Effetct of chemical compounds on quinine content of Cinchona ledgeriana in cell suspension culture
Effect of Chemical Compounds on Quinine Content of Cinchona ledgeriana in Cell Suspension Culture .
Dedi Satriawan'>, Diah Ratnadewi 1>, Sumaryono2> •
'Department of Biology, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University, Darmaga Campus, Bogor 16680, Indonesia
e-mail: dedi_nendra@yahoo.com
2
1ndonesian Biotechnology Research Institute for Estate Crops, Jalan Taman Kencana No.1, Bogor 16151, Indonesia
ABSTRACT
Quinine has been used as a tonic, appetite enhancer, antipyretic, antimalarial agent, cosmetic ingredients and tanning. Quinine can be obtained from the bar1< of seven or more years old
Cinchona tree. Cell suspension culture is an alternative means to produce secondary ｭ･ｴ｡｢ｯｬｾｳ＠
rapidly and continuously. Quinine, one of alkaloid oompounds, is expected to increase with the
addition of abscisic acid (ABA) and paclobutrazol (PBZ) as growth inhibitors, tryptophan as a precursor and mannitol as an osmotic stress inducer. ABA was used at 1 and 3 ppm, PBZ at 5 and
7 ppm combined with 5.3 gil mannitol, tryptophan at 0.2 and 2 ppm combined with 5.3 gil ｭ｡ｮｾｯｬＮ＠
ABA at 1 and 3 ppm increased the cell suspension growth but reduced the content of
quinine. PBZ at 7 ppm combined with 5.3 gil mannitol and 20 gil sucrose significantly reduced the cell suspension growth but remarkably increased quinine content, even higher than in the bar!<
of Cinchona tree in the field. Combination of mannitol and padobutrazol was effective to inhiM the cell growth but enha1ce the production of quinine.
Keywords: abscisic acid, cell suspension culture, Cinchona, mannitol, padobutrazol, quinine, tryptophan
Introduction
• Quinine has many uses especially as an antimalarial drug and as a biltering agent of
soft drinks.
• Quinine is conventionally harvested from the bar1< of Cinchona trees after 7 to 10 years
of planting.
• Cell culture can be used to produce secondary metabolites, including alkaloids, in very
short and continuous cycles.
ABA is known to enhance as well as to inhibit the growth (Finkelstein and Rock 2002).
this study ABA increased cell growth of Cinchona in A, and A, treatments.
ABA can cause impertection in the process of cell wall formation during cell division due to
impaired synthesis of cellulose. Imperfection in the cell wall increases the rate of required
nutrients to move in and out of cells make the cells to multiply more quickly. ABA induces
the accumulation of protein reserves in somatic embryos of white spruce. The availabiley
of protein reserves increases the process of multiplication and division of plant cells.
• The objective was to determine the effect of different chemical substances on the
growth and level of quinine in a cell suspension culture of Cinchona.
Materials and Methods
2
• Culture conditions: under lamps at light intensity 20 !Jmol photon/m /sec for 12 h
photoperiod and temperature 26 °C.
• Explant: young leaves of C. ledgeriana clone QRC 315.
• Callus initiation: leaf cuts were cultured on WP medium with 3.5 gil gelrite, 30 gil
s• 1crose, 151JM pikloram, 2 !JM BA, 1 !JM phloroglucinol (Sumaryono & Riyadi 2005).
• Callus proliferation: on media similar to the initiation of callus.
30 gil sucrose, 15 !JM picloram, 0.5 !JM BA and 1
• Cell culture: WP liquid medium ｷｾ＠
!JM ploroglucinol in baffle flasks. The flasks were placed on a shaker at 90 rpm
• Treatment: The cells were cultured into WP liquid medium ｷｾｨ＠
151JM picloram, 2 !JM
BA. 1 11M phloroglucinol and the treatments were: 1) 30 gil sucrose (control), 2) 30 gil
sucrose and 1 ppm ABA (A 1), 3) 30 gil sucrose and 3 ppm ABA (A3 ), 4) 20 gil suaose,
5.3 gil mannitol and 0.2 ppm tryptophan (To. 2 ) , 5) 20 gil sucrose, 5.3 gil mannitol and 2
ppm tryptophan (T2) , 6) 20 gil sucrose, ｭ｡ｮｾｯｬ＠
gll5.3 and 5 ppm paclobutrazol (P5 ), 7)
20 gil sucrose, 5.3 gil mannitol and 7 ppm paclobutrazol (P7).
