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A kind of plant tissue culture rapid propagation method using shoot tip as explant

A plant tissue culture and explant technology, applied in the field of fast propagation of plant tissue culture, can solve the problems of "ling seed" easy senescence, long plant breeding time, unstable germplasm resources, etc. Short, fast-reproducing effects

Active Publication Date: 2022-05-27
INST OF URBAN AGRI CHINESE ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a kind of stem plant for the long time of plant breeding in the prior art, unstable germplasm resources, "Lingzi" itself is easy to age and carry bacteria, and the traditional tissue culture fast propagation cycle is long and easy to be polluted. The plant tissue culture rapid propagation method with the tip of the explant, compared with the existing tissue culture rapid propagation system, this method can save 1 to 2 months of tissue culture cycle, and a large number of rapid growth in a short period of time is fast and the quality is excellent and tissue culture seedlings with low infection rate

Method used

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  • A kind of plant tissue culture rapid propagation method using shoot tip as explant
  • A kind of plant tissue culture rapid propagation method using shoot tip as explant
  • A kind of plant tissue culture rapid propagation method using shoot tip as explant

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Experimental program
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Effect test

Embodiment 1

[0034] Embodiment 1, the effect of different hormone ratios on the regeneration of adventitious roots of Chuanxiong shoot tips

[0035] (1) Material sampling: The young shoot tips of about 2 cm to 4 cm in the mature stage of Chuanxiong were directly extracted as explants, and there was no need to use a dissecting needle to peel off the young leaves and leaf primordia or cut the shoot tips under a dissecting microscope. In the prior art, what adopts the stem tip of Chuanxiong to propagate and cultivate as explant is to get the stem tip explant of Chuanxiong with 1 to 2 true leaves. The explants are then propagated through callus induction and differentiation culture, which is a cumbersome process and takes a long time.

[0036] (2) Disinfection of explants: carefully rinse the tip of Chuanxiong under running water for 1 hour, then wash it with sterile water 3 times in an ultra-clean bench, then treat it with 70% or 75% alcohol for 30s, and then add 2% sodium hypochlorite soluti...

Embodiment 2

[0050] Embodiment 2, Chuanxiong shoot tip tissue culture fast propagation method

[0051] According to the best rooting medium for the shoot tip screened in the previous stage, the direct organogenesis method was used to establish a tissue culture and fast propagation method for the shoot tip of Chuanxiong. The operation steps are as follows:

[0052] (1) Material selection: The young shoot tips of about 2 cm to 4 cm in the mature stage of Chuanxiong were selected as explants.

[0053] (2) Disinfection of explants: carefully rinse the stem tips of Chuanxiong under running water for 1 to 2 hours, then wash with sterile water 3 times in an ultra-clean bench, then treat with 70% or 75% alcohol for 30s, and then use 2 % (W / V) sodium hypochlorite solution for 4 min, and finally washed 3 times with sterile water.

[0054] (3) Shoot apex rooting medium: transfer the sterilized shoot tips of Chuanxiong to rooting medium, and cultivate under the conditions of 24-26 ° C, 12-16 hours of...

Embodiment 3

[0057] Embodiment 3, stem node tissue culture fast propagation system of Chuanxiong

[0058] Drawing lessons from the tissue culture and rapid propagation system of Chuanxiong stem tip, and using the best rooting medium screened in the early stage, a tissue culture and rapid propagation system of Chuanxiong stem node was established. The operation steps are as follows:

[0059] (1) Select the stem node explants near the tip of the stem, and the stem nodes near the ground are difficult to sterilize and have a high contamination rate.

[0060] (2) Sterilize and sterilize the stem node explants of Chuanxiong. The disinfection method adopted was as follows: first, the stem nodes were rinsed under running water for about 3 hours, then treated with 75% alcohol for 45 s and 2% sodium hypochlorite for 8 min under aseptic conditions, and finally the explants were washed with sterile water for 3- 5 times.

[0061] (3) The stem nodes of Radix Rhizoma Chuanxiong after the disinfection t...

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Abstract

The invention discloses a plant tissue culture rapid propagation method using stem tips as explants: (1) disinfection treatment of explants: take the stem tips on plants as explants and disinfect the surface; the plants are Rhizoma Chuanxiong or bean (2) rooting culture: transfer the sterilized stem tip to the rooting medium, and cultivate it at 24-26°C, 12-16 hours of light per day, and light intensity of 1000-1500Lx; (3) Seedling hardening and transplanting: When the adventitious roots regenerated at the base of the shoot tip grow to about 1-4 cm in rooting medium, open the bottle cap, harden the seedlings indoors, and transplant them into the substrate for planting. The shoot tip tissue rapid propagation technology of the present invention directly induces the shoot tip to regenerate adventitious roots in vitro, without the steps of callus induction and differentiation culture, eliminating the cumbersome process of preparing various culture media and rotating bottles, and can rapidly propagate plants. The traditional tissue culture rapid propagation method can shorten the tissue culture cycle by 1 to 2 months.

Description

technical field [0001] The invention relates to the technical field of rapid propagation of plant tissue culture, in particular to a method for rapid propagation of plant tissue culture using shoot tips as explants. Background technique [0002] Chuanxiong (scientific name: Ligusticum chuanxiong hort) is one of the most common Chinese herbal medicines in my country, and it is also an important raw material for many health products and medicines. Recently, Chuanxiong has become a star Chinese herbal medicine in the fight against the new crown (COVID-19) epidemic. "Xuebijing Injection", which uses safflower, red peony, chuanxiong, salvia, and angelica as the main raw materials, has been approved for the treatment of severe novel coronavirus pneumonia. , critical systemic inflammatory response syndrome and multiple organ failure. [0003] Chuanxiong is a biennial plant, and it is difficult for Chuanxiong to bear fruit. The traditional breeding method of Chuanxiong is to use st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/008A01H4/001
Inventor 祝婷婷任茂智李林宣方琳
Owner INST OF URBAN AGRI CHINESE ACADEMY OF AGRI SCI
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