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Rubber tree embryoid body induction medium, method for inducing embryogenic callus and method for cultivating resistant callus

A technology of embryogenic callus and induction medium, which is applied in the field of plant tissue culture and can solve the problems of low overall efficiency and different tissue transformation rates

Active Publication Date: 2019-10-01
HAINAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The regenerated plants were successively induced through rubber unfertilized ovaries, unfertilized ovules, immature anthers, and young buds. However, in the genetic transformation research of rubber trees, the transformation rates of different tissues are different, and the overall efficiency is low.

Method used

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  • Rubber tree embryoid body induction medium, method for inducing embryogenic callus and method for cultivating resistant callus
  • Rubber tree embryoid body induction medium, method for inducing embryogenic callus and method for cultivating resistant callus
  • Rubber tree embryoid body induction medium, method for inducing embryogenic callus and method for cultivating resistant callus

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preparation example Construction

[0064] The preparation method of the induction medium is not particularly limited in the present invention, and the conventional preparation method of the medium well known in the art can be used.

[0065] The invention provides a method for inducing embryogenic callus based on rubber tree anther somatic embryos, comprising the steps of:

[0066] 1) selecting yellow-green male flowers with a length of 2.81 to 3.20 mm to be sterilized, and separating anthers from the sterilized male flowers to obtain sterilized anther somatic cells;

[0067] 2) Inoculate the sterilized anther somatic cells of the step 1) on the first-generation callus induction medium, culture in dark at 26-28°C for 44-46 days, and transfer the obtained callus to the culture medium described in the above scheme. On the embryoid body induction medium, induce culture at 26-28°C for 40-50 days to obtain spherical or cotyledon-shaped somatic embryos;

[0068] 3) Scratching the somatic embryos in step 2) and inocul...

Embodiment 1

[0097] Embryoid induction medium: modified MS medium as the basic medium, including: Kt 0.55mg / L, NAA0.16mg / L, 6-BA0.04mg / L, salicylic acid 0.06mg / L, spermidine 4.5mg / L, acid hydrolyzed casein 310mg / L, coconut water volume concentration is 7%, activated carbon mass concentration is 0.11%, sucrose 55g / L and plant gel 2.4g / L; The pH value of described induction medium is 5.7.

Embodiment 2

[0099] The material is Hevea brasiliensis Müll.Arg. variety Reyan 7-33-97, and the rubber inflorescences were collected from the experimental farm of the Chinese Academy of Tropical Agricultural Sciences. Select yellow-green male flowers with a length of 2.81 mm, first disinfect them with 75% alcohol for 43 seconds, then disinfect them with 1 mg / mL mercury chloride solution for 13.5 minutes, and wash them four times with sterile water to obtain disinfected anthers. Inoculate the sterilized anthers on the first-generation callus induction medium (the first-generation callus induction medium is based on MS medium, including 2,4-D 1.0mg / L, 6-BA0.2mg / L , coconut water volume concentration 5%, sucrose 70g / L and plant gel 2.2g / L, pH value is 5.8), at 26 ℃, dark culture 46d, the callus that obtains is transferred to the embryoid body induction of embodiment 1 On the culture medium, induce at 26°C for 50 days to obtain somatic embryos with a size of 1-1.2 mm.

[0100] The obtained so...

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Abstract

The invention provides a rubber tree embryoid induction culture medium, an induction method for an embryonic callus and a culture method for regenerating resistant callus plant, and belongs to the technical field of plant tissue culture. The rubber tree embryoid induction culture medium takes an improved MS culture medium as a basic culture medium, and is prepared from Kt, NAA, 6-BA, salicylic acid, spermidine, acid hydrolyzed casein, coconut liquid endosperm coconut water, active carbon, saccharose and phytagel. The somatic embryo induction rate of the embryoid induction culture medium provided by the invention for inducing the embryonic callus of a rubber tree anther somatic embryo is increased by 2.04 to 2.16 times in comparison with that of an anther dedifferentiation callus.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a rubber tree embryoid body induction medium, a method for inducing embryogenic callus and a method for cultivating resistant callus. Background technique [0002] Brazilian rubber tree (Hevea brasiliensis Müll.Arg.) belongs to Euphorbiaceae (Euphorbiaceae), rubber tree genus (Hevea), is a perennial cross-pollinated tree, also known as rubber tree, mainly distributed in Hainan, Yunnan, Guangdong and Guangxi provinces in my country , is the main source of natural rubber. As a national strategic resource, natural rubber plays an important role in industrial development and national defense construction. As of 2016, the world's natural rubber consumption has increased for seven consecutive years. As the largest importer and consumer of natural rubber in my country, the contradiction between supply and demand has become increasingly prominent. Rubber tree di...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04A01H4/00
Inventor 陈健妙何朝族罗红丽栾林莉周广振张盛敏
Owner HAINAN UNIV
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