Results and Discussion
• Callus Culture
Callus was formed in the second week after planting. It produced a white callus slightly
browned crumbs mixed with brown compact callus (Figure 1 ).
Figure 3. The growth of C. ledgeriana cell suspension observed by CVS method.
14
12
10
ｾ＠
""
"2
ｾ＠
•
•
6
4
6"'weeks
7"'weeks
2
0
Figure 1. Callus of Cinchona ledgeriana
K
• Cell Culture
AI
A3
11l.2
12
P5
P7
Treatments
Suspension cultures were able to produce enough new cells in just one week on the
homogenization phase.
Figure 4. Quinine level in C. ledgeriana cell suspension
• Quinine Level
• Cell Growth
treatment (up to 15 times from the initial cell
The best cell growth was obtained in ｾ＠
volume) and lowest was in P7 treatment (only Slimes the initial cell volume). Cell growth in
A, and A 1 treatments were higher than the control treatment while in T02 T 2, P5 and P1
treatments were lower. Generally, the peak growth was reached at the 42;\,l day, except in
the treatment of T 2 which was still growing (Figure 2). The cell growth of Cinchona
observed by CVS method can be seen in Figure 2.
Quinine levels of all treatments at 7 weeks of culture were higher than at 6 weeks. The
highest quinine level at 10.9% was in P7 treatment at 7 weeks of cUlture and the lowest
was in treatment To.z at 6 weeks of Ct_Jiture (Figure 4).
Quinine level in all treatments (on average at 5.6%) was the higher than the yields
obtained by Robins eta/. (1986) at 0.01%, Rhodes et af. (1986) at 0.003%, and Ratnadewi
& Sumaryono (2010) at 0.12%.
The highest level of total quinoline (quinine, quinidine, cinchonine and cinchonidine) was
obtained in P7 treatment at 6 weeks of culture followed by P7 treatment at 7 weeks of
culture and the lowest was in T2 treatment at 6 weeks of culture.
20
II
16
ｾ＠
II
0
12
ｾ＠
10
tK
Conclusions
I AJ
• WP liquid medium added with 3 ppm ABA and 30 gil sucrose was the best medium for
the growth of cell suspension of C. /edgeriana .
I AJ
t 11))
ｾ＠ "
•n
&Pl
• PI
II
21
1J
ll
12
19
• Paclobutrazol at 7 ppm, 5.3 gil ｭ｡ｮｾｯｬ＠
and 20 gil sucrose at 7 weeks of culture was
the best for quinine production in Chincona cell culture.
References
Ratnadewi o. Somaryooo. 2010. Quinoline alklMoids in suspension cuftures of Cinchona ledgeriana treated
with vanous substances. HAYAn J. Biosa. 17(4): 179-182.
Rhodes MJC. Payne J, Robins RJ. 1986. cell suspension cultures of Cinchona ledgeriana; II. the effed of a
range of auxins and cytokinin on the production of quinoline alkaloids. Planta Medica: 52: 226-229.
Robins RJ, Payne J, Rhodes MJC. 1986. CeU suspension wltures at Cinchona Jedgeriana; I. growth and ...
quinoline alkaloid production. Planta Medica: 52: 226-225.
Culture Age
SumafYOOO. I. Riyadi 2005. Pertumbuhan kullur kalus dan suspensi sel kina (Ciochooa ledgeriana Moens).
Figure 2. Cell growth curve of C. ledgeriana
MenataPerlcebunan 73(1): 1-11 .
