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In-vitro high-frequency regeneration replication method of African chrysanthemum

A high-frequency regeneration and gerbera technology, applied in the field of plant tissue culture, culture and reproduction, can solve the problems of restricting the application of plant genetic engineering technology and unsatisfactory regeneration effect, and achieve the effect of less input and high output.

Inactive Publication Date: 2008-07-09
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the 1970s, tissue culture technology has been applied to the production of gerbera seedlings and genetic transformation at home and abroad, but the regeneration effect is not ideal, especially the way of direct germination of flower buds has seriously restricted the development of plant genetic engineering technology in Africa. Chrysanthemum application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] 1. Material: Gerbera (Gerbera janesonii "Sunanda").

[0012] 2. Cultivation steps:

[0013] (1) Primary culture of the receptacle: select the young receptacle of a vigorously growing gerbera as an explant during the growing season, first soak it in 70% alcohol for 10 seconds, then sterilize it with 0.1% mercury liter solution for 5 minutes, and rinse it with sterile water 4 times, inoculated with MS+6-BA 5.0mg·L -1 +NAA 0.5mg·L -1 + sucrose 20g·L -1 +Agar 5g·L -1 Complete plantlets can be formed in 50 days in the first-generation culture medium of the receptacle (pH5.5). The culture temperature is 26°C, the light intensity is 1500lx, and the light is 8h / day.

[0014] (2) Callus induction, adventitious bud differentiation and subculture proliferation: take the petiole of the leaf of the plantlet in the test tube and inoculate it with MS+TDZ 0.2mg·L -1 +NAA 0.05mg·L -1 + sucrose 20g·L -1 +Agar 5g·L -1 The primary callus induction medium (pH5.5) was used to cultur...

Embodiment 2

[0017] 1. Material: Gerbera (Gerbera janesonii "Sunanda").

[0018] 2. Cultivation steps:

[0019] (1) Primary culture of receptacles: select the young receptacles of vigorously growing gerbera as explants during the growing season, first soak them in 70% alcohol for 30 seconds, then sterilize them with 0.1% mercury liter solution for 8 minutes, and rinse them with sterile water 5 times, inoculated into MS+6-BA 8.0mg·L -1 +NAA 0.8mg·L -1 + sucrose 25g·L -1 + agar 6g·L -1 Complete plantlets can be formed in 60 days in the first-generation culture medium of the receptacle (pH5.7). The culture temperature is 28°C, the light intensity is 1600lx, and the light is 10h / day.

[0020] (2) Callus induction, adventitious bud differentiation and subculture proliferation: take the petiole of the leaf of the plantlet in the test tube and inoculate it into MS+TDZ 0.5mg·L -1 +NAA 0.1mg·L -1 + sucrose 25g·L -1 + agar 6g·L -1 The primary callus induction medium (pH 5.6) was used to cul...

Embodiment 3

[0023] 1. Material: Gerbera hybrida (Gerbera hybrida)

[0024] 2. Cultivation steps:

[0025] (1) Primary culture of the receptacle: select the young receptacle of a robust gerbera as an explant during the growing season, soak it in 75% alcohol for 30 seconds, then disinfect it with 0.1% mercury liter solution for 10 minutes, and rinse it with sterile water 5 times, inoculated with MS+6-BA 10.0mg·L -1 +NAA 1.0mg·L -1 + sucrose 30g·L -1 + agar 7g·L -1 Complete plantlets can be formed in 70 days in the first-generation medium of the receptacle (pH 5.8). The culture temperature is 30°C, the light intensity is 2000lx, and the light is 12h / day.

[0026] (2) Callus induction, adventitious bud differentiation and subculture proliferation: take the petiole of the leaf of the plantlet in the test tube and inoculate it into MS+TDZ 1.0mg·L -1 +NAA 0.2mg·L -1 + sucrose 30g·L -1 + agar 7g·L -1 The primary callus induction medium (pH5.8) was used to culture the petioles for 17 days...

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Abstract

The invention relates to an excised high frequency regeneration propagation method of an African chrysanthemum. In the invention, African chrysanthemum test-tube plantlet is the material of high frequency excised regeneration. The method comprises the culture steps of receptacle initially generation culture, callus induction, adventitious bud differentiation and subculture multiplication, rhizogenesis culture and test-tube plantlet transplantation and so on. Optical culture medium is prepared at each stage of culture and propagation which consists of receptacle initially generation culture medium, callus induction initially generation induction culture medium, callus induction culture medium, callus multiplication and differentiation culture medium, rhizogenesis culture medium and transplantation culture medium. The invention provides suitable temperature and illumination conditions, through which, the African chrysanthemum can propagate rapidly with the excised high frequency based on petiole to manufacture germchit. The invention can be applied in the genetic transformation of the African chrysanthemum, meeting the requirements of African chrysanthemum germchit market and breeding genetic engineering.

Description

technical field [0001] The invention relates to plant tissue culture, culture and propagation technology, in particular to a method for producing seedlings by using gerbera petiole as material for high-end rapid propagation of gerbera. Background technique [0002] Gerbera (Gerbera janesonii), commonly known as Gerbera japonica, is one of the five internationally famous fresh-cut flower varieties and has a large market demand. Gerbera can be propagated by sowing, branching and tissue culture. The progeny traits of seed propagation are easy to segregate, and the propagation speed of ramets is slow, so it is rarely used in commercial seedling production. Since the 1970s, tissue culture technology has been applied to the production of gerbera seedlings and genetic transformation at home and abroad, but the regeneration effect is not ideal, especially the way of direct germination of flower buds has seriously restricted the development of plant genetic engineering technology in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00A01G31/00C12N5/04
Inventor 曾宋君周俊张美吴坤林陈之林郑枫段俊
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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