This page has only limited features, please log in for full access.
Transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) have revolutionized pest control, but the benefits of this approach have been reduced by the evolution of resistance in pests. The widely adopted 'pyramid strategy' for delaying resistance entails transgenic crops producing two or more distinct toxins that kill the same pest. The limited experimental evidence supporting this strategy comes primarily from a model system under ideal conditions. Here we tested the pyramid strategy under nearly worst-case conditions, including some cross-resistance between the toxins in the pyramid. In a laboratory selection experiment with an artificial diet, we used Bt toxins Cry1Ab, Cry1F, and Cry1Ie singly or in pairs against Ostrinia furnacalis, one of the most destructive pests of corn in Asia. Under the conditions evaluated, pairs of toxins did not consistently delay the evolution of resistance relative to single toxins.
Yueqin Wang; Yudong Quan; Jing Yang; Changlong Shu; Zhenying Wang; Jie Zhang; Angharad M. R. Gatehouse; Bruce E. Tabashnik; Kanglai He. Evolution of Asian Corn Borer Resistance to Bt Toxins Used Singly or in Pairs. Toxins 2019, 11, 461 .
AMA StyleYueqin Wang, Yudong Quan, Jing Yang, Changlong Shu, Zhenying Wang, Jie Zhang, Angharad M. R. Gatehouse, Bruce E. Tabashnik, Kanglai He. Evolution of Asian Corn Borer Resistance to Bt Toxins Used Singly or in Pairs. Toxins. 2019; 11 (8):461.
Chicago/Turabian StyleYueqin Wang; Yudong Quan; Jing Yang; Changlong Shu; Zhenying Wang; Jie Zhang; Angharad M. R. Gatehouse; Bruce E. Tabashnik; Kanglai He. 2019. "Evolution of Asian Corn Borer Resistance to Bt Toxins Used Singly or in Pairs." Toxins 11, no. 8: 461.
The bacterium Bacillus thuringiensis produces several insecticidal proteins, such as the crystal proteins (Cry) and the vegetative insecticidal proteins (Vip). In this work, we report that a specific interaction between two B. thuringiensis toxins creates insecticidal synergism and unravel the molecular basis of this interaction. When applied together, the three-domain Cry toxin, Cry9Aa and the Vip, Vip3Aa, exhibited high insecticidal activity against an important insect pest, the Asiatic rice borer (Chilo suppressalis). We found that these two proteins bind specifically to brush border membrane vesicles of C. suppressalis, and that they do not share binding sites, since no-binding competition was observed between them. Binding assays revealed that the Cry9Aa and Vip3Aa proteins interacted with high affinity. We mapped their specific interacting regions by analyzing binding of Cry9Aa to overlapping fragments of Vip3Aa and by analyzing binding of Vip3Aa to individual domains of Cry9Aa. Binding to peptide arrays helped narrow the binding sites to the domain II-loop3 of Cry9Aa and to 428TKKMKTL434 in Vip3Aa. Site-directed mutagenesis confirmed that these binding regions participate in binding that directly correlates with the synergism between the two proteins. In summary, we show that the B. thuringiensis Cry9Aa and Vip3Aa toxins display potent synergy based on a specific interaction between them. Our results further our understanding of the complex synergistic activities among B. thuringiensis toxins and are highly relevant to the development of toxin combinations for effective insect control and for delaying development of insect resistance.
Zeyu Wang; Longfa Fang; Zishan Zhou; Sabino Pacheco; Isabel Gómez; Fuping Song; Mario Soberón; Jie Zhang; Alejandra Bravo. Specific binding between Bacillus thuringiensis Cry9Aa and Vip3Aa toxins synergizes their toxicity against Asiatic rice borer (Chilo suppressalis). Journal of Biological Chemistry 2018, 293, 11447 -11458.
AMA StyleZeyu Wang, Longfa Fang, Zishan Zhou, Sabino Pacheco, Isabel Gómez, Fuping Song, Mario Soberón, Jie Zhang, Alejandra Bravo. Specific binding between Bacillus thuringiensis Cry9Aa and Vip3Aa toxins synergizes their toxicity against Asiatic rice borer (Chilo suppressalis). Journal of Biological Chemistry. 2018; 293 (29):11447-11458.
Chicago/Turabian StyleZeyu Wang; Longfa Fang; Zishan Zhou; Sabino Pacheco; Isabel Gómez; Fuping Song; Mario Soberón; Jie Zhang; Alejandra Bravo. 2018. "Specific binding between Bacillus thuringiensis Cry9Aa and Vip3Aa toxins synergizes their toxicity against Asiatic rice borer (Chilo suppressalis)." Journal of Biological Chemistry 293, no. 29: 11447-11458.
Rhizosphere microorganisms contribute to the health and development of crops and these beneficial microbes are recruited to the root-zone when plants experience biotic/abiotic stress. The subterranean pests Holotrichia parallela cause severe crop loss in peanut (Arachis hypogaea L.) fields. Hypothesizing that infestation by H. parallela larva may influence the composition of rhizosphere microbial communities, deep sequencing of V3 and V4 hypervariable regions of 16S rRNA gene was used to characterize the rhizosphere bacteria of infested and uninfested peanuts. A total of 2,673,656 reads were generated and an average of 2,558 OTUs were obtained for each sample. Comparisons of rhizosphere bacterial community structure of peanuts with those infested by H. parallela larva revealed that the relative abundance of Proteobacteria and Bacteroidetes increased, while that of Actinobacteria decreased in the rhizosphere with infestation. A significant shift in bacterial communities was observed within 24 h after infestation by principal coordinate analysis. For the 332 genera identified in 24 h treatment, infestation of white grubs led to the significant changes of abundance of 67 genera. An increase in the Pseudomonas genus of infested-samples for 24 h was verified by real-time qPCR. Our results indicate H. parallela larvae infestation can quickly leads to the change of peanut rhizosphere microbiome and enrichment of specific bacterial species. But the effects were not persistent. This study provides the insight into the function of rhizosphere microbiome in the interaction between subterranean pests and crops.
