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Method for wheat tissue culture

A tissue culture, wheat technology, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of unsuccessful regeneration rate of genotypes, significant differences in cultivation effects, etc.

Inactive Publication Date: 2010-05-12
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using such a technical system, the cultivation effects of different wheat genotypes under the same conditions and the same wheat genotype in different physiological states are significantly different, and many genotypes cannot be successfully regenerated or the regeneration rate is very low, which cannot meet the needs of genetic engineering research.

Method used

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  • Method for wheat tissue culture
  • Method for wheat tissue culture
  • Method for wheat tissue culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1, carry out tissue culture with wheat immature embryo

[0039] In this example, the following wheat varieties were used as the experimental objects: Wheat Zhong 8601, Wheat HW642 and Wheat Jimai 22. The tissue culture method steps of each wheat material are as follows:

[0040] 1. Obtaining young wheat embryos

[0041] Wheat Zhong 8601, Wheat HW642 and Wheat Jimai 22 were planted in the summer propagation base in Guyuan, Hebei (the temperature during wheat growth from April to July was 10-25°C), and 13-14 days after flowering and pollination, Wheat Zhong 8601 and Wheat HW642 were collected. and immature embryos of wheat Jimai 22 (heart-shaped stage, size 1.0-1.2mm).

[0042] 2. Organizational training

[0043] 1. Induction culture to obtain embryogenic callus

[0044] The above-mentioned immature wheat embryos were inoculated on the following SD2 medium and cultivated for 8 days (darkness, 25° C.) to obtain callus I; the callus was transferred to the fol...

Embodiment 2

[0075] Embodiment 2, carry out tissue culture with wheat mature embryo

[0076] In this example, the variety CB037 was used as the experimental object. The steps of tissue culture method are as follows:

[0077] 1. Organizational training

[0078] 1. Disinfection treatment of wheat seeds

[0079] Sterilize mature wheat CB037 seeds with 70% alcohol for 10 minutes, 25% sodium hypochlorite for 25 minutes, rinse with sterile water for 5 times, soak in sterile water for 15-18 hours, and sterilize with 15% sodium hypochlorite for 15 minutes the next day , Rinse 5 times with sterile water.

[0080] 2. Induction of embryogenic callus from mature embryos

[0081] Scrape the mature embryos on the sterilized seeds with an inoculation knife, inoculate them on the following Adi medium and culture them for 8 days (dark, 25°C), induce callus, and then transfer the callus to the following medium without Dicamba Cultured on Ad0 medium for 12 days (dark, 25° C.), and then transferred the c...

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Abstract

The invention discloses a method for wheat tissue culture, which comprises the following steps: firstly, performing induction culture on a wheat germ to obtain an embryogenic callus; and secondly, performing differentiation culture on the embryogenic callus to obtain a wheat regenerated plantlet, wherein the method of the induction culture comprises the steps of successively performing induction cultures on the wheat germ with a culture medium containing auxin, a culture medium without the auxin, and the culture medium containing the auxin again to obtain the embryogenic callus. The method for the wheat tissue culture further improves the regeneration frequency of immature embryos and mature embryos of wheat, weakens the barriers caused by the conditions such as genotypes, physiological states and the like to the regeneration, provides a technical support for the cell engineering breeding, the genetic engineering breeding and the functional genomics research of the wheat, and has great significance for the biotechnology breeding and the functional genomics research of the wheat.

Description

technical field [0001] The invention relates to a method for wheat tissue culture. Background technique [0002] High-frequency regeneration of tissue culture is the basis of cell engineering breeding, genetic engineering breeding and functional genomics research. The regeneration of dicotyledonous plants such as tobacco and Arabidopsis and monocotyledonous plants such as rice and Brachypodium is very easy, so it has become the basis of genetic engineering research. pattern plants. In contrast, wheat tissue culture is still unstable, which is largely limited by factors such as explant type, physiological state, genotype and culture medium, which affects the progress of genetic engineering breeding and functional genomics research. So far, the successful explants of wheat tissue culture include young embryos, young ears, anthers, mature embryos, etc. The cultivation of young embryos is the easiest, followed by the cultivation of anthers and young ears, and the cultivation of...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 叶兴国殷桂香佘茂云陈朵朵杜丽璞徐惠君别晓敏孙伟珊李欣
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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