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Rapid preparation method of dicotyledonous plant transgenic acceptor

A dicotyledonous and transgenic technology, applied in the field of plant genetic engineering, can solve the problems of reduced cell transformation efficiency, increased chimera ratio, complicated steps, etc., and achieves the effects of stable efficiency, short transgenic cycle and high transgenic efficiency.

Active Publication Date: 2012-01-18
ZHEJIANG UNIV
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  • Application Information

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Problems solved by technology

This method not only has cumbersome steps and a long cycle, but also has poor reproducibility because the receptor material is affected by the operator's technology during the preparation process; at the same time, the relatively long incubation period also causes the cells in the receptor material to have a 5-day lifespan. During the developmental period, the transformation efficiency of cells decreased significantly, and the proportion of chimeras increased after transformation

Method used

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  • Rapid preparation method of dicotyledonous plant transgenic acceptor
  • Rapid preparation method of dicotyledonous plant transgenic acceptor
  • Rapid preparation method of dicotyledonous plant transgenic acceptor

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Embodiment 1

[0047] 1. Surface disinfection and germination of soybean seeds:

[0048] 1. Surface disinfection of soybean Heinong 37 seeds: Soybean seeds are placed in a single layer in a 100×15mm petri dish (about 150 seeds per dish), and 3-4 petri dishes with seeds are opened and placed in a 24mm diameter petri dish. 250ml beaker inside the desiccator. Add 100 ml of sodium hypochlorite solution (mass concentration 5.25%) into the beaker, then slowly add 3.5 ml of 12M HCl along the wall of the beaker, immediately cover the lid of the desiccator, and place it at room temperature (10-25°C) for 10-16 hours Sterilize. After the sterilization is completed, open the lid of the desiccator, quickly cover the petri dish, move the soybean seeds to a sterile ultra-clean bench, and open the petri dish for about 30 minutes to remove residual chlorine. Surface-sterilized seeds can be stored at room temperature for 2 weeks.

[0049] 2. Imbibition of soybean seeds: Put the sterilized soybean seeds in ...

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Abstract

The invention discloses a rapid preparation method of a dicotyledonous plant transgenic acceptor, which comprises the following steps: 1) surface sterilization of dicotyledonous plant seeds; 2) imbibition of the dicotyledonous plant seeds, that is, putting the sterilized dicotyledonous plant seeds into a sterilized differentiation tank, adding sterile water, soaking the sterilized dicotyledonous plant seeds in the dark at 18-22 DEG C for 12-15 hours to obtain the imbibed dicotyledonous plant seeds; 3) acceptor preparation, that is, putting the imbibed dicotyledonous plant seeds on sterile absorbent paper, cutting the seeds vertically along the hila to divide the cotyledon into two petals, removing seed coats to obtain the dicotyledonous plant transgenic acceptors. With the invention, the transgenic program is simple and practical, has stable efficiency, and has great significance for the improvement of dicotyledonous plant transgene efficiency.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering. Specifically, the present invention relates to a method for preparing fast, stable and efficient dicotyledonous plant (such as soybean) transgenic receptors for cultivating stable transgenic lines; meanwhile, the present invention utilizes related receptor materials to develop soybean and other Procedure for transgenic dicotyledonous plants. Background technique [0002] Soybean (Glycine max L. Merrill) is a diploid (2n=40) plant belonging to Fabaceae, Papilionaceae, and Soybean genus. It is an important food crop and economic crop in China and even the world. It occupies a very important position in domestic and foreign markets. However, due to the relatively narrow genetic basis of common cultivated soybeans, the relatively long breeding cycle, and the influence of various diseases, insect pests and various abiotic adversities, the quality and yield of soybeans cultivated only by conv...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H5/00
Inventor 寿惠霞唐桂香周练马林宋张悦杨晓凤周正剑
Owner ZHEJIANG UNIV
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