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Boletín Latinoamericano y del Caribe de Plantas Medicinales y Aromáticas 15 (4): 206 - 214
ISSN 0717 7917
www.blacpma.usach.cl

Artículo Original | Original Article

Triterpenes and β-agarofurane sesquiterpenes from tissue culture of

Maytenus boaria
[Triterpenos y β-agarofuranos sesquiterpenos desde cultivo de tejidos de Maytenus boaria]

Julio Alarcon & Carlos L. Cespedes

Laboratorio de Síntesis y Biotransformación de Productos Naturales
Departamento de Ciencias Básicas, Universidad del Bío-Bío, Chillán, Chile
Contactos | Contacts: Julio ALARCON - E-mail address: [email protected]

Abstract: Leaf explants of Maytenus boaria were induced towards callus tissue culture with different mixture of cytokinins and auxins.
MeOH extract of callus was partitioned with AcOEt and water, and through repeated chromatography procedures were isolated and
identified, four triterpenes and three β-agarofuran sesquiterpenes.

Keywords: Maytenus boaria, tissue culture, Celastraceae, terpenoids production

Resumen: Explantes de hojas de Maytenus boaria fueron inducidos a formar callos mediante diferentes mezclas de citoquininas y auxinas.
Un extracto metanólico de los callos fue fraccionado con AcOEt y agua, y mediante repetidas cromatografías fueron aislados e identificados
siete compuestos, cuatro triterpenos y 3 sesquiterpenos del tipo agarofurano.

Palabras clave: Maytenus boaria, cultivo de tejidos, Celastraceae, producción de terpenoides

Recibido | Received: July 17, 2015

Aceptado | Accepted: November 14, 2015
Aceptado en versión corregida | Accepted in revised form: February 9, 2016
Publicado en línea | Published online: July 30, 2016
Declaración de intereses | Declaration of interests: This manuscript is based in part on work supported by a grant from Comisión Nacional de Investigación Científica y
Tecnológica de Chile (CONICYT) through FONDECYT program grant # 1130463.
Este artículo puede ser citado como / This article must be cited as: J Alarcon, CL Cespedes. 2016. Triterpenes and β-Agarofurane Sesquiterpenes from tissue culture of
Maytenus boaria. Bol Latinoam Caribe Plant Med Aromat 15 (4): 206 – 214.

206
Alarcon et al. Sesquiterpenes from tissue culture of Maytenus boaria

INTRODUCTION glycoprotein-dependent multidrug resistance (MDR)

