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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, yield reduction, and vegetable quality deterioration, which seriously threatens the sustainability of facility vegetable production. development, the obstacles caused by continuous cropping have become a major technical problem to be solved urgently in vegetable cultivation and production

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

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Embodiment 1

[0028] Screening and breeding of the Bacillus subtilis M-2 bacterial strain provided by the invention.

[0029] Soak the roots of marine mangrove plants in a large amount of sterile water for about 20 minutes, wash off the soil adhering to the roots, and then rinse the remaining soil on the roots with sterile water. This part of the soil is the rhizosphere soil sample. Collect water samples in 100mL clean and sterilized wide-mouth plastic bottles, take 15mL after shaking evenly, and inoculate aseptically in 100mL ZoBell medium (medium composition: peptone 5g, yeast extract 1g, iron phosphate 0.1g, aged seawater 1000mL, pH7.6~7.8), cultured with shaking at 36°C for 6 days. Observed on the LB plate, the diameter of the colony cultivated for one day is 3-5 mm, round, with neat edges and moist surface. After mixing the 6-day enriched culture solution, take 0.1mL and spread it on the nutrient agar medium, culture at a constant temperature of 36°C for 2 days, number according to th...

Embodiment 2

[0031] Identification of the morphological characteristics and physiological and biochemical characteristics of the Bacillus subtilis M-2 strain provided by the invention.

[0032] Refer to the test method of "Bergey's Manual of Systematic Bacteriology" Vol.VIII to detect its Gram staining, cell size and shape, 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, V-P test, and utilization of mannitol, arabinose, galactose, xylose , rhamnose, sucrose, inosose and citrate tests.

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

Embodiment 3

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

[0036] The strain was inoculated in LB medium, 120 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, the genomic DNA was extracted by the phenol-chloroform method, and the positive phase primer 27F(5' -GAGAGTTTGATCCTGGCTCAG-3') and reverse primer 1541R (5'-AAGGAGGTGATCCAGCC-3'), PCR amplification of its 16S rRNA gene was performed, and the amplified product was sequenced by Beijing Sanbo Company. The PCR conditions are: 94°C, 10min; 94°C, 45s; 55°C, 45s; 72°C, 90s; 30 cycles, stored at 4°C. The length of the 16S rRNA gene sequence is 1543bp, the accession number in GenBank is EU333948, and the similarity with Bacillus subtilis (AB110598) is 99%. The results are shown in the sequence listing.

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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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