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Bacillus amyloliquefaciens Yb-2 as well as separation method and application thereof

A technology of amylolytic spores and bacillus, applied in the biological field, can solve the problems of slow effect, difference in behavior and mode of action, interference, etc., and achieve the goal of improving seed germination rate and rooting rate, significant economic benefits, and reducing soil pesticide residues Effect

Active Publication Date: 2022-03-29
海南省农业科学院植物保护研究所(海南省农业科学院农产品质量安全与标准研究中心)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Bacillus does not pollute the environment, is safe for humans, animals, and plants, and has a long-term control effect on plant diseases, but is easily interfered by chemical fungicides, and the effect is slow
Sometimes the bioassay results of Bacillus in vitro and in vivo often show inconsistencies. During the application process, due to the constant changes in field environmental factors such as temperature, humidity, and light, the colonization and reproduction of Bacillus are affected, resulting in large numbers of Bacillus in the field. Behavior and function in ecosystems can vary widely

Method used

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  • Bacillus amyloliquefaciens Yb-2 as well as separation method and application thereof
  • Bacillus amyloliquefaciens Yb-2 as well as separation method and application thereof
  • Bacillus amyloliquefaciens Yb-2 as well as separation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: A method for rapid isolation and identification of vegetable endogenous Bacillus amyloliquefaciens Yb-2

[0039] Step 1: If Figure 1-3 As shown, vegetable samples were obtained, Bacillus was isolated by grinding, dilution, and streaking methods, and the isolated Bacillus was purified and cultured for many times, and the Bacillus strain was initially isolated.

[0040] The preliminary isolation of Bacillus strains is as follows: select a vegetable sample, cut the vegetable sample into a square of about 4mm; put the square vegetable sample into a petri dish filled with 75% absolute ethanol for disinfection, and the disinfection time is 30s-2min according to different materials , and washed twice with sterile water; put the sterilized and cleaned vegetable samples into a drip dish, add sterile water to grind, absorb about 50 μL with a pipette tip, put it on an LB plate, and incubate at 30°C for 2-3 days; observe For the growth of Bacillus, the suspected coloni...

example 2

[0070] Example 2: Bacillus subtilis Yb-2 antibacterial activity identification

[0071] Four pathogenic bacteria harmful to vegetables were selected to identify the antibacterial activity of Bacillus subtilis Yb-2, and the antibacterial effect of Bacillus subtilis Yb-2 was clarified.

[0072] In this embodiment, the pathogens are four kinds of capsicum anthracnose bacteria, eggplant Verticillium dahliae, tomato Botrytis cinerea, and tomato bacterial scab.

[0073] Antagonism identification of capsicum anthracnose, eggplant Verticillium dahliae and tomato Botrytis cinerea were identified as follows: put capsicum anthracnose, eggplant Verticillium dahliae and tomato botrytis cinerea on PDA medium for 6 days at 30°C to prepare PDA medium plates ; After the PDA medium plate is dried, use a sterilized hole puncher to punch holes with a diameter of 4 mm in a triangular distribution on the plate, and dry the moisture in the holes; take 30 μl of bacillus liquid cultured to the logarit...

Embodiment 3

[0084] Embodiment 3: Bacillus amyloliquefaciens Yb-2 is to the germination promoting test of vegetable seed

[0085] Activate the Yb-2 strain on the LB plate, pick a single colony at 28°C, and culture it on a shaker at 180rpm to the logarithmic phase. According to the initial preliminary experiments, different concentration gradients were set for the strains. Take the cruciferous vegetable cabbage and water spinach, the solanaceous vegetable pepper and tomato, the cucurbit vegetable cucumber and the leguminous vegetable cowpea seeds as objects, take 20-40 seeds respectively in a petri dish according to the size of the seeds, and pad the bottom of the petri dish with absorbent filter paper , take different concentrations of bacterial solution to soak the seeds, germinate at room temperature, and moisten regularly. Each concentration was repeated three times, and sterile water was used as a control. Since the germination time of each seed is inconsistent, water spinach and cab...

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Abstract

The invention discloses bacillus amyloliquefaciens Yb-2 as well as a separation method and application thereof, and relates to the technical field of biology, the strain is preserved in China General Microbiological Culture Collection Center on June 7, 2021, and the preservation number is CGMCC No.22660. The invention further discloses a preparation method of the bacillus amyloliquefaciens Yb-2. According to the bacillus amyloliquefaciens Yb-2, the bacillus amyloliquefaciens Yb-2 capable of generating an antagonistic effect on colletotrichum capsici, verticillium wilt of eggplants, botrytis cinerea and tomato bacterial scab is separated from vegetable crops for the first time; according to a germination and growth promotion test of the bacillus amyloliquefaciens Yb-2 on the vegetable seeds, the bacillus amyloliquefaciens Yb-2 has a growth promotion effect on the bud length and root length of the seeds, and the germination rate and rooting rate of the seeds can be improved; according to the invention, the safety of bacillus to hot peppers and tomatoes is evaluated through a root irrigation method, a needle punching method and a leaf cutting method, and results show that bacillus amyloliquefaciens Yb-2 is safe to both hot peppers and tomatoes after inoculation.

Description

technical field [0001] The invention relates to the field of biological technology, more specifically, it relates to a bacillus amyloliquefaciens Yb-2 and its separation method and application. Background technique [0002] Plant disease is an important factor that threatens agricultural production. The long-term and extensive use of chemical pesticides not only makes pathogenic bacteria resistant to pesticides, but also pesticide residues are harmful to humans, storage and the environment. With the enhancement of human awareness of environmental protection and food safety, biological control strategies with good control effects have attracted great attention. Biological control is a method of using beneficial organisms to suppress or eliminate harmful organisms. It is harmless and non-toxic , non-polluting, and not easy to produce drug resistance, it may gradually replace chemical pesticides in the future and become an environmentally safe choice. There is a kind of plant ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02A01G13/00A01G22/00A01N63/22A01P1/00A01P3/00C12R1/07
CPCC12N1/20C12N1/02A01N63/22A01G13/00A01G22/00
Inventor 严婉荣赵志祥王宝肖彤斌吉训聪陈圆
Owner 海南省农业科学院植物保护研究所(海南省农业科学院农产品质量安全与标准研究中心)
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