Plants produce a wide variety of so called secondary phenotype of several human cancer cells (Spivey et
metabolites (Berenbaum, 2002). The compounds are al., 2002).
frequently accumulated by plants in smaller Many pharmaceuticals are produced from
quantities than are primary metabolites (Croteau et plant such as L-DOPA, morphine, codeine, reserpine
al., 2000; Dewick, 2009). Plant secondary and the anticancer drugs vincristine, vinblastine and
metabolites are synthetized in specific pathway and taxol. Some of these secondary metabolites are quite
sites of production can vary between kinds of expensive because of their low abundance in the
compounds as well as between plant species. plant, often less than 1% of the total carbon, or
Moreover, some molecules can be synthetized by all storage usually occurring in dedicated cell or organs.
tissues, whereas other are produced in a specific The plant cell, tissue and organ cultures are
tissue or even cell-specific fashion (Yazdani et al., considered as an alternative way to produce the
2011). Production of natural product by plants is not corresponding secondary metabolites. This progress
always satisfactory. Some plants are difficult to has notably concerned knowledge of enzyme
cultivate, or may grow very slowly. Another way is activities and regulation of biosynthetic pathways
by the synthesis chemistry, but many compounds (Kirakosyan & Kaufman, 2009).
present complex structures. For these reasons, In Chile, the genus Maytenus is represented
different biotechnological tools can be used to by fours species, M. boaria, M. chubutensis, M.
produce plant secondary metabolites of commercial disticha, and M. magellanica. In a previous paper,
interest (Verpoorte et al., 2002). Considerable have been reported on the isolation of dihydro-
advances have been made in the production of agarofuran sesquiterpenes from seeds of M. boaria,
secondary metabolites in plant cell suspension and from aerial part of M. disticha, M. chubutensis
cultures. In general, these advances were developed and M. magellanica (Gonzalez et al., 1989; Alarcon
through the manipulation of media ingredients, et al., 1993; Muñoz et al., 1993; Gonzalez et al.,
especially plant growth hormones, through the 1994; Alarcon et al., 1995; Alarcon et al., 1998). For
development of an efficient cloning procedure and another hand, the inhibition of the
the use of sensitive analytical methods. The use of acetylcholinesterase has been reported by agarofurans
medium manipulation is generally a random approach isolated from Chilean Maytenus (Cespedes et al.,
and is, therefore, time-consuming; however it has 2001; Alarcon et al., 2008).
showed good results in plant cell culture system In a continuation of our studies on these
(Heinstein, 1985; Ramachandra & Ravishankar, plants, the objectives of the present work was
2002; Matkowski, 2008; Sree et al., 2010) induced callus in first time and then evaluate the
The family Celastraceae encompasses 98 capacity by to produce characteristic compounds of
genera comprising approximately 1210 species that these plants under an exploration program forward
are widely distributed throughout Asia, Africa and production of bioactives metabolites with promissory
the Americas, these species are trees and shrubs and pharmacological applications.
sometime climbing or vining. In Chile the
Celastraceae family comprises one genus, Maytenus, MATERIAL AND METHODS
with four species. Particularly from plants of General experimental procedures
Celastraceae family a large number of highly The NMR spectra (400MHz for 1H and 100 MHz for
13
oxygenated dihydro-β-agarofuran sesquiterpenoids C) were measure in CDCl3 (which also provided the
and triterpenoids have been isolated (Brüning & lock signal) with Bruker spectrometers. The chemical
Wagner, 1978). Many are considered to be important shift was assigned with distortion-free enhancement
chemotaxonomic indicators. Members of this group of polarization transfer (DEPT) using a flip angle of
of natural products are of particular interest because 135°. IR spectra were recorder on Shimadzu FTIR-
have antifeedant, insecticidal, cytotoxic, 8400 instrument.Analytical plates (silica gel, Merck
immunosuppressive, anti-HIV, and antitumor 60G) were rendered visible by spraying with H 2SO4-
activities, as well as their ability to reverse the P- Ac2O followed by heating to 120° C.

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Alarcon et al. Sesquiterpenes from tissue culture of Maytenus boaria

