Detection method for structural change of rhizosphere soil microorganism of genetically modified paddy rice

A technology for transgenic rice and rhizosphere soil, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as restriction of transgenic soybean varieties, and achieve the effect of rich map bands, accurate measurement, and overcoming interference.

Inactive Publication Date: 2012-02-08
ZHEJIANG UNIV
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Problems solved by technology

Studies have shown that the planting of genetically modified crops will affect some specific microorganisms. For example, the planting of glyphosate-resistant soybeans will promote the growth and colonization of soybean root rot Fusarium spp., which makes the application of this genetically modified soybean variety limited. constraints

Method used

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  • Detection method for structural change of rhizosphere soil microorganism of genetically modified paddy rice
  • Detection method for structural change of rhizosphere soil microorganism of genetically modified paddy rice
  • Detection method for structural change of rhizosphere soil microorganism of genetically modified paddy rice

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Experimental program
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Embodiment Construction

[0017] rice material

[0018] Bt rice Huachi B6, parent Jiazao 935 and distant parent Zhongjiu B

[0019] Field Trials and Sampling

[0020] The field experiment was carried out in the seedling base of Jiande City, Zhejiang Province. The experimental field was divided into 9 plots, and the three rice varieties were planted respectively. Each variety had 3 repeated plots, and each plot had 26-28 rows×12 plants / row≥ There are 300 plants, and the plots of different varieties are set at intervals.

[0021] Sampling was performed at the tillering, heading, filling and maturity stages of rice. The rice and the soil at its roots were pulled out with a spatula and brought back to the laboratory, and three rice samples were taken from each plot. After the retrieved samples were moderately air-dried at room temperature, the soil around the rhizosphere was shaken off and collected. After removing debris such as plant roots and gravel, they were passed through a 2mm sieve for later use...

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Abstract

The invention discloses a detection method for structural change of a rhizosphere soil microorganism of genetically modified paddy rice. The method comprises the following steps of: (1) collecting rhizospheric soil of the genetically modified paddy rice in a tillering stage, a heading stage, a grouting stage and a maturity stage respectively, airdrying the rhizospheric soil at the temperature of between 20 and 30 DEG C, screening with a sieve of 2 to 3 mm, and dissolving the rhizosphere soil in buffer solution to obtain mixed liquor; (2) extracting the total DNA of the microorganism in the mixed liquor, amplifying 16 SrDNA V3 segments of the microorganism by a PCR (Polymerase Chain Reaction) by taking the total DNA as a template, carrying out electrophoretic separation on amplified products by using a temperature gradient gel, and dyeing to obtain a DNA fingerprint of the microorganism; and (3) analyzing the DNA fingerprint of the rhizosphere soil microorganism of the genetically modified paddy rice in all growth stages, and thus obtaining the structural change conditions of the rhizosphere soil microorganism of the genetically modified paddy rice. The detection method is easy to operate. By the detection method, the interference on soil humus is effectively overcome, and the structural change of the rhizosphere soil microorganism of the genetically modified paddy rice can be precisely determined.

Description

technical field [0001] The invention relates to the field of ecological environment protection, and mainly relates to a method for detecting microbial structure changes in transgenic rice rhizosphere soil. Background technique [0002] The development and application of transgenic crops is an important way to improve the efficiency of agricultural production and effectively solve the problem of food security in our country. The release and large-scale production of genetically modified crops may have certain impacts on the agricultural ecological environment. Objectively evaluating the possible impact of genetically modified crops on the agricultural ecological environment is an important basis and practical issue for the research and development of genetically modified crops. [0003] Soil is the carrier of agricultural production. There will be a series of interactions between the planting of transgenic crops and the soil ecosystem. The exogenous genes and their expression...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
Inventor 虞云龙董斌
Owner ZHEJIANG UNIV
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