Application of hygromycin serving as selective agent in wheat transformation
A technology of hygromycin and screening agent, which is applied in the application field of using hygromycin as a screening agent in wheat transformation, can solve the problems of few successful reports and lack of systematic research on screening doses, and achieve reduction of experimental workload, The effect of reducing experiment cost and reducing experiment cycle
Inactive Publication Date: 2016-08-10
YANGZHOU UNIV
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Problems solved by technology
[0013] In recent years, although some researchers have tried to use hygromycin as a screening agent in the practice of wheat transformation, due to the lack of systematic research on the dosage of screening, there are not many successful reports so far.
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[0039] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.
[0040] 1. Experimental materials
[0041] Transgenic homozygous lines of the T3 generation containing hygromycin selection markers and some seeds of Yangmai 14 (Y14) (Yangmai 14 came from the Agricultural Science Institute of Lixiahe District, Jiangsu Province);
[0042] Experimental reagents: hygromycin reagent, distilled water;
[0043] Experimental supplies: Petri dishes, tweezers, triangular flasks, electronic balances, flower pots, diatomaceous earth, label paper, etc.
[0044] 2. Experimental steps
[0045] (1) prepare hygromycin solutions of different concentrations for subsequent use, the concentrations of hygromycin solutions are respectively 0, 100, 150, 200, 250 and 300 mg / L;
[0046] (2) Under each hygromycin solution concentration of step (1), get respectively 30 seeds of the transgenic material and the reference material, prepa...
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The invention discloses an application of hygromycin serving as a selective agent in wheat transformation. A receptor material for wheat transformation is a T3-generation homozygous line, containing a hygromycin selection marker, of transgenic Yangmai 14, and the optimum selection concentration of a hygromycin solution is 250 mg / L; when the selection dosage of the hygromycin solution is 250 mg / L, the plant survival rate of the receptor material is 60%, and the plant survival rate of a control material is 20%. The transformed transgenic T3-generation homozygous line containing the hygromycin selection marker is taken as the material for preparation of different concentrations of hygromycin, the optimum selection concentration of the hygromycin solution is found out to be 250 mg / L with a seed soaking method and an earth culture method, the working efficiency of wheat genetic transformation is increased, and the experiment cost is saved.
Description
technical field [0001] The invention relates to a method for screening the optimum concentration of hygromycin when exogenous gene transforms wheat, in particular to the application of hygromycin as a screening agent in wheat transformation, and belongs to the field of genetic breeding of transgenic wheat. Background technique [0002] 1. Development of Wheat Transgenic Technology [0003] Plant transgenic technology is the introduction of artificially isolated and modified genes into the plant genome. Due to the expression of the introduced genes, the heritable modification of the plant's traits is caused, and the recipient plant produces new traits. In the early 1980s, scientists began to devote themselves to the research of transgenic plants: in 1983, scientists used the Agrobacterium-mediated method to obtain the world's first transgenic plant tobacco (Schell et al, 1984). Since then, transgenic technology has developed rapidly. So far, scientists have isolated more tha...
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IPC IPC(8): A01G7/06
CPCA01G7/06
Inventor 陈建民刘霜高勇刘慧君居鹏王雨竹张冬平
Owner YANGZHOU UNIV