Plant material 1464, 1189, 1035, 996. 1H NMR (400 MHz, CDCl3)
Young leaves from plant of M. boaria were δ ppm (∫, m, J(Hz); assignment): 3.13 (1H, ),
thoroughly washed with tap water, surface sterilized 4.50(1H, d), 4.65 (1H, d). 13C NMR (100 MHz,
in 70% EtOH for 1 min, rinsed twice with sterile CDCl3) δ ppm 38.4 (C-1), 27.9(C-2), 78.60(C-3),
distilled water, immersed in 1.5% sodium 38.8(C-4), 55.10(C-5), 17.8 (C-6), 35.3(C-7), 40.9(C-
hypochloride for 10 min and rinsed four times with 8), 50.3 (C-9), 37.4(C-10), 22.80(C-11), 25.10(C-12),
sterile distilled water. Callus tissue was induced on 36.40 (C-13), 42.28(C-14), 26.4(C-15), 35.6(C-16),
MS medium (Murashige & Skoog, 1962) and 48.6(C-18), 41.28(C-21), , 33.40, 31.10, 27.90 ,
Hiroshi-Harada supplemented with different 2,4- 19.10, 14.80 150.8(C-22), 110.10 (C-30). MS: m/z
dichlorophenoxy acetic acid (0, 2.5 and 5.0 mg/L), (%) [M]+ 426(14), 411(8), 383(1), 218(30), 207(8),
indol acetic acid (0.25 to 1.0 mg/L), indol butyric 189(21), 149(15), 125(10), 97(56), 69(87), 43(100).
acid (0.25 to 1.0 mg/L), kinetin (0.5 mg/L), 2-
isopentenyladenine (0.5 mg/L), 6-bencilaminopurine Betulin
(0.25 to 0.75 mg/L), giberellic acid (0,5 to 1.0 mg/L) 2: was obtained as amorphous white solid. Mp.
concentrations. The pH of the medium was adjusted 261°C. IR(KBr) νmax cm-1 3440, 2868, 1638, 1432,
to 5.7 with 1N NaOH and agar added at 7 g/L before 1388. 1H NMR (400 MHz, CDCl3) δ ppm (∫, m,
autoclaving for 20 min at 121º C. The cultures were J(Hz); assignment): 4.67 (1H,s, H-29), 4.57(1H, s, H-
maintained at 25 ± 1º C, under a 16 h photoperiod 29), 3.8 (1H, d, J= 12 Hz, H-28), 3.31 (1H, d, J=12
provided by cool white fluorescent lamps (45 Hz, H-28), 3.17 (1H, m, H-3), 1.67 (3H, s, H-30),
mmol.m-2.s-1). Calluses were routinely transferred at 1.01 (3H,s), 0.97(3H,s), 0.96(3H, s), 0.81 (3H, s),
3 week intervals. 0.73(3H, s). 13C NMR (100 MHz, CDCl3) δ 38.4 (C-
1), 27.4(C-2), 78.5 (C-3), 38.8 (C-4), 55.1 (C-5), 17.3
Isolation of products (C-6), 35.2(C-7), 40.9(C-8), 50.3(C-9), 37.4(C-10),
Fresh cells (200 g) were extracted with MeOH at 21.7 (C-11), 25.3(C-12), 38.2(C-13), 42.9(C-14),
room temperature overnight. After filtration the 26.7(C-15), 29.5(C-16), 47.8(C-17), 48.7 (C-18),
residue was further extracted with hot MeOH for 3 h. 48.3(C-19), 30.1(C-20), 34.8(C-21), 150.3(C-22),
The resulting methanol extracted was concentrated at 23.4(C-23), 23.4(C-24), 16.1(C-25), 18.5(C-26),
reduced pressure in rotatory evaporator at 40° C and 15.0(C-27), 60.5(C-28), 21.7(C-29), 110.6(C-30).
250 mb to yield a syrupy methanol extract (3.5 g). MS: m/z (%) [M]+442(2), 411(65), 222(8), 220(28),
The methanol extract was dissolved in distilled water, 203(70), 207(80), 189 (100).
diluted with methanol to a ratio 60:40
methanol:water, placed in separator funnel, and Oleanolic acid
washed with ethyl acetate (EtOAc). The EtOAc 3: was obtained as amorphous white solid. mp 293-
phases combined and concentrated under reduced 296. IR(KBr) νmax cm-1 3383, 2937,1686, 1460. 1H
pressure (1.5 g). The concentrated EtOAc was NMR(400 MHz, CDCl3), δ ppm (∫, m, J(Hz);
repeatedly chromatographed on a silica gel column assignment): 0.75(3H,s), 0.78 (3H,s), 0.92 (3H,s),
(column diameter 2.5 cm, height 55 cm, 200-425 0.94 (3H,s), 0.98 (3H,s), 1.08 (3H,s), 1.13 (3H,s),
mesh) using n-hexane, CH2Cl2:MeOH, and MeOH 4.45 (1H, dd, J=4.56 Hz, H-3), 5.25 (1H, t, J=3.76
afford 1 (0.020 g, 0.57%), 2 (0.025 g, 0.71%), 3 Hz, H-12). 13C NMR (100 MHz, CDCl3) δ ppm:
(0.030 g, 0.86%), 4 (0.032 g, 0.91%), 5 (0.006 g, 38.9(C-1), 28.4(C-2), 79.2 (C-3), 39.5(C-4), 55.4(C-
0.17%), 6 (0.003 g, 0.086%), and 7 (0.008 g, 0.23%). 5), 18.5 (C-6), 33.3(C-7), 39.8(C-8), 47.8(C-9),
The known compounds were identified by NMR 37.4(C-10), 23.8(C-11), 122.7(C-12), 144.8(C-13),
experiment and comparison with published spectral 41.8 (C-14), 28.4(C-15), 23.7(C-16), 46.8(C-17),
data. 41.2(C-18),46.6(C-19), 31.6(C-20), 34.0(C-21),
32.8(C-22), 28.8 (C-24), 16.5(C-25), 17.4(C-26),
Lupeol 26.2(C-27), 183.5(C-28), 33.4(C-29), 23.1(C-30).
1: was obtained as amorphous white solid. mp MS: m/z (%) [M]+ 456(5), 412(3), 248(100), 203(50),
215°C.IR(KBr) νmax cm-1 3311, 2946, 2870, 1638, 167(25), 44(51).

