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Serratia marcescens derived from agriphilaaeneociliella diseased insect bodies and application thereof

A technology of Serratia marcescens and bacterial suspension, applied in the field of agricultural microorganisms

Active Publication Date: 2018-06-19
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its genus Serratia entomophila has been developed as a microbial insecticide product, and is widely used in the biological control of scarab beetle, but Serratia entomophila has not been isolated from wheat pests and has no effect on insecticides. Isolation of Pathogenic Bacteria Sources and Pathogenic Mechanisms of the Wheat Pests Pest Borer and Aphid sativa

Method used

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  • Serratia marcescens derived from agriphilaaeneociliella diseased insect bodies and application thereof
  • Serratia marcescens derived from agriphilaaeneociliella diseased insect bodies and application thereof
  • Serratia marcescens derived from agriphilaaeneociliella diseased insect bodies and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: Isolation and identification of bacterial strains

[0050] 1. Isolation of strains:

[0051] (1) Soak the diseased larvae of the chrysalis moth in 75% alcohol for 10 seconds, disinfect the body surface, rinse them with sterile water for 3 times, put them into a sterile petri dish, and cut them off with sterilized surgical scissors At the end of one of the feet, use a pipette to absorb 5 μl of the overflowing body fluid, add 495 μl of sterile water to dilute, and then continue to dilute with sterile water to form a sterile water dilution of 1:10 2 , 1:10 3 , 1:10 4 , 1:10 5 diluent, and finally fully shaken;

[0052] (2) The dilution in the drawing step (1) is 1:10 2 , 1:10 3 , 1:10 4 , 1:10 5 50 μl each of the dilutions were spread on LB solid medium without anti-antibody, and cultured at 28°C for 24 hours;

[0053] (3) Pick a single colony grown after 24 hours of cultivation, and then culture it purely on LB solid medium to obtain strain JY9.

[00...

Embodiment 2

[0080] Embodiment 2: the growth curve determination of strain JY9 fermented liquid OD600 value changes with culture time

[0081] Get the single bacterium colony on the JY9 bacterial strain plate that embodiment 1 obtains and inoculate in the Erlenmeyer flask that 100ml LB medium is housed, culture temperature is 28 ℃, shaking table speed 180rpm, culture time 12h, obtain Serratia marcescens JY9 ferment liquid, add LB to dilute, and adjust the OD600 value of this fermentation liquid to 1.0 (the bacterial concentration at this time is about 6×10 8 cfu / ml), it was inoculated into 150 5ml centrifuge tubes equipped with 2ml LB according to the inoculum amount of 1%, and they were all placed in a shaker at 28°C with a rotation speed of 180rpm, shaken for 48h, and at the same time, every 0.5 Take 3 centrifuge tubes for h, and then take 3 centrifuge tubes every 2 hours, and measure their OD600 values ​​respectively. Draw the growth curve of Serratia marcescens strain JY9 fermentation...

Embodiment 3

[0083] Example 3: Determination of the control ability of Serratia marcescens strain JY9 to the weed moth

[0084] 1. Test method:

[0085] (1) Preparation of bacterial strain JY9 fermentation broth:

[0086] Inoculate a single colony of strain JY9 that was streak cultured on LB solid medium at 28°C for 16 hours into 6ml of LB liquid medium, place it on a shaker at 28°C, and cultivate it at 180rpm for 16 hours as a seed solution, and then use a pipette Take 100 μl of Serratia marcescens JY9 seed liquid with a liquid gun, inoculate it into 100 ml of sterilized LB liquid medium according to the inoculation amount of 1% (volume ratio), place it on a shaker at 28°C, and cultivate it at 180 rpm for 16 hours to obtain the strain The fermentation broth of JY9.

[0087] (2) Preparation of treatment liquid:

[0088] 1) Take 10ml of the fermentation broth of the bacterial strain JY9 prepared in (1) above, and adjust its OD600 value to 1.0 (the bacterial concentration is about 6×10 8...

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Abstract

The invention discloses serratia marcescens JY9 derived from agriphilaaeneociliella diseased insect bodies. The strain is preserved in China General Microbiological Culture Collection Center (CGMCC) on December 5, 2017 and the biological preservation number is CGMCC NO. 15020. The serratia marcescens JY9 strain provided by the invention is fermented and cultured to obtain a fermentation solution,bacterium suspension and supernatant which can be applied to prevention and control of agriphilaaeneociliella and sitobion avenae; when the bacterium concentration of the fermentation solution which is cultured in an LB culture medium for 16h is 6*10<8>cfu / ml, the fatality rate on mature larvae of the agriphilaaeneociliella can reach 55 percent or more; the fermented supernatant is directly sprayed and applied to wheat seedlings and the wheat seedlings are treated for 4d; the prevention effect on the sitobion avenae can reach 50 percent or more.

Description

technical field [0001] The invention relates to the technical field of agricultural microbes, in particular to a Serratia marcescens derived from the diseased worm body of the weed moth and its application. Background technique [0002] Lepidoptera is the second largest order of the order Insecta. Agriphila aeneociliella (Eversmann, 1844)], which belongs to this family, is a new wheat pest in recent years. Since the outbreak of the insect in 2009, the insect has spread and exploded in Laizhou City. By 2012, it had occurred on more than 6,000 mu in Laizhou, causing great damage to the wheat at the seedling stage. The lack of seedlings and ridges are formed, and more than 50% of the wheat seedlings in serious plots die, or even become extinct. In March 2013, this kind of pest was found to damage wheat in Jincheng, Shanxi. In 2014, the damage occurred in the Gaomi wheat field in Shandong Province. In 2015, it spread to Jimo, Qingdao. It causes huge economic losses every year...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P7/04C12R1/43
CPCA01N63/00A01N63/10C12N1/20C12N1/205C12R2001/43
Inventor 刘勇迟宝杰战一迪赵春青
Owner SHANDONG AGRICULTURAL UNIVERSITY
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