Soybean improvement embryonic tip transformation method

An embryo tip and soybean technology, applied in the field of soybean improvement embryo tip transformation, can solve the problems of strong genotype dependence, cumbersome culture conditions, and low regeneration rate, and achieve the effects of convenient material collection, simple operation and high regeneration rate.

Inactive Publication Date: 2014-07-09
TIANJIN UNIV
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Problems solved by technology

So far, soybean regeneration systems still generally have problems such as low regeneration rate, poor repeatability, strong genotype dependence, and cumbersome cultivation conditions, which largely limit the genetic improvement of soybeans by genetic engineering.

Method used

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  • Soybean improvement embryonic tip transformation method
  • Soybean improvement embryonic tip transformation method
  • Soybean improvement embryonic tip transformation method

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Experimental program
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Effect test

Embodiment

[0031] Strains and plasmids: The Agrobacterium used in the experiment was C58 strain, and the recombinant plasmid vector was pSOY12, which was constructed and provided by Professor Shou Huixia, Institute of Plant Science, School of Life Sciences, Zhejiang University (see the full-text database of China's excellent master's degree dissertations, Zhou Zhengjian , optimization of glyphosate-resistant transgenic soybean system [D]. Zhejiang University, 2011; CN201110303049.0), the purpose gene EPSPS and marker gene Bar were constructed on the plasmid. Take the preserved competent C58 Agrobacterium out of the -80°C refrigerator, put it on ice, take out 2 μL of pSOY12 plasmid and mix it with 200 μL of Agrobacterium, put it into a pre-cooled electric shock cup, 1500V, and transform by electric shock. Aspirate the bacterial solution into a 1.5mL EP tube, add 500μL of YEP medium, culture at 27°C for 4h in the dark with shaking, 4000r / min, 28°C, collect the bacterial cells by centrifugat...

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Abstract

The invention relates to a soybean improved embryo tip transformation method. Using Agrobacterium-mediated, vacuum infiltration assisted exogenous gene introduction, embryo tip transformation method with one cotyledon. The method comprises the following steps: obtaining a soybean explant with a cotyledon, using Agrobacterium vacuum infiltration to assist infection and co-cultivation, and transforming to obtain transgenic soybeans. The soybean explants are soaked in sterile water and germinated, then the seed coat is removed, a cotyledon and the embryo tip of the original leaf are removed; the infection method is vacuum infiltration assisted infection, 0.05MPa pressure for 5-8 minutes, 28°C Shake the infection at 150r / min for 1h in a dark environment. The invention provides a method for establishing a soybean genetic transformation system using the embryo tip with a cotyledon as a receptor, without going through the tissue culture stage, and can obtain regenerated plants in a short period of time, which is used as the basic operating system for soybean genetic engineering. For soybean Quality improvement has important theoretical and practical significance.

Description

technical field [0001] The invention relates to a method for transforming soybean improved embryo tips mediated by Agrobacterium. Background technique [0002] Soybean "Glycine max (L.)Merr." is native to my country and is one of the important economic crops. It has extremely high nutritional value, with a protein content of up to 40%. It is of good quality, easily soluble in water, and most easily absorbed by the human body Utilization, a complete protein, is the only plant product that can replace animal food. However, soybean yield and quality are easily affected by diseases, insects and weeds. People try to improve soybean by means of genetic engineering to enhance its resistance to stress. But regeneration systems for genetically modified soybeans have been one of the difficulties in this field. People's research on soybean tissue culture regeneration system can be traced back to the 1960s, and it was not until the 1980s that breakthroughs were made. The regeneration...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H5/00
Inventor 王罡季静冯远航王伟
Owner TIANJIN UNIV
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