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

A technology of Bacillus thuringiensis and YBT-008, which is applied in nematicides, applications, bacteria, etc., can solve the problems of lack of related strains and genetic resources, and achieve the effect of convenient implementation and fast onset of action

Inactive Publication Date: 2014-05-14
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Even so, so far, little research has been done on the use of Bacillus thuringiensis to control D.

Method used

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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

Experimental program
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Effect test

Embodiment 1

[0039] The preparation of embodiment 1 bacillus thuringiensis insecticidal crystal protein bioassay sample

[0040]The strain resources used in the screening process are Bacillus thuringiensis strains publicly reported by the State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, and their original numbers are YBT-007, YBT-008, YBT-017, YBT-020, YBT- 021, YBT-027, YBT-032, YBT-037 (see literature for source: Yu Zi-Quan et al., 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...

Embodiment 2

[0049] Inoculation, cultivation and collection of embodiment 2 sweet potato stem nematode

[0050] According to the lifestyle of sweet potato D. nematode, the applicant provides a simple method for artificially cultivating sweet potato D. nematode, wherein the sweet potato D. nematode is provided by Shanghai Institute of Plant Physiology, Chinese Academy of Sciences.

[0051] 1. Inoculation and cultivation of sweet potato stem nematode

[0052] Wash D. sweetpotato nematodes with sterile deionized water, suspend in sterile water, and dilute to a population density of about 5000 / ml for later use. Wash the surface of fresh sweet potatoes with uniform shape and no damage on the surface with water, then scrub twice with 75% alcohol, and then dry them in the ultra-clean bench for later use. Use a sterile scalpel to cut a small piece in the middle of the sweet potato, suck about 200 μl of nematode suspension and slowly inject it into the concave hole of the sweet potato, let it stan...

Embodiment 3

[0059] Example 3 Determination of Bacillus thuringiensis insecticidal crystal protein to the biological activity of sweet potato stem nematode

[0060] Determination procedure of Bacillus thuringiensis insecticidal crystal protein against sweet potato Dilmatode elegans References: Yu Ziquan et al., Establishment of a bioassay method for Bacillus thuringiensis parasporal crystal protein against plant parasitic nematodes and screening of highly virulent strains, Agricultural Biotechnology Journal of the Chinese Academy of Sciences, 2007(15): 867-871.

[0061] The nematodes used in this example were mixed old and young D. sweetpotato nematodes, and the bioassay process was carried out on a 96-well cell culture plate.

[0062] 1. Determination of the biological activity of Bacillus thuringiensis against Dilomata elegans

[0063] (1) Preparation of sweet potato D. nematode and insecticidal crystal protein samples. Mix the nematodes thoroughly before use, wash with sterile deioniz...

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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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Patent Type & Authority Patents(China)
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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