Polyisoprenoids Composition from Araucaria heterophylla and

Casuarina equisetifolia Leaves

Mohammad Basyuni

1,2

, Irma Deni

, Bejo Slamet

, Yuntha Bimantara

, Rahmah Hayati

, Rizka

Amelia

, Hirosuke Oku

and Hiroshi Sagami

Department of Forestry, Faculty of Forestry, Universitas Sumatera Utara, Jl. Tri Dharma Ujung No. 1 Medan, North

Sumatera 20155, Indonesia

Center of Excellence for Mangrove, Universitas Sumatera Utara, Medan, North Sumatera 20155, Indonesia

Molecular Biotechnology Group, Tropical Biosphere Research Center, University of the Ryukyus, 1 Senbaru, Nishihara,

Okinawa 903-0213, Japan

Institute of Multidisciplinary Research for Advanced Material, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, 980-

8577, Japan

ameliarizka0@gmail.com, okuhiros@comb.u-ryukyu.ac.jp, yasagami@tagen.tohoku.ac.jp

Keywords: chemotaxonomic marker, dolichol, dehydrodolichol, pine

Abstract: This current work examines the polyisoprenoids (dehydrodolichol or polyprenol and dolichol) profiling and

conformation from Norfolk Island pine Araucaria heterophylla (Araucariaceae) and Australian pine

Casuarina equisetifolia (Casuarinaceae). The pattern and structure of polyisoprenes were determined by

two-plate thin layer chromatography (2P-TLC). The polyisoprene pattern in the leaves was found and

categorized into two categories. Group-I, showing a majority of dolichols over dehydrodolichols was

detected in A. heterophylla. These dolichols showed as one longer dolichol tribe (C

–C

110

). Group-II,

exhibiting the incidence of the pair polyprenols and dolichols, was traced in C. equisetifolia. Dolichol

concentrations were faintly extra richness detected comparing to dehydrodolichols (approximately

54%:46%) in this species. Dolichols with chain length of C

–C

and shorter dehydrodolichol (C

–C

)

were detected in C. equisetifolia. This study suggested that different pattern of ficaprenols, shorter-chain

and longer dolichols are modulated in both pine species.

1 INTRODUCTION

Higher plants are renowned to generate secondary

metabolites containing polyisoprenes or

polyisoprene. The occurrence and profile of

polyisoprenoids has been demonstrated in numerous

organs tropical and subtropical plants (Jankowski et

al., 1994; Tateyama et al., 1999; Skorupinska-Tudek

et al., 2008; Surmacz and Swiezewska, 2011;

Basyuni et al., 2016, 2017, 2018a; Arifiyanto et al.,

2017; Basyuni and Wati, 2017; Sagami et al., 2018).

These papers showed the ubiquitous composition

and occurrence of polyisoprenoids in the flora.

Several works have been revealed for biological

and pharmacological properties of plant species such

as Norfolk Island pine Araucaria heterophylla

(Araucariaceae) and Australian pine

Casuarina equisetifolia (Casuarinaceae) (Aslam et

al., 2013; Elkady et al., 2018). It has been reported

that the resin isolate of A. Heterophylla depicted

antiulcerogenic activity in opposition to ethanol-

activated stomach ulcers in Sprauge Dawely rats

(Abdel-Sattar et al., 2009). Furthermore, this extract

showed variable cytotoxic activities contrary to the

breast (MCF7) and colon (HCT116) row of cancer

cells (Abdel-Sattar et al., 2009). Elkady et al., (2018)

reported that chemical mark and antiproliferative

influence of important oils of A. heterophylla.

Likewise, C. equisetifolia has been shown to have

potential antibacterial activity (Parekh et al., 2006).

The physiological significance of C. equisetifolia

has been playing an essential role in reply to cold

pressure (Li et al., 2017).

To get more understanding into the critical role

of polysioprenoids in plant species, the feature data

on the pattern and distribution of polyisoprenoids

from plant species are entirely needed. Thus the

current report proposed to determine the

108

Basyuni, M., Deni, I., Slamet, B., Bimantara, Y., Hayati, R., Amelia, R., Oku, H. and Sagami, H.

Polyisoprenoids Composition from Araucaria heterophylla and Casuarina equisetifolia leaves.

DOI: 10.5220/0008526201080111

In Proceedings of the International Conference on Natural Resources and Technology (ICONART 2019), pages 108-111

ISBN: 978-989-758-404-6

dehydrodolichol and dolichol pattern and

conformation from A. heterophylla and C.

equisetifolia extending our prior studies.

2 MATERIALS AND METHODS

2.1 Chemicals

A standard combination of dolichols (C

-C

) and

dehydrodolichols (C

-C

) as prior applied (Basyuni

et al., 2016) was applied to classify the

polyisoprenes in the experiment. The recognition of

the tribe linking to dehydrodolichols or dolichols

was performed with three triplicates.

