Assessing the limitations to terpenoid indole alkaloid biosynthesis in <i>Catharanthus roseus</i> hairy root cultures through gene expression profiling and precursor feeding

Goklany, Sheba; Loring, Ralph H.; Glick, James; Lee‐Parsons, Carolyn W. T.

doi:10.1002/btpr.204

Cited by 47 publications

(36 citation statements)

References 74 publications

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“…Therefore, up-regulation of TDC alone is evidently not sufficient to explain the large increase of serpentine induced by CrWRKY1. In addition, feeding the precursor, tryptamine, to C. roseus hairy roots does not lead to significant increases in serpentine or ajmalicine production (Goklany et al, 2009); simply increasing the concentration of tryptamine cannot explain how CrWRKY1 preferentially activates the serpentine branch of the pathway. We speculate that CrWRKY1 genetically interacts with other genes to regulate the TIA pathway (Fig.…”

Section: Overexpression or Repression Of Crwrky1 Affects The Accumulamentioning

confidence: 98%

The Transcription Factor CrWRKY1 Positively Regulates the Terpenoid Indole Alkaloid Biosynthesis in Catharanthus roseus

et al. 2011

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Catharanthus roseus produces a large array of terpenoid indole alkaloids (TIAs) that are an important source of natural or semisynthetic anticancer drugs. The biosynthesis of TIAs is tissue specific and induced by certain phytohormones and fungal elicitors, indicating the involvement of a complex transcriptional control network. However, the transcriptional regulation of the TIA pathway is poorly understood. Here, we describe a C. roseus WRKY transcription factor, CrWRKY1, that is preferentially expressed in roots and induced by the phytohormones jasmonate, gibberellic acid, and ethylene. The overexpression of CrWRKY1 in C. roseus hairy roots up-regulated several key TIA pathway genes, especially Tryptophan Decarboxylase (TDC), as well as the transcriptional repressors ZCT1 (for zinc-finger C. roseus transcription factor 1), ZCT2, and ZCT3. However, CrWRKY1 overexpression repressed the transcriptional activators ORCA2, ORCA3, and CrMYC2. Overexpression of a dominant-repressive form of CrWRKY1, created by fusing the SRDX repressor domain to CrWRKY1, resulted in the downregulation of TDC and ZCTs but the up-regulation of ORCA3 and CrMYC2. CrWRKY1 bound to the W box elements of the TDC promoter in electrophoretic mobility shift, yeast one-hybrid, and C. roseus protoplast assays. Up-regulation of TDC increased TDC activity, tryptamine concentration, and resistance to 4-methyl tryptophan inhibition of CrWRKY1 hairy roots. Compared with control roots, CrWRKY1 hairy roots accumulated up to 3-fold higher levels of serpentine. The preferential expression of CrWRKY1 in roots and its interaction with transcription factors including ORCA3, CrMYC2, and ZCTs may play a key role in determining the root-specific accumulation of serpentine in C. roseus plants.

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Section: Overexpression or Repression Of Crwrky1 Affects The Accumulamentioning

confidence: 98%

The Transcription Factor CrWRKY1 Positively Regulates the Terpenoid Indole Alkaloid Biosynthesis in Catharanthus roseus

et al. 2011

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“…However, the process would become costly if the chemical or natural obtainment of the precursor molecule itself is difficult. To circumvent limitations like precursor unavailability or surplus availability that creats negative feedback, designing the metabolic engineering strategies to amend the expression of related genes may optimize the adequate biosynthesis of the precursor molecule (Morgan and Shanks 2000; Goklany et al 2009). Another judicious alternative and relatively simple method is the co-cultured hairy root systems.…”

Section: Precursor Feedingmentioning

confidence: 99%

Hairy root biotechnology—indicative timeline to understand missing links and future outlook

et al. 2015

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Agrobacterium rhizogenes-mediated hairy roots (HR) were developed in the laboratory to mimic the natural phenomenon of bacterial gene transfer and occurrence of disease syndrome. The timeline analysis revealed that during 90 s, the research expanded to the hairy root-based secondary metabolite production and different yield enhancement strategies like media optimization, up-scaling, metabolic engineering etc. An outlook indicates that much emphasis has been given to the strategies that are helpful in making this technology more practical in terms of high productivity at low cost. However, a sequential analysis of literature shows that this technique is upgraded to a biotechnology platform where different intra- and interdisciplinary work areas were established, progressed, and diverged to provide scientific benefits of various hairy root-based applications like phytoremediation, molecular farming, biotransformation, etc. In the present scenario, this biotechnology research platform includes (a) elemental research like hairy root-mediated secondary metabolite production coupled with productivity enhancement strategies and (b) HR-based functional research. The latter comprised of hairy root-based applied aspects such as generation of agro-economical traits in plants, production of high value as well as less hazardous molecules through biotransformation/farming and remediation, respectively. This review presents an indicative timeline portrayal of hairy root research reflected by a chronology of research outputs. The timeline also reveals a progressive trend in the state-of-art global advances in hairy root biotechnology. Furthermore, the review also discusses ideas to explore missing links and to deal with the challenges in future progression and prospects of research in all related fields of this important area of plant biotechnology.

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“…To know the activity of the TIAs biosynthetic pathway genes, such as tryptophan decarboxylase (TDC), geraniol 10-hydroxylase (G10H), stritosidine synthase (STR), desacetoxyvindole 4-hydroxylase (D4H), and acetyl-CoA:4-O-deacetylvindoline (DAT) [26,27], qRT-PCR analysis was carried out in cultured Catharanthus cells.…”

Section: Qrt-pcr Analysis Of Cmcs and Ddcsmentioning

confidence: 99%

Differential induction of meristematic stem cells of Catharanthus roseus and their characterization

Moon

Venkatesh

et al. 2015

Comptes Rendus. Biologies

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Plant cell culture technology has been introduced for the mass production of the many useful components. A variety of plant-derived compounds is being used in various fields, such as pharmaceuticals, foods, and cosmetics. Plant cell cultures are believed to be derived from the dedifferentiation process. In the present study, an undifferentiated cambial meristematic cell (CMCs) of Catharanthus is isolated using histological and genetic methods, and compared with dedifferentiation-derived callus (DDCs) cultures. Furthermore, differential culture conditions for both DDCs- and CMCs-derived cell lines were established. A suitable media for the increased accumulation of terpenoid indole alkaloids (TIAs) was also standardized. Compared with DDCs, CMCs showed marked accumulation of TIAs in cell lines grown on media with 1.5 mg·mL(-1) of NAA and 0.5 mg·mL(-1) of kinetin. CMCs-derived cultures of Catharanthus, as a source of key anticancer drugs (viblastine and vincristine), would overcome the obstacles usually associated with the production of natural metabolites through the use of DDCs. Cell culture systems that are derived from CMCs may also provide a cost-effective and eco-friendly basis for the sustainable production of a number of important plant natural products.

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Assessing the limitations to terpenoid indole alkaloid biosynthesis in Catharanthus roseus hairy root cultures through gene expression profiling and precursor feeding

Cited by 47 publications

References 74 publications

The Transcription Factor CrWRKY1 Positively Regulates the Terpenoid Indole Alkaloid Biosynthesis in Catharanthus roseus

The Transcription Factor CrWRKY1 Positively Regulates the Terpenoid Indole Alkaloid Biosynthesis in Catharanthus roseus

Hairy root biotechnology—indicative timeline to understand missing links and future outlook

Differential induction of meristematic stem cells of Catharanthus roseus and their characterization

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