Li-Li Geng; Gao-Xiang Shao; Ben Raymond; Mei-Ling Wang; Xiao-Xiao Sun; Chang-Long Shu; Jie Zhang. Subterranean infestation by Holotrichia parallela larvae is associated with changes in the peanut (Arachis hypogaea L.) rhizosphere microbiome. Microbiological Research 2018, 211, 13 -20.
AMA StyleLi-Li Geng, Gao-Xiang Shao, Ben Raymond, Mei-Ling Wang, Xiao-Xiao Sun, Chang-Long Shu, Jie Zhang. Subterranean infestation by Holotrichia parallela larvae is associated with changes in the peanut (Arachis hypogaea L.) rhizosphere microbiome. Microbiological Research. 2018; 211 ():13-20.
Chicago/Turabian StyleLi-Li Geng; Gao-Xiang Shao; Ben Raymond; Mei-Ling Wang; Xiao-Xiao Sun; Chang-Long Shu; Jie Zhang. 2018. "Subterranean infestation by Holotrichia parallela larvae is associated with changes in the peanut (Arachis hypogaea L.) rhizosphere microbiome." Microbiological Research 211, no. : 13-20.
Genetically modified crops that express insecticidal Bacillus thuringiensis (Bt) proteins have become a primary approach for control of lepidopteran (moth) and coleopteran (beetle) pests that feed by chewing the plants. However, the sap-sucking insects (Hemiptera) are not particularly susceptible to Bt toxins. In this study, we describe two Cry toxins (Cry64Ba and Cry64Ca) from Bt strain 1012 that showed toxicity against two important hemipteran rice pests, Laodelphax striatellus and Sogatella furcifera . Both of these proteins contain an ETX/MTX2 domain and share common sequence features with the β-pore-forming toxins. Coexpression of cry64Ba and cry64Ca genes in the acrystalliferous Bt strain HD73 − resulted in high insecticidal activity against both hemipteran pests. No toxicity was observed on other pests such as Ostrinia furnacalis , Plutella xylostella , or Colaphellus bowringi . Also, no hemolytic activity or toxicity against cancer cells was detected. Binding assays showed specific binding of the Cry64Ba/Cry64Ca toxin complex to brush border membrane vesicles isolated from L. striatellus . Cry64Ba and Cry64Ca are Bt Cry toxins highly effective against hemipteran pests and could provide a novel strategy for the environmentally friendly biological control of rice planthoppers in transgenic plants. IMPORTANCE In Asia, rice is an important staple food, whose production is threatened by rice planthoppers. To date, no effective Bacillus thuringiensis (Bt) protein has been shown to have activity against rice planthoppers. We cloned two Bt toxin genes from Bt strain 1012 that showed toxicity against small brown planthoppers ( Laodelphax striatellus ) and white-backed planthoppers ( Sogatella furcifera ). To our knowledge, the proteins encoded by the cry64Ba and cry64Ca genes are the most efficient insecticidal Bt Cry proteins with activity against hemipteran insects reported so far. Cry64Ba and Cry64Ca showed no toxicity against some lepidopteran or coleopteran pests. These two proteins should be able to be used for integrated hemipteran pest management.
Yonglei Liu; Yinglong Wang; Changlong Shu; Kejian Lin; Fuping Song; Alejandra Bravo; Mario Soberón; Jie Zhang. Cry64Ba and Cry64Ca, Two ETX/MTX2-Type Bacillus thuringiensis Insecticidal Proteins Active against Hemipteran Pests. Applied and Environmental Microbiology 2018, 84, e01996-17 .
AMA StyleYonglei Liu, Yinglong Wang, Changlong Shu, Kejian Lin, Fuping Song, Alejandra Bravo, Mario Soberón, Jie Zhang. Cry64Ba and Cry64Ca, Two ETX/MTX2-Type Bacillus thuringiensis Insecticidal Proteins Active against Hemipteran Pests. Applied and Environmental Microbiology. 2018; 84 (3):e01996-17.
Chicago/Turabian StyleYonglei Liu; Yinglong Wang; Changlong Shu; Kejian Lin; Fuping Song; Alejandra Bravo; Mario Soberón; Jie Zhang. 2018. "Cry64Ba and Cry64Ca, Two ETX/MTX2-Type Bacillus thuringiensis Insecticidal Proteins Active against Hemipteran Pests." Applied and Environmental Microbiology 84, no. 3: e01996-17.
The Bacillus thuringiensis strain HBF-18 (CGMCC 2070), containing two cry genes ( cry8 -like and cry8Ga ), is toxic to Holotrichia oblita larvae. Both Cry8-like and Cry8Ga proteins are active against this insect pest, and Cry8-like is more toxic. To analyze the characteristics of the binding of Cry8-like and Cry8Ga proteins to brush border membrane vesicles (BBMVs) in H. oblita larvae, binding assays were conducted with a fluorescent DyLight488-labeled Cry8-like toxin. The results of saturation binding assays demonstrated that Cry8-like bound specifically to binding sites on BBMVs from H. oblita , and heterologous competition assays revealed that Cry8Ga shared binding sites with Cry8-like. Furthermore, Cry8-like-binding proteins in the midgut from H. oblita larvae were identified by pulldown assays and liquid chromatography-tandem mass spectrometry (LC-MS/MS). In addition, the H. oblita midgut transcriptome was assembled by high-throughput RNA sequencing and used for identification of Cry8-like-binding proteins. Eight Cry8-like-binding proteins were obtained from pulldown assays conducted with BBMVs. The LC-MS/MS data for these proteins were successfully matched with the H. oblita transcriptome, and BLASTX results identified five proteins as serine protease, transferrin-like, uncharacterized protein LOC658236 of Tribolium castaneum , ATPase catalytic subunit, and actin. These identified Cry8-like-binding proteins were different from those confirmed previously as receptors for Cry1A proteins in lepidopteran insect species, such as aminopeptidase, alkaline phosphatase, and cadherin. IMPORTANCE Holotrichia oblita is one of the main soil-dwelling pests in China. The larvae damage the roots of crops, resulting in significant yield reductions and economic losses. H. oblita is difficult to control, principally due to its soil-dwelling habits. In recent years, some Cry8 toxins from Bacillus thuringiensis were shown to be active against this pest. Study of the mechanism of action of these Cry8 toxins is needed for their effective use in the control of H. oblita and for their future utilization in transgenic plants. Our work provides important basic data and promotes understanding of the insecticidal mechanism of Cry8 proteins against H. oblita larvae.
