Bacillus pumilus NMCC 46 and application thereof

A technology of Bacillus pumilus, nmcc46cgmccno.3378, applied in application, bacteria, fungicides, etc., can solve problems such as environmental pollution and soil fertility decline

Inactive Publication Date: 2010-06-30
NANJING HOUJI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For a long time, chemical pesticides have been an important means of preventing and controlling plant diseases, insects, weeds and rodents. However, a series of problems such as soil fertility decline, environmental pollution, and pathogenic bacteria resistance caused by the abuse of pesticides have continuously prompted people to look for low-toxicity pesticides in the biological field. , efficient an

Method used

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  • Bacillus pumilus NMCC 46 and application thereof
  • Bacillus pumilus NMCC 46 and application thereof
  • Bacillus pumilus NMCC 46 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] 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 and 45ml of normal 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 colonies grew on the plate, single colonies were picked, streaked and purified, and numbered and stored, and screened to obtain Bacillus pumilus NMCC46C GMCC No.3378.

Embodiment 2

[0021] The Bacillus pumilus (Bacillus pumilus) NMCC46CGMCC No.3378 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). Not only can grow at 10°C, but also can grow at 50°C, can tolerate 7% NaCl culture, but can not tolerate pH 5.7 culture, contact enzyme test, citrate utilization test, V-P test, glucose fermentation and casein The tests were all positive, and the starch hydrolysis test, nitrate reduction test, anaerobic growth test and propionate utilization test were all negative.

Embodiment 3

[0023] The Bacillus pumilus (Bacillus pumilus) NMCC46CGMCC No.3378 strain of Example 1 was inoculated in liquid LB medium, 180 rpm, 37 ° C shaking culture for 10 hours, centrifuged to collect the bacteria, after suspension, lysozyme and SDS were added to break the wall, and the phenol - Chloroform method to extract genomic DNA. 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 482bp. The sequencing result was shown in the sequence table, and the similarity with Bacillus pumilusEF157741.1 in GenBank was 94%.

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Abstract

The invention relates to Bacillus pumilus NMCC 46, which is preserved in CGMCC with preservation date as November 2, 2009, and preservation number as CGMCC No.3378. The Bacillus pumilus NMCC 46 CGMCC No.3378 in the invention is a biological control bacillus separated from grass root surrounding soil near Namco Lake in Tibet, which can grow at 10 DEG C, is favorable for exerting growth promotion and biological control effects, can restrain soybean root pythium myriotylum and saccharomycetes, can boost growth of Arabidopsis and soybean, has a prevention and control effects on soybean root pythium myriotylum as high as 85%, can increase yield of soybean to 24%, and has good application prospect.

Description

technical field [0001] The invention relates to a bacillus pumilus, specifically, a low-temperature-suitable bacillus pumilus (Bacillus pumilus) NMCC46 isolated from Tibet with growth-promoting and disease-preventing functions and its application in preventing and controlling soybean root rot. Background technique [0002] Soybean root rot occurs in all soybean producing areas in my country. For example, in the northeast region of my country's main soybean producing areas, the rate of diseased plants in the field is 30% to 40%, and the rate of diseased plants in low-lying land and heavy cropping land is over 70%. Seedling rate is over 15%; while in the eastern region with humid soil, the rate of diseased plants in the early stage of soybean growth (before flowering) in general years is about 75%, the disease index is 35% to 50%, and the disease index in rainy years can reach more than 60%. , a serious threat to the safety of my country's soybean production. For a long time, c...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P21/00A01P3/00C12R1/07
CPCY02A40/10
Inventor 高学文薛鹏琦刘芳伍辉军沈波
Owner NANJING HOUJI BIO TECH
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