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Bacillus siamese yj15 and its application

A Bacillus, cgmccno.15952 technology, applied in the field of microorganisms, can solve problems such as the prevention and control of bilberry botrytis, large genetic variation, and fast reproduction speed with antagonistic bacteria, achieve a good development prospect of biological pesticides, and optimize fermentation conditions. , the effect of good control effect

Active Publication Date: 2022-02-18
SHANXI AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Chemical pesticides are effective measures to prevent and control Botrytis cinerea. However, Botrytis cinerea has fast reproduction speed, large genetic variation, and high fitness, and it is easy to develop resistance to chemical pesticides.
However, there is no research report on the use of antagonistic bacteria to control lingonberry botrytis

Method used

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  • Bacillus siamese yj15 and its application
  • Bacillus siamese yj15 and its application
  • Bacillus siamese yj15 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Isolation and screening of Bacillus siamese YJ15.

[0035] The lingonberry fruit came from Xiaodian District, Taiyuan City, Shanxi Province, the variety was Lanfeng, and it was stored at 4°C.

[0036] Wash the surface of the lingonberry fruit with tap water, and then rinse it once with sterile water. Soak the clean cranberry fruit in 70% ethanol for 3 min, 2% NaClO for 5 min, 70% ethanol for 30 s, and finally rinse with sterile water for 5 times, and dry the residual water with sterile absorbent paper.

[0037] Put the surface-sterilized lingonberry fruit into a sterilized mortar, add a small amount of quartz sand and 10 mL of sterile water, grind thoroughly, take 200 μL to NA medium, spread it evenly, and culture it at 28 °C for 2 days to obtain the inner Bacterial strains.

[0038] According to the shape, color, size, edge, protrusion, transparency and other characteristics of the colony, pick a single colony and continue to streak and purify.

[0039]Th...

Embodiment 2

[0043] Example 2: Morphological characteristics and physiological and biochemical characteristics of Bacillus siamese YJ15.

[0044] The antagonistic bacterial strains screened in Example 1 were inoculated on NA medium, and cultured at 28° C. for 48 hours. According to the morphological characteristics of the colonies grown by bacterial strains, including size, luster, bulge, shape, transparency, uniformity, edges, etc., as well as the morphological characteristics of bacterial cells observed by Gram staining reaction and transmission electron microscope, and the physiological and biochemical characteristics of bacterial strains, refer to "Common Bacterial System Identification Manual" [Dong Xiuzhu, Cai Miaoying. 2001. Beijing: Science Press.] for identification.

[0045] figure 2 It is the result of morphological characteristics of Bacillus siamese YJ15 cultured on NA medium at 28°C for 48 hours. The colony of strain YJ15 on NA medium is nearly round, with neat edges, opaq...

Embodiment 3

[0048] Example 3: Molecular identification of Bacillus siamese YJ15.

[0049] The strain YJ15 was inoculated in LB liquid medium, and cultured with shaking at 28°C and 180r / min for 10h. The DNA of antagonistic bacteria was extracted using BioFlux BSC12S1 Bacterial Genomic DNA Extraction Kit (Hangzhou Bioer Technology Co., Ltd.).

[0050] 16S rRNA gene sequence analysis uses 25µL PCR reaction system: 10×PCR Buffer 2.5µL, dNTP (10mM) 2µL, MgCl 2 1 µL, 1 µL each of universal primers 27F and 1492r at a concentration of 10 mM, 1 µL template DNA, 0.5 µL Taq DNA polymerase, ddH 2 O to make up to 25 µL.

[0051] The PCR reaction conditions were: pre-denaturation at 95°C for 5 min, 34 cycles (94°C for 45 s, 45°C for 1 min, 72°C for 90 s), and 72°C for 10 min. PCR amplification products were purified and sequenced.

[0052] The PCR reaction conditions were: pre-denaturation at 95°C for 5 min, 34 cycles (94°C for 45 s, 45°C for 1 min, 72°C for 90 s), and 72°C for 10 min. PCR amplif...

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Abstract

The invention relates to a bacillus siamese YJ15 and its application in biological control of botrytis cinerea. Bacillus siamese of the present invention ( Bacillus siamensis ) strain YJ15 belongs to the genus Bacillus ( Bacillus sp.), preserved in the General Microorganism Center (CGMCC) of China Committee for Culture Collection of Microorganisms, with the culture preservation number CGMCC No.15952. Inoculate the cultivated Bacillus siamese YJ15 in LB liquid medium, cultivate and prepare the fermentation seed liquid, then inoculate the fermented seed liquid in the fermentation culture liquid, and obtain the biological bacteria by culturing the obtained fermentation liquid or the supernatant of the fermentation liquid agent. The biological bacterial agent prepared by the invention is used to prevent and control lingonberry gray mold, and a good control effect comparable to that of carbendazim is obtained.

Description

technical field [0001] The invention belongs to the technical field of microbes, and relates to microorganisms for biological control of agricultural crop diseases, in particular to a bacillus siamese used for preventing and treating lingonberry gray mold. Background technique [0002] Bilberry gray mold is caused by Botrytis cinerea ( Botrytis cinerea ) infection, which is an important disease in cranberry production. Botrytis cinerea has the characteristics of common occurrence, serious damage, rapid disease expansion and multiple damage sites. Especially under protected cultivation conditions, the occurrence of gray mold is more serious, and the incidence rate can reach more than 30%, which seriously affects the yield and fruit quality of cranberry. [0003] In 1924, Botrytis cinerea was first discovered on lingonberries grown in New Jersey, USA. At present, Canada, South Korea, Chile and other countries have reported the occurrence of lingonberry botrytis. In 2012, C...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P3/00C12R1/07
CPCA01N63/00C12N1/20C12R2001/07C12N1/205
Inventor 王春伟王燕张曦倩刘倩如侯普兴王建明张作刚王美琴
Owner SHANXI AGRI UNIV
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