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Banana endophytic bacillus velezensis strain and application thereof

A technology of Bacillus and Velez, which is applied in the field of endogenous Bacillus Velez in bananas, can solve the problems of accelerating the resistance of pathogenic microorganisms, achieve good development and application prospects, reduce the occurrence of symptoms, and increase the speed of reproduction

Active Publication Date: 2020-04-21
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The extensive use of chemical pesticides not only brings potential risks to the drug residue problem in bananas, but also accelerates the emergence of pathogenic microorganisms' drug resistance

Method used

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  • Banana endophytic bacillus velezensis strain and application thereof
  • Banana endophytic bacillus velezensis strain and application thereof
  • Banana endophytic bacillus velezensis strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The isolation and purification of embodiment 1 Veles bacillus strain BS

[0031] (1) Preparation of PDA medium: first wash and peel the potatoes, then weigh 200g of potatoes and cut them into small pieces, boil them in water (boil for 20-30 minutes, and it can be pierced by a glass rod), and use eight layers of gauze filter. Add 20g of glucose, add water to 1000mL, stir and mix evenly, add 15g of agar, fully heat and dissolve, then dispense into 250mL Erlenmeyer flasks, sterilize with damp heat at 121°C for 20min, and store for later use.

[0032] (2) Preparation of LB medium: Weigh 10g of tryptone, 5g of yeast extract, and 10g of sodium chloride, add water to 1000mL, stir and mix, add 15g of agar, fully heat and dissolve, then pack into 250mL Erlenmeyer flasks, Sterilize with damp heat at 121°C for 20 minutes, and store for future use.

[0033](3) The samples of banana tissue cultured seedlings were collected in Huadu District, Guangzhou City, Guangdong Province. Aft...

Embodiment 2

[0034] The screening of embodiment 2 Veles bacillus strain BS

[0035] (1) Determination of the activity of Bacillus BS by plate confrontation method

[0036] With No. 4 physiological race FOC4 of banana fusarium wilt as target, place the No. 4 physiological race FOC4 bacterium cake of banana fusarium wilt with a diameter of 7mm on the PDA plate medium, dip a little embodiment 1 step (3) with a sterilized toothpick The bacterium liquid of the obtained strain was streaked about 5 cm away from the fungus cake, and cultured upside down in a 28° C. incubator for 4-5 days. A strain with obvious antibacterial effect on the target bacteria was screened out and named BS. The results showed that Bacillus velesii BS had obvious inhibitory effect on Fusarium wilt fungus FOC4 ( figure 1 ). According to the above method, it is effective against Peronophthora litchii, Colletotrichum siamense, Phomopsis longanae, Neofusicoccum parvum and Nalanthamala psidii These pathogenic bacteria were...

Embodiment 3

[0040] The identification of embodiment 3 Veles bacillus strain BS

[0041] Pick a single colony in 3mL LB liquid medium, place it on a 28°C, 180rpm shaker for overnight culture, extract the DNA of the bacteria according to the method of "OMEGAD3350-1 Bacteria DNA Kit", and store it at -20°C for later use. 16S rRNA general primer 27F / 1492R was used for PCR amplification. After passing the test, Sangon Bioengineering (Shanghai) Co., Ltd. was commissioned to perform sequencing. The nucleotide sequence of 16S rRNA is shown in SEQ ID No.1.

[0042] 27F:5'-AGAGTTTGATCCTGGCTCAG-3';

[0043] 1492R: 5'-GGTTACCTTGTTACGACTT-3'.

[0044] The 16S rRNA sequencing results were submitted to NCBI for sequence comparison. The results showed that the strain BS had high homology with Bacillus velezensis K2, B. velezensis BH21, B. velezensis DH82, Bacillus amyloliquefaciens TUL308, B. amyloliquefaciens WH1G and other strains. Both are as high as 99%. Therefore, it is difficult to judge the ta...

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Abstract

The invention discloses a banana endophytic bacillus velezensis strain and an application thereof. The name of the bacillus velezensis strain is bacillus velezensis BS (Bacillus velezensis). The preservation number is GDMCC No: 60930, and the strain is preserved in Guangdong Province Microbial Culture Collection Center of 59 # building of 100 # courtyard of Xianlie Middle Road of Guangzhou City onDecember 11, 2019. The strain disclosed by the invention belongs to banana endophytic bacteria, has no pathogenicity and has a growth promoting effect on bananas. The bacterial strain and the biological agent thereof have a remarkable inhibition effect on banana wilt pathogen FOC4, the banana wilt symptom is obviously relieved, and the biological agent is environment-friendly and non-toxic in source and has small influence on the ecological environment; the strain has different degrees of inhibition effects on plant pathogenic fungi of different genera; the culture conditions are simple, a propagation speed is fast, thalli and a bacterial liquid can serve as main bodies for prevention and control, storage and transportation are easy, and good development and application prospects are achieved.

Description

technical field [0001] The invention belongs to the technical field of biological control of plant diseases, and in particular relates to a banana endophytic Bacillus velaisi and its application. Background technique [0002] Banana (Musa spp.) is a plant of the genus Musa of the Musa family, and its plants are large herbaceous plants. It is planted in about 130 countries and regions around the world, with an annual output of more than 100 million tons. my country's banana production areas are mainly concentrated in Guangdong, Guangxi, Yunnan, Hainan, Fujian, Taiwan and other places, with a planting area of ​​more than 36hm 2 , is an important economic crop in subtropical regions of my country. [0003] Diseases are the main factors restricting the development of the global banana industry. There are many diseases that affect the development of the banana industry, and one of the most important diseases is banana wilt. Banana wilt, also known as banana Panama disease, was ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/22A01P3/00C12R1/07
CPCC12N1/20A01N63/00C12R2001/07C12N1/205
Inventor 李敏慧曾敬苑曼琳王猛习平根李华平姜子德
Owner SOUTH CHINA AGRI UNIV
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