Presented at the IGN TTRC International Conference, 17-18 July 2012, Bogar, Indonesia
THE 1st iiC
I GN-TTRC
I NTERNATIONAL
C ONFERENCE
"BIOTECHNOLOGY FOR HUMAN LIFE"
/
PT. ELO KARSA UTAMA
PREFACE
It is always a great time to meet with scientists to talk about science and
technology since what they say is only the facts of phenomena . This first international
conference held by the IGN-TIRC, the Indonesia-German Network in Teaching, Training
and Research Collaboration, is expected to have pictures of research progresses on
biomolecule and biotechnology in the world, in particular in the Asia region as well as in
Indonesia. The conference also invites students to involve scientifically as they will be
the next developer of this world. That is the reason of having the theme of the
conference that covers large scope of biotechnology researches.
In this two day-conference, 25 research results will be presented orally and 31
are in form of posters. They were sent by scientists/academicians/students from
Germany, Malaysia, Riau, West Sumatra, East Kalimantan, South Sulawesi and North
Sulawesi, East Java, Yogyakarta, West Java, Banten and Jakarta. Six senior scientists have
been invited to give their talk about recent development of their focus of research, five
come from Germany and one from Indonesia.
Although this book contains only abstracts, we hope that the presentations
would be the main focus of the conference that enlighten the discussion among all the
participants. A list of participants along with their email address will be useful to start,
maintain or develop interaction among the participants.
Have a fruitful conference.
Organizing Committee Chairwoman
Diah Ratnadewi
FOREWORD
Since March 2011, seven Indonesian universities have engaged to collaborate
with Kassel University. This collaboration aims at developing academic and scientific
matters, especially for post-garduate studies. Directorate General of Higher Education of
the Ministry of National Education of Republic Indonesia hasbeen promoting as well as
supporting any Indonesian university to increase its quality of education and researches
to international level. Despite much efforts have been carried out at university level,
collaboration with other universities having international reputation in some areas of
study is believed to speed up the target.
Collaboration among the in-country universities is also encouraged since
scientific communication among the scientists would make the researches going faster
to rearch outputs which are needed to help developing this country, i.e. for industries,
medical or agriculture purposes. Fragmented researches in every research laboratoria
are wasting in term of time, money, as well as resources. Therefore the ign-ttrc was
designed to pioneer such collaboration.
Biotechnology is a vast field of study. The mondial progresses and achievement
of this kind of research has proven solving many human problems. Although controversy
on the application of biotechnology results remains strong, the use of biotech products
in some extent has given benefit to human being particularly in the _third world.
Indonesia has developed some regulations dealing with genetically modified organisms
and their products that the acceptance has to be taken with careful and wise.
Several researches on molecular biology and biotechnology have been
developing at IPB. The focuses are to be mentioned: rice plants for resistance to
aluminum or diseases, Jatropha plant for more seed-productive, wheat plant for
adaptation to tropical area, and more others. Almost all the research groups are working
together with partners in Indonesia and in foreign countries.
In this occasion I would like to express that it is a great honor for IPB to be the
host of the first international conferenceof ign-ttrc. We welcome all of you, senior and
junior scientists to our university compound, and wish you all success.
Bogar, the 17th of July 2012
Vice Rector for Research and Collaboration,
· Anas M. Fauzi
TABLE OF CONTENTS
Page
PREFACE ................................................... ... ........................................................................
FOREWORD............................................................... ........................................................
TABLE OF CONTENTS............................................. ...........................................................
AGENDA OF THE CONFERENCE..................................................... .................................
IS 1
Epigenetics:Rapid Evolution, Biodiversity, Environmental Challenges
(Wolfgang Nellen, Benjamin Boesler, Sara Miller, Doreen Meier,
lndra Windhof, Christian Joppich,) ........................ ................... .. .......... ....
IS 2
Metagenome Analysis of Tempeh, a Unique Indonesian Fermented Food
(Cecillia A. Seumahu, Efriwati Sabron, Tati Barus, and Antonius
Suwanto)...... ............ .................................... ............... .................. ........ ............. ....
IS 3
3
4
5
9
12
13
Biomolecular Interaction Analysis in Functional Proteomics
(Prof. Dr. Friedrich W. Herberg).......................................... ..............................
Properties and Functions of dsRNA Binding Proteins in miRNA Mediated
Gene Silencing (Doreen Meier)................................. ......... ... ......... .....................