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Alarcon et al. Sesquiterpenes from tissue culture of Maytenus boaria

β-amyrin J=1.0 Hz, H-6), 2.38 (1H, d, J=3.0 Hz, H-7), 5.32
4: was obtained as amorphous white solid.mp 186°C. (1H, d, J=4.0 Hz, H-8), 5.90 (1H, d, J=1.0 Hz, H-9),
IR νmax 3450, 2945, 2850, 1460, 1385. 1H NMR (400 1.44 (3H, s, H-12), 1.40 (3H, s, H-13), 1.23 (3H, s,
MHz, CDCl3) (∫, m, J(Hz); assignment): δ 5.18 (1H, H-14), 4.32(1H, d, J=12.5 Hz, H-15), 4.24 (1H, d,
t, J = 3.2 Hz, H-12), 3,22 (1H, dd, J = 4.4,10.8 Hz, J=12.5 Hz, H-15). 13C NMR(100 MHz, CDCl3) δ
H-3),1.93(1H, dd, J=4.0, 13.7 Hz, H-19β), 1.89(1H, 69.02 (C-1), 71.42 (C-2), 73.70 (C-3), 41.61 (C-4),
td, J=4.0, 14.0 Hz, H-15β), 1.80 (1H, m, H-22), 91.45 (C-5), 79.87 (C-6), 48.11(C-7), 32.10 (C-8),
1.70(1H, td, J =4.0, 14 Hz, H-16β) 1.13 (3H, s), 73.33 (C-9), 53.38(C-10), 85.59 (C-11), 30.65 (C-
0.99(3H,s), 0.96(3H,s), 0.92 (3H, s), 0.86 (6H,s), 12), 26.34 (C-13), 20.22 (C-14), 16.22 (C-15). MS :
0.83(3H, s), 0.78(3H, s),0.68(1H, d, J=11 Hz, H-5). m/z (%) [M]+ 590(2), 548(2), 530(4), 488(2), 485(1),
13
C NMR(100 MHz, CDCl3) 38.27(C-1), 26.49, 468(1), 428(2), 408(2), 348(3), 306(6), 261(2),
78.54(C-3), 38.14(C-4), 54.75(c-5), 17.96(C-6), 202(3), 105(100).
30.78(C-7), 39.34(C-8), 47.28(C-9), 36.46(C-10),
23.04(C-11), 121.26(C-12), 144.68(C-13), 41.62(C- 1α,2α,6β,8α,15-pentaacetoxy-9β-benzoyl-dihydro-β-
14), 27.64(C-15), 26.16(C-16), 31.99(C-17), agarofuran
47.18(C-18), 46.37(C-19), 30.57(C-20), 33.26(C-21), 7: was obtained by crystallization from n-hexane.Mp
36.68( (C-22), 27.89(C-23), 15.12(C-24), 15.12(C- 250-251°C. IR(nujol) νmax cm-1 2950,
1
25), 16.34(C-26), 25.26(C-27), 27.64(C-28), 1725,1715,1460, 1380, 720. H NMR (400 MHz,
32.83(C-29), 22.78(C-30). MS: m/z (%) [M]+426(6), CDCl3), δ ppm (∫, m, J(Hz); assignment): 5.71(1H, d,
218(100), 207(29), 203(47), 189(25). J=3.5 Hz, H-1), 5.59(1H, dd, J=2.35, 3.5 Hz, H-2),
1.77 (1H, m, H-3α), 2.49 (1H, m, H-3β), 2.39 (1H,
2β,6β-diacetoxy-1α,9β-dibenzoyl-3β-hydroxy- ddq, J=1.2, 6.5, 7.5 Hz, H-4), 6.38 (1H, d, J=1.0, H-
dihydro-β-agarofuran 6), 2.38 (1H, brd, J=3.0 Hz, H-7), 5.27 (1H, d, J=3.0
5: was obtained as white amorphous powder. M.p. Hz, H-8), 5.52(1H, s, H-9), 1.56(3H, s, H-12),
