Bacillus thuringiensis YBT-008 for killing ditylenchus destructor and application thereof

A YBT-008, Bacillus thuringiensis technology, applied in nematicides, applications, bacteria and other directions, can solve the problems of lack of related strains and genetic resources, and achieve the effect of convenient implementation method and fast effect

Inactive Publication Date: 2012-10-03
HUAZHONG AGRI UNIV
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Problems solved by technology

Even so, so far, little research has been done o

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  • Bacillus thuringiensis YBT-008 for killing ditylenchus destructor and application thereof
  • Bacillus thuringiensis YBT-008 for killing ditylenchus destructor and application thereof
  • Bacillus thuringiensis YBT-008 for killing ditylenchus destructor and application thereof

Examples

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Example Embodiment

[0039] Example 1 Preparation of Bacillus thuringiensis insecticidal crystal protein bioassay sample

[0040] The strain resources used in the screening process were the Bacillus thuringiensis strains publicly reported by the State Key Laboratory of Agricultural Microbiology of Huazhong Agricultural University. The original numbers were YBT-007, YBT-008, YBT-017, YBT-020, YBT- 021, YBT-027, YBT-032, YBT-037 (source see literature: Yu Zi-Quan, etc., Bacillus thuringiensis crystal protein toxicity against plant-parasitic nematodes. Chinese Journal of Agricultural Biotechnology. 2008, 5(1): 13- 17) and Bacillus thuringiensis subspecies Israel and Kustak subspecies, and two Bacillus thuringiensis engineering strains BMB171 / pHT304-cry5Ba2 (Suxia Guo et al., New strategy for isolating novel nematicidal crystal protein genes from Bacillus thuringiensis strain YBT1518. Applied and Environment Microbiology.2008, 74: 6997-7001) and BMB171 / pHT304-cry6Aa2 (Yu Ziquan et al. Cloning and express...

Example Embodiment

[0049] Example 2 Inoculation, Cultivation and Collection of Ditylenchus sweetpotato

[0050] According to the lifestyle of Ditylenchus sweetpotato, the applicant provided a simple and convenient method for artificially cultivating Ditylenchus sweetpotato, wherein Ditylenchus sweetpotato was provided by Shanghai Institute of Plant Physiology, Chinese Academy of Sciences.

[0051] 1. Inoculation and cultivation of Ditylenchus sweetpotato

[0052] Wash Ditylenchus sweet potato with sterile deionized water, suspend it in sterile water, and dilute to a density of about 5,000 pieces / ml for later use. Wash the surface of fresh sweet potatoes with uniform shape and no damage on the surface with water, and then scrub them with 75% alcohol twice, then blow dry in the ultra-clean table for use. Use a sterile scalpel to cut a small piece in the middle part of the sweet potato, suck about 200μl of the nematode suspension slowly into the sweet potato cavity, let it stand for a while, wait for the...

Example Embodiment

[0059] Example 3 Determination of the biological activity of Bacillus thuringiensis insecticidal crystal protein against Ditylenchus sacchari

[0060] Procedure for the determination of biological activity of Bacillus thuringiensis insecticidal crystal protein against Ditylenchus sweetpotato Reference: Yu Ziquan et al., Establishment of bioassay method for plant parasitic nematodes by Bacillus thuringiensis parasporial crystal protein and screening of highly virulent strains, agricultural biotechnology Journal, 2007(15): 867-871.

[0061] The test nematode used in this example was a mixture of old and young Ditylenchus spp. The bioassay process was performed on a 96-well cell culture plate.

[0062] 1. Determination of the biological activity of Bacillus thuringiensis against Ditylenchus sweetpotato

[0063] (1) Preparation of Ditylenchus spp. and insecticidal crystal protein samples. Mix the nematodes thoroughly before use, wash them with sterile deionized water for several times, a...

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Abstract

The invention belongs to the technical field of microbial insecticides, and specifically relates to the screening for a bacillus thuringiensis strain with high virulence for ditylenchus destructors and the application for the prevention and treatment of ditylenchus destructors. The invention screens a bacterial strain YBT-008 with high virulence from the bacillus thuringiensis for the ditylenchus destructors. The bacterial strain is stored in China Center for Type Culture Collection, and the collection number is CCTCC NO: M2011039. The invention also includes the application effects of a microbial agent prepared by the YBT-008 on controlling the ditylenchus destructors, the types of insecticidal crystal proteins generated by the YBT-008, the verification of Cyt2Ba, Cry11Aa, Cry4Aa and Cry4Ba, and the verification of that the microbe and the microbial agent of the invention belong to a novel microbe for killing the ditylenchus destructors.

Description

technical field [0001] The invention belongs to the technical field of microbial insecticidal pesticides, and in particular relates to the screening and application of a strain of Bacillus thuringiensis YBT-008 which has high toxicity to the agricultural production pest Diloma thuringiensis. technical background [0002] Sweet potato is an important grain, feed and industrial raw material, widely planted, in the world's grain production, the total output ranks seventh, and in my country's grain production, it is second only to rice, wheat and corn, ranking fourth (Zhu Jinting, Gao Jianwei .Summary of sweet potato production at home and abroad [J].Agricultural and Animal Husbandry Product Development, 1995, 7:20-21). At present, the annual planting area of ​​sweet potato in my country is about 6 million hectares, accounting for about 65% of the world's total area, and the annual total output accounts for about 85% of the world's total output (FAO bulletin of statistics. 2002, ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02A01P5/00C12R1/07
Inventor 李明顺喻子牛何进孙明刘金辉班虎栋侯艳飞王慧
Owner HUAZHONG AGRI UNIV
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