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Induction Culture of Wheat Mature Embryo Callus and Construction Method of Plant Regeneration System

A callus induction and construction method technology, applied in the field of agricultural biotechnology engineering, can solve the problems of susceptible bacteria, difficulty in controlling the degree of disinfection of immature embryos, and reducing the induction rate.

Inactive Publication Date: 2017-09-01
TIANJIN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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  • Induction Culture of Wheat Mature Embryo Callus and Construction Method of Plant Regeneration System
  • Induction Culture of Wheat Mature Embryo Callus and Construction Method of Plant Regeneration System
  • Induction Culture of Wheat Mature Embryo Callus and Construction Method of Plant Regeneration System

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Embodiment 1

[0079] A method for constructing a callus induction culture and plant regeneration system of wheat mature embryos is carried out according to the following steps:

[0080] (1) Plant material: select winter wheat cultivar Jing 411 with uniform color and size, treated with 70% (w / w) alcohol for 5 minutes, rinsed with sterile water twice, sterilized with 50% (w / w) sodium hypochlorite for 30 minutes, sterile Rinse with water 3 times, take it out after 2 hours of refrigeration at 4°C and rinse once with sterile water; the winter wheat cultivar mentioned refers to: Jing411;

[0081] (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 the formation of regenerated shoots is: callus induction media MS+2.0mg·L -1 2,4-D+200mg·L -1 CH+100mg·L -1 MI+250mg·L -1 Glu,

[0082] Differentiation and plant regeneration medium is MS+10mg·L -1 ZT +...

Embodiment 2

[0090] A method for constructing a callus induction culture and plant regeneration system of wheat mature embryos is carried out according to the following steps:

[0091] (1) Plant material: select winter wheat cultivar Jing 411 with uniform color and size, treated with 70% (w / w) alcohol for 5 minutes, rinsed with sterile water twice, sterilized with 50% (w / w) sodium hypochlorite for 30 minutes, sterile Rinse with water 3 times, take it out after 2 hours of refrigeration at 4°C and rinse once with sterile water; the winter wheat cultivar mentioned refers to: Jing411;

[0092] (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 the formation of regenerated shoots is: callus induction media MS+2.0mg·L -1 2,4-D+200mg·L -1 CH+100mg·L -1 MI+250mg·L -1 Glu,

[0093] Differentiation and plant regeneration medium is MS+10mg·L -1 KT +...

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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) and Tianjin Science and Technology Commission Youth Fund Project (14JCQNJC14900) Tianjin Science and Technology Support Key (11ZCKFNC00700). Technical field [0002] The invention belongs to agricultural biotechnology engineering, and involves a series of processes of wheat mature embryo callus induction, redifferentiation, plant regeneration, field transplantation 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 diseases and insect pests restrict the yield and quality of wheat. Cultivating 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, cell engineering bre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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