Wheat mature embryo callus induction culture and plant regeneration system construction method

A technology of callus induction and construction method, which is applied in the field of agricultural biotechnology engineering, and can solve the problems of reduced induction rate, easy bacterial infection, low redifferentiation rate of immature embryos, etc.

Inactive Publication Date: 2016-03-02
TIANJIN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Due to the strong regeneration ability of callus derived from immature embryos, most scholars use immature embryos as explants for induction and research, but the acquisition of wheat immature embryos is limited by seasons, and the degree of disinfection of immature embryos is not easy to control: disinfection intensity Small, easy to be infected with bacteria in the subsequent induction culture; strong disinfection will inhibit the growth of callus and reduce the induction rate. Compared with immature embryos, mature embryos are explants, which are easy to disinfect, convenient to obtain materials, and are not affected by seasons. Limitation and other advantages, but the redifferentiation rate is lower than that of immature embryos

Method used

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  • Wheat mature embryo callus induction culture and plant regeneration system construction method
  • Wheat mature embryo callus induction culture and plant regeneration system construction method
  • Wheat mature embryo callus induction culture and plant regeneration system construction method

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Effect test

Embodiment 1

[0078] A method for inducing callus induction culture of mature wheat embryos and a plant regeneration system is carried out according to the following steps:

[0079] (1) Plant material: select winter wheat cultivar Jing 411 with uniform color and size, treat with 70% (w / w) alcohol for 5 minutes, rinse with sterile water twice, disinfect with 50% (w / w) sodium hypochlorite for 30 minutes, and sterile Rinse with water for 3 times, refrigerate at 4°C for 2 hours, take it out and rinse with sterile water once; the cultivar of winter wheat mentioned above refers to: Jing 411;

[0080] (2) Screening of induction and regeneration media: Through the screening of 6 induction media and 8 differentiation media, the system for obtaining mature embryos from dedifferentiation to redifferentiation and forming regenerated shoots is: callus induction medium MS+2.0mg·L -1 2,4-D+200mg·L -1 CH+100mg·L -1 MI+250mg·L -1 Glu,

[0081] Differentiation and plant regeneration medium is MS+10mg·L ...

Embodiment 2

[0089] A method for inducing callus induction culture of mature wheat embryos and a plant regeneration system is carried out according to the following steps:

[0090] (1) Plant material: select winter wheat cultivar Jing 411 with uniform color and size, treat with 70% (w / w) alcohol for 5 minutes, rinse with sterile water twice, disinfect with 50% (w / w) sodium hypochlorite for 30 minutes, and sterile Rinse with water for 3 times, refrigerate at 4°C for 2 hours, take it out and rinse with sterile water once; the cultivar of winter wheat mentioned above refers to: Jing 411;

[0091] (2) Screening of induction and regeneration media: Through the screening of 6 induction media and 8 differentiation media, the system for obtaining mature embryos from dedifferentiation to redifferentiation and forming regenerated shoots is: callus induction medium MS+2.0mg·L -1 2,4-D+200mg·L -1 CH+100mg·L -1 MI+250mg·L -1 Glu,

[0092] Differentiation and plant regeneration medium is MS+10mg·L ...

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Abstract

The present invention is as follows: wheat mature embryos of four kinds of winter wheat cultivated variety Beijing 411, Brock and spring wheat S10 jian-6 and spring wheat strong Tianjin No. 5 are taken as explants for callus induction, differentiation and regeneration system optimization for building efficient dedifferentiation regeneration plants and regeneration grain systems; the mature embryos adopt longitudinal cutting processing, a callus induction medium is MS + 2.0mg.L<-1>2,4-D + 200mg. L<-1>CH + 100mg.L<-1>MI + 250mg.L<-1>Glu, a differentiation and plant regeneration culture medium is MS + 10mg.L<-1>ZT + 0.1mg.L<-1>IAA, the differentiation rate is 100%, the seedling rate is 49.33%, a rooting medium is 1 / 2MS, and regeneration grains can be obtained by hardening, field transplanting and management after the transplanting of rooted test-tube plantlets. Compared with reported Beijing 411 regeneration system, the plant regeneration rate in the study is significantly improved. The Beijing 411 mature embryo tissue culture and regeneration system established in the study provides a good acceptor material for subsequent transgenic studies.

Description

[0001] Supported by the National Natural Science Foundation of China (No. 31071671), the Youth Fund Project of Tianjin Science and Technology Commission (14JCQNJC14900), and the Tianjin Science and Technology Support Key Project (11ZCKFNC00700). technical field [0002] The invention belongs to agricultural biotechnology engineering, and relates to a series of processes of wheat mature embryo callus induction, redifferentiation, plant regeneration, field transplanting and seed setting, and has important applications in wheat molecular breeding and wheat gene transformation. Background technique [0003] Wheat is one of the main food crops in my country. In terms of production, adversity stress and pests and diseases restrict the yield and quality of wheat. Breeding stress-resistant, disease-resistant and high-quality wheat varieties is one of the effective ways to solve this problem. At present, the main technologies of wheat breeding include genetic engineering breeding, cel...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/008
Inventor 王振英刘晓颖范宝莉陈宏刘君蒋鲁亚王君雅
Owner TIANJIN NORMAL UNIVERSITY
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