176-178°C. IR (CHCl3) νmax cm-1 3520, 3480, 3010, 1.43(3H, s, H-13), 1.16 (3H, 6, J=7.5 Hz, H-14), 5.10
2950, 2910, 1750, 1740, 1720, 1600, 1400, 1270, (1H, d, J=12.5 Hz, H-15), 4.53 (1H, d, J=12.5 Hz, H-
1112, 1049. 1H NMR (400 MHz, CDCl3) δ ppm (∫, m, 15).13C NMR (100 MHz, CDCl3) δ 71.6(C-1),
J(Hz); assignment): 1.19 (3H, d, J=7.6 Hz, H-14), 69.2(C-2), 31.9(C-3), 32.7(C-4), 89.8(C-5), 74.9(C-
1.46 (3H, s, H-12), 1.48 (3H, s, H-13), 1.57(3H, s, H- 6), 53.1(C-7), 77.8(C-8), 72.9(C-9), 52.6(C-10,
15), 1.86(3H, s), 2.15(3H, s), 2.24 (1H, m, H-8), 2.42 77.5(C-11), 25.8(C-12), 30.3(C-13), 10.9(C-14),
(1H, m, H-4), 4.0(1H, t, H-3), 5.01(1H, d, J=6.4 Hz, 65.3(C-15).MS: m/z (%) [M]+632, 617[M-Me](2),
H-9), 5.36(1H, dd, J=2.6,11 Hz, H-2), 5.44 (1H, s, H- 105(100), 77(2).
6), 6.36 (d, J=11.0 Hz, H-1). 13C NMR (100
MHz,CDCl3) δ 69.01 (C-1), 71.40(C-2), 73.70 (C-3), RESULTS AND DISCUSSION
41.61(C-4), 91.45(C-5), 79.85(C-6), 48.11(C-7), Many plant species that provide medicinal herbs have
32.10(C-8), 73.30(C-9), 53.38(C-10), 85.59(C-11), been scientifically evaluated for their possible
30.65(C-12), 26.34(C-13), 20.22(C-14), 16.22(C-15). medical applications. It has been mentioned that
MS: m/z (%) 579.2[M-Me](6), 552(19), 472(2), natural habitats for medicinal plants are disappearing
457(3), 416(10), 397(2), 352(2), 248(5), 105(100). fast and together with environmental and geopolitical
instabilities; it is increasingly difficult to acquire
1α,2α,6β,8α-tetraacetoxy-9β-benzoyl-15-hydroxy- plant-derived compounds. This has prompted
dihydro-β-agarofuran industries, as well as scientists to consider the
6: was obtained as oil. IR(CHCl3) νmax cm-1 3565, possibilities of investigation into cell cultures as
3020, 2920, 2840, 1735,1365, 1270, 1230, 1090, 710. alternative supply for the production of plant
1
H NMR (400 MHz, CDCl3) δ ppm (∫, m, J(Hz); pharmaceuticals. The tissue culture cells typically
assignment): 5.75 (1H, d, J=3.30, H-1), 5.50 (1H, dd, accumulate large amounts of secondary compounds
J=4.0, 6.8 Hz, H-2β), 1.83 (1H, ddd, J=1.2, 2.5, 15.0 only under specific conditions.
Hz, H-3α), 2.49(1H, ddd, J=3.9, 6.5, 15.0 Hz, H-3β), For many years, synthetic chemist have been
2.39(1H, ddq, J=1.2, 6.4, 7.5 Hz, H-4), 5.58(1H, d, afforded the challenge of developing synthesis of

