, limitations of tissue culture technology application, etc., and achieve the effect of solving the shortage

Inactive Publication Date: 2019-02-26
张世燊
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, some progress has been made in the tissue culture research of Yizhiren, but there are still some problems, such as serious browning of explants, low shoot rate, difficulty in rooting, and low survival rate in transplanted fields. The application in Yizhiren is very limited

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] (1) Seed disinfection: select immature fruits at the end of August, soak in washing powder water for 12 minutes after removing leaves, rinse with tap water for 3 hours after brushing, take out immature milky white or slightly brown seeds, and peel them in an ultra-clean workbench. Remove the outer testa, first sterilize with 75% ethanol for 7 seconds, then wash 5 times with sterile water, then sterilize with 0.1% mercuric chloride solution for 12 minutes, rinse with sterile water 6 times and set aside, the statistical pollution rate can be as low as 3% one week after inoculation .

[0014] (2) Embryo callus induction: the immature seed embryos treated in step (1) were cut into 0.6cm×0.6cm×0.6cm and inoculated on the induction medium, and cultured at 28°C in total darkness for 32 days. Then placed in light for 12 hours a day, the light intensity was 1700x, until embryogenic callus was induced, and the callus induction rate was 69%. The induction medium is ER+6-BA1.2mg / L...

Embodiment 2

[0019] (1) Seed disinfection: select the fruit, remove the leaves, soak in washing powder water for 18 minutes, rinse with tap water for 4 hours after brushing, take out the immature milky white or slightly brown seeds, peel off the outer testa in an ultra-clean workbench, first Disinfect with 75% ethanol for 10 seconds, wash with sterile water for 5 times, then disinfect with 0.1% mercury liter solution for 14 minutes, rinse with sterile water for 6 times and set aside. The statistical pollution rate can be as low as 5% one week after inoculation.

[0020] (2) Embryo callus induction: the immature embryos treated in step (1) were cut into 0.5cm×0.5cm×0.5cm and inoculated on the induction medium, and cultured at 28°C in total darkness for 32 days. Then placed in light for 14 hours a day, the light intensity was 1800x, until embryogenic callus was induced, and the callus induction rate was 64%. The induction medium is ER+6-BA1.0mg / L+NAA0.2mg / L+GA 3 3 mg / L+ sucrose 18g / L+ agar ...

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Abstract

The invention discloses a method for constructing a fructus alpiniae oxyphyllae tissue culture regeneration system, and relates to a technical method for obtaining fructus alpiniae oxyphyllae seedlings with stable properties through a vegetative rapid propagation technology. According to the method, by taking an immature embryo of fructus alpiniae oxyphyllae as an explant, the fructus alpiniae oxyphyllae tissue culture regeneration system is constructed by steps of seed disinfection, embryo callus induction, differentiation culture, rooting culture, seedling cultivation, transplanting and thelike, thereby solving the problem of the lack of high-quality seedlings of the fructus alpiniae oxyphyllae and laying a foundation for future development of genetic transformation of the fructus alpiniae oxyphyllae.

Description

technical field [0001] The invention relates to a method for cultivating plant tissue in agricultural biotechnology, in particular to a method for constructing a tissue culturing and regeneration system for cultivars. Background technique [0002] Yizhiren is a ginger plant, which is the dry and mature fruit of Yizhi. Born in Guangdong, Guangxi and Hainan. From May to June of the next year, the fruit is brown, and the fruit skin is picked when the hairs are reduced, and the fruit stalk is removed and dried in the sun. Spicy and slightly bitter. There is warming the spleen and arresting diarrhea, taking saliva, warming the kidney, consolidating essence and reducing urination. For spleen cold diarrhea, cold pain in the abdomen, excessive salivation, frequent urination, enuresis due to kidney deficiency. The conventional breeding technology of Yizhi kernel has limitations, such as the difficulty of distant hybridization, low frequency of beneficial mutations, long time-cons...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 张世燊
Owner 张世燊
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