Bacillus subtilis and application thereof in prevention and treatment of continuous cropping obstacle of greenhouse vegetable

A technology of Bacillus subtilis and strains, applied in the fields of biotechnology and agricultural disease prevention and control, can solve the problems of high multiple cropping index, reduced yield, serious vegetable diseases and insect pests, etc., and achieve strong inhibitory effect and growth promotion effect

Inactive Publication Date: 2009-09-16
TIANJIN MODERN VOCATIONAL TECH COLLEGE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, facility vegetables have the characteristics of high intensification, high multiple cropping index and single species. Continuous cropping for many years has caused a series of adverse phenomena such as soil environment deterioration, serious vegetable diseases and insect pests,

Method used

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  • Bacillus subtilis and application thereof in prevention and treatment of continuous cropping obstacle of greenhouse vegetable
  • Bacillus subtilis and application thereof in prevention and treatment of continuous cropping obstacle of greenhouse vegetable
  • Bacillus subtilis and application thereof in prevention and treatment of continuous cropping obstacle of greenhouse vegetable

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0027] Example 1:

[0028] The invention provides the screening and breeding of Bacillus subtilis M-2 strain.

[0029] Soak the roots of marine mangrove plants in a large amount of sterile water for about 20 minutes to wash away the soil adhering to the roots, and then rinse the residual soil at the roots with sterile water. This part of the soil is the rhizosphere soil. Collect the water sample in a 100mL clean, sterilized wide-mouth plastic bottle, shake it evenly, take 15mL, aseptically inoculate it in 100mL ZoBell medium (medium composition: peptone 5g, yeast extract 1g, iron phosphate 0.1g, Chen sea water 1000mL, pH 7.6~7.8), shake culture at 36°C for 6 days. Observed on the LB plate, the diameter of the colony cultured for one day is 3~5mm, it is round, the edges are neat, and the surface is moist. After mixing the enriched culture solution for 6 days, spread 0.1mL on nutrient agar medium, culture at 36°C for 2 days, number according to colony morphology, and use Myxococcus ...

Example Embodiment

[0030] Example 2:

[0031] The invention provides the identification of the morphological characteristics and physiological and biochemical characteristics of the Bacillus subtilis M-2 strain.

[0032] Refer to the test method of "Bergey’s Mannual of Systematic Bacteriology" Vol.VIII to detect its Gram stain, cell size and morphology, presence or absence of flagella and spores, growth temperature, and pH range. NaCL tolerance. Catalase, H 2 S test, starch hydrolysis, cellulose hydrolysis, MR test, phenylalanine deamination, urease test, oxidase, gelatin liquefaction, tyrosine degradation, VP test, and the use of mannitol, arabinose, galactose, and xylose , Rhamnose, sucrose, inosose and citrate test.

[0033] The results showed that the bacterium is Gram-positive, rod-shaped, with a size of 0.5μm~1.0μm×3.5μm~6.0μm, spores are terminal or subterminal, and the perinatal flagella can move (see figure 1 ). The growth temperature is 27~45℃, the pH is 4~9, and the NaCL tolerance is 0~2...

Example Embodiment

[0034] Example 3:

[0035] The present invention provides PCR amplification and sequence determination of the 16S rRNA gene of Bacillus subtilis M-2 strain.

[0036] The strain was inoculated in LB medium, cultured at 120 rpm, shaking at 37°C for 10 hours, and collected by centrifugation. After suspension, lysozyme and SDS were added to break the wall. Genomic DNA was extracted by phenol-chloroform method, and normal phase primer 27F(5' -GAGAGTTTGATCCTGGCTCAG-3') and reverse primer 1541R (5'-AAGGAGGTGATCCAGCC-3'), PCR amplification of its 16S rRNA gene, the amplified products were sequenced by Beijing Sanbo Company. The PCR conditions were: 94°C, 10min; 94°C, 45s; 55°C 45s; 72°C, 90s; 30 cycles, 4°C storage. The 16S rRNA gene sequence is 1543 bp in length, the accession number in GenBank is EU333948, and the similarity with Bacillus subtilis (AB110598) is 99%. The results are shown in the sequence table.

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Abstract

The invention relates to Bacillus subtilis and the application thereof in prevention and treatment of continuous cropping obstacle of greenhouse vegetable. The Bacillus subtilis is M-2 strain with the preservation number of CGMCC No. 2271. The M-2 strain is plant growth-promoting rhizobacteria separated from oceanic mangrove plant, which can restrain the growth of various harmful fungi such as fusarium wilt of cucumber, root rot of tomato, the root rot of soybean and the like, and can effectively prevent and treat the continuous cropping obstacle of the greenhouse vegetable and increase the quality and output of the greenhouse vegetable.

Description

technical field [0001] The invention belongs to the technical field of biotechnology and agricultural disease prevention and control, and in particular relates to a Bacillus subtilis and its application in preventing and controlling obstacles to facility vegetable continuous cropping. Background technique [0002] According to the survey data of the Food and Agriculture Organization of the United Nations, crops taken away by diseases and insect pests in the world account for 20% to 30% of the harvest every year, and the resulting economic losses amount to 120 billion US dollars. At present, it is mainly controlled by spraying chemical reagents. However, due to the extensive use of chemical pesticides, air, water, soil and food are polluted to varying degrees, and even poisonous substances gradually accumulate in domestic animals and humans to cause poisoning. Around the world, about 2 million people are poisoned by chemical pesticides every year, and about 40,000 of them di...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01P3/00C12R1/125
Inventor 孙勇民范延辉任超王芃傅明范树生权
Owner TIANJIN MODERN VOCATIONAL TECH COLLEGE
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