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Method for wheat tissue culture
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A technique of tissue culture and wheat, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of genotypes that cannot be successfully regenerated, regeneration rate, and significant differences in culture effects
Inactive Publication Date: 2012-06-20
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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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.
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Embodiment 1
[0038] Example 1. Tissue culture with immature wheat embryos
[0039] In this example, the following wheat varieties were used as experimental objects: Wheat Zhong 8601, Wheat HW642 and Wheat Jimai 22. The steps of the tissue culture method for each wheat material are as follows:
[0040] First, the acquisition of wheat embryos
[0041] Wheat Zhong 8601, Wheat HW642 and Wheat Jimai 22 were planted in the summer breeding base of Guyuan, Hebei (the temperature during the wheat growing period from April to July was 10-25°C), and 13-14 days after flowering and pollination, the Wheat Zhong 8601, Wheat HW642 were collected. and immature embryos of wheat Jimai 22 (heart-shaped stage, 1.0-1.2 mm in size).
[0042] 2. Tissue culture
[0043] 1. Induction culture to obtain embryogenic callus
[0044] The above wheat embryos were inoculated on the following SD2 medium for 8 days (dark, 25°C) to obtain callus I; the callus was transferred to the following SDO medium for 3, 6, 9 or 12 ...
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 the tissue culture method are as follows:
[0077] 1. Tissue culture
[0078] 1. Disinfection of wheat seeds
[0079] The mature wheat CB037 species were sterilized with 70% alcohol for 10 minutes, sterilized with 25% sodium hypochlorite for 25 minutes, rinsed with sterile water for 5 times, soaked in sterile water for 15-18 hours, and sterilized 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] Mature embryos were scraped on sterilized seeds with an inoculation knife, inoculated on Adi medium described below for 8 days (dark, 25°C), callus induced, and then transferred to the following Dicamba-removed cells. Cultured on Ad0 medium for 12 days (dark, 25°C), and then transferred the callus to Adi...
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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 present invention relates to a method for tissue culture of wheat. Background technique [0002] High-frequency regeneration of tissue culture is the basis of cell engineering breeding, genetic engineering breeding and functional genomics research. Dicotyledonous plants such as tobacco and Arabidopsis and monocotyledonous plants such as rice and Brachypodium are very easy to regenerate, so it has become a genetic engineering research. pattern plant. In contrast, wheat tissue culture is still unstable and is largely limited by factors such as explant type, physiological state, genotype and medium, which affect 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 young embryos are the easiest to culture, followed by anther culture and young ear culture, and mature embryos are the most difficult. Ther...
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