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A strain of Aspergillus niger with rice blast antagonism and its application
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A technology for Aspergillus niger and rice blast, applied in the research field of biological control of rice blast, can solve problems such as human and animal poisoning, environmental pollution, etc.
Active Publication Date: 2019-08-13
TRUST CROP PROTECTION TECH CO LTD
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Embodiment 1
[0017] Embodiment one: the separation and preservation of Aspergillus niger (Aspergillus niger) JS-1
[0018] In 2012, the leaves of Cypress twigs were collected in Yanzhiba, Yichang, Hubei, in the Three Gorges River Basin, and were obtained during separation and purification after disinfection;
[0019] Separation and purification: Inoculate the treated sample into PDA (fresh potato 200g / L, sucrose 20g / L, agar 20g / L, the rest is water, pH7.2-7.5, sterilized at 121°C for 20min) solid medium (add concentration 25 μg / mL penicillin inhibits the growth of bacteria and actinomycetes), placed in a constant temperature incubator at 28 ° C for cultivation, when mycelia grow obviously around the sample, the tip mycelium picking method is used to pick strains with different shapes, Transfer to PDA solid medium, successively subculture several times to obtain purified bacterial strains.
[0020] Cultivation: Take the purified strain, inoculate it on the PDA solid plate medium, and culti...
[0024] 18S rDNA sequence analysis: 18S rDNA sequence was amplified by PCR using universal primers. Upstream primer ITS1: 5'-TCC GTA GGT GAA CCT GCG G-3' and universal downstream primer ITS4: 5'-TCC TCC GCT TAT TGA TAT GC-3'. The reaction system is as follows:
[0025]
[0026] Reaction conditions: pre-denaturation at 94°C for 5 minutes, denaturation at 94°C for 1 minute, annealing at 55-58°C for 30 seconds, extension at 72°C for 1.5 minutes, 33 cycles, and finally extension at 72°C for 10 minutes. The PCR product was detected by 1% agarosegel electrophoresis, and a band of about 750 bp was picked for gel recovery and sent to Shanghai Sangon for sequencing.
[0027] Submit the measured sequence to GenBank and obtain the accession number KX822145. The sequence was compared by Blast in GenBank, and the sequence of the strain with higher homology was selected to construct the 18SrDNA phylogen...
Embodiment 3
[0028] Embodiment three: the preparation of Aspergillus niger (Aspergillus niger) JS-1 thalline
[0029] Strain activation: Take Aspergillus niger (Aspergillus niger) JS-1 stored in a refrigerator at 4°C, inoculate a small amount of bacteria on PDA solid plate medium, and culture at 28°C for 4 days.
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Abstract
The invention discloses a strain of Aspergillus niger and its application. The Aspergillus niger deposit name is Aspergillus niger JS‑1; depository unit: China Center for Type Culture Collection; deposit address: Wuhan University, Wuhan, China; deposit date: 2017.1.11; deposit number: CCTCC M 2017024. The present invention proves through experiments that the Aspergillus niger strain has a strong antagonistic effect on rice blast, and its fermentation liquid has a strong antibacterial effect on rice blast, which can be used to prepare rice blast fungi inhibitors, and provide a new method for the development of rice blast biocontrol. Bacterial extracts and fungicides provide new options and have good economic and social benefits.
Description
technical field [0001] The invention relates to a strain of Aspergillus niger which has a strong inhibitory effect on rice blast and its application, and is mainly used in the research of biological control of rice blast. Background technique [0002] Biological control refers to a control method that uses beneficial organisms or other organisms to suppress or eliminate harmful organisms. Common biological control methods include microbial-derived pesticides and botanical-derived pesticides. [0003] Rice blast is a worldwide rice disease, which is listed as the three major diseases of rice together with sheath blight and bacterial blight. Grain loss caused by rice blast severely restricts the development of rice. The current methods of controlling rice blast and sheath blight mainly include chemical control, biological control and cultivation management. Chemical control has quick results and low cost, but it is easy to cause environmental pollution, human and animal poi...
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