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Quantification of Taxol and 10-Deacetyl Baccatin III in the Leaf and Cell Suspension Cultures of Two Taxus L. Species

H. Ahadi; M.H. Mirjalili; M. Farzaneh; H. Rezadoost

Volume 41, Issue 3 , December 2018, , Pages 95-104

https://doi.org/10.22055/ppd.2018.20838.1440

Abstract

Abstract Background and Objectives Taxanes are a family of diterpene alkaloids which is currently known as the most important anticancer compounds to suppress and inhibit cell growth, ... Read More Abstract Background and Objectives Taxanes are a family of diterpene alkaloids which is currently known as the most important anticancer compounds to suppress and inhibit cell growth, differentiation and proliferation in indefinitely known cancer cell lines. The most well recognized member of the taxane family is taxol (paclitaxel), a minor cytotoxic component derived originally from yew bark (Taxus brevifolia). 10-Deacetylbaccatin III (10-DAB), which is a homolog of taxol, can be extracted at high yields from the needles of Taxus baccata and is currently considered the most appropriate precursor for semi-synthesizing taxol and its analogue, taxotere (docetaxel). The natural source of both compounds is the inner bark and needles of several Taxus species, especially T. baccata and T. brevifolia. Plant cell culture is an environmentally sustainable source of taxol and offers several advantages as it is not subjected to weather, season or contamination. The aim of the present study was to evaluate taxol and 10-DAB content of two Taxus species and their ability in production of both taxanes through cell culture. Materials and Methods The plant materials of T. baccata and T. berevifolia were collected from Shahid Beheshti University (35° 48' N, 51° 23' E at an altitude of 1785 m) and botanical garden of Faculty of Agriculture, University of Tehran, Karaj (35° 48' N, 50° 59' E at an altitude of 1238 m), respectively. Callus culture of both species was initiated from internodal segments on B5 medium supplemented with plant growth reaulators. Cell suspension culture was established on the medium supplemented with 2.0 mg L−1 naphtalene acetic acid (NAA) and 0.1 mg L−1 benzyl amino purine (BAP). Cell viability was determined by Evan’s blue staining test. After 21 days, the amount of taxol and 10-DAB was analysed by HPLC. Results The results showed that mother stocks of T. baccata and T. brevifolia contain 45 and 27 µg/g DW of taxol and 40 and 4 µg/g DW of 10-DAB, respectively. Cell growth was estimated by measuring fresh weight (FW) and dry weight (DW). Results showed that the maximum growth was reached after 21 days of cultivation. Cell cultures of T. baccata and T. brevifolia produced 540 and 20 µg/l taxol and 1 and 110 µg/l of 10-DAB, respectively. Taxol production in T. baccata cell culture was 26-fold higher than in T. berevifolia cell culture. Discussion As far as our literature survey could ascertain, growth and taxol production in T. baccata cell cultures have previously been reported. Although, T. baccata cells are more potent for the production of taxol, T. brevifolia cell culture can be considered for the production of 10-DAB, a homolog of taxol, as an appropriate precursor for semi-synthesizing taxol and its analogue, taxotere (docetaxel). However, it is essential to study and quantify the effect of selected key medium components on growth as well as product accumulation and strike a balance between the two to enhance the yield and productivity.

Essential Oil Quantity and Quality of Different Plant Organs from Perovskia abrotanoides Karel in Natural Habitat of North Khorasan Province

S.H. Pourhosseini; M.H. Mirjalili; S. Nejad Ebrahimi; A. Sonboli

Volume 40, Issue 4 , March 2018, , Pages 53-62

https://doi.org/10.22055/ppd.2018.13446

Abstract

Abstract Background and Objectives Chemical variability among populations according to their geographical and bio climatic distribution imposes that conservation strategies of populations ... Read More Abstract Background and Objectives Chemical variability among populations according to their geographical and bio climatic distribution imposes that conservation strategies of populations should be made appropriately, taking into account these factors. Perovskia, with the common Persian name of "Brazemble", a small genus from Lamiaceae family, is distributed in various regions of Asia, as Iran, Afghanistan and Pakistan. The genus is represented in Iran by only three species as P. abrotanoides, P. atriplicifolia and P. artemisoides. The plant is an aromatic shrub which mainly grow in mountains at an altitude of 2200 to 4200 m from Northeastern across center to Southeastern of Iran. All parts of the plant are aromatic, but Perovskia is not edible. The plant is used in Iranian folk medicine as an analgesic in rheumatic pains, treatment of leishmaniasis, fever and headache. The present study, was planned to evaluate variations in essential oil composition and contents of different Plant Organs from P. abrotanoides. Materials and Methods In this study, different plant organs (leaf, flower and stalk) of P. abrotanoides were collected from natural habitat in North Khorasan province and their essential oil content and composition were studied. The experiment was arranged in a completely randomized design with three replications for the essential oil contents. Essential oils were analyzed by Gas chromatography (GC) and Gas chromatography–mass spectrometry (GC-MS) in respect to their chemical composition. Results Essential oil contents of leaf, flower and stalk were 1.0, 2.3 and 0.7% (w/w), respectively. The total number of compounds identified and quantified was thirty-nine in leaf, thirty-two in flower, and thirty-five in stalk, representing 99.4, 99.8, and 98.7 % of the total essential oil, respectively. The major compounds of the essential oil were α-bisabolol (2.6-29 %), 1,8-cineole (11.4-16.5 %), camphor (10.1-21 %), α-pinene (1.9-16.2 %) and δ-3-carene (7.4-11.3 %). Results showed that leaf essential oils of the plant characterized with high content of oxygenated sesquiterpenes (41 %). Oxygenated monoterpenes, were rich in flower (42.1 %) and stalk (49.5 %). The main volatile compound identified in the leave was α-bisabolol, which reached a concentration of 29%. The content of essential oil in the flower was higher than that in the leave and stalk. Discussion Results of the current study show that the main compounds and content percentage of essential oil were different in each plant parts. By using the hydrodistillation, 32 to 39 compounds were identified representing 98.7% to 99.4 % of the total oil components. The essential oil analysis revealed that α-bisabolol, 1, 8-cineole, camphor, α-pinene and δ-3-carene, were main essential oil constituents in all organs. The variation observed for essential oil content was 0.7% to 2.3% in stalk and flower, respectively. Chemical variation of P. abrotanoides essential oils from different plant parts, which is significant for conservation and breeding programs, can be considered by medicinal plants breeders and pharmaceutical industries for breeding and processing uses.

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Author = Mirjalili, M.H.

Quantification of Taxol and 10-Deacetyl Baccatin III in the Leaf and Cell Suspension Cultures of Two Taxus L. Species

Abstract

Essential Oil Quantity and Quality of Different Plant Organs from Perovskia abrotanoides Karel in Natural Habitat of North Khorasan Province

Abstract