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Bacterial strain and application thereof

A strain and culture medium technology, applied in the field of microorganisms, can solve the problems of lack of wild-type biocontrol bacillus, difficult genetic manipulation, low genetic transformation efficiency, etc., and achieve the effect of effectively inhibiting pepper blast bacteria

Active Publication Date: 2018-03-06
SPICE & BEVERAGE RES INST CHINESE ACAD OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the genetic manipulation of these strains is still difficult, and their genetic transformation efficiency is much lower than that of "B.subtilis 168"
Wild-type biocontrol bacilli that are easy to genetically manipulate are extremely scarce

Method used

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  • Bacterial strain and application thereof
  • Bacterial strain and application thereof
  • Bacterial strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] A strain of Bacillus subtilis (Bacillus subtilis) VR19 for controlling pepper blast was isolated from the root system of vanilla orchid in Qionghai City, Hainan Province.

[0063] Bio-control bacteria isolation method: collect fresh vanilla orchid root, wash off the soil, grind it with a sterile mortar, add sterile water to dilute 10,000 times, take 100 μl of coated PDA plate, incubate at 30°C for 12 hours, pick A single colony was streaked and purified on a new PDA plate, and a single colony was picked again and inoculated on a PDA medium for later use.

[0064] Screening method for biocontrol bacteria: the strains passed through "indoor plate antagonism test" and "greenhouse potted seedling biocontrol test" (respectively see Examples 2 and 3), and finally screen out biocontrol strains that have good control effects on pepper blast.

[0065] Preparation of bio-control bacteria treatment solution:

[0066] The strains were inoculated in LB medium, and cultured with sha...

Embodiment 2

[0067] Example 2 Laboratory plate antagonism test

[0068] Pepper blast fungus (Phytophthora capsici, Sang Liwei et al., 2011, Pathogen Identification and Occurrence Regularity of Pepper Blast in Hainan Province) was inoculated on PDA medium and cultured at 26°C for 5 days, and the colonies were punched out with a sterile puncher. Circular mycelium block with a diameter of 0.5 cm, inoculate the VR19 mycelium block into the center of the PDA plate, place 4 sterile filter paper sheets with a diameter of 0.5 cm around the inoculation point, and inoculate 5 μl of biocontrol treatment solution on each filter paper sheet, Put the plates into a 26°C incubator and incubate them for 7 days to observe the results. All the conditions of the control plate were the same, only the biocontrol treatment solution was replaced with sterile water.

[0069] The results showed that the control group grew rapidly on the PDA plate, the mycelium covered the whole plate, the mycelium was dense, and t...

Embodiment 3

[0070] Example 3 Greenhouse potted seedling biocontrol test

[0071] Use 1 × 10 7 ~5×10 7 CFU / ml biocontrol bacteria VR19 treatment solution for root irrigation treatment, 50ml per pot, and pepper seedlings treated with clear water and 25% metalaxyl 500 times solution respectively as controls. Pepper blast fungus was cultured on PDA medium at 26°C for 5 days, and was punched into mycelial blocks with a diameter of about 0.5 cm for later use. 24 hours after the second biocontrol treatment, pierce the center of the 3rd and 4th true leaves of pepper seedlings with a pin, inoculate mycelium blocks on the wound and moisturize with absorbent cotton. After the disease was inoculated, the disease incidence of the plants was monitored every day, and the incidence of the pepper seedlings was counted 7 days after the inoculation, and the area of ​​the diseased spots was counted using a plant image analyzer. Disease severity grading standard: 0 grade, lesion area 2 ; Grade 1, 0.5cm 2 ...

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Abstract

The invention relates to the field of microorganisms, in particular to a bacterial strain and application thereof, and provides a bacterial strain used for preventing and / or treating phgtophora palmivora butler. Flat antagonistic experiment results show that bacillus subtilis VR19 has good inhibitory effect on the growth of hyphae of phytophthora capsici. Greenhouse pot culture experiment resultsshow that the incidence of phgtophora palmivora butler and the average lesion area of treated plants are respectively and obviously reduced after root-irrigation treatment is carried out on the bacillus subtilis VR19, and the control effect reaches 68%. Genetic transformation experiment results show that the bacillus subtilis VR19 can be stably, efficiently and genetically transformed after a matched genetic transformation method is used and the transformation efficiency can reach 200-300CFU / microg.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to strains and applications thereof. Background technique [0002] Pepper blast is the number one disease on peppers. Caused by Phytophthora capsici, it mainly damages the base of pepper main vines and causes rapid wilting and death of the plants. It spreads rapidly and is a very destructive disease. The disease occurs widely in pepper planting areas in my country, and can cause economic losses of more than 50%, and even destroy the whole pepper field in severe cases. At present, the control of pepper blast mainly relies on chemical pesticides. However, the large-scale application of chemical agents has hidden dangers such as drug resistance, pesticide residues, and environmental damage, which is not in line with the current green ecological development strategy. [0003] The use of biocontrol Bacillus to control pepper blast has the characteristics of good broad-spectrum, long-last...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P3/00C12R1/125
CPCA01N63/00A01N63/10C12N1/205C12R2001/125
Inventor 高圣风桑利伟刘爱勤苟亚峰孙世伟王政孟倩倩
Owner SPICE & BEVERAGE RES INST CHINESE ACAD OF TROPICAL AGRI SCI
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