Pittosporum tobira somatic embryogenesis and plant regeneration method

A technology of embryogenesis and somatic cells, which is applied in the field of plant propagation, can solve the problems of difficult germination of Pittosporum seeds and low seedling rate of seeds, and achieve the effects of shortening the cultivation period, uniform emergence of seedlings, and easy management

Inactive Publication Date: 2016-03-30
GUANGXI BOTANICAL GARDEN OF MEDICINAL PLANTS
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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 Pittosporum somatic embryogenesis and plant regeneration, w

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Pittosporum somatic embryogenesis and plant regeneration method of the present invention, comprises the following steps:

[0016] (1) Selection and disinfection of explants: Take Pittosporum fresh seeds as explants, soak them in 2v / v% detergent solution for 5min, rinse them with tap water for 15-30min, add 2-3 drops of Tween -20 0.1v / v% mercuric chloride 100mL was sterilized for 8-10min, rinsed with sterile water for 3-5 times, and finally removed the surface moisture with sterile filter paper to obtain explants. Wherein, sterile water is distilled water sterilized by high temperature and high pressure.

[0017] (2) Induction of explants to obtain sterile test-tube plantlets: place the explants obtained in step (1) in an ultra-clean workbench, inoculate them into MS medium, and culture them at a temperature of 23-27°C under dark conditions Cultivate for 20 days to obtain sterile test-tube plantlets. Among them, GA30.1mg / L, sucrose30g / L and agar5g / L were added to the M...

Embodiment 2

[0023] (1) Selection and disinfection of explants: Take Pittosporum fresh seeds as explants, soak them in 2v / v% detergent solution for 5min, rinse them with tap water for 15-30min, add 2-3 drops of Tween -20 0.1v / v% mercuric chloride 100mL was sterilized for 8-10min, rinsed with sterile water for 3-5 times, and finally removed the surface moisture with sterile filter paper to obtain explants. Wherein, sterile water is distilled water sterilized by high temperature and high pressure.

[0024] (2) Induction of explants to obtain sterile test-tube plantlets: place the explants obtained in step (1) in an ultra-clean workbench, inoculate them into MS medium, and culture them at a temperature of 23-27°C under dark conditions Cultivate for 20 days to obtain sterile test-tube plantlets. Among them, GA30.1mg / L, sucrose30g / L and agar5g / L were added to the MS medium, and the pH value of the medium was 5.8.

[0025] (3) Induction of embryogenic callus: the cotyledon of the aseptic test-...

Embodiment 3

[0030](1) Selection and disinfection of explants: Take Pittosporum fresh seeds as explants, soak them in 2v / v% detergent solution for 5min, rinse them with tap water for 15-30min, add 2-3 drops of Tween -20 0.1v / v% mercuric chloride 100mL was sterilized for 8-10min, rinsed with sterile water for 3-5 times, and finally removed the surface moisture with sterile filter paper to obtain explants. Wherein, sterile water is distilled water sterilized by high temperature and high pressure.

[0031] (2) Induction of explants to obtain sterile test-tube plantlets: place the explants obtained in step (1) in an ultra-clean workbench, inoculate them into MS medium, and culture them at a temperature of 23-27°C under dark conditions Cultivate for 20 days to obtain sterile test-tube plantlets. Among them, GA30.1mg / L, sucrose30g / L and agar5g / L were added to the MS medium, and the pH value of the medium was 5.8.

[0032] (3) Induction of embryogenic callus: the cotyledon of the aseptic test-t...

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Abstract

The invention discloses a pittosporum tobira somatic embryogenesis and plant regeneration method. The pittosporum tobira somatic embryogenesis and plant regeneration method comprises the following steps: (1) by taking fresh pittosporum tobira seeds as explants, sterilizing the explants; (2) placing the sterilized explants in an MS minimal medium to induce sterile test-tube seedlings; (3) placing the cotyledons of the sterile test-tube seedlings in an MS embryonic callus induction medium to induce embryogenic calli; (4) placing the embryogenic calli in an MS embryoid germination medium to induce regeneration plants; (5) placing the regeneration plants in an MS strong seedling culture medium for culturing to obtain complete small seedlings; and (6) uncovering the vessel caps of tissue culture vessels of the complete small seedlings, hardening the seedlings for 4 to 7 days at room temperature, taking out the young seedlings, cleaning the culture medium at the roots of the young seedlings, transplanting the young seedlings into a vermiculite medium which is watered thoroughly, transplanting the young seedlings into a large field after the young seedlings grow for one month at room temperature, and spraying once every morning and evening. By adopting the pittosporum tobira somatic embryogenesis and plant regeneration method, a large amount of high-quality pittosporum tobira seedlings can be obtained within a short time; the transplanting survival rate is 95% or above; the problem about scale seedling is effectively solved; basis is also provided for transgenic research, biotechnological breeding and the like of the pittosporum tobira.

Description

technical field [0001] The invention relates to a method for plant propagation, in particular to a method for Pittosporum somatic embryogenesis and plant regeneration. Background technique [0002] Pittosporumtobira (Thunb.) Ait) is mainly distributed in the provinces and regions south of the Yangtze River Basin. Pittosporumtobira (Thunb.) Ait is an evergreen shrub or small tree, which has strong resistance to harmful gases such as sulfur dioxide, chlorine, and hydrogen fluoride. The landscaping plants with strong allelopathic effects were found in all parts. Pittosporum branches and leaves, synonymous with Qilixiangye (Taiwan), have the functions of detoxification and insecticide, and are mainly used to treat scabies and swollen toxins (edited by the editorial board of "Zhonghua Materia Medica" of the State Administration of Traditional Chinese Medicine, Zhonghua Materia Medica 4, Shanghai Science and Technology Press, 1999:65-66). [0003] Modern studies have shown that ...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/008
Inventor 李刚缪剑华唐美琼李正文周琼
Owner GUANGXI BOTANICAL GARDEN OF MEDICINAL PLANTS
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