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Method for screening nitrogen fixing blue green algae with biofertilizer application potential

A technology of nitrogen-fixing cyanobacteria and screening method, applied in the field of microorganisms, can solve the problems of increasing effective nitrogen content and having only reference value and the like

Inactive Publication Date: 2017-01-04
SHANGHAI ADVANCED RES INST CHINESE ACADEMY OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the problems in laboratory screening have the following aspects: 1) the growth evaluation in the laboratory is generally only carried out on a single algae species, but in practical applications, mixed algae species are often used; 2) nitrogenase in the laboratory The activity determination is generally carried out under the condition of nitrogen deficiency or nitrogen limitation. In actual application, the soil will not be in a state of strict nitrogen deficiency, and the nitrogenase activity also changes during the life cycle of nitrogen-fixing cyanobacteria. The inspection index in the laboratory It is only of reference value; 3) The mechanism of cyanobacteria to increase crop yield is not limited to the increase of soil available nitrogen content brought about by nitrogen fixation

Method used

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  • Method for screening nitrogen fixing blue green algae with biofertilizer application potential
  • Method for screening nitrogen fixing blue green algae with biofertilizer application potential
  • Method for screening nitrogen fixing blue green algae with biofertilizer application potential

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Symbiotic experiment of embodiment 1 nitrogen-fixing cyanobacteria and two kinds of bacteria

[0035] Nitrogen-fixing cyanobacteria Anabaena B1611 (purchased from the UTEX algae bank of the University of Texas), Nostoc F280 and Anabaena F243 (respectively purchased from the FACHB freshwater algae bank of the Chinese Academy of Sciences), three soil microorganisms Bacillus thuringiensis Kenya species (Btsk), Pseudomonas fluorescens (Pf) (purchased from China General Microorganism Culture Collection Center (CGMCC) respectively), with numbers 1.1012 and 1.867, respectively.

[0036] According to the principle of high-throughput experiments, a total of 31 experiments were designed for the symbiosis experiment between nitrogen-fixing cyanobacteria and bacteria. Cycle (light:dark=14:10), medium is BG11 medium, culture time is 7 days, the change of chlorophyll a content of three kinds of cyanobacteria, the number of CFU of two kinds of bacteria are measured. The experimental ...

Embodiment 2

[0048] Embodiment 2 pot experiment

[0049] For judging the impact of single cyanobacteria and different cyanobacteria combinations on soil fertility in Example 1, potted soil experiments were carried out, 8 groups of experiments were designed, the culture temperature was 22°C, the light intensity was 64.33 ± 17.62mol / m2 / s, and the light-dark cycle (light : dark=14:10), the content of available phosphorus, soluble organic carbon, and total soluble nitrogen in the soil was measured on the 6th day, and the experimental design scheme is shown in Table 2.

[0050] The determination of soil available phosphorus refers to methods such as Watanabe [Watanabe F.S., S.R.Olsen, Test of an Ascorbic Acid Method for Determining Phosphorus in Water and NaHCO Extracts from Soil. Soil Science Society of America Journal, 1965.29 (6): 677-678]; The determination of carbon and total soluble nitrogen content refers to the method of Techtmann et al [TechtmannS.M., J.L.Fortney, K.A.Ayers, et al., Th...

Embodiment 3

[0058] Symbiotic experiment of embodiment 3 nitrogen-fixing cyanobacteria and two kinds of bacteria

[0059] Nitrogen-fixing cyanobacteria Anabaena B1611 (purchased from the UTEX algae bank of the University of Texas), Nostoc F280 and Anabaena F243 (respectively purchased from the FACHB freshwater algae bank of the Chinese Academy of Sciences), three soil microorganisms Bacillus thuringiensis Kenya species (Btsk), Pseudomonas fluorescens (Pf) and Bacillus subtilis (Bs) (purchased from China General Microorganism Culture Collection Center (CGMCC) respectively), with numbers 1.1012, 1.867 and 1.4255, respectively.

[0060] According to the principle of high-throughput experiments, a total of 63 groups of experiments were designed for the symbiosis experiment of nitrogen-fixing cyanobacteria and bacteria. The culture volume of each group of experiments was 3 mL, the culture temperature was 20 °C, the light intensity was 87.63±3.44 μmol / m2 / s, and the light-dark cycle ( Light: dark...

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Abstract

The invention discloses a method for screening nitrogen fixing blue green algae with biofertilizer application potential. The method is characterized by at least comprising the following steps: (1) preparing 3-10 kinds of candidate nitrogen fixing blue green algae strains, at least two bacteria in soil and a microporous cell culture plate, and designing an alga-alga symbiotic experiment and an alga-bacterium symbiotic experiment to inoculate algal strains and bacteria on a microporous cell culture medium for culturing so as to evaluate the interaction between different nitrogen fixing blue green algae and between nitrogen fixing blue green algae and bacteria; and (2) detecting the chlorophyll a of nitrogen fixing blue green algae and the number of CFU of bacteria in each group of experiment. By adopting the experimental scheme, the nitrogen fixing blue green algae with biofertilizer efficacy in accordance with production application can be effectively screened out.

Description

technical field [0001] The invention relates to a screening method, in particular to a screening method for nitrogen-fixing cyanobacteria with biological fertilizer application potential, and belongs to the field of microorganisms. Background technique [0002] Cyanobacteria, also known as blue-green algae and cyanobacteria, are the simplest and most primitive microalgae among algal organisms. Nitrogen-fixing cyanobacteria, such as Nostoc, Anabaena, and Tolypothrix, can reduce free molecular nitrogen in the atmosphere to nitrogen available for plants through the action of nitrogenase in cells At the same time, nitrogen compounds and active substances such as polysaccharides, amino acids, and polypeptides are continuously secreted during its growth process, which can promote crop growth while improving soil fertility. [0003] Karthikeyan et al. studied the effects of three kinds of cyanobacteria (Calothrix ghosei, Hapalosiphon intricatus, Nostoc sp.) isolated from the wheat...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/01
CPCC12N1/20
Inventor 孙予罕薛垂召赵权宇王丽波汪靓唐涛
Owner SHANGHAI ADVANCED RES INST CHINESE ACADEMY OF SCI
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