Radix curcumae endophytic bacteria strain ZJU-C612-2 and application thereof

A technology of use and germs, applied in the field of endophytic bacteria in curcuma, which can solve the problems of under-exploitation of important resources

Active Publication Date: 2018-11-23
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Curcuma curcuma contains abundant endophytic bacteria resources, and the reports on endophytic bacteria of Curcuma curcuma mostly focus on the isolation of endophytic fungi and the research on secondary metabolites of endophytic bacteria (Yan et al., 2014). So far, 7 families have been isolated and a total of 3...

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  • Radix curcumae endophytic bacteria strain ZJU-C612-2 and application thereof
  • Radix curcumae endophytic bacteria strain ZJU-C612-2 and application thereof
  • Radix curcumae endophytic bacteria strain ZJU-C612-2 and application thereof

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Experimental program
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Effect test

Embodiment 1

[0024] Embodiment 1, the screening of bacterial strain ZJU-C612-2

[0025] Healthy Wenyujin tubers are collected from Yunhe County, Lishui City, Zhejiang Province. The isolation of endophytic bacteria adopts the conventional tissue separation method (refer to "A Method for Separating Endophytic Bacteria in Plants" already published in CN103122331A), and a total of 25 strains of endophytic bacteria were isolated. .

[0026] The screening of antagonistic strains adopts the plate confrontation method, and the specific steps are:

[0027] (1) Activate the endophytic bacteria on the LB solid plate, select a ring of single colonies and inoculate them into 4ml of LB liquid medium, and place them in a 200r / min shaking incubator at 37°C for 24h as a strain, with 1% (Volume %) Inoculum amount Inoculate the endophytic bacteria into a new LB liquid medium, culture at 37° C. with shaking at 200 r / min for 24 hours, and obtain endophytic bacteria fermentation liquid for later use.

[0028]...

Embodiment 2

[0031] Embodiment 2, identification of bacterial strain ZJU-C612-2

[0032] The 16S rDNA gene sequence of strain ZJU-C612-2 was amplified and sequenced with universal primers F27 and R1522, and a sequence with a size of 1487bp was obtained. The sequence was uploaded to GenBank to obtain the gene accession number: KY078349.1. Blast comparison analysis was performed on the sequence on the NCBI website, and the results showed that the similarity between it and Bacillus Velez (CP023414.1), Bacillus amyloliquefaciens (KT961125.1) and Bacillus subtilis (HQ711983.1) was average. 99%.

[0033] After the bacterial strain ZJU-C612-2 was cultured on the LB plate for 24 hours, the single colony formed was similar to a circle, with irregular edges, bulges, wrinkled surfaces, opaque, and the colonies were white ( figure 2A). The strain B-11 observed under the transmission electron microscope was rod-shaped or cylindrical, with multiple flagella around it, and the size was (0.94-1.18) μm×...

Embodiment 3

[0038] Embodiment 3, bacterial strain ZJU-C612-2 produce the mensuration of bacteriostatic substance

[0039] 1. Determination of Protease Production Ability

[0040] Spot the strain ZJU-C612-2 on the protease detection plate (A: Dissolve 8g of skimmed milk powder in 300mL of distilled water, sterilize at 115°C for 10min, B: add 8g of agar with distilled water to 300mL, sterilize at 121°C for 20min, A and B respectively sterilized and then mixed), cultured in the dark at 30°C for 2 days, and it was found that the edge of the strain ZJU-C612-2 produced an obvious transparent circle ( image 3 A), showing that strain ZJU-C612-2 can produce protease.

[0041] 2. Determination of the ability to produce chitinase

[0042] Bacterial strain ZJU-C612-2 was spot-inoculated on the culture medium (ammonium dihydrogen phosphate 1.0g, potassium chloride 0.2g, magnesium sulfate hydrate 0.2g, w=1% colloidal chitin) with colloidal chitin as the only carbon source. Butin 100mL, agar 20g, di...

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Abstract

The invention discloses Bacillus velezensis ZJU-C612-2, and a preservation number thereof is CCTCC NO: M 2018312. The Bacillus velezensis ZJU-C612-2 is capable of inhibiting growth of dendrobium officinale colletotrichum orbiculare, saffron corm valsa ceratosperma, walnut twig wilt pathogenic bacteria, sclerotinia sclerotiorum, cotton rhizoctonia solani pathogenic bacteria, and didymella bryoniae,and promoting growth of a dendrobium officinale protocorm and a tissue culture seedling.

Description

technical field [0001] The invention belongs to the field of microorganisms, in particular to an endophytic bacterium ZJU-C612-2 of Curcuma mellifera with antibacterial and growth-promoting functions and its application. Background technique [0002] Endophytes refer to microorganisms that parasitize in plant tissues during the whole life cycle or at a certain stage in the life cycle, and do not cause obvious visible diseases and functional changes to the plants themselves. Studies have shown that many plant endophytes have biocontrol and growth-promoting effects. The biocontrol mechanism of endophytes mainly includes nutrient and space site competition, production of antibacterial substances and induction of plant disease resistance. There are two main reasons why endophytes produce growth-promoting effects. On the one hand, many endophytes can produce plant growth hormones such as auxins, gibberellins, and cytokinins, which can effectively promote the growth and developme...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P1/00C12R1/07
CPCA01N63/00C12N1/205C12R2001/07
Inventor 毛碧增曾欣迟惠荣张亚惠陈卫良
Owner ZHEJIANG UNIV
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