Effect of differentiation on the regulation of indole alkaloid production in Catharanthus roseus hairy roots

Moreno‐Valenzuela, Oscar A.; Galáz-Ávalos, Rosa M.; Minero-García, Yereni; Loyola‐Vargas, Víctor M.

doi:10.1007/s002990050539

Cited by 30 publications

(14 citation statements)

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“…In the case of TIAs analysis, there is no consensus on mobile phase selection. An effective separation can be achieved using a saline buffer with potassium phosphate or ammonium acetate as aqueous phase, and methanol or acetonitrile as organic phase (Bhadra et al 1993;Moreno-Valenzuela et al 1998;Choi et al 2002;. These methods seem to be inspired from the early works of Renaudin (1984) on the separation of ajmalicine, catharanthine, serpentine, vindoline and vinblastine.…”

Section: Hplc Methods For the Analysis Of Tiasmentioning

confidence: 98%

Analysis of Catharanthus roseus alkaloids by HPLC

Hisiger

Jolicœur

2007

Phytochem Rev

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Catharanthus roseus is a medicinal plant from which secondary metabolites used in chemotherapy to treat diverse cancers are extracted. The well known high value metabolites vincristine and vinblastine are just 2 of 130 alkaloids that can be found in C. roseus. However, only few (~11) of this high number of chemical entities are frequently analyzed and even fewer (~8) are available commercially. For more than 30 years, different analytical techniques have been developed to isolate and identify C. roseus metabolites, and then allowing revealing the therapeutic potential of C. roseus metabolites. Among few approaches, high performance liquid chromatography (HPLC) technique is still widely used for the separation and analysis of secondary metabolites such as those from C. roseus. This article thus reviews the most recent developments in HPLC analysis of alkaloids from C. roseus. Diverse considerations that are crucial to the efficiency of secondary metabolites separation and identification steps, such as biomass manipulation, extraction phase and protocols, HPLC separation and analysis protocols are reviewed in details.Examples of spectra obtained using the most common detectors are also shown and suggestions are made on how to proceed in developing efficient separation and identification methods at the analytical and semi-preparative scales.

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Section: Hplc Methods For the Analysis Of Tiasmentioning

confidence: 98%

Analysis of Catharanthus roseus alkaloids by HPLC

Hisiger

Jolicœur

2007

Phytochem Rev

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“…Many factors are responsible for this, but a lack of cellular or tissue differentiation in cell suspension cultures is considered as one of the important reasons (Moreno et al, 1995). Differentiated cultures, such as shoot or root cultures, transformed hairy root cultures and some green callus cultures of C. roseus synthesize higher amounts of alkaloids than undifferentiated cell suspension cultures, and some of them synthesize vindoline and vinblastine, which cannot be detected in cell suspension cultures (Moreno et al, 1995;Misawa and Goodbody, 1996;O'Keefe et al, 1997;Moreno-Valenzuela et al, 1998).…”

Section: Introductionmentioning

confidence: 97%

Compact callus cluster suspension cultures of Catharanthus roseus with enhanced indole alkaloid biosynthesis

Zhao

Zhu

et al. 2001

In Vitro Cell.Dev.Biol.-Plant

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Compact callus clusters showing a certain level of cellular or tissue differentiation were established from Catharanthus roseus stem and leaf explants in a modified MS liquid induction medium supplemented with 5.37 mM a-naphthaleneacetic acid and 4.65 mM kinetin. In the induction medium most leaf explants developed into friable half-closed hollow callus clusters, whereas in the same medium containing 2,4-dichlorophenoxyacetic acid instead of a-naphthaleneacetic acid, most leaf explants were induced to form dispersed cell suspension cultures. Characteristics of these different types of suspension cultures were compared, and the results showed that the compact callus clusters could synthesize indole alkaloids 1.9-and 2.4-fold higher than the half-closed hollow callus clusters and dispersed cell cultures, respectively. The degree of compaction expressed by the ratio of fresh weight to dry weight of these suspension cultures was correlated to indole alkaloid production. Our studies also postulated that the level of cellular/tissue differentiation might be responsible for these different alkaloid synthesis capabilities. Sucrose regime affected some properties (the size, degree of compaction, differentiation level) of the compact callus cluster cultures and therefore influenced alkaloid production.

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“…The antiserum anti-G10H represents an important biochemical tool in the study of the regulation of this P450 enzyme as it has the ability to recognize G10H in C. roseus and other Apocynaceae. G10H activity was monitored during growth of the C. roseus hairy root J1 line, a stable root culture that accumulates indole alkaloids, particularly ajmalicine, catharanthine and yohimbine (Islas-Flores et al, 1994;Moreno-Valenzuela et al, 1998;Vázquez-Flota et al, 1994). G10H activity was almost undetectable at the beginning of the culture cycle and it then rose to 8-9 pKat mg microsomal protein À1 from day 6 to 12 (Fig.…”

Section: Resultsmentioning

confidence: 99%