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Endophytic bacillus subtilis

A technology of Bacillus subtilis and bacterial liquid, applied in the field of microorganisms, can solve the problems of complex composition groups, reduced inhibitory activity, poor genetic stability, etc., and achieve the effect of high genetic stability

Inactive Publication Date: 2011-11-23
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 constituent groups of pepper blast strains are very complex, and typical or single pepper blast strains are used as indicator bacteria in existing screening, so the microorganisms screened have certain limitations, and their control effect is far inferior to that of chemical control.
In addition, as the number of passages increases, the inhibitory activity of the currently screened microorganisms against the spores of Pepper blast is gradually reduced, and the genetic stability is poor.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: the isolation method of endophytic Bacillus subtilis of the present invention

[0021] Healthy leaves were collected from areas with severe pepper blast disease, cut into tissue pieces of about 2cm×2cm size with a sterile scalpel, rinsed with sterile water several times, soaked in 75% alcohol for 1min, rinsed with sterile water for 3 Next, place the tissue block in a sterile mortar, cut it into pieces with scissors, add sterile water to grind, let it stand for 30 minutes, and dilute it to 10 -1 、10 -2 、10 -3 For the 3 gradients, pipette 0.3 mL each of the last rinsing solution and grinding solution, and smear it on the PDA medium plate. The last rinsing solution was used as CK, cultured at 28°C, and observed daily. Select a strain that has no colonies in CK but grows colonies in the grinding liquid, and picks a single colony according to the characteristics of the colony shape, color, etc. After purification and numbering, it is cultured in a plate to t...

Embodiment 2

[0022] Example 2: Determination of Physiological and Biochemical and 16S DNA Sequences of Endophytic Bacillus subtilis in the present invention

[0023] 1. Physiological and biochemical detection

[0024] The growth of strain Z on the NA plate, colony shape and color, Gram staining, spore staining and flagella staining were detected by referring to the method on page 182 of "Plant Disease Research Methods" edited by Fang Zhongda.

[0025] Detect the physiological and biochemical characteristics of strain Z, including: litmus milk, catalase, V.P. reaction, starch hydrolysis, nitrate reduction reaction, gelatin liquefaction reaction, oxidase, M.R., phenylalanine deaminase, citrate utilization , urease test, utilization of propionate, whether it can grow on a medium containing 7% NaCl and pH5.7, etc., the method refers to the method on pages 353-385 of "Common Bacteria System Identification Manual" edited by Dong Xiuzhu and Cai Miaoying .

[0026] The test results are as follow...

Embodiment 3

[0037] Embodiment 3: Inhibition test of endophytic Bacillus subtilis of the present invention to spore germination of pepper blast pathogen

[0038] Streak inoculation of the activated endophytic Bacillus subtilis of the present invention in a PDA culture dish, culture at 28° C., transfer the dish once every 24 hours, and transfer 40 times in total. The inhibitory effect of the endophytic Bacillus subtilis strains of the 1st generation, 10th generation, 20th generation, 30th generation and 40th generation on the spore germination of pepper blast pathogen was determined by the hanging drop method. The results are shown in Table 2.

[0039] Table 2 Inhibition test of endophytic bacillus subtilis of the present invention to pepper blast pathogen spore germination

[0040]

[0041] As can be seen from the results in Table 2, the endophytic Bacillus subtilis described in the present invention suppresses the EC of pepper blast pathogen spore germination from the 1st generation to...

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Abstract

The invention specifically discloses an endophytic bacillus subtilis, which relates to the field of microbes. The endophytic bacillus subtilis provided in the invention is conserved in the CCCCM common micro-organism center and is assigned an accession number of CGMCC No.4844. The 40th generation of the endophytic bacillus subtilis still has high activity in inhibiting spores of phytophthora footrot strains, high genetic stability, an effect on controlling sudden death disease similar to the effect of chemical controlling and no pollution to environment, is ecological and safe, and can be widely applied in controlling sudden death disease and in preparing preparations for controlling sudden death disease.

Description

technical field [0001] The present invention relates to the field of microorganisms, and more specifically relates to an endophytic Bacillus subtilis. Background technique [0002] Pepper (Piper nigrum L.) is a perennial evergreen climbing vine of the family Piperaceae. It is an important tropical spice crop in the world. It is one of the most commonly used and favorite condiments for people. It is widely used in food, medical care and other fields. . [0003] Pepper blast is a climate-dependent disease that often occurs during the rainy season. It is a highly contagious soil-borne disease caused by Phytophthora infection. Since the introduction of pepper, pepper blast has been the primary disease that endangers pepper production in my country. When the disease is severe, the loss of pepper gardens can reach more than 90%, or even destroy the whole garden, which has brought immeasurable losses to my country's pepper planting industry. The control technology of pepper blast ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/02A01P3/00C12R1/125
Inventor 桑利伟刘爱勤谭乐和邬华松孙世伟
Owner SPICE & BEVERAGE RES INST CHINESE ACAD OF TROPICAL AGRI SCI
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