沙漠研究 22-1, 53 -56 (2012 )"ICAL 1 / DT X"

Journal of Arid Land Studies－Refereed Paper－Valorization of Bio-Resources in Semi-Arid and Arid Land- Functional Analysis Group of STREPS Project in Tunisia Junkyu HAN1) and Hiroko ISODA*1)
Abstract: Olive, halophyte and aromatic plants are undeniable natural resources especially in semi-arid and arid land of Tunisia. In
this research, we used the multifaceted bioassay and the molecular biology approaches in valorizing medicinal plants, proteomics andgenomics in particular, to decipher the specific signal transduction that occur in mammalian cells following treatment with components
or extract from olive, halophyte and aromatic plants. The molecular biology approaches provide reliable and quick results inthe determination of the effect of components or extract from olive, halophyte and aromatic plants at the molecular level in mammaliancells to complement the known traditional uses of olive, halophyte and aromatic plants. Molecular biology approach, therefore,
provides opportunities for the establishment of scientific evidence regarding the medicinal properties or therapeutic use of olive,
halophyte and aromatic plants.
Key Words: Aromatic plants, Halophytes, Molecular biology approach, Multifaceted bioassay, Olive

1. Introduction
This project targets the arid land in Tunisia in northern
Africa, and we will deploy the investigation of useful
bio-resources based on food culture and traditional medicinal
information, investigation of vegetation distribution,
investigation of temperature/soil saline characteristics that arefactors of environmental deterioration in arid land,
investigation of habitat environment, perform exhaustive
component analysis and functionality analysis of bio-resourcesthat were selected based on these investigations, and create a
database that encompasses the species/habitat environment
information and functionality/element/chemical compound
information of arid land bio-resources, as well as a barcode
management library of seeds and functional elements.
Furthermore, the project will develop the technologies forcultivation/breeding methods that aim at adapting to theenvironment, and will seek a high-degree utilization of arid
land bio-resources by introducing food utilization andprocessing technologies.
The characteristics of the northern African region is that
unlike the continental type of arid land, the distance between
the Mediterranean Sea and the Sahara Desert is short, resulting
in a large aridity gradient that produces a unique biodiversity.
It is the only holarctic floral region in the Gondwana land
(South America / Africa / Australia / India / Antarctica), andunique flora exist from the plant geography point of view aswell. We are investigating useful biological activities fromthe traditional medicinal plants in North African region.
* Corresponding Author: isoda.hiroko.ga@u.tsukuba.ac.jp
Tennoudai 1-1-1, Tsukuba city, Ibaraki, Japan1) Alliance of Research on North Africa, University of Tsukuba

We seek anti-oxidative activities from local plant resources
which have been used as traditional medicines. Plants thatgrow on arid land possess functions that protect them from thestress of harsh habitat, and it is thought that they contain uniquefunctional components that play such roles. To take an
example from recent studies, it has been reported that olives
and medicinal plants that grow in Sahara desert oases containantioxidant materials (such as polyphenol) that are several
times higher than those found in species growing in theMediterranean region (Omri et al., 2010; Han et al., 2009a;
Abaza et al., 2008) and that these materials possess antitumor
activity, anti-allergic activity, neuronal cell death repressor
activity, skin function activity, etc (Villareal et al., 2010; Han etal., 2009b; Omri et al., 2009; Matsuyama et al., 2009; Kawanoet al., 2009; Han et al., 2007; Yamada et al., 2008; Kashiwagiet al., 2009).
From aromatic plants and olives, we also explore forbioactivities effective against various diseases such as allergy,
nervous disease like Alzheimer's disease, and cancer. In
order to elucidate molecular mechanisms of their action,fractionation of the plant extracts and identification of theactive compounds are performed. Moreover, we investigate
their effect on intracellular signal transduction systems related
to the diseases.

2. Materials and Methods
To evaluate the functional molecular of bio-resources, thevarious methods were carried out. First, we used thebioassays to perform the screening of bioactivity ofTable 1. The bioassay list for functional analysis of bio-resources

Fig. 1. The process of functional analysis from bio-resources.

bio-resources. We performed the basic assay (DPPH assay,Total polyphenol assay, MTT assay, LDH assay, Intracellular
ROS assay, Intracellular F-actin assay, Intracellular ATP assay),
Neuronal assay (Acetylcholinesterase assay, Neurite outgrowth
assay, Neuroprotection assay), Intestinal assay (TER assay,
α-Glucosidase assay, Cholesterol assay, Epithelium
permeability assay), Immunity assay (β-hexosaminidase assay,
Histamin assay, Intracellular Ca ion assay, Immunoactivity
assay), Anti-cancer assay (Leukemia differentiation assay,Cancer migration assay), Lifestyle-related disease assay(Adipogenesis assay, Adiponectin assay, ACE II assay,
Alkaline phosphatase activity assay, Stress recovery assay),Skin assay (Melanogenesis assay, Dermal papilla growth assay,
Epithelium permeability assay) to find a new functional
molecular from olives, halophytes and aroma plants (Table 1).
After bioassay, we performed the molecular biology
experiments to make clear the mechanism of these effects fromolives, halophytes and aroma plants. We carried out thewestern blotting, Real time-PCR, Micro array, Cell cycle,
Proteomics, DNA fragmentation test, Immunostaining, ELISA
to evaluated the effect of functional molecular on proteins andgenes expression (Table 2).

