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Plant organic matter-bacillus subtils mixed inoculum and its preparation method and use

A technology of Bacillus subtilis and plant organic matter, applied in the directions of botanical equipment and methods, biochemical equipment and methods, plant growth regulators, etc., can solve problems such as no relevant reports, achieve broad market prospects, good control effects, Good control effect

Active Publication Date: 2015-01-14
深圳华农中绿生物科技研究有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Based on the above ideas and technical requirements, the present invention adopts an adsorption carrier and a production process that are different from traditional biological agents, and uses plant organic matter such as neem residue, tea dead, and palm kernel meal as the raw material of the culture medium for biocontrol bacteria and the production process of the agent. The adsorption carrier, after secondary fermentation and decomposing, is prepared into a plant organic matter-Bacillus subtilis mixed bacterial agent (abbreviated as a plant organic bacterial agent) with the ability to prevent and control plant diseases and insect pests. This has not been reported in the study of biological pesticide bacterial agents.

Method used

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  • Plant organic matter-bacillus subtils mixed inoculum and its preparation method and use
  • Plant organic matter-bacillus subtils mixed inoculum and its preparation method and use
  • Plant organic matter-bacillus subtils mixed inoculum and its preparation method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Enrichment Screening and Identification of Example 1 Bacillus subtilis HN09

[0047] 1. Enrichment screening of Bacillus subtilis HN09

[0048] Soil samples were collected from the herbarium of insecticidal plants of South China Agricultural University in Guangzhou City, Guangdong Province. Weigh 10g of soil and put it into a triangular flask filled with sterilized plant organic matter medium (30g neem residue, 30g tea seed meal, 30g palm kernel meal, 5g peptone, 3g yeast powder, natural pH) for enrichment and cultivation for one week. Then, 10 g of the enriched culture was weighed and put into a Erlenmeyer flask filled with 90 mL of sterile water, shaken sufficiently and left to stand for 10 min. Take 1mL supernatant and dilute to 10 -4 、10 -5 、10 -6 Concentration, take 0.1mL dilution respectively, spread on LB medium (peptone 10g, yeast powder 5g, NaCl 10g, agar powder 15g, distilled water 1L, pH natural, sterilized at 121°C for 30min) plate, and cultured upside d...

Embodiment 2

[0054] Embodiment 2 Bacillus subtilis HN09 produces volatile gas experiment with antibacterial activity

[0055] The activated Bacillus subtilis HN09 was inoculated in the nutrient broth medium, and cultured on a shaker at 30° C. at 180 rpm / min for 24 hours. Take 200 μl strain HN09 bacteria solution and spread evenly on the pre-prepared gas producing medium (peptone 10g, yeast powder 5g, beef extract 3g, glucose 15g, K 2 HPO 4 2g, FeSO 4 0.2g, agar 20g, distilled water 1L, pH6.8~7.2) on the flat plate; FOC4 bacteria cake with a diameter of 5mm was inserted in the center of another flat PDA medium, and then buckled upside down on the gas-producing medium flat plate containing bacterial strain HN09 ( Treatment group), using the plate without bacterial solution as a control, seal the gap between the upper and lower plates with a sealing film, and place it at 28°C for dark cultivation. Set up 3 repetitions. After the control FOC4 mycelium grows to 1 cm from the edge of the pla...

Embodiment 3

[0056] The preparation of the plant organic matter bacterial agent of embodiment 3 bacillus subtilis HN09

[0057] (1) Preparation of primary seed solution

[0058] The seed liquid medium is nutrient broth liquid medium: beef extract 0.5%, peptone 0.5%, NaCl 0.5%, the rest is distilled water, pH 6.5-7.3. The culture medium was sterilized at 121°C for 20 minutes, and activated Bacillus HN09 was inserted into it, and cultured in shake flasks. Culture conditions: rotation speed 180rpm, 30°C, culture time 24h.

[0059] (2) Secondary seed solution preparation

[0060] The secondary seed medium formula is 1-3% of corn flour, 0.5-2% of soybean flour, 0.1-0.3% of yeast powder, K 2 HPO 4 0.05~0.15%, MgSO 4 0.01~0.03%, CaCO 4 0.1-0.2%, the rest is water, pH6.5-7.3. The medium was sterilized at 121°C for 20 minutes, and when the temperature dropped to about 30°C, the first-grade seed solution was put into the seed tank at a volume ratio of 5-15% for cultivation. The ventilation r...

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Abstract

The invention discloses a plant organic matter-bacillus subtils mixed inoculum and its preparation method and use. The preparation method comprises that one or more of plant organic matters of azadirachta residue, tea cake and palm kernel meal as bacillus subtils HN09 medium main raw materials and an inoculums adsorption carrier are subjected to two-step fermentation decomposition so that the plant organic matter-bacillus subtils mixed inoculum is obtained. Active components of the plant organic matter-bacillus subtils mixed inoculum comprise the plant organic matters, bacillus subtils HN09 and fermentation metabolite of the bacillus subtils HN09. The bacillus subtils HN09 is preserved in the China general microbiological culture collection center (CGMCC) on April 26, 2013 and has a preservation number of CGMCC NO. 7524. The bacillus subtils HN09 and the plant organic matter-bacillus subtils mixed inoculum can be used for preventing and controlling plant diseases and soil insects, can promote plant growth and nitrogen absorption of plants, have important meaning for enlarging an insect disease prevention and control scope and promoting agricultural waste recycle and have a wide application prospect in production practice.

Description

technical field [0001] The invention belongs to the field of biological pesticides and plant protection, and in particular relates to a preparation method of bacillus subtilis strain HN09 and its plant organic matter-biocontrol bacteria mixed bacterial agent, and its application in plant disease and insect pest control. Background technique [0002] Neem slag, tea seed and palm kernel meal are the leftovers of agricultural by-products in the processing of neem seeds, tea seeds and palm kernels. Due to the low utilization rate, they are usually discarded as waste. This is obviously not in line with the purpose of sustainable agricultural development, so it is necessary to find a way to effectively reuse these agricultural by-product waste. In view of the fact that these plant organic matter still contains a large amount of organic matter and plant protein and other nutrients, it can be used as a medium material for microorganisms. Adding biocontrol bacteria to these plant org...

Claims

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Application Information

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IPC IPC(8): A01N63/02A01P3/00A01P7/04A01P21/00C12N1/20C12R1/125
Inventor 徐汉虹赖多康向辉
Owner 深圳华农中绿生物科技研究有限公司
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