Supplementary Information. Deciphering the Venomic Transcriptome of Killer- Wasp Vespa velutina
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1 Supplementary Information Deciphering the Venomic Transcriptome of Killer- Wasp Vespa velutina Zhirui Liu, Shuanggang Chen, You Zhou, Cuihong Xie, Bifeng Zhu, Huming Zhu, Shupeng Liu, Wei Wang, Hongzhuan Chen, Yonghua Ji 1
2 Supplementary Figure legends: Fig. S1: Annotation of 454 FLX+ Unigenes in GO, COG and KEGG database. A) GO categories of the unigenes: BR: biological regulation; CCO: cellular 2
3 component organization; CP: cellular process; Death: death; DP: developmental process; EOL: establishment of localization; Growth:growth; Localization: localization; MP: metabolic process; MOP: multicellular organismal process; Pigmentation: pigmentation; Reproduction: reproduction; RTS: response to stimulus; VR: viral reproduction; Cell: cell; CP: cell part; Envelope: envelope; ER: extracellular region; ERP: extracellular region part; MC: macromolecular complex; ML: membrane-enclosed lumen; Organelle: organelle; OP:organelle part; AX: antioxidant activity; Binding: binding; CL: catalytic activity; EC: electron carrier activity; ER: enzyme regulator activity; MT: molecular transducer activity; SMA: structural molecule activity; Transcription RL: transcription regulator activity; Translation RL: translation regulator activity; TP: transporter activity; B) COG classification of the unigenes. A: RNA processing and modification; B: Chromatin structure and dynamics; C: Energy production and conversion; D: Cell cycle control, cell division, chromosome partitioning; E: Amino acid transport and metabolism; F: Nucleotide transport and metabolism; G: Carbohydrate transport and metabolism; H: Coenzyme transport and metabolism; I: Lipid transport and metabolism; J: Translation, ribosomal structure and biogenesis; K: Transcription; L: Replication, recombination and repair; M: Cell wall/membrane/envelope biogenesis; N: Cell motility; O: Posttranslational modification, protein turnover, chaperones; P: Inorganic ion transport and metabolism; 3
4 Q: Secondary metabolites biosynthesis, transport and catabolism; R: General function prediction only; S: Function unknown; T: Signal transduction mechanisms; U: Intracellular trafficking, secretion, and vesicular transport; V: Defense mechanisms; W: Extracellular structures; Y: Nuclear structure; Z: Cytoskeleton C) KEGG pathways of the unigenes CMb: Carbohydrate Metabolism; EM: Energy Metabolism; LM: Lipid Metabolism; NM: Nucleotide Metabolism; AAM: Amino Acid Metabolism; MOAA: Metabolism of Other Amino Acids; GBM: Glycan Biosynthesis and Metabolism; MCV: Metabolism of Cofactors and Vitamins; MTP: Metabolism of Terpenoids and Polyketides; XBM: Xenobiotics Biodegradation and Metabolism; TC: Transport and Catabolism; CM: Cell Motility; CGD: Cell Growth and Death; CC: Cell Communication; IM: Immune System; END: Endocrine System; CL: Circulatory System; DT: Digestive System; EXC: Excretory System; NS: Nervous System; Development: Development; Translation: Translation; FSD: Folding, Sorting and Degradation; ST: Signal Transduction. 4
5 Fig. S2: Anotation of Illumina Unigenes annotation in GO COG and KEGG database. 5
6 The percentage of unigenes in Wa1p and Wa2p were presented in deep and light colors, respectively. A. GO categories of the unigenes. ASF: anatomical structure formation; BA: biological adhesion; BR: biological regulation; CCB: cellular component biogenesis; CCO: cellular component organization; CP: cellular process; DP: developmental process; EL: establishment of localization; Localization: localization; MP: metabolic process; MOP: multicellular organismal process; Pigmentation: pigmentation; RTS: response to stimulus Cell: cell; CP: cell part; Envelope: envelope; ER: extracellular region ERP: extracellular region part; MC: macromolecular complex; ML: membrane-enclosed lumen; Organelle: organelle; OP: organelle part; Binding: binding; CL: catalytic activity; EC: electron carrier activity; ER: enzyme regulator activity; MT: molecular transducer activity; SMA: structural molecule activity; Transcription RL: transcription regulator activity; Translation RL: translation regulator activity; TP: transporter activity. B.COG classification of the unigenes. A: RNA processing and modification; B: Chromatin structure and dynamics; C: Energy production and conversion; D: Cell cycle control, cell division, chromosome partitioning; E: Amino acid transport and metabolism; F: Nucleotide transport and metabolism; G: Carbohydrate transport and metabolism; H: Coenzyme transport and 6