Jian Jiang; Ying Huang; Changlong Shu; Mario Soberón; Alejandra Bravo; Chunqing Liu; Fuping Song; Jinsheng Lai; Jie Zhang. Holotrichia oblita Midgut Proteins That Bind to Bacillus thuringiensis Cry8-Like Toxin and Assembly of the H. oblita Midgut Tissue Transcriptome. Applied and Environmental Microbiology 2017, 83, e00541-17 .
AMA StyleJian Jiang, Ying Huang, Changlong Shu, Mario Soberón, Alejandra Bravo, Chunqing Liu, Fuping Song, Jinsheng Lai, Jie Zhang. Holotrichia oblita Midgut Proteins That Bind to Bacillus thuringiensis Cry8-Like Toxin and Assembly of the H. oblita Midgut Tissue Transcriptome. Applied and Environmental Microbiology. 2017; 83 (12):e00541-17.
Chicago/Turabian StyleJian Jiang; Ying Huang; Changlong Shu; Mario Soberón; Alejandra Bravo; Chunqing Liu; Fuping Song; Jinsheng Lai; Jie Zhang. 2017. "Holotrichia oblita Midgut Proteins That Bind to Bacillus thuringiensis Cry8-Like Toxin and Assembly of the H. oblita Midgut Tissue Transcriptome." Applied and Environmental Microbiology 83, no. 12: e00541-17.
Bacillus thuringiensis Cry1Ah protein is highly toxic against Helicoverpa armigera but shows no toxicity against Bombyx mori larvae. In contrast, the closely related Cry1Ai toxin showed the opposite phenotype: high activity against B. mori but no toxicity against H. armigera. Analysis of binding of Cry1Ah to brush border membrane vesicle (BBMV) proteins from H. armigera and B. mori by surface plasmon resonance revealed association of toxin binding with insect specificity. Pulldown experiments identified aminopeptidase N1 (APN1) as a Cry1Ah binding protein that was not observed in the assays using B. mori BBMV proteins. The APN1 Cry1Ah binding region was narrowed to the region from A 548 to S 798 (fragment H3) by expressing four different APN1 fragments in Escherichia coli and analyzing Cry1Ah binding by ligand blot. Binding competition experiments of Cry1Ah to APN1 fragment H3 using synthetic peptides corresponding to four predicted domain II loop regions showed that loop 2 and loop 3 have additive effects on binding to APN1 fragment H3. Moreover, switching of loop 2 and loop 3 regions from Cry1Ah to Cry1Ai toxins showed that loop 2 and loop 3 are both involved in specificity and toxicity against H. armigera . IMPORTANCE Domain II loop regions have been shown to be involved in binding to larval gut proteins mediating insect specificity. The modification of loop regions is a direct and effective method to construct new Cry toxin variants to increase toxicity or modify specificity. Our results show that the exchange of loop regions from one toxin into another is a successful scheme for modification of B. thuringiensis Cry toxin specificity.
Zishan Zhou; Yuxiao Liu; Gemei Liang; Yongping Huang; Alejandra Bravo; Mario Soberón; Fuping Song; Xueping Zhou; Jie Zhang. Insecticidal Specificity of Cry1Ah to Helicoverpa armigera Is Determined by Binding of APN1 via Domain II Loops 2 and 3. Applied and Environmental Microbiology 2017, 83, e02864-16 .
AMA StyleZishan Zhou, Yuxiao Liu, Gemei Liang, Yongping Huang, Alejandra Bravo, Mario Soberón, Fuping Song, Xueping Zhou, Jie Zhang. Insecticidal Specificity of Cry1Ah to Helicoverpa armigera Is Determined by Binding of APN1 via Domain II Loops 2 and 3. Applied and Environmental Microbiology. 2017; 83 (4):e02864-16.
Chicago/Turabian StyleZishan Zhou; Yuxiao Liu; Gemei Liang; Yongping Huang; Alejandra Bravo; Mario Soberón; Fuping Song; Xueping Zhou; Jie Zhang. 2017. "Insecticidal Specificity of Cry1Ah to Helicoverpa armigera Is Determined by Binding of APN1 via Domain II Loops 2 and 3." Applied and Environmental Microbiology 83, no. 4: e02864-16.
Endophytic bacterial communities play a key role in promoting plant growth and combating plant diseases. However, little is known about their population dynamics in plant tissues and bulk soil, especially in transgenic crops. This study investigated the colonization of transgenic maize harboring the Bacillus thuringiensis (Bt) cry1Ah gene by Bacillus subtilis strain B916‐gfp present in plant tissues and soil. Bt and nontransgenic maize were inoculated with B916‐gfp by seed soaking, or root irrigation under both laboratory greenhouse and field conditions. During the growing season, B916‐gfp colonized transgenic as well as nontransgenic plants by both inoculation methods. No differences were observed in B916‐gfp population size between transgenic and nontransgenic plants, except at one or two time points in the roots and stems that did not persist over the examination period. Furthermore, planting transgenic maize did not affect the number of B916‐gfp in bulk soil in either laboratory or field trials. These results indicate that transgenic modification of maize with the cry1Ah gene has no influence on colonization by the endophytic bacteria B916‐gfp present in the plant and in bulk soil.
Chongsi Sun; Lili Geng; Meiling Wang; Gaoxiang Shao; Yongfeng Liu; Changlong Shu; Jie Zhang. No adverse effects of transgenic maize on population dynamics of endophyticBacillus subtilisstrain B916-gfp. MicrobiologyOpen 2016, 6, e00404 .