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Alarcon et al. Sesquiterpenes from tissue culture of Maytenus boaria

such components but often due structural complexity applications such as antitumoral, insecticidal and
the resulting multi-step synthesis rarely find other (Alarcón et al., 1998). It is very important
application in large scale production as required. On consider that the synthesis conventional of
this way, our goal is to know the condition for the agarofurans it has only allowed a few compounds
production of promissory secondary metabolites from (Spivey et al., 2002; Ishiyama et al., 2013). Therefore
Celastraceae plants, since these have shown potential is necessary development new alternatives for have
pharmacological properties for pharmacological molecules by biological studies.

Table 1
The callus inducing media (CIM) with Gamborg’s B5 salts, vitamins and plant growth regulators.
Callus
CIM Growth regulators (mg/L)
formation

CIM -1 IAA(0.25); IBA (0.25);2-iP (0.5) -

CIM- 2 IAA(0.50); IBA (0.50);2-iP (0.5) +
CIM-3 IAA(0.75); IBA (0.75);2-iP (0.5) +
CIM-4 IAA(1.00); IBA (1.00);2-iP (0.5) +

CIM 5 IAA(0.50):BAP(0.5):GA3(0.75) -

CIM-6 IAA (0.75); BAP(0.5); GA3 (0,75) +

CIM-7 IAA (1.0); BAP(0.5); GA3 (0,75) +
CIM-8 IAA (1.5); BAP(0.5); GA3 (0,75) +
CIM-9 IAA (1.0);IBA (1.0); GA3 (0.50) +
CIM-10 IAA (1.5);IBA (1.5); GA3 (0.50) +

CIM-11 IAA (1.0);BAP (1.0); GA3 (0.75) +

CIM-11 IAA (1.0);IBA (1.0); GA3 (1.0) +

IAA = indol acetic acid; IBA = indol butyric acid; 2-iP = 2-isopentenyladenine;
BAP= bencyl aminopurine; GA3 = giberellic acid

Plant cell suspension cultures are the most maintained under sterile condition was used to
popular strategy among the in vitro plant culture establish callus cultures. They treated with IBA, IAA,
techniques- it is simple, provides homogenous and 2-iP, and GA3 in different proportion resulted into
fast growing material, and is easy to scale up. good callus formation and maximum callus biomass
However, the biosynthetic machinery necessary to with friable features after 8 weeks of culture (Figure
generate the various target metabolites may not be 2 and Figure 3). The concentration used in the
available or may afford only small amounts of the induction of callus formation (Table 1) did not
desired compounds. We verified initially formation induced organogenesis. The callus material was
from the leaf explants from M. boaria plants subcultures monthly and after 8 months appeared

