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Method for rapidly evaluating nitrogen fixing capacity of root nodule nitrogen fixation symbionts of leguminous plants

A technology of leguminous plants and symbiotic systems, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of isotope application, real-time detection, long detection cycle, etc., and achieve the effect of simple operation steps

Active Publication Date: 2016-02-03
SHENYANG INST OF APPL ECOLOGY CHINESE ACAD OF SCI
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Problems solved by technology

However, because nitrogenase itself is not easy to extract and is easy to inactivate, and is limited by the physiological characteristics of rhizobia, the related gene encoding nitrogenase is only expressed in a symbiotic state, so the current detection of nitrogen fixation efficiency mostly uses indirect methods
[0004] Currently, 15 The N isotope dilution method is a common method to determine the nitrogen fixation capacity of leguminous plants and rhizobia nitrogen-fixing symbionts. This method has a direct detection index, but the detection cycle is long, and the isotope needs to be applied in advance, so it cannot be used for field detection, nor can it be done in real time. detection

Method used

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  • Method for rapidly evaluating nitrogen fixing capacity of root nodule nitrogen fixation symbionts of leguminous plants
  • Method for rapidly evaluating nitrogen fixing capacity of root nodule nitrogen fixation symbionts of leguminous plants
  • Method for rapidly evaluating nitrogen fixing capacity of root nodule nitrogen fixation symbionts of leguminous plants

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Experimental program
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Embodiment 1

[0048] In this experiment, the expression efficiency of the rhizobia nitrogenase gene was detected in situ, and the method for evaluating the nitrogen fixation ability was carried out in the following steps:

[0049] Control group: sterile vermiculite pot experiment;

[0050] Test group: the same sterile vermiculite pot experiment as the control group, adding chlorimuron-methyl herbicide A) normal field dosage (20ug / kg), B) 5 times the field dosage (100ug / kg), C) 25 Times field dosage (500ug / kg).

[0051] Each group consisted of 3 samples, each sample was taken from 3 soybean plants.

[0052] Take 100 mg of soybean root nodules from each sample, wash them with tap water, and immediately put them into liquid nitrogen refrigeration; sterilize the mortar and pestle at 121 ° C for 20 minutes, dry them, and pre-cool them with liquid nitrogen before grinding; when grinding them into fine powder, Immediately add RNA extraction solution TrizolA+1ml, continue to grind into a homogena...

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Abstract

The invention relates to a method for rapidly evaluating nitrogen fixing capacity of root nodule nitrogen fixation symbionts of leguminous plants. The method mainly includes steps: 1) co-extraction of plant-rhizobium transcript information; 2) in-situ testing of nitrogenase expression; 3) evaluation of the nitrogen fixing capacity of soybean root nodule nitrogen fixation symbionts according to relative transcription abundance of nitrogenase gene nifH. The method has the advantages that firstly, operation steps are simple and convenient; secondly, the method can be used for real-time rapid field detection of nitrogen fixing level.

Description

technical field [0001] The invention relates to a rapid evaluation method for the nitrogen-fixing ability of the root nodule nitrogen-fixing symbiont of leguminous plants. Background technique [0002] The proportion of nitrogen in the atmosphere accounts for 79%, but all animals and plants, as well as most microorganisms, cannot use molecular nitrogen as a nitrogen source, and all life needs nitrogen. Biological nitrogen fixation is the process by which microorganisms reduce nitrogen to ammonia. According to a rough estimate by the Food and Agriculture Organization of the United Nations (FAO) in 1995, the amount of nitrogen fixed by biological nitrogen fixation in the world is close to 2×10 6 tons, accounting for about 3 / 4 of the world's plant nitrogen requirements. According to the relationship between nitrogen-fixing microorganisms and higher plants and other organisms, they can be divided into three categories: authigenic nitrogen-fixing systems, symbiotic nitrogen-fix...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 徐明恺邹谨张惠文李旭张成刚
Owner SHENYANG INST OF APPL ECOLOGY CHINESE ACAD OF SCI
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