Enhancement of Vindoline Production in Suspension Culture of the Catharanthus roseus Cell Line C20hi by Light and Methyl Jasmonate Elicitation

He, Lihong; Yang, Li; Tan, Ronghui; Zhao, Shujuan; Zhi-bi, HU

doi:10.2116/analsci.27.1243

Cited by 18 publications

(4 citation statements)

References 24 publications

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“…Since CYP71D12 (T16H1) was initially isolated from C. roseus cell cultures, we first analyzed transcript levels of both enzymes in this system by quantitative real-time PCR. As expected, CYP71D12 transcripts were easily detectable in cell cultures and were responsive to methyl jasmonate (MeJa) treatment, as reported previously (Schröder et al, 1999;He et al, 2011;Fig. 3A).…”

Section: Cyp71d351 (T16h2) and Cyp71d12 (T16h1) Display Different Trasupporting

confidence: 85%

A Pair of Tabersonine 16-Hydroxylases Initiates the Synthesis of Vindoline in an Organ-Dependent Manner inCatharanthus roseus

et al. 2013

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Hydroxylation of tabersonine at the C-16 position, catalyzed by tabersonine 16-hydroxylase (T16H), initiates the synthesis of vindoline that constitutes the main alkaloid accumulated in leaves of Catharanthus roseus. Over the last decade, this reaction has been associated with CYP71D12 cloned from undifferentiated C. roseus cells. In this study, we isolated a second cytochrome P450 (CYP71D351) displaying T16H activity. Biochemical characterization demonstrated that CYP71D12 and CYP71D351 both exhibit high affinity for tabersonine and narrow substrate specificity, making of T16H, to our knowledge, the first alkaloid biosynthetic enzyme displaying two isoforms encoded by distinct genes characterized to date in C. roseus. However, both genes dramatically diverge in transcript distribution in planta. While CYP71D12 (T16H1) expression is restricted to flowers and undifferentiated cells, the CYP71D351 (T16H2) expression profile is similar to the other vindoline biosynthetic genes reaching a maximum in young leaves. Moreover, transcript localization by carborundum abrasion and RNA in situ hybridization demonstrated that CYP71D351 messenger RNAs are specifically located to leaf epidermis, which also hosts the next step of vindoline biosynthesis. Comparison of high-and low-vindoline-accumulating C. roseus cultivars also highlights the direct correlation between CYP71D351 transcript and vindoline levels. In addition, CYP71D351 down-regulation mediated by virus-induced gene silencing reduces vindoline accumulation in leaves and redirects the biosynthetic flux toward the production of unmodified alkaloids at the C-16 position. All these data demonstrate that tabersonine 16-hydroxylation is orchestrated in an organ-dependent manner by two genes including CYP71D351, which encodes the specific T16H isoform acting in the foliar vindoline biosynthesis.

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Section: Cyp71d351 (T16h2) and Cyp71d12 (T16h1) Display Different Trasupporting

confidence: 85%

A Pair of Tabersonine 16-Hydroxylases Initiates the Synthesis of Vindoline in an Organ-Dependent Manner inCatharanthus roseus

et al. 2013

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“…The results of gene expression investigation have demonstrated that upregulation of tryptophan decarboxylase (TDC), D4H, and DAT has been observed in C. roseus cultures after light expression. [ 12 13 14 15 16 17 ] Researches have also shown that the content of catharanthine, vindoline, and vinblastine is markedly increased by ultraviolet (UV)-B radiation in C. roseus . [ 13 18 ]…”

Section: Regulation Factors Of Dimeric Indole Alkaloids Dias and Theimentioning

confidence: 99%

Biosynthesis and regulation of terpenoid indole alkaloids in Catharanthus roseus

et al. 2015

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Catharanthus roseus produces a wide range of terpenoid indole alkaloids (TIA). Many of them, such as vinblastine and vincristine, have significant bioactivity. They are valuable chemotherapy drugs used in combination with other drugs to treat lymphoma and leukemia. The TIA biosynthetic pathway has been investigated for many years, for scientific interest and for their potential in manufacturing applications, to fulfill the market demand. In this review, the progress and perspective of C. roseus TIA biosynthesis and its regulating enzymes are described. In addition, the culture condition, hormones, signaling molecules, precursor feeding on the accumulation of TIA, and gene expression are also evaluated and discussed.

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“…quality control, fingerprinting, authentication, standardization or identification of biomarkers. Other approaches include microfractionation bioactivity-based analysis (Hou et al, 2012), chromatographic profiling (Xu et al, 2012), monitoring alkaloid production in cell suspension cultures (He, Yang, Tan, Zhao, & Hu, 2011; He, Yang, Xiong, et al, 2011), metabonomic approaches (Wang et al, 2010), toxicological studies (Liu, Zhu, Li, Yan, & Lei, 2011) or in functional studies (Lorenz, Olsovska, Sulc, & Tudzynski, 2010) using UHPLC-MS where alkaloids are include identified by their retention time, UV spectra, fragmentation pattern data and high-resolution MS data and in some cases confirmed by NMR experiments.…”

Section: Ultra High-pressure Lc-msmentioning

confidence: 99%

New Methods of Analysis and Investigation of Terpenoid Indole Alkaloids

Goldhaber-Pasillas

Choi

Verpoorte

2013

New Light on Alkaloid Biosynthesis and Future Prospects

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Introduction 235 2. Extraction and Purification Methods 236 2.1 Solvent extraction 237 2.2 Ionic liquids 239 2.3 Molecular imprinted polymers 239 2.4 Supercritical CO 2 240 3. Analytical Methods 242 3.1 HPLC and hyphenated techniques 242 3.2 Ultra high-pressure LC-MS 243 3.3 Gas chromatography coupled to mass spectroscopy 248 3.4 High-speed counter-current chromatography 251 3.5 Capillary electrophoresis 253 3.6 Quantitative nuclear magnetic resonance 259 4. Applications in Fingerprint Analysis 260 4.1 NMR and LC-MS-based metabolic fingerprinting on TIAs 260 5. Conclusions 262 References 263

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Enhancement of Vindoline Production in Suspension Culture of the Catharanthus roseus Cell Line C20hi by Light and Methyl Jasmonate Elicitation

Cited by 18 publications

References 24 publications

A Pair of Tabersonine 16-Hydroxylases Initiates the Synthesis of Vindoline in an Organ-Dependent Manner inCatharanthus roseus

A Pair of Tabersonine 16-Hydroxylases Initiates the Synthesis of Vindoline in an Organ-Dependent Manner inCatharanthus roseus

Biosynthesis and regulation of terpenoid indole alkaloids in Catharanthus roseus

New Methods of Analysis and Investigation of Terpenoid Indole Alkaloids

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