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Propagation method for tissue culture and propagation of cardiocrinum giganteum by taking seeds as explants

An explant and seed technology, applied in the field of plant tissue culture, can solve the problems of slow plant growth, restrictions on the selection and propagation of high-quality provenance of Dalilia, virus accumulation, etc.

Active Publication Date: 2021-10-01
GUANGXI FORESTRY RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Because no matter whether it is ball propagation, or cutting propagation, and the use of scales for tissue culture propagation, it is limited to the use of large lily bulbs, the perennial vegetative propagation will lead to the accumulation of viruses, and at the same time the pollution rate is extremely high, and the plant growth is slow (Lu Yan , 2005; Zhou Yanping et al., 2007; Wang Xiaofei et al., 2014; Xiao Yufei et al., 2018), at the same time, the range of germplasm resources of bulb bulbs is narrow, which limits the selection and propagation of high-quality provenances of Dalilia

Method used

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  • Propagation method for tissue culture and propagation of cardiocrinum giganteum by taking seeds as explants
  • Propagation method for tissue culture and propagation of cardiocrinum giganteum by taking seeds as explants
  • Propagation method for tissue culture and propagation of cardiocrinum giganteum by taking seeds as explants

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Experimental program
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Embodiment 1

[0057] The propagation method of carrying out tissue culture and multiplying Lilium japonicus with seeds as explants comprises the following steps:

[0058] 1. Aseptic seedlings: the treated germinated Dalilia seeds ( figure 1 ) with 75% (v / v) alcohol (prepared with sterile water, add 1 drop of Tween-20 to 100ml) soak for 30s, wash with sterile water for 3 times; then use available chlorine 2% sodium hypochlorite NaClO (prepared with sterile water, 100ml Add 1 drop of Tween-20) to sterilize for 5 minutes, wash with sterile water 3 times, and obtain sterile seedlings.

[0059] 2. Callus and adventitious bud induction:

[0060] Inoculate the sterile seedlings into the induction medium, the formula is as shown in the following 1-10, wherein the acidity and alkalinity is pH5.8; the solvent is deionized water.

[0061] Culture conditions: dark culture, temperature 25 ℃, culture 60 days, observation statistics (results see Table 1 and figure 2 ).

[0062] (1) MS+4.0mg / L6-BA+2.0...

Embodiment 2

[0102] The propagation method of carrying out tissue culture and multiplying Lilium japonicus with seeds as explants comprises the following steps:

[0103] 1. Sterile seedlings: Soak the treated and germinated Dalily seeds with 75% (v / v) alcohol (prepared with sterile water, add 2 drops of Tween-20 to 100ml) for 30s, wash them with sterile water for 3 times; Disinfect with available chlorine 2.5% NaClO (prepared with sterile water, add 2 drops of Tween-20 to 100 ml) for 15 minutes, rinse with sterile water for 5 times to obtain sterile seedlings.

[0104] 2. Callus and adventitious bud induction:

[0105] Inoculate the sterile seedlings in the induction medium, the formula is as follows: MS+1.0mg / L6-BA+2.0mg / L2,4-D+0.5mg / L NAA+30g / L sucrose+7.0g / L Agar+0.2g / LPVP-40, wherein the pH value is pH5.8; the solvent is deionized water.

[0106] Culture conditions: culture in the dark, temperature 22°C, culture for 50 days, observe statistics.

[0107] 3. Proliferation of adventiti...

Embodiment 3

[0116] The propagation method of carrying out tissue culture and multiplying Lilium japonicus with seeds as explants comprises the following steps:

[0117] 1. Sterile seedlings: Soak the treated and germinated Dalilia seeds with 75% (v / v) alcohol (prepared with sterile water, add 1 drop of Tween-20 to 100ml) for 30s, wash them with sterile water for 3 times; Disinfect with available chlorine 2% NaClO (prepared with sterile water, add 1 drop of Tween-20 to 100 ml) for 10 minutes, rinse with sterile water for 4 times to obtain sterile seedlings.

[0118] 2. Callus and adventitious bud induction:

[0119] Inoculate the sterile seedlings in the induction medium, the formula is as follows: MS+1.0mg / L6-BA+2.0mg / L2,4-D+0.5mg / L NAA+30g / L sucrose+7.0g / L Agar+0.2g / LPVP-40; wherein, the pH is pH5.8; the solvent is deionized water.

[0120] Culture conditions: culture in the dark, temperature 23°C, culture for 55 days, observe statistics.

[0121] 3. Proliferation of adventitious buds...

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Abstract

The invention relates to the technical field of plant tissue culture, and particularly discloses a propagation method for tissue culture and propagation of cardiocrinum giganteum by taking seeds as explants. The cardiocrinum giganteum induction proliferation method comprises the following steps: carrying out induction culture on cardiocrinum giganteum seed aseptic seedlings by using an induction culture medium to form calluses and adventitious buds, carrying out further proliferation culture on the adventitious buds by using a proliferation culture medium to obtain amplified adventitious buds, and carrying out induced rooting in a rooting induction culture medium. The method is high in induced differentiation rate, rich in adventitious bud root hair and beneficial to subsequent utilization. According to the method, the sexual reproduction technology and the tissue culture technology are combined, so that the germplasm resources of cardiocrinum giganteum can be well preserved, the genetic stability is effectively guaranteed, the reproduction speed can be greatly increased, and a new strategy is provided for solving the bottleneck of industrial development of cardiocrinum giganteum.

Description

technical field [0001] The invention belongs to the field of plant tissue culture, and relates to a method for tissue culture and multiplication by using plant seeds. Background technique [0002] The lily belongs to the genus Cardiocrinum of the Liliaceae (Liliaceae), and includes three species of the lily (C. giganteum), the buckwheat leaf lily (C. cathayanum), and the Japanese lily (C. cordatum). Lilium and Lilium are closely related, but the tall plant type and large reticulated veined leaves of Lilium are obviously different from those of Lilium (Yu Hong et al., 2005). Dalily is a perennial ball herbaceous plant, which is mainly distributed in the southwestern provinces and autonomous regions of my country, where it grows at the edge of forests, under forests, grasslands or valleys at an altitude of 600-3200m. [0003] Dalily is a precious plant integrating ornamental, food and medicinal materials (Qin Jingjing et al., 2019; Pan Zhengwei et al., 2019). The lily plant ...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 赵志珩陈玉珍卢存福安家成马锦林梁文汇黄晓露廖健明
Owner GUANGXI FORESTRY RES INST