OP 2
Property of Myoglobin Protein in Rattus norvegicus- Normoxic Lung
Tissue (Rini Puspitaningrum, Guenter Lochnit)... ............................................
OP 3
Identification of New Interaction Partners of PKA (Jennifer Hermann).... ..
OP 4
Molecular Analysis of the Type 2 Diabetes Mellitus-Associated Region of Mitochondrial tRNA DNA in Gorontalonese Cases (Amien R. Ishak,
Risma Dwi Untari, Rini Puspitaningrum, Tri Murtiati, Chris Adhiyanto,
Takenori Nitta, Hattori Yukio, Yasuhiro Yamashiro).........................................
OP 5
Identification and Analysis of Single Nucleotide Polymorphism of Growth
Hormone Gene Exon 3, lntron 3 in Local Cattle Breeds in West Sumatera
Province of Indonesia (Yurnalis, Sarbaini, Arnim, Wolfgang Nellen,
Jamsari)...... ................................. ............ ..............................................................
OP 6
Polymorphisms Study on Several Specimen of Indonesian Coelacanth,
Latimeria menadoensis (Jxchel Feibie Mandagi, Yoshitaka Abe, Masamitsu
Iwata, Sinya Yamauchi, Fransisko Pangalila, Hefry Veibert Dien, Laurentius TX
Lalamentik, Kawilarang Warouw Alex Masengi, Kanako Koyanagi,Hiroki
Arimura and Hidemi Watanabe)............... ..........................................................
OP 7
Production of Double Haploid Orchids through Microspore Embryogenesis
(Ari lndrianto, Chairani Siregar, Sutikno Linuhung, Mekartinita,
Tri Sartikoningsih)...................................................................................................
OP 8
In 'Qtro Flowering of Orange Subfamily as an Alternative Way for Breeding
Techniques (Hasan Basri Jumin)... ......................................................... .............
OP 9
Effect of Aeration on Biomass and Saponin Content of Talinuin ｰ｡ｨｩ｣ｵｬｾｭ＠
Gaertn. Hairy Roots in Balloon-Type Bubble Bioreactor (ArifVachya, Alfinda
Novi Kristanti, Yosephine Sri Wulan Manuhara)............... ...................................
OP 10 The Effects of Saccharomyces cerevisiae and CuS0 4 on the Production of
Saponin in Talinum paniculatum (Jacq) Gaertn Callus (Agus Muji Santoso,
Alfinda Novi Kristanti, Yosephine Sri Wulan Manuhara)......................................
OP 11 Transformation of Aluminum Tolerant Gene Candidate in Rice
(Oryza sativa L.) (Arief Pambudi, Miftahudin, Tetty Chaidamsari)....................
OP 12 Modification of DNA Isolation Protocol from Dried-Leaf Tissues of
Threatened Species "Jernang" (Daemonorops draco (Willd.) Blume)
14
OP 1
(Revis Asra)....... ................................... ............................................. ... ... ...................
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IS 1
IS 2
IS 3
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(Jens Gebauer) ..................... ........................... ... ...... ...........................................
Genes Characterization of Non-aggressive and Aggressive Strain of
PepYLCV from Chilli in West Sumatera (Jamsari)..................... ............ .... ........
Cloning C1 Gene (Rep) of Geminivirus to pGem-T Easy Vector In Escerichia
coli BL21 (EIIy Syafriani, lrwan Darfis, Jamsari).................................................
Cloning of Geminivirus Coat Protein Gene (V1) From Capsicum annuum
(Ester Kristin Natalia, Jamsari, Sutoyo)....................................... ......................
Cloning of V2 Gene (Pre Coat Protein) Pepper Yellow Leaf Curl Virus
(Pepylcv) from Chili Pepper (Capsicum annuum L.) (Siti Nur Aisyah, Ujang
Khairul, Jamsari)............... ................................. ......................................................
Draft Genome Sequence of Streptomyces sp. strain GMR22, Isolated
from Cajupuut Plantation, Wanagama, Yogyakarta, Indonesia (Jaka Widada,
Alimuddin, and Camelia Herdini)..........................................................................
