摘要
目的克隆出浙贝母法呢基焦磷酸合酶(Farnesyl pyrophosphate synthase,FPS)基因,并对其表达量与生物碱积累量之间的相关性进行分析,以探究FPS基因在浙贝母生物碱合成代谢过程中的调控作用。方法本研究采用RACE技术克隆出浙贝母FPS基因的全长序列;以浙贝母狭叶、宽叶和新梅园3个品种6个组织及10个不同发育时期的新鳞茎为材料,采用RT-qPCR和HPLC-ELSD技术分别测定了FPS基因的表达量与生物碱(浙贝甲素和浙贝乙素)的量,并对其进行相关性分析。结果 FPS基因cDNA序列全长1 629 bp,开放阅读框为1 056 bp,编码351个氨基酸;3个不同浙贝母品种中FPS基因组织特异性表达趋势相同,花中的表达量最高,其次为新鳞茎;不同发育时期新鳞茎FPS基因表达量与生物碱量相关性分析显示,在宽叶和新梅园品种中,FPS基因表达量与浙贝甲素、浙贝乙素量均具有明显的正相关性(相关系数分别为0.289、0.613,0.427、0.622),但在狭叶品种中,该相关性相对较低(0.054、0.236);狭叶、宽叶和新梅园3个品种不同组织中FPS基因表达量与生物碱量具有正相关性(相关系数分别为0.057、0.476,0.085、0.495,0.375、0.432)。结论本研究成功克隆出浙贝母FPS基因的全长序列,FPS基因参与了浙贝母生物碱的合成代谢调控。
Objective Farnesyl pyrophosphate synthase（FPS） gene of Fritillaria thunbergii was cloned and the correlation between the expression content of gene and alkaloid accumulation was analyzed to explore the role of FPS gene in the regulation of alkaloid synthesis and metabolism. Methods In this study, the new bulbs and different tissues of Fritillaria thunbergii varieties ＂Narrow leaf＂ ＂Broad leaf＂ and ＂New Meiyuan＂ were used to clone the full-length sequence of FPS gene via RACE technology. RT-q PCR and HPLC-ELSD technology was used to determine the expression of FPS in 10 different developmental stages and the content of alkaloid（Peimine and Verticinone）, respectively. Finally, the correlation was analyzed. Results The full-length c DNA sequence of FPS gene was 1 629 bp. Open Reading Frame（ORF） was 1 056 bp and encoded 351 amino acids. Among them, the tissue specific expression trends of FPS gene were same, flowers had the highest expression level, followed by the new bulb. The results of correlation analysis showed thatthe content Peimine and Verticinone had significant positive correlation with the expression level of FPS gene in ＂Broad leaf＂ and ＂New Meiyuan＂（The correlation coefficients were 0.289 and 0.613, 0.427 and 0.622, respectively）;FPS gene expression and alkaloid content in different tissues had a positive correlation（The correlation coefficients were 0.057 and 0.476, 0.085 and 0.495, 0.375 and 0.432, respectively）. Conclusion The full-length sequence of FPS gene was successfully cloned in this study. The FPS gene involved in the regulation of alkaloid synthesis and metabolism.
作者
冯亚斌
李沫宇
吴秋丽
江建铭
沈晓霞
王忠华
FENG Ya-bin LI Mo-yu WU Qiu-li JIANG Jian-ming SHEN Xiao-xia WANG Zhong-hua(Institute of Biotechnology, Zhejiang Wanli University, Ningbo 315100, China Zhejiang Institute of Chinese Meteria Medica, Hangzhou 310023, China)
出处
《中草药》
CAS
CSCD
北大核心
2017年第5期971-978,共8页
Chinese Traditional and Herbal Drugs
基金
浙江省科技厅育种专项(2016C02058)
浙江省重点研发项目(2007C02011)
宁波市公益类重大专项(2015C110007)
浙江万里学院重中之重学科开放基金项目(KF2016001)
关键词
浙贝母
FPS基因
基因表达
生物碱
相关性
代谢调控
Fritillaria thunbergii
FPS gene
gene expression
Alkaloid
Correlation
Metabolic regulation
作者简介
冯亚斌（1990-），男，河南新乡人，硕士研究生，研究方向为浙贝母分子生物学。E-mail：1547247657@qq．com
通信作者 王忠华（1972-），男，浙江开化人，教授，硕士生导师，研究方向为药用植物分子生物学。E-mail：wang1972@zwu．edu．cn
