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Genetic transformation method for soybean

A genetic transformation method and soybean technology, applied in the field of soybean genetic transformation, can solve the problems of poor repeatability, difficult regeneration of soybean tissue culture, and large differences in transformation frequency.

Inactive Publication Date: 2011-04-06
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This is because the regeneration of soybean tissue culture is difficult, the reproducibility is poor, and the transformation frequency varies greatly among genotypes, which limits the genetic improvement of soybean by genetic engineering.

Method used

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  • Genetic transformation method for soybean
  • Genetic transformation method for soybean
  • Genetic transformation method for soybean

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1, the influence factor that soybean hypocotyl regeneration plant obtains

[0055] 1. The effect of BAP concentration on the frequency of hypocotyl regenerated adventitious buds

[0056] 1. Obtaining recipient soybean hypocotyls

[0057] Test material: Soybean (Glycine max) Heinong 44, recorded in Man Weiqun, Du Weiguang, Chen Yi, Luan Xiaoyan, Liu Xinlei, Wang Fengli, Breeding of a new soybean variety Heinong 44 and the effects of different planting methods on its yield and quality, Heilongjiang Agricultural Sciences, 2004, 5:1-3, in, publicly available from Institute of Botany, Chinese Academy of Sciences.

[0058] Surface sterilization of soybean seeds: select disease-free and plump soybean seeds and spread them on an open petri dish, put them into a desiccator with a beaker of 100mL sodium hypochlorite stock solution, then slowly add 4mL concentrated hydrochloric acid along the beaker, and seal the desiccator , placed in a fume hood to sterilize for 18 ...

Embodiment 2

[0246] Embodiment 2: transfect pCAMBIA2301 plasmid soybean plant and the expression of GUS gene in hypocotyl

[0247] 1. Obtaining of soybean plants transformed with pCAMBIA2301

[0248] method one:

[0249] 1. Obtaining recipient soybean hypocotyls

[0250] 1) Test material: soybean (Glycine max) variety: Heinong 44

[0251] 2) Surface sterilization of soybean seeds: select disease-free and plump soybean seeds and spread them on an open petri dish, put them into a built-in desiccator containing a beaker of 100mL sodium hypochlorite stock solution, then slowly add 4mL of concentrated hydrochloric acid along the beaker, and seal it tightly. Desiccator, placed in a fume hood to sterilize for 18h. The sterilized seeds were placed in the germination medium (1 / 2 MS basic medium + 15,000mg / L sucrose + 7,500mg / L agar, pH 5.8), placed at 25°C, 120μEm -2 S -1 Under the conditions of light intensity (light cycle 18 / 6h), cultured for 5 days.

[0252] 3) Preparation of explants

[...

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Abstract

The invention discloses a genetic transformation method for soybean. The method provided by the invention comprises the following steps of: delivering an exogenous gene into hypocotyl of the soybean to obtain transgenic soybean, wherein the hypocotyl of the soybean contains apical meristem of 0.5 to 1cm; and preparing the hypocotyl of the soybean according to the following steps of: germinating soybean seeds for 5 to 6 days, removing seed coats, cotyledons and terminal buds to obtain the hypocotyl of the soybean containing apical meristem of 0.5 to 1cm. The invention provides the method for establishing a genetic transformation system of the soybean by using the hypocotyl as an receptor, so that adventitious buds can be quickly and efficiently produced in a shorter time, and the method is used for a basic operating system of genetic engineering of the soybean and has important theory and practical significance for improvement on quality of the soybean.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to a soybean genetic transformation method. Background technique [0002] Soybean [Glycine max(L.)Merr.] is an annual leguminous herb plant. China is the origin of soybean and has a history of planting soybean for more than 4,700 years. Soybean is an important oil crop and food crop, and an important source of edible oil and vegetable protein in the world. However, the difficulty of soybean regeneration and genetic transformation hinders the further improvement of soybean agronomic traits and the progress of functional genomics research. To date, more than 300,000 expressed sequence tags (ESTs) obtained from the soybean expression library are waiting for further gene cloning and functional verification. A large class of genes and so on. Soybean genetic resources urgently need to be developed and used to improve soybean cultivation. [0003] Compared with crops such as co...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N1/21A01H4/00A01H5/00
Inventor 许亦农吴韩英王戈亮刘超武旺赵明明
Owner INST OF BOTANY CHINESE ACAD OF SCI
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