AMA StyleChongsi Sun, Lili Geng, Meiling Wang, Gaoxiang Shao, Yongfeng Liu, Changlong Shu, Jie Zhang. No adverse effects of transgenic maize on population dynamics of endophyticBacillus subtilisstrain B916-gfp. MicrobiologyOpen. 2016; 6 (1):e00404.
Chicago/Turabian StyleChongsi Sun; Lili Geng; Meiling Wang; Gaoxiang Shao; Yongfeng Liu; Changlong Shu; Jie Zhang. 2016. "No adverse effects of transgenic maize on population dynamics of endophyticBacillus subtilisstrain B916-gfp." MicrobiologyOpen 6, no. 1: e00404.
Bacillus thuringiensis produces a variety of insecticidal crystal proteins (ICPs). Genome sequencing is a promising strategy for detecting and identifying B. thuringiensis ICPs, which are of great interest to the biocontrol field. In this study, a novel ICP gene was cloned from B. thuringiensis BRC-ZYR2 based on genomic data from 454 GS-FLX Titanium sequencing and an analysis of the results using the B. thuringiensis Toxin_Scanner ( http://bcam.hzaubmb.net/BtToxin_scanner/index.php ). cry1Na3 designated by the B. thuringiensis Toxin Nomenclature Committee, encoded a 601-amino acid, 68.0-kDa protein that exhibited 95% identity with Cry1Na1 and 99% identity with Cry1Na2. Cry1Na3 contained three conserved domains commonly found in three-domain ICPs. Cry1Na3 was toxic to Plutella xylostella (L.) and Ostrinia furnacalis (Guenée), with LC 50 values of 3.69 μg/ml and 31.30 μg/ml, respectively. However, Laodelphax striatellus (Fallén) nymphs were unaffected when fed purified Cry1Na3 (250 μg/ml) in their diet. Spodoptera exigua (Hübner) and Colaphellus bowringi (Baly) larvae survived even when the concentration of Cry1Na3 protein reached 500 μg/ml. Cry1Na3 is a promising agent for the control of lepidopteran insect pests.
Ruiqing Dai; Xiaoyu Su; Xin Jin; Jie Zhang; Xiong Guan; Chen Chen; Changlong Shu; Tianpei Huang. Cloning, Expression, Purification, and Insecticidal Activity of a Novel Cry1Na3 Toxin FromBacillus thuringiensisBRC-ZYR2. Journal of Economic Entomology 2016, 109, 1064 -1070.
AMA StyleRuiqing Dai, Xiaoyu Su, Xin Jin, Jie Zhang, Xiong Guan, Chen Chen, Changlong Shu, Tianpei Huang. Cloning, Expression, Purification, and Insecticidal Activity of a Novel Cry1Na3 Toxin FromBacillus thuringiensisBRC-ZYR2. Journal of Economic Entomology. 2016; 109 (3):1064-1070.
Chicago/Turabian StyleRuiqing Dai; Xiaoyu Su; Xin Jin; Jie Zhang; Xiong Guan; Chen Chen; Changlong Shu; Tianpei Huang. 2016. "Cloning, Expression, Purification, and Insecticidal Activity of a Novel Cry1Na3 Toxin FromBacillus thuringiensisBRC-ZYR2." Journal of Economic Entomology 109, no. 3: 1064-1070.
During evolution the creation of single crossover chimeras between duplicated paralogous genes is a known process for increasing diversity. Comparing the properties of homologously recombined chimeras with one or two crossovers is also an efficient strategy for analyzing relationships between sequence variation and function. However, no well-developed in vitro method has been established to create single-crossover libraries. Here we present an in vitro template-change polymerase change reaction that has been developed to enable the production of such libraries. We applied the method to two closely related toxin genes from B. thuringiensis and created chimeras with differing properties that can help us understand how these toxins are able to differentiate between insect species.
Changlong Shu; Jianqiao Zhou; Neil Crickmore; Xianchun Li; Fuping Song; Gemei Liang; Kanglai He; Dafang Huang; Jie Zhang. In vitro template-change PCR to create single crossover libraries: a case study with B. thuringiensis Cry2A toxins. Scientific Reports 2016, 6, 23536 .
AMA StyleChanglong Shu, Jianqiao Zhou, Neil Crickmore, Xianchun Li, Fuping Song, Gemei Liang, Kanglai He, Dafang Huang, Jie Zhang. In vitro template-change PCR to create single crossover libraries: a case study with B. thuringiensis Cry2A toxins. Scientific Reports. 2016; 6 (1):23536.
Chicago/Turabian StyleChanglong Shu; Jianqiao Zhou; Neil Crickmore; Xianchun Li; Fuping Song; Gemei Liang; Kanglai He; Dafang Huang; Jie Zhang. 2016. "In vitro template-change PCR to create single crossover libraries: a case study with B. thuringiensis Cry2A toxins." Scientific Reports 6, no. 1: 23536.
Bacillus thuringiensis Cry toxins bind with different insect midgut proteins leading to toxin oligomerization, membrane insertion and pore formation. However, different Cry toxins had been shown to readily form high molecular weight oligomers or aggregates in solution in the absence of receptor interaction. The role of Cry oligomers formed in solution remains uncertain. The Cry9A proteins show high toxicity against different Lepidoptera, and no-cross resistance with Cry1A. Cry9Aa655 protein formed oligomers easily in solution mediated by disulfide bonds, according to SDS-PAGE analysis under non-reducing and reducing conditions. However, oligomerization is not observed if Cry9Aa655 is activated with trypsin, suggesting that cysteine residues, C14 and C16, located in the N-terminal end that is processed during activation participate in this oligomerization. To determine the role of these residues on oligomerization and in toxicity single and double alanine substitution were constructed. In contrast to single C14A and C16A mutants, the double C14A–C16A mutant did not form oligomers in solution. Toxicity assays against Plutella xylostella showed that the C14A–C16A mutant had a similar insecticidal activity as the Cry9Aa655 protein indicating the oligomers of Cry9Aa formed in solution in the absence of receptor binding are not related with toxicity. The aggregation of Cry9Aa655 polypeptides was mediated by disulfide bonds. Cry9Aa655 C14 and C16C are involved in oligomerization in solution. These aggregate forms are not related to the mode of action of Cry9Aa leading to toxicity.
