Bacillus siamensis YJ15 and application thereof

A technology of bacillus, cgmccno.15952, applied in the field of microbiology, can solve the problems that have not yet been seen in the prevention and control of lingonberry gray mold, fast reproduction speed, large genetic variation, etc., and achieve good development prospects and good control effects

Active Publication Date: 2019-01-15
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 siamensis YJ15 and application thereof
  • Bacillus siamensis YJ15 and application thereof
  • Bacillus siamensis YJ15 and application thereof

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 siamensis YJ15 and an application thereof in biological control of blueberry gray mold. Bacillus siamensis strain YJ15 of that invention belong to the genus Bacillus sp., Deposited in the Common Microorganism Center of China Microorganism Culture Collection Management Committee (CGMCC), strain collection No. CGMCC No. 15952. Bacillus siamensis YJ15 is inoculatedin LB liquid culture medium to obtain fermentation seed liquid, and then the fermentation seed liquid is inoculated in fermentation culture liquid to obtain fermentation broth or supernatant of fermentation broth to obtain biological microbial agent. The biological bacterial agent prepared by the invention can control the blueberry gray mold and has good control effect equivalent to carbendazim.

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 Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P3/00C12R1/07
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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