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Cloning and characterization of an elicitor-responsive gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase involved in 20-hydroxyecdysone production in cell cultures of Cyanotis arachnoidea

文献类型： 外文期刊

作者： Wang, Qiu Jun 1 ; Zheng, Li Ping 2 ; Zhao, Pei Fei 3 ; Zhao, Yi Lu 1 ; Wang, Jian Wen 1 ;

作者机构： 1.Soochow Univ, Coll Pharmaceut Sci, Suzhou 215123, Peoples R China

2.Soochow Univ, Sch Architecture & Urban Environm, Suzhou 215123, Peoples R China

3.Yunnan Acad Agr Sci, Inst Hort Crops, Kunming 650205, Peoples R China

关键词： Cyanotis arachnoidea;Elicitation;Gene expression;20-Hydroxyecdysterone;3-Hydroxy-3-methylglutaryl coenzyme A;reductase

期刊名称：PLANT PHYSIOLOGY AND BIOCHEMISTRY （ 影响因子：4.27； 五年影响因子：4.816 ）

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收录情况： SCI

摘要： Cyanotis arachnoidea contains a rich source of bioactive phytoecdysteroids (i.e. analogues of insect steroid hormones). 3-Hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) supplies mevalonate for the synthesis of many secondary metabolites including 20-hydroxyecdysone (20E), one of metabolismenhancing phytoecdysteroids. In this study, in order to develop a sustainable source of 20E, cell suspension cultures were established from shoot cultures of C. arachnoidea, and a full length cDNA encoding HMGR (designated as CaHMGR) was cloned and characterized. The cDNA contained 2037 nucleotides with a complete open reading frame (ORF) of 1800 nucleotides, which was predicted to encode a peptide of 599 amino acids. Expression analysis by real-time PCR revealed that CaHMGR mRNA was abundant in C. arachnoidea stems, roots and leaves. When cultivated in Murashige & Skoog medium supplemented with 0.2 mg L~(-1) 1-naphthlcetic acid (NAA) and 3.0 mg L~(-1) 6-benzyladenine (6-BA), C. arachnoidea cells in suspension culture grew rapidly, yielding 20E (124.14 μg L~(-1)) after 12 days. The content of 20E in cell cultures elicited by 0.2 mM methyl jasmonate (MeJA), 100 mg L_1 yeast elicitor (YE) or 25 μM AgNO_3 was increased 8-, 2-, and 6-fold over the control, respectively. Quantitative real-time PCR analysis showed that CaHMGR was expressed at a higher level under the treatment of MeJA or Ag~+ elicitor. Our results suggested that 20E accumulation may be the result of the expression up-regulation of CaHMGR involved in the biosynthesis under the treatment of various elicitors.

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