Journal
PLANT PHYSIOLOGY AND BIOCHEMISTRY
Volume 84, Issue -, Pages 1-9Publisher
ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER
DOI: 10.1016/j.plaphy.2014.08.021
Keywords
Cyanotis arachnoidea; Elicitation; Gene expression; 20-Hydroxyecdysterone; 3-Hydroxy-3-methylglutaryl coenzyme A reductase
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Funding
- Graduate Program of Higher Education in Jiangsu Province [CXLX12-0851]
- Suzhou Scholar Program [14317363]
- NNSF [81273487]
- PAPD
- SRF for ROCS [K513201011]
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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 metabolism-enhancing 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-naphthIcetic acid (NAA) and 3.0 mg L-1 6-benzyladenine (6-BA), C. arachnoidea cells in suspension culture grew rapidly, yielding 20E (124.14 mu g 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 mu M AgNO3 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. (C) 2014 Elsevier Masson SAS. All rights reserved.
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