Longfa Fang; Bo Wang; Zishan Zhou; Sujuan Yang; Changlong Shu; Fuping Song; Alejandra Bravo; Mario Soberón; Jie Zhang. Oligomerization of Cry9Aa in solution without receptor binding, is not related with insecticidal activity. Electronic Journal of Biotechnology 2016, 21, 54 -57.
AMA StyleLongfa Fang, Bo Wang, Zishan Zhou, Sujuan Yang, Changlong Shu, Fuping Song, Alejandra Bravo, Mario Soberón, Jie Zhang. Oligomerization of Cry9Aa in solution without receptor binding, is not related with insecticidal activity. Electronic Journal of Biotechnology. 2016; 21 ():54-57.
Chicago/Turabian StyleLongfa Fang; Bo Wang; Zishan Zhou; Sujuan Yang; Changlong Shu; Fuping Song; Alejandra Bravo; Mario Soberón; Jie Zhang. 2016. "Oligomerization of Cry9Aa in solution without receptor binding, is not related with insecticidal activity." Electronic Journal of Biotechnology 21, no. : 54-57.
Cry1Ac toxin‐binding proteins from Helicoverpa armigera brush border membrane vesicles were identified by an improved pull‐down method that involves coupling Cry1Ac to CNBr agarose combined with liquid chromatography–tandem mass spectrometry (LC‐MS/MS). According to the LC‐MS/MS results, Cry1Ac toxin could bind to six classes of aminopeptidase‐N, alkaline phosphatase, cadherin‐like protein, ATP‐binding cassette transporter subfamily C protein (ABCC2), actin, ATPase, polycalin, and some other proteins not previously characterized as Cry toxin‐binding molecules such as dipeptidyl peptidase or carboxyl/choline esterase and some serine proteases. This is the first report that suggests the direct binding of Cry1Ac toxin to ABCC2 in H. armigera.
Zishan Zhou; Zeyu Wang; Yuxiao Liu; Gemei Liang; Changlong Shu; Fuping Song; Xueping Zhou; Alejandra Bravo; Mario Soberón; Jie Zhang. Identification of ABCC2 as a binding protein of Cry1Ac on brush border membrane vesicles from Helicoverpa armigera by an improved pull‐down assay. MicrobiologyOpen 2016, 5, 659 -669.
AMA StyleZishan Zhou, Zeyu Wang, Yuxiao Liu, Gemei Liang, Changlong Shu, Fuping Song, Xueping Zhou, Alejandra Bravo, Mario Soberón, Jie Zhang. Identification of ABCC2 as a binding protein of Cry1Ac on brush border membrane vesicles from Helicoverpa armigera by an improved pull‐down assay. MicrobiologyOpen. 2016; 5 (4):659-669.
Chicago/Turabian StyleZishan Zhou; Zeyu Wang; Yuxiao Liu; Gemei Liang; Changlong Shu; Fuping Song; Xueping Zhou; Alejandra Bravo; Mario Soberón; Jie Zhang. 2016. "Identification of ABCC2 as a binding protein of Cry1Ac on brush border membrane vesicles from Helicoverpa armigera by an improved pull‐down assay." MicrobiologyOpen 5, no. 4: 659-669.
Genes with different functions are originally generated from some ancestral genes by gene duplication, mutation and functional recombination. It is widely accepted that orthologs are homologous genes evolved from speciation events while paralogs are homologous genes resulted from gene duplication events.With the rapid increase of genomic data, identifying and distinguishing these genes among different species is becoming an important part of functional genomics research. Using 35 plant and 6 green algal genomes from Phytozome v9, we clustered 1,291,670 peptide sequences into 49,355 homologous gene families in terms of sequence similarity. For each gene family, we have generated a peptide sequence alignment and phylogenetic tree, and identified the speciation/duplication events for every node within the tree. For each node, we also identified and highlighted diagnostic characters that facilitate appropriate addition of a new query sequence into the existing phylogenetic tree and sequence alignment of its best matched gene family. Based on a desired species or subgroup of all species, users can view the phylogenetic tree, sequence alignment and diagnostic characters for a given gene family selectively. PlantOrDB not only allows users to identify orthologs or paralogs from phylogenetic trees, but also provides all orthologs that are built using Reciprocal Best Hit (RBH) pairwise alignment method. Users can upload their own sequences to find the best matched gene families, and visualize their query sequences within the relevant phylogenetic trees and sequence alignments. PlantOrDB (http://bioinfolab.miamioh.edu/plantordb) is a genome-wide ortholog database for land plants and green algae. PlantOrDB offers highly interactive visualization, accurate query classification and powerful search functions useful for functional genomic research. The online version of this article (doi:10.1186/s12870-015-0531-4) contains supplementary material, which is available to authorized users.
Lei Li; Guoli Ji; Congting Ye; Changlong Shu; Jie Zhang; Chun Liang. PlantOrDB: a genome-wide ortholog database for land plants and green algae. BMC Plant Biology 2015, 15, 161 .
AMA StyleLei Li, Guoli Ji, Congting Ye, Changlong Shu, Jie Zhang, Chun Liang. PlantOrDB: a genome-wide ortholog database for land plants and green algae. BMC Plant Biology. 2015; 15 (1):161.
Chicago/Turabian StyleLei Li; Guoli Ji; Congting Ye; Changlong Shu; Jie Zhang; Chun Liang. 2015. "PlantOrDB: a genome-wide ortholog database for land plants and green algae." BMC Plant Biology 15, no. 1: 161.
