A method for detoxification of clover green vitrification with ultra-low temperature therapy

A technology of ultra-low temperature vitrification and clover, applied in horticultural methods, botany equipment and methods, plant regeneration, etc., can solve the problems of low plant tissue survival rate and low virus clearance rate, and improve the survival rate of recovery culture, damage reduction effect

Active Publication Date: 2022-08-02
SHANGRAO NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But at present, this method still has the problems of low survival rate of treated plant tissue and low virus clearance rate.

Method used

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  • A method for detoxification of clover green vitrification with ultra-low temperature therapy

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The steps for detoxification of clover green vitrification ultra-low temperature therapy are as follows:

[0033] (1) Sterile seedling culture: Take the tested stems with TMV clover with buds, rinse them with water, soak them in 0.1% mercuric chloride solution for 30 minutes under aseptic conditions, and then transfer them to sodium hypochlorite solution for disinfection. After washing with sterile water, use sterile paper to absorb the water on the surface of the stem section of cloverleaf with buds, and transfer it into the MS medium containing 6-BA 1.0mg / L and NAA 0.5mg / L. is 2000Lux, and the illumination time is 12h·d -1 , the temperature is 25 ± 1 ℃ under the conditions of 5 days of culture, to obtain three leaf green sterile seedlings.

[0034] (2) Pre-cultivation: the aseptic seedlings of Trifolium chinensis cultivated in step (1) were subjected to a light intensity of 2000Lux and a light time of 12h·d. -1 , and continued to culture for 7 days at a temperature ...

Embodiment 2

[0039] The steps for detoxification of clover green vitrification ultra-low temperature therapy are as follows:

[0040](1) Sterile seedling culture: Take the tested stems with TMV clover with buds, rinse them with water, soak them in 0.1% mercuric chloride solution for 30 minutes under aseptic conditions, and then transfer them to sodium hypochlorite solution for disinfection. After washing with sterile water, use sterile paper to absorb the water on the surface of the stem section of cloverleaf with buds, and transfer it into the MS medium containing 6-BA 1.0mg / L and NAA 0.5mg / L. is 2000Lux, and the illumination time is 12h·d -1 , the temperature is 25 ± 1 ℃ under the conditions of 5 days of culture, to obtain three leaf green sterile seedlings.

[0041] (2) Pre-cultivation: the aseptic seedlings of Trifolium chinensis cultivated in step (1) are 1800Lux in the light intensity and 13h·d for the light time. -1 , the temperature was 40 ± 2 ℃ for 8 days, and the medium was MS ...

Embodiment 3

[0046] The steps for detoxification of clover green vitrification ultra-low temperature therapy are as follows:

[0047] (1) Sterile seedling culture: Take the tested stems with TMV clover with buds, rinse them with water, soak them in 0.1% mercuric chloride solution for 30 minutes under aseptic conditions, and then transfer them to sodium hypochlorite solution for disinfection. After washing with sterile water, use sterile paper to absorb the water on the surface of the stem section of cloverleaf with buds, and transfer it into the MS medium containing 6-BA 1.0mg / L and NAA 0.5mg / L. is 2000Lux, and the illumination time is 12h·d -1 , the temperature is 25 ± 1 ℃ under the conditions of 5 days of culture, to obtain three leaf green sterile seedlings.

[0048] (2) Pre-cultivation: the aseptic seedlings of Trifolium chinensis cultivated in step (1) were subjected to a light intensity of 2000Lux and a light time of 12h·d. -1 , the temperature was 40 ± 2 ℃ and the culture was cont...

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Abstract

The invention discloses a method for detoxification by vitrification ultra-low temperature therapy of cloverleaf, belonging to the technical field of plant detoxification. The disclosed specific steps include: (1) pre-culturing the aseptic seedlings of cloverleaf; (2) cutting the shoot tips of the aseptic clover seedlings after the pre-cultivation in step (1), and performing differentiation culture to obtain cloverleaf green seedlings; (3) cutting the shoot tips of the three-leaf green seedlings obtained in step (2), first using PVS with a concentration of 50-60% 2 The solution loads the shoot tips and reuses PVS containing nano-calcium carbonate 2 The solution dehydrates the shoot tips and will be loaded with PVS 2 The solution and the container of the shoot tip are frozen at ultra-low temperature; (4) the shoot tip after the ultra-low temperature freezing in step (3) is thawed, and then resumed and cultured to obtain a cloverleaf virus-free seedling. The stem tips of Cloveria chinensis treated by the vitrification ultra-low temperature therapy detoxification method provided by the invention not only have a high survival rate, but also have a high detoxification rate of the surviving seedlings.

Description

technical field [0001] The invention relates to the technical field of plant detoxification, in particular to a method for detoxification by vitrification ultra-low temperature therapy of cloverleaf. Background technique [0002] Raidxe Tetrastigmae, also known as golden thread hanging gourd, is a herbaceous vine of the Vitiaceae plant Tetrastigma hemsleyanum Diels et Gilg. The whole plant can be used as medicine, and the medicinal effect of the underground tuberous root is the best. Its main components are flavonoids, phenolic acids, triterpenes, steroids and stilbenes. Pharmacological activity studies have shown that cloverleaf has a variety of biological activities such as anti-tumor, antioxidant, immune regulation, anti-viral, anti-inflammatory, analgesic, antipyretic, and hepatoprotective. It is clinically used to treat febrile convulsions, abdominal pain, pneumonia, asthma, hepatitis and other diseases, and has a wide range of applications in the folk. [0003] In t...

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 尹明华王文霞王艳敏文心雨谢莲熊艺馨熊梓伊
Owner SHANGRAO NORMAL UNIV
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