Indole alkaloid production by transformed and non-transformed root cultures ofCatharanthus roseus

Ciau-Uitz, Aldo; Miranda-Ham, M. L.; Coello-Coello, J.; Chi, Bo-ram; Pacheco, Luz; Loyola‐Vargas, Víctor M.

doi:10.1007/bf02632126

Cited by 64 publications

(25 citation statements)

References 25 publications

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“…Subsequently, until the 30th day, conductance decreased in concert with growth and could be quantitatively correlated to hairy root biomass. This relationship between biomass of C. roseus hair roots and conductance corroborates the findings of Toivonen et al (1990) and Ciau-Uitz et al (1994). Trends of FW/DW and medium conductance were similar in 4% sucrose cultures (data not shown).…”

Section: Growth Of Hairy Rootssupporting

confidence: 91%

“…These levels of initial sucrose in culture medium are in the range for optimal growth of hairy root clones of C. roseus as well as optimal accumulation of ajmalicine, serpentine, and tabersonine (Shanks and Bhadra, 1997). The growth rates of C. roseus hairy root clone LBE-6-1 obtained in this study-3.6 and 4.1 days-are faster than the published values of 6.5 days by Ciau-Uitz et al (1994) and 5 days by Jung et al (1992) and Sim et al (1994) and comparable to 4 days by Toivonen et al (1989) for hairy roots of C. roseus. The basal salt composition in most cases was Gamborg's B5, but inocula were not optimized for growth rate; optimized inoculum developed by Bhadra and Shanks (1995) was employed in this study.…”

Section: Discussioncontrasting

confidence: 52%

“…Published reports of hairy root cultures of C. roseus have focused largely on ajmalicine, serpentine, and/or catharanthine in a variety of short-and medium-term studies (Ciau-Uitz et al, 1994;Islas et al, 1994;Parr et al, 1988;Toivonen et al, 1989Toivonen et al, , 1990Vasquez-Flota et al, 1994). Though tabersonine is known to occur in hairy roots (Parr et al, 1988;Toivonen et al, 1989), its kinetics of accumulation has not been studied.…”

Section: Introductionmentioning

confidence: 96%

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Transient studies of nutrient uptake, growth, and indole alkaloid accumulation in heterotrophic cultures of hairy roots ofCatharanthus roseus

Bhadra

Shanks

1997

Biotechnol. Bioeng.

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The kinetics of growth, the uptake of macronutrients, and the accumulation of indole alkaloids were investigated in long-term, heterotrophically cultured transgenic (''hairy'') roots of Catharanthus roseus. Tabersonine, ajmalicine, and serpentine were monitored over a 70-day period. The doubling time [dry-weight (DW) basis] of C. roseus hairy roots in B5/2 nutrients supplemented with 3% sucrose was 3.6 days. NH 4 + , NO 3 , − and P i were depleted sequentially from culture medium by hairy roots, while sugars remained undepleted. The growth-limiting nutrient was inorganic nitrogen, NH 4 + and NO 3 − , with exponential-phase overall biomass yields of 34.1 and 5.0 g DW/g nutrient, respectively. Extracellular pH decreased to 4.8 in early exponential phase of culture growth from the initially adjusted value of 5.7, increased subsequently to a maximum of 7.7 in late exponential phase of growth coincident with the maximum of fresh weight (FW)/DW ratio, before decreasing to 5.5-5.0. The organic acids, pyruvate, formate, lactate, and succinate were excreted by hairy roots starting in late phase of exponential growth, possibly resulting in the late-culture pH decrease. Tabersonine accumulation was distinctly growth associated with maximum specific and total yields of 1.15 mg/g DW and 5.6 mg/L, respectively, in late-exponential phase of growth. Serpentine accumulation was non-growth associated with increasing specific and total levels in stationary growth phase: 1.3 mg/g DW and 10.5 mg/L, respectively. The accumulation of ajmalicine also appeared growth associated.

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Section: Growth Of Hairy Rootssupporting

confidence: 91%

Section: Discussioncontrasting

confidence: 52%

Section: Introductionmentioning

confidence: 96%

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Transient studies of nutrient uptake, growth, and indole alkaloid accumulation in heterotrophic cultures of hairy roots ofCatharanthus roseus

Bhadra

Shanks

1997

Biotechnol. Bioeng.

