Bletilla striata germplasm resource in-vitro preservation method

A technology for in vitro preservation and resources, which is applied in the field of in vitro preservation of bletilla germplasm resources, can solve the problems of not being able to meet the needs of in vitro preservation of bletilla and germplasm resources, and shorten the storage time of bletilla and germplasm resources in vitro, so as to prolong the in vitro preservation time , Solve the problem of germplasm resource preservation and large-scale seedling breeding, and the effect of high survival rate

Pending Publication Date: 2022-04-15
GUANGXI BOTANICAL GARDEN OF MEDICINAL PLANTS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the prior art, the storage time of bletilla and in vitro is relatively short, which cannot meet the needs of in vitro preservation of bacilli.

Method used

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  • Bletilla striata germplasm resource in-vitro preservation method
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  • Bletilla striata germplasm resource in-vitro preservation method

Examples

Experimental program
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Effect test

Embodiment 1

[0023] A method for in vitro preservation of P. chinensis germplasm resources, comprising the following steps:

[0024] Step 1, place Pseudobulb in the first culture medium as explant, obtain clump bud, before placing Pseudomonas Pseudobulb in the first medium, first place Pseudobulb at 75 ℃. Soak in ethanol for 45s, then rinse 3 times with sterile water, and put into 0.1% Hgcl with 2% Tween-20 2 The solution was soaked for 4 minutes, then rinsed with sterile water for 3 times, and then put into 0.1% Hgcl with 2% Tween-20 again. 2 The solution was soaked for 4min, finally used sterile water for 3 times, then peeled off the 3 layers of outer skin of Pseudobulb, and inoculated in the first medium for cultivation;

[0025] Step 2, treat the clump buds to grow to 2~3cm, place the clump buds in the second medium and cultivate to have 4~6 leaves (the test tube seedlings have taken root at this moment), obtain the test tube seedlings;

[0026] Step 3: Inoculate the test tube seedli...

Embodiment 2

[0034] A method for in vitro preservation of P. chinensis germplasm resources, comprising the following steps:

[0035] Step 1, place Pseudobulb in the first culture medium as explant, obtain clump bud, before placing Pseudomonas Pseudobulb in the first medium, first place Pseudobulb at 75 ℃. Soak in ethanol for 45s, then rinse 3 times with sterile water, and put into 0.1% Hgcl with 2% Tween-20 2 The solution was soaked for 4 minutes, then rinsed with sterile water for 3 times, and then put into 0.1% Hgcl with 2% Tween-20 again. 2 The solution was soaked for 4min, finally used sterile water for 3 times, then peeled off the 3 layers of outer skin of Pseudobulb, and inoculated in the first medium for cultivation;

[0036] Step 2, treat the clump buds to grow to 2~3cm, place the clump buds in the second medium and cultivate to have 4~6 leaves (the test tube seedlings have taken root at this moment), obtain the test tube seedlings;

[0037] Step 3: Inoculate the test tube seedli...

Embodiment 3

[0045] A method for in vitro preservation of P. chinensis germplasm resources, comprising the following steps:

[0046] Step 1, place Pseudobulb in the first culture medium as explant, obtain clump bud, before placing Pseudomonas Pseudobulb in the first medium, first place Pseudobulb at 75 ℃. Soak in ethanol for 45s, then rinse 3 times with sterile water, and put into 0.1% Hgcl with 2% Tween-20 2The solution was soaked for 4 minutes, then rinsed with sterile water for 3 times, and then added 0.1% Hgcl with 2% Tween-20 2 Soak in the solution for 4 minutes, and finally use sterile water for 3 times, then peel off the three layers of skin of the white and pseudobulbs, and inoculate them in the first medium for cultivation;

[0047] Step 2, when the clustered buds grow to 2-3 cm, the clustered buds are placed in the second culture medium and cultivated until there are 4-6 leaves (the test-tube plantlets have taken root at this time), to obtain the test-tube plantlets;

[0048] S...

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Abstract

The invention discloses a rhizoma bletillae germplasm resource in-vitro preservation method which comprises the following steps: step 1, putting rhizoma bletillae pseudobulb as an explant into a first culture medium for culture to obtain multiple shoots; step 2, putting the cluster buds into a second culture medium for culturing to obtain test-tube plantlets; 3, the test-tube plantlet is inoculated into a third culture medium for in-vitro preservation, and the sucrose content in the third culture medium is 40-100 g / L. In the third step, the third culture medium further comprises 1 / 2MS, 0.3 mg.L <-22 >-BA, 0.2 mg / L NAA and 6g / L agar, and the pH value of the third culture medium is 5.8. In the step 3, the in-vitro preservation method is as follows: the temperature is 22 + / -1 DEG C, the illumination intensity is 1200-2200Lx, and the illumination time is 6-8h / d. By changing the content of cane sugar in the third culture medium, the in-vitro preservation time of rhizoma bletillae is prolonged.

Description

technical field [0001] The invention relates to the technical field of seedling cultivation. More specifically, the present invention relates to a method for in vitro preservation of P. chinensis germplasm resources. Background technique [0002] Bletilla striata (Thunb.) Reichb.f. is a national second-class protected plant. It is a perennial herb that grows in the humid places of the mountains and valleys. Distributed in Fujian, Guangdong, Guangxi, Jiangxi and other places. It is a medicinal and edible plant with the functions of astringent, hemostasis, swelling and muscle regeneration. Used for hemoptysis, hematemesis, traumatic hemorrhage, sore swelling and poison, chapped skin. In recent years, a large number of people have excavated and purchased wild resources, which has led to the shortage of wild resources, and the resources of wild white fungus are on the verge of depletion. With the massive excavation and acquisition of wild resources on the verge of depletion,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 李翠韦莹张占江郭晓云陈晓英雷明黄浩
Owner GUANGXI BOTANICAL GARDEN OF MEDICINAL PLANTS
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