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Method for mycorrhization of bletilla striata tissue culture seedling

The technology of Bletilla striata group and mycorrhizalization is applied in the directions of horticultural methods, botanical equipment and methods, horticulture, etc., which can solve the problems of poor stress resistance, low seedling survival rate, low germination rate, etc. The effect of more sprouts tillering and large growth

Inactive Publication Date: 2019-02-15
JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Bletilla striata is a perennial herbaceous orchid. Its root structure is a typical mycorrhizal structure. Its seeds have no endosperm. Under natural conditions, they need symbiotic fungi to germinate. Due to the rise of tissue culture and rapid propagation technology, the problem of seed The problem of germination, tissue culture and rapid propagation of Bletilla striata seedlings has become one of the main ways to cultivate seedlings, but because the root system of tissue culture seedlings does not have the corresponding mycorrhizal fungi to coexist with it, resulting in the process of domestication and transplanting, Seedling survival rate, germination rate are low, and stress resistance is poor, therefore, a kind of development of the tissue culture method of the bletilla striata tissue culture seedling that can provide high-quality domesticated seedling for field cultivation of bletilla striata is very necessary

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Isolation and identification of strains

[0021] The strain JSNLBJ-001 was isolated in May 2015 from the plants with better growth conditions in the Bletilla striata planting area of ​​Jiangsu Maoshan Daodi Medicinal Materials Co., Ltd. in Jurong City, Jiangsu Province.

[0022] The separation method is as follows: take wild fresh Bletilla striata roots, rinse the substrate soil on the surface of the root section with flowing tap water, and dry the water on the surface with filter paper on an ultra-clean workbench. Then rinse it with sterilized deionized water, and then immerse it in 75% alcohol for 45 seconds and 2% sodium hypochlorite solution for 5 minutes for disinfection. After the disinfection is completed, rinse with sterilized deionized water for 5-6 times to completely remove sodium hypochlorite, then use sterilized filter paper to absorb the residual water droplets on the root surface, and then use a sterilized scalpel to cut the root segment into about 0.5mm ...

Embodiment 2

[0031] The method for the mycorrhization of Bletilla striata tissue cultured seedlings of the present invention comprises the following steps:

[0032] Step 1: Activate the P. lilacensis JSNLBJ-001 strain from the slant test tube stored at low temperature, inoculate it into a PDA petri dish, and culture it at a constant temperature of 28°C for 6-8 days, and then punch holes at the edge of the colony to form a fungus cake. Standby; prepare liquid medium according to the following components: oat 30g / L+glucose 30g / L+calcium chloride dihydrate 2.5g / L+potato 120g / L, medium pH value 6.0; among them, potato uses its extract; liquid medium preparation After that, it is divided into triangular flasks or culture bottles for fungi, with a dispensing volume of 100mL / bottle, and then placed in an autoclave for high-pressure sterilization. After cooling, it is inserted into the bacteria cake; the sterilized liquid medium is inserted into the bacteria Put the root fungus cake into a constan...

Embodiment 3

[0036] The method for the mycorrhization of Bletilla striata tissue cultured seedlings of the present invention comprises the following steps:

[0037] Step 1: Activate the P. lilacensis JSNLBJ-001 strain from the slant test tube stored at low temperature, inoculate it into a PDA petri dish, and culture it at a constant temperature of 28°C for 6-8 days, and then punch holes at the edge of the colony to form a fungus cake. Standby; prepare liquid medium according to the following components: oat 20g / L+glucose 15g / L+calcium chloride dihydrate 2.5g / L+potato 80g / L, medium pH value 6.0; among them, potato uses its extract; liquid medium preparation After that, it is divided into triangular flasks or culture bottles for fungi, with a dispensing volume of 100mL / bottle, and then placed in an autoclave for high-pressure sterilization. After cooling, it is inserted into the bacteria cake; the sterilized liquid medium is inserted into the bacteria Put the root fungus cake into a constant...

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Abstract

The invention discloses a method for mycorrhization of a bletilla striata tissue culture seedling. The method comprises the following steps: step 1: preparing a liquid microbial agent by utilizing a mycorrhizal fungus JSNLBJ-001 of bletilla striata; step 2, adding a suitable concentration of the liquid microbial agent into a culture medium of the bletilla striata tissue culture seedling in the plantlet hardening stage before transplanting the bletilla striata tissue culture seedling for mycorrhizal culture; step 3, taking out the mycorrhized tissue culture seedling of bletilla striata from a tissue culture flask when a basal stem of the tissue culture seedling is covered with hyphae and the hyphae can be observed to attached to the root, removing the culture medium from the root and the basal stem, and then transplanting the mycorrhized tissue culture seedling of bletilla striata into a bletilla striata seedling substrate for domestication. The method provided by the invention can provide high-quality domesticated seedlings for the field cultivation of bletilla striata; according to the method, the liquid microbial agent prepared from the mycorrhizal fungus of bletilla striata is added to enable the root system of the tissue culture seedling to be mycorrhized before the bletilla striata tissue culture seedling is transplanted, after the tissue culture seedling is transplanted,the survival rate can reach 99% or more than 99%, the growth amount is large, tillers of sprouts are many, the growth vigor is strong, and the stress resistance is strong.

Description

technical field [0001] The invention relates to a method for mycorrhizing bletilla striata tissue cultured seedlings, and belongs to the technical field of seedling cultivation. Background technique [0002] Under natural conditions, orchids must establish a symbiotic relationship with mycorrhizal fungi. After the formation of mycorrhizae, they can complete their normal growth and development. The formation of mycorrhizae is essential for orchids, and mycorrhizal fungi Plants provide carbohydrates and other possible secondary metabolites, such as vitamins and hormones. Therefore, in a sense, orchids are parasitic on fungi. Bletilla striata is a perennial herbaceous orchid. Its root structure is a typical mycorrhizal structure. Its seeds have no endosperm. Under natural conditions, they need to be symbiotic with suitable fungi to germinate. Due to the rise of tissue culture and rapid propagation technology, the problem of seed The problem of germination, tissue culture and r...

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 席刚俊酒亚东李傲伟顾陈恩孙明宇陈雪萍
Owner JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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