2.2 Plant Materials

The leaves of Norfolk Island pine Araucaria

heterophylla (Araucariaceae) and Australian pine

Casuarina equisetifolia (Casuarinaceae) were

collected from Universitas Sumatera Utara campus

in October 2017. Both species usually are rising in

forthright sunlight. In the month of compilation, the

average temperature was 31 ºC with ordinary

moisture of 76%. samples were placed at -20ºC prior

to utilization.

2.3 Extraction of Polyisoprene

A protocol for the isolation of polyisoprene as

already depicted (Basyuni et al., 2018a,b). Simplely,

the leaves of two species were placed on oven at

75°C for 1-2 days. The drained organ (4-6 g) was

blended in and suppressed in chloroform/methanol

then saponified and re-suspended in hexane.

2.4 Determination by Two-plate Thin

Layer Chromatography (2P-TLC)

To determine polyisoprenoid profile, two

approaches were carried out: first-plate TLC (1P-

TLC) and two-plate TLC (2P-TLC) as formerly

reported (Basyuni et al., 2017). The polyisoprenoid

tribe was analyzed by the evaluation of progress on

TLC with the accurate examples of dolichol or

dehydrodolichol. The polyisoprenoids were

calculated using ImageJ ver. 1.46r (Schneider et al.,

2012).

3 RESULTS AND DISCUSSION

3.1 Polyisoprene Pattern and

Distribution

The investigation for polyisoprenes derived the

leaves of A. heterophylla and C. equisetifola, from

North Sumatra, Indonesia was done using 2D-TLC

(Basyuni et al., 2016; Basyuni et al., 2017) lead to

the vibrant parting of dehydrodolichols and

dolichols involving to the carbon chain length.

Tables 1-2 recapitulate the quantitative

determination of polysioprenoids and

dehydrodolichols and dolichols pattern and

configuration with the carbon-chain lengths

provided every species. The amount of TL was the

major in C. equisetifolia leaves and the lowest in A.

heterophylla. In contrast to this observation, the

measure of PI was the uppermost in A. heterophylla

(8.8 mg g

-1

dw), the bottommost concentration of PI

was in C. equisetifolia (6.3 mg g

-1

dw).

The analogous outcomes for TL and PI

concentrations were described for North Sumatran

coastal leaves (Basyuni et al., 2018a). On the other

hand, The TL and PI contents in true and associate

mangrove forests reported in the work were lower

than that studied derived from true and mangrove

associate species (Basyuni et al., 2016; Basyuni et

al., 2017; Basyuni et al., 2018a, b).

Table 1: Pattern and composition of polyisoprenes in two leaves

Plant

Organ

Pol

Dol

% in TL

% in PI

Group

(mg/g dw)

(mg/g)

Pol

Dol

Pol

Dol

A. heterophylla

Leaves

9.0

8.8

7.8

100

C. equisetifolia

Leaves

9.1

6.3

2.9

3.4

2.8

4.0

3.4

46.0

54.0

nd= not found, TL = Total lipids, PI = Polyisoprenes, Pol = Polyprenols/Dehydrodolichols, Dol = Dolichols. Results are

displayed as an average of three repeatation examines.

Polyisoprenoids Composition from Araucaria heterophylla and Casuarina equisetifolia leaves

109

Tabel 2: Dehydrodolichol and dolichol carbon-chain lengths in two plant photosynthetic organs

Plant

Organ

Dehydrodolichol

Dolichol

A. heterophylla

Leaves

40 45 50 55 60 65 70 75 80 85 90 95 100 105 110

C. equisetifolia

Leaves

50 55

75 80 85 90 95

3.2 Analysis Polyisoprene by 2P-TLC

The physical categories of dehydrodolichols and

dolichols in both species were grouped as earlier

termed (Basyuni et al., 2016, 2017, 2018a) into two

groups (I and II). Group-I, possessing dominance of

dolichols over dehydrodolichols (>90%) was

discerned in A. heterophylla leaves. Group-II is

depicting the occurrance of the pair dehydrodolichol

and dolichols, was verified in the leaves of C.

equisetifolia.

Dolichols occurred one dolichol family (C40-

C110) detected in A. heterophylla, while shorter

dehydrodolichol type (as called ficaprenol)

happened in C. equisetifolia (C50-C55) and dolichol

with a carbon chain length of C75-C95 (Table 2). 

Figure 1. 2P-TLC chromatograms of samples from the

leaves of A. heterophylla (A) and C. equisetifolia (B). The

carbon numbers indicate the polyisoprene carbon chain

length.

It is important to observe that significant of

dolichols over dehydrodolichols have been described

in coastal plant and mangroves plants (Basyuni et

al., 2016, 2017, 2018a). The presence and

distribution of both compounds namely

dehydrodolichols and dolichols were characterized

of oil palm (Elaeis guineensis) leaves and non-

mangrove plant species (terrestrial plants or dry land

forests) (Basyuni et al., 2018b; Basyuni and Wati,

2017, 2018).

