Lan Truong Thi Phuong, Huy Nguyen Duc, Luong Nguyen Ngoc, Nghi Nguyen Van, Tan Trinh Huu, Quan Le Viet, Loc Nguyen Hoang
Institute of Bioactive Compounds, College of Sciences, Hue University, 77 Nguyen Hue St., Hue, Vietnam.
College of Medicine and Pharmacy, Hue University, 6 Ngo Quyen St., Hue, Vietnam.
Curr Pharm Biotechnol. 2018;19(10):839-846. doi: 10.2174/1389201019666181008112244.
Curcuminoid genes have an important role in the biosynthesis of curcumin, a valuable bioactive compound, in Curcuma species. However, there have not been any reports of these genes in Curcuma zedoaria.
The present work reports on the isolation of genes encoding enzymes in curcuminoid metabolic pathway and their expression in C. zedoaria.
The primers were designed from untranslation regions of DCS, CURS1, CURS2 and CURS3 genes which are involved in curcuminoid biosynthesis in C. longa to isolate the corresponding fulllength genes in C. zedoaria. RT-PCR amplification and HPLC analysis are used to estimate the expression of genes and biosynthesis of curcumin in both rhizome and callus.
The results showed that all four genes from C. zedoaria (named CzDCS, CzCURS1, CzCURS2 and CzCURS3) and C. longa have a high identity (approximately 99%) and lengths of genes from C. zedoaria are 1382, 1240, 1288 and 1265 nu, respectively. CzCURS1, 2 and 3 genes have one intron while CzDCS has two introns. RT-PCR amplification indicated that curcuminoid genes expressed mRNA in rhizome and callus of C. zedoaria. Curcumin, a major component of curcuminoids, was also found in callus by HPLC analysis.
The sequence information of DCS and CURS1-3 genes in C. zedoaria will be very valuable for a subsequent study on the effects of elicitors on the transcription of genes involved in curcuminoid biosynthesis pathway.
姜黄素类基因在姜黄属植物中姜黄素(一种有价值的生物活性化合物)的生物合成中起重要作用。然而,莪术(Curcuma zedoaria)中尚未有这些基因的相关报道。
本研究报道了姜黄素类代谢途径中编码酶的基因的分离及其在莪术中的表达。
根据参与姜黄(Curcuma longa)姜黄素类生物合成的DCS、CURS1、CURS2和CURS3基因的非翻译区设计引物,以分离莪术中相应的全长基因。采用RT-PCR扩增和HPLC分析来评估根茎和愈伤组织中基因的表达以及姜黄素的生物合成。
结果表明,莪术的所有四个基因(命名为CzDCS、CzCURS1、CzCURS2和CzCURS3)与姜黄的基因具有高度同源性(约99%),莪术基因的长度分别为1382、1240、1288和1265个核苷酸。CzCURS1、2和3基因有一个内含子,而CzDCS有两个内含子。RT-PCR扩增表明姜黄素类基因在莪术的根茎和愈伤组织中表达mRNA。通过HPLC分析在愈伤组织中也发现了姜黄素类的主要成分姜黄素。
莪术中DCS和CURS1-3基因的序列信息对于后续研究诱导子对姜黄素类生物合成途径中相关基因转录的影响将非常有价值。