caa a22 4500 605505047 CHVBK 20210128100619.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00253-014-5966-2 doi (NATIONALLICENCE)springer-10.1007/s00253-014-5966-2 Genomic sequencing identifies novel Bacillus thuringiensis Vip1/Vip2 binary and Cry8 toxins that have high toxicity to Scarabaeoidea larvae [Elektronische Daten] [Yang Bi, Yanrui Zhang, Changlong Shu, Neil Crickmore, Qinglei Wang, Lixin Du, Fuping Song, Jie Zhang] The Bacillus thuringiensis strain HBF-18 (CGMCC 2070), which has previously been shown to encode the cry8Ga toxin gene, is active against both Holotrichia oblita and Holotrichia parallela. Recombinant Cry8Ga however is only weakly toxic to these insect pests suggesting the involvement of additional toxins in the native strain. We report that through the use of Illumina sequencing three additional, and novel, genes, namely vip1Ad1, vip2Ag1, and cry8-like, were identified in this strain. Although no protein corresponding to these genes could be identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the HBF-18 proteome, reverse transcription (RT)-PCR indicated that all three genes were transcribed in the native strain. The two vip genes were cloned and expressed and, as with other Vip1/2 toxins, appeared to function as a binary toxin and showed strong activity against H. oblita, H. parallela and Anomala corpulenta. This is the first report to demonstrate that the Vip1/Vip2 binary toxin is active against these Scarabaeoidea larvae. The cry8-like gene appeared to be a C-terminally truncated form of a typical cry8 gene and was not expressed in our usual recombinant Bt expression system. When however the missing C-terminal region was replaced with the corresponding sequence from cry8Ea, the resulting hybrid expressed well and the toxin was active against the three test insects. Springer-Verlag Berlin Heidelberg, 2014 Bacillus thuringiensis nationallicence Holotrichia oblita nationallicence Holotrichia parallela nationallicence Anomala corpulenta nationallicence Vip1/Vip2 binary toxin nationallicence Bi Yang State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, China aut Zhang Yanrui State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, China aut Shu Changlong State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, China aut Crickmore Neil School of Life Sciences, University of Sussex, BN1 9QG, Brighton, UK aut Wang Qinglei Cangzhou Academy of Agricultural and Forestry Sciences, 061001, Cangzhou, China aut Du Lixin Institute of Plant Protection, Hebei Academy of Agricultural and Forestry Sciences, 071000, Baoding, China aut Song Fuping State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, China aut Zhang Jie State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/2(2015-01-01), 753-760 0175-7598 99:2<753 2015 99 253 https://doi.org/10.1007/s00253-014-5966-2 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-5966-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Bi Yang State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, China aut NATIONALLICENCE 700 1- Zhang Yanrui State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, China aut NATIONALLICENCE 700 1- Shu Changlong State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, China aut NATIONALLICENCE 700 1- Crickmore Neil School of Life Sciences, University of Sussex, BN1 9QG, Brighton, UK aut NATIONALLICENCE 700 1- Wang Qinglei Cangzhou Academy of Agricultural and Forestry Sciences, 061001, Cangzhou, China aut NATIONALLICENCE 700 1- Du Lixin Institute of Plant Protection, Hebei Academy of Agricultural and Forestry Sciences, 071000, Baoding, China aut NATIONALLICENCE 700 1- Song Fuping State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, China aut NATIONALLICENCE 700 1- Zhang Jie State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/2(2015-01-01), 753-760 0175-7598 99:2<753 2015 99 253