Method for detecting bacillus subtilis Y13 volatile bacteriostatic substances based on HS-SPME and GC-MS

A technology of Bacillus subtilis and antibacterial substances, which is applied in the field of detection and can solve problems such as unstable inhibition effects

Inactive Publication Date: 2020-04-10
HAINAN KEDA FORESTRY CO LTD +1
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Problems solved by technology

[0004] Bacillus subtilis (B.subtilis) Y13 is an endophytic bacterium isolated from healthy leaves of camellia oleifera by our research group and has good control effect on camellia oleifera anthracnose. Strong inhibitory effect, but the inhibitory effect is unstable

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  • Method for detecting bacillus subtilis Y13 volatile bacteriostatic substances based on HS-SPME and GC-MS
  • Method for detecting bacillus subtilis Y13 volatile bacteriostatic substances based on HS-SPME and GC-MS
  • Method for detecting bacillus subtilis Y13 volatile bacteriostatic substances based on HS-SPME and GC-MS

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[0042] Below in conjunction with accompanying drawing, table and embodiment the present invention will be further described

[0043] Headspace solid-phase microextraction (HS-SPME) was used to collect the volatile gas of Bacillus subtilis Y13. The specific method was: the prepared Y13 bacterial liquid (1.0×10 9 CFU / mL) at a ratio of 1:20 (v / v) mixed with NB liquid medium heated and cooled to 45°C to 50°C, and then pipette 5mL of NB liquid culture after mixing Y13 bacterial liquid based on 20mL IM In the empty sampling bottle, the control group was 5 mL of NB liquid medium without Y13 bacterial solution added to the headspace sampling bottle, and the headspace bottle was placed in a constant temperature shaker at 28 °C and 140 r / min for 3 days, and then the samples of Y13 were collected. volatile gas. Volatile gases were manually extracted and equipped with a 50 / 30 μm divinylbenzene / carbon molecular sieve / polydimethylsiloxane micro-extraction head (DVB / CAR / PDMS, gray). Put th...

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Abstract

The invention belongs to the field of testing, and particularly discloses a method for detecting bacillus subtilis Y13 volatile bacteriostatic substances based on HS-SPME technology and GC-MS. According to the method, volatile substances of bacillus subtilis Y13 are collected by utilizing the HS-SPME technology, and GC-MS is used for detecting the components and amounts of the volatile substances.The method can comprehensively and accurately determine the components and amounts of the bacillus subtilis Y13 volatile substances, also can be completed in a normal-temperature range, and avoids the generation of a new compound in an analysis process. The working conditions of the HS-SPME technology provided by the invention can effectively enrich each component of the Y13 volatile substances;GC-MS detection adopts sectional heating, volatile substance components can be effectively separated, and the volatile substance components can be rapidly and accurately identified through corresponding mass spectrum parameters; in addition, colletotrichum fructicola is used as an indicator bacterium to identify the effective components of the bacillus subtilis Y13 volatile substances with strongbacteriostatic effect.

Description

technical field [0001] The invention belongs to the detection field, and in particular relates to a detection method for volatile antibacterial substances of Bacillus subtilis (Bacillus subtilis) Y13. [0002] technical background [0003] Camellia oleifera is a unique woody oil plant in my country. It has good economic benefits and a large planting area. However, due to extensive management and management, pests and diseases are becoming more and more rampant. Camellia oleifera anthracnose (Camellia oleifera anthracnose) is a major disease that occurs very commonly in Camellia oleifera producing areas in my country. It has a wide range of hosts, rapid onset, and serious damage. It is also an important disease of tropical and subtropical economic crops. For a long time, camellia oleifera anthracnose was mainly prevented and controlled by chemical agents. This makes pathogenic bacteria have serious drug resistance, and also pollutes the environment, making the pesticide resid...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72G01N1/22
CPCG01N30/02G01N30/06G01N30/7233G01N1/2226G01N2030/025G01N2030/062G01N2030/009G01N2001/2229
Inventor 倪源周国英徐睿冯福山
Owner HAINAN KEDA FORESTRY CO LTD
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