Method for measuring nitrogenase activity of azotobacteria in sugarcane

A determination method and technology of nitrogenase, applied in the determination/inspection of microorganisms, biochemical equipment and methods, measuring devices, etc., can solve the problems of inaccurately reflecting the activity of nitrogenase, and achieve low cost, simple composition, and easy preparation Effect

Inactive Publication Date: 2009-03-11
广西壮族自治区甘蔗研究所
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  • Abstract
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  • Application Information

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Problems solved by technology

Isolation, identification and related characteristics of bacteria with nitrogenase activity in sugarcane [J], Southwest Agricultural Journal, 2007, 5), so the results of nitrogenase activity determination cannot accurately reflect the nitrogenase activity of nitrogenase bacteria in sugarcane

Method used

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  • Method for measuring nitrogenase activity of azotobacteria in sugarcane
  • Method for measuring nitrogenase activity of azotobacteria in sugarcane
  • Method for measuring nitrogenase activity of azotobacteria in sugarcane

Examples

Experimental program
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Embodiment 1

[0091] 1. Composition and preparation of SH medium

[0092] Weigh 100g of sucrose, add distilled water to make the volume 850ml, adjust the pH to 5.8 with 1N hydrochloric acid, adjust the volume to 1000ml, divide it, and heat it at 121°C for 15min for later use.

[0093] 2). Sampling and sample processing

[0094] Take the cane stems of ROC25 perennial root cane +5 to +7 leaves back to the room, rinse with tap water, distilled water, sterile water, dewater with sterile paper, remove the cane skin, take the middle part of the +6 leaf cane stems and chop them finely and evenly. Weigh the sample and repeat it three times. The weight of the sample is 2.0g, 1.9g, and 2.1g, respectively, and transfer it to a 60ml culture bottle, add culture medium to a total volume of 10ml, and seal it with a rubber stopper;

[0095] 3). Incubation

[0096] Transfer the sealed culture bottle to a shaker with a temperature of 28°C and a rotation speed of 60-70 rpm. After culturing for one day, inje...

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Abstract

The invention discloses a method for determining the nitrogenase activity of azotobacteria in sugar cane. The method comprises the following steps: distilled water and the cane sugar are prepared into a culture medium which contains 10 percent of the cane sugar and the pH value of which is 5.8, and the culture medium is named as SH culture medium for short; and the nitrogenase activity value of a sample is calculated through the steps of sampling, sample treatment, incubation, manufacture of a standard curve for determining the content of ethylene, determination of the content of the ethylene, calculation of the nitrogenase activity and so on. The method has the advantages that: firstly, the SH culture medium is simple in compositions, is easy to prepare and has low cost; secondly, the selectivity of the SH culture medium on the azotobacteria in the sugar cane is small; and thirdly, the SH culture medium can more accurately reflect the difference of the nitrogenase activity of different varieties.

Description

technical field [0001] The invention relates to a method for measuring nitrogenase activity of nitrogen-fixing bacteria, in particular to a method for measuring nitrogenase activity of nitrogen-fixing bacteria in sugarcane. Background technique [0002] The determination of nitrogenase activity is an index to identify the nitrogen-fixing ability or nitrogen-fixing efficiency of nitrogen-fixing microorganisms. The commonly used methods for determining the biological nitrogen fixation capacity of sugarcane are acetylene reduction method, nitrogen balance method, 15 N isotope dilution method, 15 N isotope natural abundance method, etc. Among them, the acetylene reduction method is commonly used to measure nitrogenase activity, which has the characteristics of high sensitivity, simple method and fast speed. Its reaction formula is as follows: [0003] [0004] [0005] Theoretically, the reduction of 3 gram molecules of acetylene is equivalent to the reduction of 1 gra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/25G01N30/02
Inventor 李杨瑞杨荣仲方锋学桂意云周会刘惜辉
Owner 广西壮族自治区甘蔗研究所
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