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Method for culturing and rapidly propagating tissue culture seedlings of lotus

A fast technology for tissue culture seedlings, applied in the field of plant tissue culture, can solve the problems of easy contamination of explants, low proliferation efficiency, and low survival rate, and achieve the effects of high survival rate, low pollution rate, and simple operation

Active Publication Date: 2020-08-21
WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a method for the cultivation and rapid propagation of lotus tissue culture seedlings, so as to solve the problems of easy pollution of explants, low survival rate, low proliferation efficiency and long cycle in the existing lotus tissue culture technology

Method used

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  • Method for culturing and rapidly propagating tissue culture seedlings of lotus
  • Method for culturing and rapidly propagating tissue culture seedlings of lotus
  • Method for culturing and rapidly propagating tissue culture seedlings of lotus

Examples

Experimental program
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Effect test

Embodiment 1

[0026] The screening of lotus tissue culture seedling explant is carried out as follows:

[0027] (1) The young leaves, terminal buds of underground stems, mature lotus seeds and fresh lotus seeds 15-18 days after pollination of 'Qiu Hongyang' were selected as explants to cultivate aseptic seedlings. Wherein the leaves (after removing the veins) and the terminal buds of the underground stems (after peeling off the sheaths) are directly sterilized, and the ripe lotus seeds and fresh lotus seeds are sterilized after removing the outer testa, and finally the germs are taken out with sterile scissors as explants. The sterilization method is as follows: sterilize with 75% ethanol for 30 seconds, rinse once with sterile water; then sterilize with 0.1% mercuric chloride for 8 minutes, and rinse with sterile water for 5 times.

[0028] (2) Under aseptic conditions, four kinds of lotus explants were inoculated in the aseptic seedling medium, the photoperiod was 16h / 8h (day / night), the ...

Embodiment 2

[0033] The optimization of the aseptic seedling medium of lotus tissue culture is carried out according to the following steps:

[0034] (1) Fresh lotus seeds picked 15-18 days after pollination in 'Qiu Hongyang' were taken, the outer testa was removed, rinsed with tap water for 15 minutes, and then transferred to a sterile workbench. Disinfect with 75% ethanol for 30 seconds and rinse with sterile water. Then disinfect with 0.1% mercury liter for 8 minutes and rinse with sterile water for 5 times. The embryos were finally removed as explants using sterile scissors.

[0035] (2) Under sterile conditions, inoculate lotus germs in sterile seedling medium with a photoperiod of 16h / 8h (day / night), light intensity of 2000Lx, and temperature control of 25±2°C. After 1 month of cultivation, the aseptic seedlings take root, and after 3 months of cultivation, the aseptic seedlings with rooted stems are obtained. The aseptic seedling medium has been designed with the following propor...

Embodiment 3

[0047] The optimization of the subculture medium of lotus tissue culture is carried out according to the following steps:

[0048] (1) Fresh lotus seeds picked 15-18 days after pollination in 'Qiu Hongyang' were taken, the outer testa was removed, rinsed with tap water for 15 minutes, and then transferred to a sterile workbench. Disinfect with 75% ethanol for 30 seconds and rinse with sterile water. Then disinfect with 0.1% mercury liter for 8 minutes and rinse with sterile water for 5 times. The embryos were finally removed as explants using sterile scissors.

[0049] (2) Under sterile conditions, inoculate lotus germs in sterile seedling medium with a photoperiod of 16h / 8h (day / night), light intensity of 2000Lx, and temperature control of 25±2°C. After 1 month of cultivation, the aseptic seedlings take root, and after 3 months of cultivation, the aseptic seedlings with rooted stems are obtained. The sterile seedling medium uses MS as the basic medium, adds 0.5mg / L 6-BA+0....

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Abstract

The invention discloses a method for culturing and rapidly propagating tissue culture seedlings of lotus. The method comprises the following steps of (1) taking lotus germs on the 15th-18th day afterpollination as a material, inoculating the lotus germs into an aseptic seedling culture medium under an aseptic condition, and sealing and putting the aseptic seedling culture medium into a tissue culture room; (2) culturing for 3 months to obtain rooted stolon aseptic seedlings; (3) cutting each section of the stolon seedling into new buds, inoculating the new buds into a subculture medium for subculture, obtaining a large number of rooted stolon aseptic seedlings after one month, and repeating the steps until 5 times of subculture multiplication culture is carried out; and (4) transplantingthe stolon seedlings obtained in the step (3) to a greenhouse for acclimatization and domestication, and transplanting to a natural condition for growth after 3 weeks. According to the method, the embryo of the fresh lotus seed is taken as the explant in a breakthrough manner, the stolon seedling is successfully cultured, the success rate is up to 98%, the pollution rate is lower than 5%, the propagation coefficient of each subculture can reach 9.6, the period is short, the propagation efficiency is high, large-scale production of the lotus seedling is facilitated, and a new way is provided for meeting the market demand of the lotus seedling.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for cultivating and rapidly multiplying lotus tissue culture seedlings. Background technique [0002] The lotus is a plant of the genus Nelumbo, which is the largest aquatic economic crop cultivated in my country. It has extremely important edible, ornamental and medicinal values. At the same time, lotus is also rich in a large number of secondary metabolites including alkaloids, flavonoids and antioxidants. , has antibacterial, anti-inflammatory, anti-oxidant and anti-cancer effects, and has become one of the most promising first-choice medicinal materials for the development of natural and safe health care drugs. Generally, lotus has two propagation methods: seed propagation and lotus root propagation. Seed propagation has higher requirements on time and season, and lotus is mostly a cross-pollinated plant. The offspring of seed propagation ofte...

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 杨美邓显豹熊雅倩杨东刘艳玲王云梦孙恒刘娟
Owner WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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