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“…In addition, the maximum production of both alkaloids coincides with the late exponential biomass growth phase. This direct correlation between biomass growth and production of alkaloids is also noted in other plant species (Ciau-Uitz et al 1994;Almagro et al 2011), where the highest level of secondary metabolite production was correlated with the cell division activity (Hirano & Komamine 1994). However, Harfi et al (2013) reported an inverse relationship in Datura hairy root lines, in which the maximum production of alkaloids was observed in the stationary phase of hairy root growth.…”

Section: Correlation Between Biomass and Alkaloids Accumulationmentioning

confidence: 71%

“…Among three selected lines, the fastest growing line was LP10, which showed 24.48 times increase in biomass production as compared with control. Other studies in C. roseus have observed that hairy root growth rate increased 20 times compared with the initial inoculums in a period of 30 d (Ciau-Uitz et al 1994). The potential of biomass production was found to be dependent on hairy root line and its adaptation to liquid medium (Toivonen et al 1989;Hughes et al 2004).…”

Section: Selection Of Efficient Hairy Root Linesmentioning

confidence: 89%

Screening and kinetic studies of catharanthine and ajmalicine accumulation and their correlation with growth biomass inCatharanthus roseushairy roots

Benyammi¹,

Paris

Khelifi-Slaoui³

et al. 2016

Pharmaceutical Biology

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Context Catharanthus roseus (L.) G. Don (Apocynaceae) is still one of the most important sources of terpene indole alkaloids including anticancer and hypertensive drugs as vincristine and vinblastine. These final compounds have complex pathway and many enzymes are involved in their biosynthesis. Indeed, ajmalicine and catharanthine are important precursors their increase can lead to enhance levels of molecules of interest. Objective This study aims at selecting the highest yield of hairy root line(s) and at identifying best times for further treatments. We study kinetics growth and alkaloids (ajmalicine and catharanthine) accumulation of three selected hairy root lines during the culture cycle in order to determine the relationship between biomass production and alkaloids accumulation. Materials and methods Comparative analysis has been carried out on three selected lines of Catharanthus roseus hairy roots (LP10, LP21 and L54) for their kinetics of growth and the accumulation of ajamalicine and catharanthine, throughout a 35-day culture cycle. The methanolic extract for each line in different times during culture cycle is analyzed using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Results Maximum accumulation of the alkaloids is recorded for LP10 line in which the peak of ajmalicine and catharanthine accumulation reached to 3.8 and 4.3 mg/g dry weight (DW), respectively. This increase coincides with an exponential growth phase. Discussion and conclusion Our results suggest that the evolution of accumulation of ajmalicine and catharanthine are positively correlated with the development of the biomass growth. Significantly, for LP10 line the most promising line to continue optimizing the production of TIAs. Additionally, the end of exponential phase remains the best period for elicitor stimuli. ARTICLE HISTORY

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Quantification of metabolites in the indole alkaloid pathways ofCatharanthus roseus: Implications for metabolic engineering

Shanks

Bhadra

Morgan

et al. 1998

Biotechnol. Bioeng.

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In this article, we present a review of the current state of metabolic engineering in Catharanthus roseus. A significant amount of research has contributed to characterization of several individual steps in the biosynthetic pathway of medicinally valuable alkaloids. However, knowledge of the regulation of these pathways is still sparse. Using hairy root cultures, we studied the responses of alkaloid metabolism to environmental stimulation such as light and elicitation. Through precursor feeding studies, the putative rate‐limiting steps of the terpenoid pathway in hairy root cultures also have been examined. Relating this knowledge to specific events at the molecular level, and the cloning of corresponding genes are the next key steps in metabolic engineering of the C. roseus alkaloids. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:333–338, 1998.

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Indole alkaloid production by transformed and non-transformed root cultures ofCatharanthus roseus

Cited by 64 publications

References 25 publications

Transient studies of nutrient uptake, growth, and indole alkaloid accumulation in heterotrophic cultures of hairy roots ofCatharanthus roseus

Transient studies of nutrient uptake, growth, and indole alkaloid accumulation in heterotrophic cultures of hairy roots ofCatharanthus roseus

Screening and kinetic studies of catharanthine and ajmalicine accumulation and their correlation with growth biomass inCatharanthus roseushairy roots

Quantification of metabolites in the indole alkaloid pathways ofCatharanthus roseus: Implications for metabolic engineering

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