Pyramiding of diverse cry toxin genes from Bacillus thuringiensis with different modes of action is a desirable strategy to delay the evolution of resistance in the European corn borer ( Ostrinia nubilalis ). Considering the dependency of susceptibility to Cry toxins on toxin binding to receptors in the midgut of target pests, a diverse mode of action is commonly defined as recognition of unique binding sites in the target insect. In this study, we present a novel cry1Ie toxin gene ( cry1Ie2 ) as a candidate for pyramiding with Cry1Ab or Cry1Fa in corn to control Ostrinia species larvae. The new toxin gene encodes an 81-kDa protein that is processed to a protease-resistant core form of approximately 55 kDa by trypsin digestion. The purified protoxin displayed high toxicity to Ostrinia furnacalis and O. nubilalis larvae but low to no activity against Spodoptera or heliothine species or the coleopteran Tenebrio molitor . Results of binding assays with 125 I-labeled Cry1Ab toxin and brush border membrane vesicles from O. nubilalis larvae demonstrated that Cry1Ie2 does not recognize the Cry1Ab binding sites in that insect. Reciprocal competition binding assays with biotin-labeled Cry1Ie2 confirmed the lack of shared sites with Cry1Ab or Cry1Fa in O. nubilalis brush border membrane vesicles. These data support Cry1Ie2 as a good candidate for pyramiding with Cry1Ab or Cry1Fa in corn to increase the control of O. nubilalis and reduce the risk of resistance evolution.
Can Zhao; Juan Luis Jurat-Fuentes; Heba M. Abdelgaffar; Hongyu Pan; Fuping Song; Jie Zhang. Identification of a New cry1I -Type Gene as a Candidate for Gene Pyramiding in Corn To Control Ostrinia Species Larvae. Applied and Environmental Microbiology 2015, 81, 3699 -3705.
AMA StyleCan Zhao, Juan Luis Jurat-Fuentes, Heba M. Abdelgaffar, Hongyu Pan, Fuping Song, Jie Zhang. Identification of a New cry1I -Type Gene as a Candidate for Gene Pyramiding in Corn To Control Ostrinia Species Larvae. Applied and Environmental Microbiology. 2015; 81 (11):3699-3705.
Chicago/Turabian StyleCan Zhao; Juan Luis Jurat-Fuentes; Heba M. Abdelgaffar; Hongyu Pan; Fuping Song; Jie Zhang. 2015. "Identification of a New cry1I -Type Gene as a Candidate for Gene Pyramiding in Corn To Control Ostrinia Species Larvae." Applied and Environmental Microbiology 81, no. 11: 3699-3705.
Bacillus thuringiensis GabR is a Sigma 54-dependent transcriptional activator containing three typical domains, an N-terminal regulatory domain Per-ARNT-Sim (PAS), a central AAA+ (ATPases associated with different cellular activities) domain and a C-terminal helix-turn-helix (HTH) DNA binding domain. GabR positively regulates the expression of the gabT gene of the gab gene cluster, which is responsible for the γ-aminobutyric acid (GABA) shunt. Purified GabR was shown to specifically bind to a repeat region that mapped 58 bp upstream of the gabT start codon. The specific signal factors GABA and succinic semialdehyde (SSA) activated gabT expression, whereas GABA- and SSA-inducible gabT transcription was abolished in sigL and gabR mutants. GABA and SSA did not induce the expression of either SigL or GabR. Deletion of the PAS domain of GabR resulted in increased gabT transcriptional activity, both in the presence and absence of GABA. This study identified the GabR-binding site on the gabT promoter; however, GabR does not bind to its own promoter. gabT transcription is induced by GABA and SSA, and inducible expression is dependent on SigL and activated by GabR. The PAS domain in GabR is repressing its enhancer transcriptional activity on the gabT promoter. Repression is released upon GABA addition, whereupon transcription is induced.
Qi Peng; Min Yang; Wei Wang; Lili Han; Guannan Wang; Pengyue Wang; Jie Zhang; Fuping Song. Activation of gab cluster transcription in Bacillus thuringiensis by γ-aminobutyric acid or succinic semialdehyde is mediated by the Sigma 54-dependent transcriptional activator GabR. BMC Microbiology 2014, 14, 1 -12.
AMA StyleQi Peng, Min Yang, Wei Wang, Lili Han, Guannan Wang, Pengyue Wang, Jie Zhang, Fuping Song. Activation of gab cluster transcription in Bacillus thuringiensis by γ-aminobutyric acid or succinic semialdehyde is mediated by the Sigma 54-dependent transcriptional activator GabR. BMC Microbiology. 2014; 14 (1):1-12.
Chicago/Turabian StyleQi Peng; Min Yang; Wei Wang; Lili Han; Guannan Wang; Pengyue Wang; Jie Zhang; Fuping Song. 2014. "Activation of gab cluster transcription in Bacillus thuringiensis by γ-aminobutyric acid or succinic semialdehyde is mediated by the Sigma 54-dependent transcriptional activator GabR." BMC Microbiology 14, no. 1: 1-12.
Peanut (Arachis hypogaea L.), one of the most important oil legumes in the world, is heavily damaged by white grubs. Tissue-specific promoters are needed to incorporate insect resistance genes into peanut by genetic transformation to control the subterranean pests. Transcriptome sequencing is the most effective way to analyze differential gene expression in this non-model species and contribute to promoter cloning. The transcriptomes of the roots, seeds and leaves of peanut were sequenced using Illumina technology. A simple digital expression profile was established based on number of transcripts per million clean tags (TPM) from different tissues. Subsequently, 584 root-specific candidate transcript assembly contigs (TACs) and 316 seed-specific candidate TACs were identified. Among these candidate TACs, 55.3% were root-specific and 64.6% were seed-specific by semi-quantitative RT-PCR analysis. Moreover, the consistency of semi-quantitative RT-PCR with the simple digital expression profile was correlated with the length and TPM value of TACs. The results of gene ontology showed that some root-specific TACs are involved in stress resistance and respond to auxin stimulus, whereas, seed-specific candidate TACs are involved in embryo development, lipid storage and long-chain fatty acid biosynthesis. One root-specific promoter was cloned and characterized. We developed a high-yield screening system in peanut by establishing a simple digital expression profile based on Illumina sequencing. The feasible and rapid method presented by this study can be used for other non-model crops to explore tissue-specific or spatially specific promoters.
