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A rape endophyte Bacillus amyloliquefaciens 4‑3 and its application method

A technology for dissolving starch spores and rape sclerotinia, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, and applications, and can solve the problems of polluting agricultural products and the environment, and lack of chemical agents for disease-resistant varieties of resources.

Active Publication Date: 2017-06-06
HUNAN INST OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, disease-resistant varieties and chemical agents are used for prevention and control in production, but there are problems such as lack of resources of disease-resistant varieties and pollution of agricultural products and the environment by chemical agents, while biological control has the advantages of less environmental pollution, low cost, and long-lasting effects. are getting more and more attention

Method used

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  • A rape endophyte Bacillus amyloliquefaciens 4‑3 and its application method
  • A rape endophyte Bacillus amyloliquefaciens 4‑3 and its application method
  • A rape endophyte Bacillus amyloliquefaciens 4‑3 and its application method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Isolation of plant endophytes

[0030] Sturdy rapeseed plants were collected from Changsha County, Wangcheng, Hunan Academy of Agricultural Sciences and other places, the roots were taken to wash the sediment, and the white, strong and spotless roots and phloem were cut off. After absorbing the surface water with filter paper, soak in 75% alcohol, carry out surface sterilization treatment for 12 minutes, soak and wash in sterile water for 6 times, and then use 0.2% HgCl 2 Soak, carry out surface sterilization for 18 minutes, soak and wash in sterile water 9 times, leave the 9th soak in washing water as a control. Then put the rape root phloem into a sterile grinding bowl and add an appropriate amount of sterile water to grind into a slurry, and then take the slurry to perform gradient dilution to level 5. Extract 0.5ml rape plant slurry from each dilution gradient to add potatoes plus agar-rich medium (potato 200.0g glucose 20.0g yeast extract 2.0g protein build...

Embodiment 2

[0031] Example 2: Screening of endogenous antagonistic bacteria

[0032] Preliminary Screening

[0033] Take the sclerotium sclerotium slant strain (with agar block): pick the sclerotium sclerotium block and place it in the center of the petri dish (diameter 9 cm) containing the potato enriched medium, with the sclerotium block as the center, Connect the endophytic bacteria to the ring 3cm apart from the center of the circle at 3cm intervals (make each place equal to the center of the circle), and do three repetitions. Place in a constant temperature incubator at 23°C for 3-7 days. It was observed that endophytes 4-3 had obvious antagonism to Sclerotinia sclerotiorum.

[0034] Cup and saucer method

[0035] Pick the sclerotium clumps and place them in the center of the biochemical experiment petri dish (12 cm in diameter) filled with potatoes and enriched medium, and incubate them in a thermostat at 23°C for 2 days. The sclerotia colonies have expanded to about 2-3 cm in diameter. ...

Embodiment 3

[0036] Example 3: The antagonistic test of rape against Sclerotinia sclerotiorum after treatment of rape with endogenous antagonistic antimicrobial 4-3

[0037] 1. Cultivation of antagonistic antibacterial solution

[0038] The antagonistic endophyte 4-3 was inoculated into 100ml optimized medium (mannitol 1.8g, tryptone 1.6g, NaCl 0.4g, MgSO 4 ·7H 2 O 0.3g, KH 2 PO 4 0.4g, 1% MnSO 4 0.2ml, 1% FeC 6 H 5 O 7 0.5ml, 1% Na 2 MoO 4 ·2H 2 O 0.4ml, deionized water 100ml, pH 7.2, culture temperature 32°C. ) In the Erlenmeyer flask, shaker for 3 days. Culture conditions: the rotating speed of the shaker is 180 rpm.

[0039] 2. Treat rapeseed plants

[0040] Take 2 bottles of antagonistic antibacterial liquid and dilute it to about 1500ml with water, and water the rape plants for 3 times with an interval of about 10 days. Watering the bacterial liquid is better in the evening on a sunny day. Rape plants watered with water were used as controls. (12 rapeseed for each test sample) routine ...

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Abstract

The invention relates to a rapeseed endophyte Bacillus amyloliquefaciens 4-3 and an application method thereof. According to the morphological characteristics, colony characteristics and physiological and biochemical characteristics of the strain, combined with the analysis of 16S rDNA sequence results, the endophytic strain 4‑3 was preliminarily determined to be Bacillus amyloliquefaciens. The strain has a strong ability to antagonize the sclerotinia sclerotiorum. The test results showed that the biological control effect of rapeseed endophytic strain 4‑3 on S. sclerotiorum reached 100%. At the same time, the rapeseed endophytic bacterium 4‑3 also has an excellent function of dissolving phosphorus and potassium, and it can be made into biological bacterial fertilizer, which can reduce the use of chemical fertilizers, and has great application prospects for my country's green agriculture.

Description

[0001] Technical field: [0002] The invention relates to a rape endophytic bacterium Bacillus amyloliquefaciens 4-3 and its application. [0003] Background technique: [0004] Rape is the main oil crop in my country. Sclerotinia sclerotiorum (Sclerotinia sclerotiorum) is one of the three major diseases in rape production. The disease is especially serious in the rape production areas in southern my country. The incidence is generally 10%-30%, and the serious More than 80%, resulting in huge economic losses (Qiu Yexian, Wang Jinlian, Wang Weifang, etc.. Screening of rapeseed sclerotinia resistance inducers[J]. Journal of Jiangxi Agricultural University, 1996,12(18):395-398.). The occurrence is particularly prominent in the main oilseed rape production areas in the middle and lower reaches of the Yangtze River and the southeast coast, including Jiangsu, Anhui, Shanghai, Zhejiang, Hunan, Hubei, Jiangxi and other places, which has seriously affected my country's rape production. At pre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C05F11/08A01N63/00A01P3/00C12R1/07
CPCA01N63/00C05F11/08C12N1/20C12N1/205C12R2001/07
Inventor 刘冬华伍善东郭照辉单世平
Owner HUNAN INST OF MICROBIOLOGY