Effects of Adding Vindoline and MeJA on Production of Vincristine and Vinblastine, and Transcription of their Biosynthetic Genes in the Cultured CMCs of Catharanthus roseus

Abstract

Vincristine and vinblastine were found by Liquid Chromatography-Mass Spectrometry (LC-MS) in Catharanthus roseus cambial meristem cells (CMCs) jointly treated with 0.25 mM vindolineand methyl jasmonate (MeJA), suggesting that C. roseus CMCs contain a complete set of the enzymes which are in response to convert vindoline into vincristine and vinblastine. Based on the facts that the transcript levels of vindoline-biosynthetic genes ( STR, SGD and D4H) were up-regulated instead of being down-regulated by adding itself to the culture, and that the transcriptional factor ORCA3 was up-regulated simultaneously, we further confirmed that the transcription of STR, SGD, D4H was manipulated by ORCA3.

Stable Internal Reference Genes for Normalizing Real-Time Quantitative PCR in Baphicacanthus cusia under Hormonal Stimuli and UV Irradiation, and in Different Plant Organs

“…B. cusia is still a less-studied species at the molecular level. Terpenoid indole alkaloids (TIAs), derivatives of shikimate and terpenoid pathways, are important medicinal ingredients in B. cusia , and they are activated in response to hormonal stresses (Schluttenhofer et al, 2014 ; Zhang W. et al, 2015 ). Shikimate kinase and 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) are the main enzymes involved in the synthesis of TIAs (Veau et al, 2000 ; Kasai et al, 2005 ).…”

Interest of cellular differentiation in the production of vincristine and vinblastine in suspension cultures of Catharanthus roseus (L.) G Don.

“…This fact implies that cells in suspension cultures globally contained the machinery for VC and VB synthesis provided some degree of differentiation within cell aggregates existed. Indeed, it was recently discovered that in vitro cultured cambial meristem cells contained complete sets of enzymes that are responsible for the production of VC and VB from vindoline, though vindoline supplementation was required (Zhang et al, 2015). Most enzymes responsible for vindoline synthesis from tabersonine are already elucidated (tabersonine 16-hydroxylase (T16H), O-methyltransferase (OMT), N-methyltransferase (NMT), desacetoxyvindoline-4-hydroxylase(D4H), and deacetylvindoline-4-Oacetyltransferase (DAT)) (Yu, Zhu, Wang, & Wen, 2015).…”

“…These reports have claimed that production capability is closely related to cell differentiation, though no further details are provided. Moreover, suspension cultures with VC and VB production capability have also been reported (Taha, Shams, Nazif, & Seif-El-Nasr, 2014;Zhang et al, 2015), although no inferential about this capacity was provided.…”

“…On the other hand, there have been reports of vindoline producing cultures under heterotrophic conditions (Naaranlahti et al, 1989;Scott, Mizukami, Hirata, & Lee, 1980). Indeed, it was recently discovered that suspension cultures of cambial meristem cells contained complete sets of enzymes that are responsible for the production of VC and VB (Zhang et al, 2015) from vindoline. Thus, there must be alternative metabolic pathways in in vitro heterotrophic cultures for vindoline, VC and VB, which could be activated by cell differentiation.…”

Artemisinic Acid Serves as a Novel ORCA3 Inducer to Enhance Biosynthesis of Terpenoid Indole Alkaloids in Catharanthus roseus Cambial Meristematic Cells

“…TIAs were extracted and quantified according to the reported method by HPLC-DAD and HPLC-MS [11,14]. Commercial standards were used to identify and quantify the corresponding alkaloids.…”