7 metabolism; I: Lipid transport and metabolism; J: Translation, ribosomal structure and biogenesis; K: Transcription; L: Replication, recombination and repair; M: Cell wall/membrane/envelope biogenesis; N: Cell motility; O: Posttranslational modification, protein turnover, chaperones; P: Inorganic ion transport and metabolism; Q: Secondary metabolites biosynthesis, transport and catabolism; R: General function prediction only; S: Function unknown; T: Signal transduction mechanisms; U: Intracellular trafficking, secretion, and vesicular transport; V: Defense mechanisms; W: Extracellular structures; Y: Nuclear structure; Z: Cytoskeleton. C.KEGG pathways of the unigenes CMb: Carbohydrate Metabolism; EM: Energy Metabolism; LM: Lipid Metabolism NM: Nucleotide Metabolism; AAM:Amino Acid Metabolism; MOAA: Metabolism of Other Amino Acids; GBM: Glycan Biosynthesis and Metabolism; MCV: Metabolism of Cofactors and Vitamins; MTP: Metabolism of Terpenoids and Polyketides; BSM: Biosynthesis of Other Secondary Metabolites; XBM: Xenobiotics Biodegradation and Metabolism; TC: Transport and Catabolism; CM: Cell Motility; CGD: Cell Growth and Death; CC: Cell Communication; IM: Immune System; END: Endocrine System; CL: Circulatory System; DT: Digestive System; EXC: Excretory System; NV: Nervous System; SS: Sensory System; Development: Development; EA: Environmental Adaptation; Transcription: Transcription; Translation: Translation FSD: Folding, Sorting and Degradation; RR: Replication and Repair; MT: Membrane 7
8 Transport; ST: Signal Transduction; SMI: Signaling Molecules and Interaction. Fig. S3: Relative quantification analysis of 36 differentially expressed unigenes in Wa1p and Wa2p. Log 2 Fold of changes for the 26 putative toxins (A) and 10 enzymes (B) considered to be differential expressed after venom extraction; (C-D) qrt-pcr validation for expression level of the above unigenes. 8
9 Fig. S4: Reads quality and length distributions of transcriptome of V. velutina venom sequenced by 454 GS-FLX+. 9
10 Fig. S5: Quality control and sequence content for reads obtained from Hiseq
11 Fig. S6 Electrophoretic analyses of venom proteins. Venom reservoirs of V. velutina were dissectedd in PBS buffer supplemented with a protease inhibitors cocktail (PI; Roche) and residual tissues were removed by centrifugation. The total protein content of 20 μgg venom reservoirs was run on a 10% SDS PAGE under reducing conditions (right). The separated proteins were visualized by Colloidal Coomassie staining. Molecular weight standards (left) were in kda. 11
12 Supplementary table: Table S1: Primers used for qrt-pcr validation of putative toxin unigenes (No.1-26)and enzymes for toxins synthesis or performances (No.27-36) No. Primer Sequence(5-3 ) Location(5 ) Product(bp) 1 S CTCATTTTGTGCATACATGACATAC A TTCTTTTGGTTCTGGATGTGC S GGAGGGATTATAGTCGAAGCTAG A AGCAACCTTGGCCCATAGAA S TTCAAACGACGATACCTATGAC A CCTACACGGCATACCGCTATT S CACAAGGTAGTTTTTCCTGTCTT A CTCCAGCAATATTTCTACACTCTTC S AGAGGATTGCCGAGACATAGAC A TTTGTATCCTATTCGCATGAG S GCAGAGGGGAGGGATTATAGTC A AGACCTTGTTCGACTCTAGGA S CACAAGGTAGTTTTTCCTGTCTTA A CACTTAATATCACACTCATAAGCTC S AACATGATGAAACTCATTCTCTTCG A ATTACTTGAAGTTAAACCCCATTGC S AACGAACAAGAAACGCATAATG A AACAATGAAGCGCAGTAATGAC S CCTTCCTCTCGATCAAACAAAC A ATTCCTATTTGAGTGCGATTGA
13 11 S CTTGGACGACCGTGTATAACT A GACGAAGCATAGAATGAGAATC S AACGACGAGGACGACAATCTT A TTGGAACGGATCTCAGTCTCAC S AGGCAAAGGAAACATCGACTTAG A TTGTTGGGGGGATAGATGACGA S CCATAATATGAAGTAAAGAGAATCC A GGCAAGGTCTTACTGGAAATG S TACCTTCGCAAAGATCCACTC A TGATGAAAACTTCAACTTGGG S ACACATACGATTACATTGGGACC A CGACCGATTAGTACAGATTTGATT S CAAGAGATACGAGGTGGCGTTT A CACGTCTGTGATCATCGTTCGA S ACCTTGTTCTATCAAATACTCTGC A CTTTCTATAGTAAAGTGTCTTGTGG S TGCTATAATTACATGTTTGACAAAG A CTTTCTATAGTAAAGTGTCTTGTGG S AAGTTTTAGTGCTGTTCAATGC A ACATTTGTACCACCTTATAGATACG S GTTCACTTTAACGACCTGACC A GATCTCCACAGAGATTTCCAC S CTTTTTCTTCTCAAGTTGTCAAGG A GCTGGCATATTTATTCATCCC S ACCAGTTGCCTCAAACGATTA A GATCCATTCGAGATTAAGGAAAA
14 25 S GATCGAGACGAAGTTTGGATC A TGAGTTGTAGGCACGGCATAG S ATGTCTATACTCCGCAGATTCC A ACAGTGACGACTACGATGTCCT S AAAGATGGCAAACGCTACGAG A TTCAGGCATATCTCCGTTCAA S CAACGAAGGTTTTTACGGTAAA A AAAAGACTTCCTGCGGGTTC S AATGTCAAGAAAGTGGTTCGC A GTATCGCAATATCGATGGTGG S AAAAGACGAACAATAAGGACAA A AGGGAACCAGACCATAAGAGT S TAACAACCCCAAAAGTAACAGC A CGTCTAGCGAGATAGATTTAGAAG S ATTTATGCTGTGGGACAACTC A GAAAGCTAGATATTAGGGAGCC S CTGTGATTATGGAAGACCTGC A TGGAAGAGTTTTAGTGTCGTTATA S GATTCGATCTGATTTATAGCGA A CAATATCTGAACCTGCACTGAC S AAGACCAAGTTGCGAATAGTT A TTCCTTAGTTACAGATACCAGC S GTTCCTAACAAGAATGACCGT A TTTATCCGCAACTAGGTATGC
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