4 CONCLUSION

This report shed light on the pattern and distribution

of polyisoprenes in A. heterophylla and C.

equisetifolia from North Sumatra province,

Indonesia. The obtainable work specified that the

formation of shorter-chain dehydrodolichols,

shorter-chain dolichols and longer dolichols are

conducted in higher plants.

ACKNOWLEDGMENTS

Sincere thank was due to Universitas Sumatera

Utara for TALENTA grant 2018.

REFERENCES

Abdel-Sattar, E., Monem, A. R. A., Ezzat, S. M., El-

Halawany, A. M., and Mouneir, S. M. 2009. Chemical

and biological investigation of Araucaria heterophylla

Salisb. Resin. Zeitschrift für Naturforschung C, 64(11-

12), 819-823.

Arifiyanto, D., Basyuni, M., Sumardi, Putri, L.A.P.,

Siregar, E.S., Risnasari, I., and Syahputra, I. 2017.

Occurrence and cluster analysis of palm oil (Elaeis

guineensis) fruit type using two-dimensional thin layer

chromatography. Biodiversitas, 18, 1487-1492.

Aslam, M. S., Choudhary, B. A., Uzair, M., and Ijaz, A. S.

2013. Phytochemical and ethno-pharmacological

review of the genus Araucaria–review. Tropical

Journal of Pharmaceutical Research, 12(4), 651-659.

Basyuni, M., Sagami, H., Baba, S., Iwasaki, H., and Oku,

H. 2016. Diversity of polyisoprenoids in ten Okinawan

mangroves. Dendrobiology, 75, 157-175

Basyuni, M., and Wati, R. 2017 Distribution and

occurrence of polysioprenoids in rambutan

(Nepheliumlappaceum). IOP Conference Series: Earth

Environmental Science, 101, 012001.

Basyuni, M., Sagami, H., Baba, S., and Oku, H. 2017.

Distribution, occurrence, and cluster analysis of new

polyprenyl acetones and other polyisoprenoids from

North Sumatran mangroves. Dendrobiology, 78, 18-31.

Basyuni M, Wati R, Sagami H, Sumardi, Baba S, and Oku

H., 2018a. Diversity and abundance of polyisoprenoid

composition in coastal plant species from North

Sumatra, Indonesia. Biodiversitas, 19, 1–11.

Basyuni, M., Wati, R., Tia, A. R., Deni, I., and Prabuanisa,

A. N. 2018b. The occurrence of polyisoprenoids from

the leaves of selected Fabaceae family. Journal of

Physics: Conference Series, 1116, 052010.

Basyuni, M., and Wati, R. 2018. Polyisoprenoids profile

and composition from selected plant Sapotaceae

family. IOP Conference Series: Materials Science and

Engineering, 434, 012104.

Elkady, W. M., and Ayoub, I. M. (2018). Chemical

profiling and antiproliferative effect of essential oils of

ICONART 2019 - International Conference on Natural Resources and Technology

110

two Araucaria species cultivated in Egypt. Industrial

Crops and Products, 118, 188-195.

Jankowski, W. J., Swiezewska, E., Sasak, W., and

Chojnacki, T. 1994. Occurrence of dehydrodolichols

and dolichols in plants. Journal of Plant Physiology,

143(4-5), 448-452.

Li, H. B., Li, N., Yang, S. Z., et al., 2017. Transcriptomic

analysis of Casuarina equisetifolia L. in responses to

cold stress. Tree Genetics & Genomes, 13(1), 7.

Parekh, J., Jadeja, D., and Chanda, S. 2006. Efficacy of

aqueous and methanol extracts of some medicinal

plants for potential antibacterial activity. Turkish

Journal of Biology, 29(4), 203-210.

Schneider, C.A., Rasband, W.S. and Eliceiri, K.W., 2012.

NIH Image to ImageJ: 25 years of image analysis.

Nature methods, 9(7), 671.

Skorupinska-Tudek, K., Wojcik, J., and Swiezewska, E.

2008. Polyisoprenoid alcohols—recent results of

structural studies. The Chemical Record, 8(1), 33-45.

Surmacz, L. and Swiezewska, E., 2011. Polyisoprenoids–

secondary metabolites or physiologically important

superlipids. Biochemical and biophysical research

communications, 407(4), 627-632.

Tateyama, S., Wititsuwannakul, R., Wititsuwannakul, D.,

Sagami, H., and Ogura, K. 1999. Dolichols of rubber

plant, ginkgo and pine Phytochemistry. 51. 11–15.

Polyisoprenoids Composition from Araucaria heterophylla and Casuarina equisetifolia leaves

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