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Alarcon et al. Sesquiterpenes from tissue culture of Maytenus boaria

green, yellow less and white. The combination used 4). The compounds 5 was previously isolated aerial
in callus inducing medium (CMI), CIM 6, 7, 8 and 9 part of Maytenus magellanica with 0.032% yield
have the best biomass formation. The other CIM (Gonzalez et al., 1992), and the compound 6 and 7
present formation of callus but the velocity of were isolated from M. chubutensis with 0.13% and
upgrowth biomass formation is slowly. The crude 0.12% respectively. Our results indicate that the yield
methanol extract of the callus was partitioned find with our procedures show a production five time
between ethyl acetate and water. Repeated higher than occur in plant, thus leaf explant cultures
chromatography of ethyl acetate fraction yield four of M. boaria are a promising source of terpenoids
triterpene identified as lupeol (1), betuline (2), because the yield are greater in the cell cultures than
oleanolic acid (3), and β-amyrine (4), and three in the plant, and a base for scaling-up adaptation in
sesquiterpene agarofuran (5-7) (Figure 1 and Figure view of its future production in bioreactors.

CH2
CH2
H3C H3C CH3
H3C

CH3 CH3 CH3

CH3 CH3
CH3 CH3 R
OH
CH3
CH3
CH3
HO
HO
H3C CH3 HO
H3C CH3 H3C CH3
1 Lupeol 2 Betulin
3 R = COOH, oleanolic acid
4 R = Me, amyrin

OH OAc
OBz OBz OAc OBz OAc OBz
CH3
AcO AcO OAc AcO OAc

HO
CH3 CH3 CH3
O O O
CH3 OAc CH3 CH3 OAc CH3 CH3 OAc CH3
5 6 7
Figure 1
Compounds isolated from callus of Maytenus boaria

Previously, best callogenic response have 1981; Corsino et al., 1998; Antunez et al., 2013).
been observed on NAA in combination with BAP, These studies have been oriented to the production of
using explant of Maytenus ilicifolia, Peritassa QMTs therefore developing tissue cultures have from
campestris, or Tripterygium wilfordii (Nakano et al., roots, as these compounds accumulate in the roots of
1997; Buffa et al., 2004; Antunez et al., 2013) which, plants belonging to this family. The results shows
in accordance with our results, show the combination that it is possible to achieve concentrations of
of cytokinins and auxins is a good mixture by metabolites to 8 times larger than it is possible to
inducing friable callus in leaf explant of M. boaria. obtain from the roots (Coppede et al., 2014). Other
Tissue cultures of some species of the family works has been oriented to the resource conservation
Celastraceae seem to be excellent sources of (Chen, 2009; Sanchez-Chuquicusma et al., 2015).
quinonemethide triterpenoids (QMTs) (Kutney et al.,
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Alarcon et al. Sesquiterpenes from tissue culture of Maytenus boaria

Figure 2
Growth kinetics of callus culture of Maytenus boaria on MS medium
supplemented with IBA, IAA, 2-iP, and GA3.

Figure 3
Callus from explants of leaf M. boaria

The major advantage of the cell cultures secondary metabolites under a large-scale would
include synthesis of bioactive secondary metabolites, open new possibilities for development of novel
running in controlled environment, independently of pharmaceuticals with several biological activities.
climate and soil conditions. The use of in vitro plant
cell culture for the production of chemicals and ACKNOWLEDGEMENTS
pharmaceutical has made great strides building on This manuscript is based in part on work supported
advances in plant science. by a grant from Comisión Nacional de Investigación
From this study a best response was obtained Científica y Tecnológica de Chile (CONICYT)
with mixture of BAP and NAA in vitro production of through FONDECYT program grant # 1130463.

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Alarcon et al. Sesquiterpenes from tissue culture of Maytenus boaria

Figure 4
Comparative TLC between apolar extract of leaves of M. boaria and fraction from extract of callus of M.
boaria.

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