Manjunatha B L, Singh H R, Ravikanth G, Nataraja Karaba N, Shankar Ravi, Kumar Sanjay, Shaanker R Uma
Department of Crop Physiology and School of Ecology and Conservation, University of Agricultural Sciences, GKVK, Bangalore 560 065, India.
J Biosci. 2016 Mar;41(1):119-31. doi: 10.1007/s12038-016-9591-3.
Camptothecin (CPT), a monoterpene indole alkaloid, is a potent inhibitor of DNA topoisomerase I and has applications in treating ovarian, small lung and refractory ovarian cancers. Stem wood tissue of Nothapodytes nimmoniana (Graham) Mabb. (family Icacinaceae) is one of the richest sources of CPT. Since there is no genomic or transcriptome data available for the species, the present work sequenced and analysed transcriptome of stem wood tissue on an Illumina platform. From a total of 77,55,978 reads, 9,187 transcripts were assembled with an average length of 255 bp. Functional annotation and categorization of these assembled transcripts unraveled the transcriptome architecture and also a total of 13 genes associated with CPT biosynthetic pathway were identified in the stem wood tissue. Four genes of the pathway were cloned to full length by RACE to validate the transcriptome data. Expression analysis of 13 genes associated with CPT biosynthetic pathway in 11 different tissues vis-a-vis CPT content analysis suggested an important role of NnPG10H, NnPSLS and NnPSTR genes in the biosynthesis of CPT. These results indicated that CPT might be synthesized in the leaves and then perhaps exported to stem wood tissue for storage.
喜树碱(CPT)是一种单萜吲哚生物碱,是DNA拓扑异构酶I的有效抑制剂,可用于治疗卵巢癌、小细胞肺癌和难治性卵巢癌。印度马比木(Nothapodytes nimmoniana (Graham) Mabb.)(茶茱萸科)的茎木组织是CPT最丰富的来源之一。由于该物种没有可用的基因组或转录组数据,本研究在Illumina平台上对茎木组织的转录组进行了测序和分析。从总共7755978条 reads 中,组装得到了9187条转录本,平均长度为255 bp。对这些组装转录本的功能注释和分类揭示了转录组结构,并且在茎木组织中总共鉴定出13个与CPT生物合成途径相关的基因。通过RACE将该途径的四个基因克隆到全长,以验证转录组数据。对11种不同组织中与CPT生物合成途径相关的13个基因的表达分析以及CPT含量分析表明,NnPG10H、NnPSLS和NnPSTR基因在CPT生物合成中起重要作用。这些结果表明,CPT可能在叶片中合成,然后可能转运到茎木组织中储存。