Table 2. The mechanism analysis methods list for functional
molecular.

3. Results and Discussion
In this year, we investigated the 10 varieties of olive, 12species of halophyte and 13 species of aroma plant (Table 3).
The traditional usage information of Tunisia wascontributed to obtain the positive results on screening using
various bioassays. We found that the olive extracts and its
components have anti-oxidant, anti-allergy and anti-cancer
effects. Some of halophyte extracts have anti-oxidant,anti-cancer, anti-fat and activation of energy metabolism
effects. The some of aroma plant extracts and its components
have anti-oxidant, anti-stress and melanogenesis regulation
effects (Table 4). We expect that these finding will be a basic

Table 3. The species number of annual research (2010-2011) on eachbio-resources.

research for the application of functional food, medicine andcosmetic. Moreover, we hope that our research cancontribute to the development of bio-resources region.
The obtained results of SATREPS project were used ascontents of the SATREPS database. About new finding, weTable 4. The results of bio-resources (Olive, Halophyte, Aromaplant) in 2010.

and aroma plants that grow in Tunisia have anti-oxidant
activity, anti-cancer activity, anti-allergic activity, anti-fatactivity, melanogenesis regulation activity, energy metabolism
activity and anti-stress activity, etc.
Acknowledgement
This study was supported by SATREPS project from JSTand JICA.

ReferencesFig. 2. The road map of SATREPS functional analysis group work.

will apply to patent, then present by international journal andconference.
Figure 2 shows that the road map of SATREPS functionalgroup work. To develop the possibilities of these seeds, wehave planning to do the isolation of functional materials andthe clinical test.

4. Conclusion
We investigated anti-oxidative activities from local
bio-resources that have been used as traditional medicines.
Plants that grow on arid land possess functions that protect
them from the stress of harsh habitat, and it is thought that they
contain unique functional components that play such roles.
From our SATREPS project, we found that olives, halophytes

Abaza L., Talorete T.P.N., Yamada P., Kurita Y., Zarrouk M.,
Isoda H. (2008): Induction of Growth Inhibition andDifferentiation of Human Leukemia HL-60 Cells by aTunisian Gerboui Olive Leaf Extract. Bioscience,Biotechnology, Biochemistry, 71 (5): 1306-1312.
Han J., Talorete T., Yamada P., Isoda H. (2009a):
Anti-proliferative and apoptotic effects of olive polyphenols
on human breast cancer MCF-7 cells. Journal of Arid Land
Studies, 19: 367-370.
Han J., Talorete T., Yamada P., Isoda H. (2009b):
Anti-proliferative and apoptotic effects of oleuropein andhydroxytyrosol on human breast cancer MCF-7 cells.
Cytotechnology, 59: 45-53.
Han J., Inoue S., Isoda H. (2007): Effects of silkworm powder
on glucose absorption by human intestinal epithelial cell line
Caco-2. Journal of Natural Medicines, 61: 387-390.
Kashiwagi K., Isoda H., Han J., Irie M., Nabetani H.,
Nakajima M. (2009): Advanced processing of useful foodmaterial for the establishment of a regional development
model in arid areas. Journal of Arid Land Studies, 19:375-378.
Kawano M., Han J., Kchouk M., Isoda H. (2009): Hair growth
regulation by the extract of aromatic plant Erica multiflora.
Journal of Natural Medicines, 63: 335-339.
Matsuyama K., Miyamae Y., Seki Y., Han J., Abderrabba M.,
Morio T., Shigemori H., Isoda H. (2009): Effect ofMediterranean medicinal plant extract on melanogenesis
regulation. Journal of Arid Land Studies, 19: 387-390.
Omri A., Han J., Yamada P., Kawada K., Abdrabbah M., IsodaH. (2010): Rosmarinus officinalis polyphenols activatecholinergic activities in PC12 cells through phosphorylationof ERK1/2. Journal of Ethnopharmacology, 131: 451-458.
Omri A., Han J., Abdrabbah M., Isoda H. (2009):
Anti-neuronal stress effect of Rosmarinus officinalis extract.
Journal of Arid Land Studies, 19: 117-120.
Villareal M., Han J., Yamada P., Shigemori H., Isoda H.
(2010): Hirseins inhibit melanogenesis by regulating the gene
expressions of Mitf and melanogenesis enzymes.
Experimental Dermatology, 19: 450-457.

Yamada P., Zarrouk M., Kawasaki K., Isoda H., (2008):
Inhibitory effect of various Tunisian olive oils on chemical

mediator release and cytokine production by basophilic cells.
Journal of Ethnopharmacology, 116(2): 279-287.