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
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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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