Lili Geng; Xiaohong Duan; Chun Liang; Changlong Shu; Fuping Song; Jie Zhang. Mining Tissue-specific Contigs from Peanut (Arachis hypogaea L.) for Promoter Cloning by Deep Transcriptome Sequencing. Plant and Cell Physiology 2014, 55, 1793 -1801.
AMA StyleLili Geng, Xiaohong Duan, Chun Liang, Changlong Shu, Fuping Song, Jie Zhang. Mining Tissue-specific Contigs from Peanut (Arachis hypogaea L.) for Promoter Cloning by Deep Transcriptome Sequencing. Plant and Cell Physiology. 2014; 55 (10):1793-1801.
Chicago/Turabian StyleLili Geng; Xiaohong Duan; Chun Liang; Changlong Shu; Fuping Song; Jie Zhang. 2014. "Mining Tissue-specific Contigs from Peanut (Arachis hypogaea L.) for Promoter Cloning by Deep Transcriptome Sequencing." Plant and Cell Physiology 55, no. 10: 1793-1801.
The entomopathogen Bacillus thuringiensis is used to control various pest species of scarab beetle but is not particularly effective. Gut bacteria have diverse ecological and evolutionary effects on their hosts, but whether gut bacteria can protect scarabs from B. thuringiensis infection remains poorly understood. To investigate this, we isolated 32 cultivable gut bacteria from Holotrichia oblita Faldermann, Holotrichia parallela Motschulsky, and Anomala corpulenta Motschulsky, and analyzed their effect on B. thuringiensis multiplication and Cry toxin stability. 16S rDNA analysis indicated that these gut bacteria belong to the Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes phyla. A confrontation culture analyses of the 32 isolates against three scarab-specific B. thuringiensis strains showed that the majority of the scarab gut bacteria had antibacterial activity against the B. thuringiensis strains. The Cry toxin stability analysis results showed that while several strains produced proteases capable of processing the scarab-specific toxin Cry8Ea, none were able to completely degrade it. These results suggest that gut bacteria can potentially affect the susceptibility of scarabs to B. thuringiensis and that this should be considered when considering future control measures.
Yueming Shan; Changlong Shu; Neil Crickmore; Chunqin Liu; Wensheng Xiang; Fuping Song; Jie Zhang. Cultivable Gut Bacteria of Scarabs (Coleoptera: Scarabaeidae) InhibitBacillus thuringiensisMultiplication. Environmental Entomology 2014, 43, 612 -616.
AMA StyleYueming Shan, Changlong Shu, Neil Crickmore, Chunqin Liu, Wensheng Xiang, Fuping Song, Jie Zhang. Cultivable Gut Bacteria of Scarabs (Coleoptera: Scarabaeidae) InhibitBacillus thuringiensisMultiplication. Environmental Entomology. 2014; 43 (3):612-616.
Chicago/Turabian StyleYueming Shan; Changlong Shu; Neil Crickmore; Chunqin Liu; Wensheng Xiang; Fuping Song; Jie Zhang. 2014. "Cultivable Gut Bacteria of Scarabs (Coleoptera: Scarabaeidae) InhibitBacillus thuringiensisMultiplication." Environmental Entomology 43, no. 3: 612-616.
The cry1 -type genes of Bacillus thuringiensis represent the largest cry gene family, which contains 50 distinct holotypes. It is becoming more and more difficult to identify cry1 -type genes using current methods because of the increasing number of cry1 -type genes. In the present study, an improved PCR-restriction fragment length polymorphism (PCR-RFLP) method which can distinguish 41 holotypes of cry1 -type genes was developed. This improved method was used to identify cry1 -type genes in 20 B. thuringiensis strains that are toxic to lepidoptera. The results showed that the improved method can efficiently identify single and clustered cry1 -type genes and can be used to evaluate cry1 -type genes in novel strain collections of B. thuringiensis . Among the detected cry1 -type genes, we identified four novel genes, cry1Ai , cry1Bb , cry1Ja , and cry1La . The bioassay results from the expressed products of the four novel cry genes showed that Cry1Ai2, Cry1Bb2, and Cry1Ja2 were highly toxic against Plutella xylostella , whereas Cry1La2 exhibited no activity. Moreover, Cry1Ai2 had good lethal activity against Ostrinia furnacalis , Hyphantria cunea , Chilo suppressalis , and Bombyx mori larvae and considerable weight loss activity against Helicoverpa armigera .
Changlong Shu; Dongming Liu; Zishan Zhou; Jilin Cai; Qi Peng; Jiguo Gao; Fuping Song; Jie Zhang. An Improved PCR-Restriction Fragment Length Polymorphism (RFLP) Method for the Identification of cry1 -Type Genes. Applied and Environmental Microbiology 2013, 79, 6706 -6711.
AMA StyleChanglong Shu, Dongming Liu, Zishan Zhou, Jilin Cai, Qi Peng, Jiguo Gao, Fuping Song, Jie Zhang. An Improved PCR-Restriction Fragment Length Polymorphism (RFLP) Method for the Identification of cry1 -Type Genes. Applied and Environmental Microbiology. 2013; 79 (21):6706-6711.
Chicago/Turabian StyleChanglong Shu; Dongming Liu; Zishan Zhou; Jilin Cai; Qi Peng; Jiguo Gao; Fuping Song; Jie Zhang. 2013. "An Improved PCR-Restriction Fragment Length Polymorphism (RFLP) Method for the Identification of cry1 -Type Genes." Applied and Environmental Microbiology 79, no. 21: 6706-6711.
