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Method for fast genetic transformation of rice

A technology of genetic transformation and rice, applied in horticultural methods, botanical equipment and methods, using vectors to introduce foreign genetic material, etc., can solve problems such as complicated operation steps, lack of acceptance and promotion by researchers, difficulty in indica rice transformation, etc. Cycle, the effect of simplifying the process

Inactive Publication Date: 2011-09-07
ANHUI AGRICULTURAL UNIVERSITY
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Problems solved by technology

However, most of this genetic transformation is aimed at some varieties that are relatively easy to transform, and the experimental methods are limited to a few varieties, especially the transformation of indica rice is difficult, or the operation steps are complicated, and have not been accepted and promoted by the majority of researchers.

Method used

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Experimental program
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Effect test

Embodiment Construction

[0019] 1. Disinfection method of seeds

[0020] Use 2.5% sodium hypochlorite (containing 0.1% Tween 20), pour off the disinfectant, then disinfect with 70% alcohol for 2 minutes, and wash with sterile water 4-6 times. The seeds sterilized by different disinfection methods were placed on the same YN induction medium, and then cultured at 30°C under full light. The number of contaminated bottles was counted after 5 days, and the induction of callus was observed and recorded after 10 days. For the seeds sterilized by mercury chloride, the buds grew vigorously, but the callus grew very small. Some seeds did not form callus, but green shoots appeared. However, after sodium hypochlorite disinfection, most of the seeds have formed enlarged callus, and the sprouts germinated from the seeds have shrunk and become very small. At the same time, the induction rate was counted, and 200 seeds were inoculated at the same time. The callus induction rate after mercury chloride disinfection wa...

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Abstract

The invention discloses a method for fast genetic transformation of rice, comprising the following steps: culturing rice ripe seeds to induce callus; selecting primary callus to serve as a receptor infected by agrobacterium; screening obtained resistive callus directly on a selective media; enabling the resistive callus to be differentiated into a resistive plant; and carrying out domestication and transplantation to obtain the genetically modified rice. According to method provided by the invention, the transgenic cycle of rice is greatly shortened by around two months compared with that obtained by the original method; the steps of genetic transformation of rice are simplified, full exposure culture at 30 DEG C is adopted, culture conditions is not replaced, and the process is simplified, thus saving time and labour; and the dependence on rice varieties is not strong, and the method is not only applicable to transformation of japonica rice but also to the transformation of nonglutinous rice.

Description

technical field [0001] The invention belongs to the field of rice transgenic technology, specifically a method for using the primary callus tissue of mature rice embryos to transform with Agrobacterium, thereby greatly shortening the rice transgenic cycle. Background technique [0002] Rice is not only an important crop, but also a model organism in plant research. Since the success of the world's first transgenic tobacco in 1984, the transgenic work of rice has been the goal of scientists until the first case in 1994 The success of Agrobacterium transformation of rice has been confirmed by conclusive evidence. The reason why Agrobacterium transformation of rice started very late is mainly because monocotyledonous plants such as rice and corn are not natural hosts of Agrobacterium, and Agrobacterium cannot infiltrate under natural conditions. Rice, but after decades of hard work, the continuous optimization of transformation methods and systems, especially the addition of th...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H4/00A01H5/00
Inventor 江海洋张小辉朱苏文唐秀丽周玉琼
Owner ANHUI AGRICULTURAL UNIVERSITY
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