TY - BOOK AU - Zhang,Jun AU - Li,Jintian AU - Huang,Zebo AU - Yang,Bing AU - Zhang,Xiaojie AU - Li,Dehua AU - Craik,David J AU - Baker,Alan J M AU - Shu,Wensheng AU - Liao,Bin TI - Transcriptomic screening for cyclotides and other cysteine-rich proteins in the metallophyte Viola baoshanensis SN - 0176-1617 AV - QK711.2 JOU PY - 2015/// CY - Amsterdam PB - Elsevier GmbH KW - CrpExcel KW - Cyclotides KW - Cysteine-rich proteins N2 - Cysteine (Cys)-rich proteins (CRPs) are frequently associated with plant defense and stress resistance. Viola baoshanensis is a cadmium (Cd) hyper-accumulating plant whose CRPs-based defense systems are so far poorly understood. Next generation sequencing (NGS) techniques and a specialist searching tool, CrpExcel, were employed for identifying CRPs in V. baoshanensis. The transcriptome sequences of V. baoshanensis were assembled primarily from 454FLX/Hiseq2000 reads of plant cDNA sequencing libraries. CrpExcel was then used to search the ORFs and 9687 CRPs were identified, and included zinc finger (ZF) proteins, lipid transfer proteins, thaumatins and cyclotide precursors. Real-time PCR results showed that all CRP genes tested are constitutively expressed, but the genes of defensive peptides showed greater up-regulated expression than those of ZF-proteins in Cd- and/or wounding (Wd) treatments of V. baoshanensis seedlings. The NGS-derived sequences of cyclotide precursor genes were verified by RT-PCR and ABI3730 sequencing studies, and 32 novel cyclotides were identified in V. baoshanensis. In general, the metal-binding sites of ZF-containing CRPs also represented the potential vulnerable targets of toxic metals. This study provides broad insights into CRPs-based defense systems and stress-vulnerable targets in V. baoshanensis. It now brings the number of cyclotide sequences in V. baoshanensis to 53 and based on projections from this work, the number of cyclotides in the Violaceae is now conservatively estimated to be >30000 UR - https://doi.org/10.1016/j.jplph.2015.01.017 ER -