A kind of endophytic bacteria of Atractylodes macrocephala and its application

An endophytic bacteria, Atractylodes macrocephala technology, applied in bacteria, applications, biocides, etc., can solve the problems of Atractylodes root rot that have not been reported.

Active Publication Date: 2022-03-11
ZHEJIANG CHINESE MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is no report on the application of controlling Atractylodes rhizome root rot through the endophytic bacteria of Atractylodes rhizome

Method used

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  • A kind of endophytic bacteria of Atractylodes macrocephala and its application
  • A kind of endophytic bacteria of Atractylodes macrocephala and its application
  • A kind of endophytic bacteria of Atractylodes macrocephala and its application

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Effect test

Embodiment 1

[0027] The endophytic bacteria are isolated and obtained according to the following steps: take the rhizome of Zhejiang Yuqian healthy fresh Atractylodes Rhizoma Rhizoma Rhizoma Rhizome and wash it under clean water to remove surface impurities. Cut rhizomes of appropriate size, wrap them in gauze and carry out three-step surface disinfection treatment, 75% ethanol solution for 1 min, 1% sodium hypochlorite solution for 3 min, and 75% ethanol solution for 30 s. Disinfect and take it out, add quartz sand and 20mL PBS buffer, fully grind to a slurry, take 100μL of supernatant and spread it on the surface of NA medium, and place it in a mold incubator at 28°C for 2-3 days in dark. Purify the endophytic bacteria of Atractylodes macrocephala by streaking method, observe regularly until a single colony is isolated, and finally obtain the endophytic bacterial strain AM201 of the present invention, such as figure 1 As shown, the overall color of the colony is yellowish-white, and the ...

Embodiment 2

[0030] Take the cryopreservation tube of the AM201 strain, and under aseptic conditions, pick an appropriate amount of bacteria into NB medium, and culture it for 48 hours at 26°C, in the dark, and at 180rpm to reach the OD of the bacteria solution 600 =0.6~1.0; the three kinds of pathogenic fungi of Atractylodes macrocephala root rot were made into bacterial blocks with a sterile puncher with a diameter of 5mm, respectively connected to the center of the freshly prepared PDA medium, and three equidistant ones were placed at the center of the circle 2cm Use a sterile filter paper piece (6 mm in diameter), and use a pipette gun to draw 5 μL of the bacterial suspension to soak the filter paper piece. 5 μL NB culture solution was used as a negative control, and each treatment was repeated 3 times. Put it into a mold incubator, and measure the relative bacteriostatic rate after cultivating in a dark environment at 26±2°C for 5-7 days. The results showed that AM201 endophytic bact...

Embodiment 3

[0032] Take the cryopreservation tube of the AM201 strain, under aseptic conditions, pick a small amount of bacteria with an inoculation needle, insert it into the sterilized NB liquid medium, and cultivate it in the dark at 26°C, 65% humidity, and 180rpm for 48 hours; further Under sterile conditions, draw the bacterial suspension and adjust the bacterial suspension to OD 600 = 0.6~1.0, the diseased plants of Atractylodes macrocephala were applied by root irrigation, the application amount was 5mL / time / 3d, and continued for 21 days, and an equal volume of distilled water was used as the control group, n=30. Investigate and count the number of Atractylodes macrocephala diseased plants, incidence rate, damage area, and death rate of diseased plants, detect the number of pathogenic bacteria with blood counting method, and calculate the disease index (DI) = [∑ (number of disease-level plants × representative value) / sum of the number of plants × the highest level of disease inciden...

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Abstract

The invention discloses an endophytic bacterium of Atractylodes macrocephala and its application, and belongs to the technical field of plant endophytic bacterium resource utilization and plant disease prevention and control. The endophytic bacterium is isolated from the living body of Atractylodes Rhizoma Atractylodes Rhizoma Atractylodes Rhizoma Atractylodes Rhizoma Atractylodes by endophytic bacterium separation and purification technology, and is classified as Actinomycetes Rhodococcus AM201 (Rhodococcus sp.), and the strain preservation number is CGMCCNo. 22155. The invention also discloses the use of the endophytic bacteria: it can inhibit the growth of Atractylodes rhizome root rot pathogens, induce systemic disease resistance of Atractylodes rhizome, down-regulate the expression of BXL2, At1g65750, and BZIP43 genes of Atractylodes rhizome, and reduce the disease index of Atractylodes rhizome root rot by 35.71 %. The endophytic bacteria of the present invention have obvious control effect on Atractylodes rhizome root rot, and bring broad application prospects to the field of biological control of Atractylodes rhizome fungal diseases.

Description

technical field [0001] The invention relates to the technical field of utilization of plant endophytic bacteria resources and prevention and control of plant diseases, in particular to an endophytic bacterium of Atractylodes macrocephala and its application in the prevention and treatment of root rot of Atractylodes macrocephala. Background technique [0002] The traditional Chinese medicine Atractylodes macrocephala is the dried rhizome of Atractylodes macrocephala Koidz., a plant of the genus Atractylodes atractylodes. It is one of the famous authentic medicinal materials in Zhejiang Province, "Zhe Bawei". It is warm in nature, bitter in taste and sweet in taste. Qi, dampness and diuresis, antiperspirant, anti-abortion and other effects. Atractylodes macrocephala is widely used clinically, and is known as "the first essential medicine for invigorating the spleen and invigorating qi". [0003] Root rot is the main disease in the cultivation of Atractylodes macrocephala. It...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/20A01P3/00C12R1/01
CPCC12N1/20A01N63/20
Inventor 朱波秦路平吴威刘世禹何玟叶张轲杰
Owner ZHEJIANG CHINESE MEDICAL UNIVERSITY
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