Cry8Ea1 from Bacillus thuringiensis strain Bt185 has insecticidal activity against Holotrichia parallela. Cry8Ca2 from strain HBF-1 is effective against Anomala corpulenta. Cry8Ga1 from strain HBF-18 is toxic to H. oblita. Given the need to broaden the spectrum of B. thuringiensis, a broad-spectrum coleopteran active strain of B. thuringiensis, BIOT185, engineered to express multiple cry genes, including cry8Ea1, cry8Fa1 and cry8Ca2, was created by homologous recombination introducing the cry8Ca2 into the B. thuringiensis strain Bt185 by Liu et al. (Appl Microbiol Biotechnol 87:243–249, 2010). To further broaden the spectrum, an engineered B. thuringiensis strain BIOT1858G was constructed by introducing the recombinant plasmid pSTK-8G containing cry8Ga1 into the engineered B. thuringiensis strain BIOT185. PCR and Southern blotting demonstrated that the cry8Ga1 gene was transferred to the novel strain BIOT1858G. SDS-PAGE and RT-PCR confirmed the normal expression and transcription of the cry8Ga1 gene in addition to the cry8Ea1, cry8Fa1 and cry8Ca2 genes. Bioassays of BIOT1858G indicated that the recombinant strain broadened the spectrum against not only H. parallela susceptible to the Cry8E protein and A. corpulenta susceptible to the Cry8C protein but also H. oblita susceptible to the Cry8G protein. The pesticide could also decrease the cost of production and field labor.
Yanhua Jia; Can Zhao; Qinglei Wang; Changlong Shu; Xiaojie Feng; Fuping Song; Jie Zhang. A genetically modified broad-spectrum strain of Bacillus thuringiensis toxic against Holotrichia parallela, Anomala corpulenta and Holotrichia oblita. World Journal of Microbiology and Biotechnology 2013, 30, 595 -603.
AMA StyleYanhua Jia, Can Zhao, Qinglei Wang, Changlong Shu, Xiaojie Feng, Fuping Song, Jie Zhang. A genetically modified broad-spectrum strain of Bacillus thuringiensis toxic against Holotrichia parallela, Anomala corpulenta and Holotrichia oblita. World Journal of Microbiology and Biotechnology. 2013; 30 (2):595-603.
Chicago/Turabian StyleYanhua Jia; Can Zhao; Qinglei Wang; Changlong Shu; Xiaojie Feng; Fuping Song; Jie Zhang. 2013. "A genetically modified broad-spectrum strain of Bacillus thuringiensis toxic against Holotrichia parallela, Anomala corpulenta and Holotrichia oblita." World Journal of Microbiology and Biotechnology 30, no. 2: 595-603.
In Bacillus thuringiensis , a novel N -acetylmuramoyl- l -alanine amidase gene (named cwlB ) was detected, and the CwlB protein was purified and characterized. Reverse transcription-PCR (RT-PCR) results indicated that cwlB and an upstream gene (named cwlA ) formed one transcriptional unit. 5′ rapid amplification of cDNA ends (5′-RACE)-PCR and transcriptional fusions with the lacZ gene indicated that transcription of the operon was directed by a promoter, P cwlA , which is located upstream from the cwlA gene and that the transcription start site is a single 5′-end nucleotide residue T located 25 nucleotides (bp) upstream from the cwlA translational start codon. Moreover, the activity of P cwlA was controlled by σ K . Morphological analysis suggested that the mutation of cwlB could delay spore release compared to the timing of spore release in the wild-type strain. Western blot assay demonstrated that purified CwlB bound to the B. thuringiensis cell wall. Observations with laser confocal microscopy and a green fluorescent protein-based reporter system demonstrated that the CwlB protein localizes to the cell envelope. All results suggest that the CwlB protein is involved in mother cell lysis in B. thuringiensis .
Jingni Yang; Qi Peng; Zhen Chen; Chao Deng; Changlong Shu; Jie Zhang; Dafang Huang; Fuping Song. Transcriptional Regulation and Characteristics of a Novel N -Acetylmuramoyl- l -Alanine Amidase Gene Involved in Bacillus thuringiensis Mother Cell Lysis. Journal of Bacteriology 2013, 195, 2887 -2897.
AMA StyleJingni Yang, Qi Peng, Zhen Chen, Chao Deng, Changlong Shu, Jie Zhang, Dafang Huang, Fuping Song. Transcriptional Regulation and Characteristics of a Novel N -Acetylmuramoyl- l -Alanine Amidase Gene Involved in Bacillus thuringiensis Mother Cell Lysis. Journal of Bacteriology. 2013; 195 (12):2887-2897.
Chicago/Turabian StyleJingni Yang; Qi Peng; Zhen Chen; Chao Deng; Changlong Shu; Jie Zhang; Dafang Huang; Fuping Song. 2013. "Transcriptional Regulation and Characteristics of a Novel N -Acetylmuramoyl- l -Alanine Amidase Gene Involved in Bacillus thuringiensis Mother Cell Lysis." Journal of Bacteriology 195, no. 12: 2887-2897.
Bacillus thuringiensis is a Gram-positive bacterium that produces intracellular protein crystals toxic to a wide variety of insect larvae. We report the complete genome sequence of Bacillus thuringiensis subsp. kurstaki strain HD73 from the Centre OILB (Institut Pasteur, France), which belongs to serotype 3ab and is toxic to lepidopteran larvae.
Guiming Liu; Lai Song; Changlong Shu; Pinshu Wang; Chao Deng; Qi Peng; Didier Lereclus; Xumin Wang; Dafang Huang; Jie Zhang; Fuping Song. Complete Genome Sequence of Bacillus thuringiensis subsp. kurstaki Strain HD73. Genome Announcements 2013, 1, e00080-13 .
AMA StyleGuiming Liu, Lai Song, Changlong Shu, Pinshu Wang, Chao Deng, Qi Peng, Didier Lereclus, Xumin Wang, Dafang Huang, Jie Zhang, Fuping Song. Complete Genome Sequence of Bacillus thuringiensis subsp. kurstaki Strain HD73. Genome Announcements. 2013; 1 (2):e00080-13.
Chicago/Turabian StyleGuiming Liu; Lai Song; Changlong Shu; Pinshu Wang; Chao Deng; Qi Peng; Didier Lereclus; Xumin Wang; Dafang Huang; Jie Zhang; Fuping Song. 2013. "Complete Genome Sequence of Bacillus thuringiensis subsp. kurstaki Strain HD73." Genome Announcements 1, no. 2: e00080-13.