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Bacillus pumilus NMCN1 and application thereof

A technology of Bacillus pumilus and application method, applied in application, bacteria, fungicide, etc., can solve problems such as serious disease of young fruit, fruit fall off, air, water, soil and food pollution, etc.

Inactive Publication Date: 2010-12-29
NANJING HOUJI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Botrytis cinerea is caused by the fungus of the subphylum Deuteromycota and Botrytis cinerea. It mainly damages flowers and fruits, and can also damage leaves and stems; young fruits are more seriously affected, and stigmas and petals are mostly infected first. Afterwards, it moves to the fruit, and the fruit lesions mostly spread from the fruit stem to the fruit surface. The diseased pericarp is off-white and soft rot, and a large number of gray-green mold layers grow out of the disease. In severe cases, the fruit falls off, which seriously affects the growth of protected vegetables, especially tomatoes. Production
At present, the control of tomato gray mold is mainly through spraying chemical reagents, but the extensive use of chemical pesticides not only causes air, water, soil and food pollution, but more importantly, the directional selection of chemical pesticides makes pathogenic bacteria resistant to pesticides , people are eager to find new pesticides with low toxicity, high efficiency and environmental friendliness in the biological field
Plant Growth-Promoting Rhizobacteria (PGPR) has a good symbiotic relationship with plants, which can not only induce plant disease resistance, but also promote plant growth and increase crop yield, so it has become a popular An eye-catching class of biocontrol bacteria, but there are no reports of biocontrol bacteria specifically used to control tomato cinerea

Method used

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  • Bacillus pumilus NMCN1 and application thereof
  • Bacillus pumilus NMCN1 and application thereof
  • Bacillus pumilus NMCN1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Using the five-point sampling method, use a small shovel to remove the topsoil, take dozens of grams of soil samples at a distance of 5-25 mm, put them into a sampling bag, and mark the sampling time, location, and vegetation. Weigh 5g of soil sample, put it into 45ml of physiological saline, shake it on a shaker for 30 minutes, and let it stand for clarification. Dilute the supernatant with sterile water for 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 Five concentration gradients, 80°C water bath for 10min. Aspirate 200μl coated LB plates and culture overnight at 37°C. After the colony grows on the plate, pick a single colony, streak and purify, and save it by number. The screening obtained Bacillus pumilus NMCN1 CGMCCNo.3377.

Embodiment 2

[0020] The Bacillus pumilus (Bacillus pumilus) NMCN1CGMCC No.3377 of embodiment 1 is carried out morphological characteristic observation and physiological and biochemical characteristic identification, the result shows, this bacterium is Gram-positive, rod-shaped, produces spore (as figure 1 shown). Can grow at 10°C, but not at 50°C, can tolerate pH5.7 and 7% NaCl culture, contact enzyme test, V-P test, citrate utilization test, glucose fermentation and casein test are all positive, starch Hydrolysis test, nitrate reduction test, anaerobic growth test and propionate utilization test were all negative.

Embodiment 3

[0022] The Bacillus pumilus (Bacillus pumilus) NMCN1CGMCC No.3377 of Example 1 was inoculated in liquid LB medium, 180 rpm, 37 ° C shaking culture for 10 hours, the bacteria were collected by centrifugation, after suspension, lysozyme and SDS were added to break the wall, and phenol- Genomic DNA was extracted by chloroform method. Using the extracted total DNA as a template, the 16S rDNA gene of the strain was amplified by PCR with forward primer PRB1601 (5'-GGATCCTAATACATGCAAGTCGAGCGG-3') and reverse primer PRB1602 (5'-GGATCCACGTATTACCGCGGCTGCTGGC-3'). The PCR conditions are: 95°C, 4min; 94°C, 1min; 50°C, 1min; 72°C, 2min; 34cycle; 72°C, 10min, stored at 4°C. After the PCR product was recovered, it was sequenced, and its length was confirmed to be 484bp. The sequencing result was shown in the sequence table, and the similarity with Bacillus pumilusGQ911554.1 in GenBank was 99%.

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Abstract

The invention relates to bacillus pumilus NMCN1, which is collected at China General Microbiological Culture Collection Centre (CGMCC) on November 2, 2009 with a collection number of CGMCC No. 3377. The bacillus pumilus NMCN1CGMCC No. 3377 is biocontrol bacillus which is separated from lakeside soil of Tibet Namsto Lake; and the biocontrol bacillus can grow at the temperature of 10 DEG C, is favorable for playing a role in growth promotion and biological control, can suppress pathogenic bacteria and yeast of tomato gray mold, promote the growth of arabidopsis thaliana and tomato, particularly has a prevention and control effect of 75 percent on the tomato gray mold and can also increase the output of the tomato by 12 percent.

Description

technical field [0001] The invention relates to a bacillus pumilus, specifically a low-temperature-suitable bacillus pumilus (Bacillus pumilus) NMCN1 isolated from Tibet and capable of promoting growth and preventing diseases, and its application in preventing and treating gray mold diseases of tomatoes. Background technique [0002] Botrytis cinerea is caused by the fungus of the subphylum Deuteromycota and Botrytis cinerea. It mainly damages flowers and fruits, and can also damage leaves and stems; young fruits are more seriously affected, and stigmas and petals are mostly infected first. Afterwards, it transfers to the fruit, and the fruit lesions mostly extend from the stem to the fruit surface. The diseased pericarp is off-white and soft rot, and a large number of gray-green mold layers grow out of the disease. In severe cases, the fruit falls off, which seriously affects the protection of vegetables in protected areas, especially tomatoes. Production. At present, the ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P21/00A01P3/00C12R1/07
Inventor 高学文薛鹏琦刘芳伍辉军沈波
Owner NANJING HOUJI BIO TECH
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