The Localization of Two Genes for Erucic Acid Content using lntervarietal
Substitution Lines in Rapeseed (Brassica napus L.) (Nurhasanah, Berisso
Kebede, Rubens Marschalek, Heiko C. Becker, and Wolfgang Ecke).............
Analysis of Genetic Diversity of Rodent Tuber Cultivars in Indonesia
Based on RAPD Marker (Nesti F. Sianipar, Danny Laurent, Chelen, Listiarini,
and Ariandana Wantho)............ ............ ............ ................................................ ...
Screening of Antifungal Product from Bacteria for Control of Anthracnose
OP8
Diseases Caused by Col/etotrichum gluosporioides In Chilli (Maizar,
I. Suliansyah, U. Khiarul, and Jamsari)...............................................................
Characterization of Glucanase Produced by Bacteria which Inhibit
OP9
the Growth of Pyricularia Oryzae and Rhizoctonia so/ani (Nurul Hikmawati,
Nisa Rachmania Mubarik, Yadi Suryadi, M. Sc)............... ....................................
OP 10 Test of Bacterial Consortium Formulation to Reduce Neck-Blast Disease on
Rice Plants (Zuhay Ratuz Zaffan, Nisa Rachmania Mubarik, Yadi Suryadi)....
OP' 11 Potency of Brevundimonas Sp. Hgp11solated from Gold Mining in Pongkor
Village, Bogor as Mercury Bioremediation Agent (Wahyu lrawati, Patricia,
Yenny Soraya, and Abyatar Hugo Baskoro).........................................................
OP 12 Isolation of Proteolytic Bacteria from GiliMeno Lake- West Nusa Tenggara
fnd Characterization of the Protease Enzyme (Susana Sagitha Raunsay, Ern in
Hidayati, Nisa Rachmania Mubarik) ........................ ............... :........ :....................
OP 13 Isolation and Characterization of Proteolytic Bacteria f rom Cricula
Trifenestrata Cocoon that Influence Quality of Goldsilk (Abdi Gunawan,
Dedy Duryadi Solihin, Nisa Rachmania Mubarik)..........................................
POSTER.................................... .......................................................................................... ...
P1
Determination of Sex of Salacca (Salacca edu/is L.) Based on
PCR-RPD (Ediwirman, Jamsari, I. Suliansyah, Gustian)......................................
P2
Effect of Pretreatmenton the Rice Straw as Substrate on Solid State
FermentatiQn of Trichoderma viride Strain T051 to the Cellulase Activity Sri
(Sugiwati, Siswati Setiasih, Nenci and Haznan Abimayu) ................................-.
P3
Starch Characterization and Yield Variation of "Gebang" Cassava
Regenerated from Irradiated In Vitro Shoots (Supatmi, Enny Sudarmonowati
@
ce
J
Molecular Diversity and Evolution of South-East Asian Ant-Plants (Genus
Macaranga, Euphorbiaceae) (Dr. Daniela Guicking)........................ ...............
Plant Genetic Resources and Their Preservation via Biotechnological and
Other Methods (Klaus J. Dehmer).................................... .................................
Mountain Oases of Northern Oman: Hotspots of Agrobiodiversity
27
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Pl 9
and N.Sri Hartati).. .................................................................................................
Cloning Geminivirus DNA-satellite Sequence of Pepper Plant
(Capsicum annum L.) (Dila Febria, Yusniwati, Herviyanti, Jamsari)............ ..
Spesific Primer Desain Related Potential High Catechin Content on
Gambier Plant (Uncaria gambir (Hunter) Roxb) (lstino Ferita, lrfan
Suliansyah, Gustian, Hamda Fauza, dan Jamsari)............ ........................ ........
DNA Finger print of Several Cassava Genotypes Comprising Different
Carotene Content and Drought Responses (Ahmad Fathoni, N. Sri Hartati,
Enny Sudarmonowati)..................... .....................................................................
Analysis of Aedes aegypti Microsatelite DNA in Endemic Regions of Dengue
Hemorragic Fever (DHF) Padang, Pariaman and Solok City, West Sumatra
(Hasmiwati, Djong Hon Tjong and Rita Maliza)................................................
Influence of Plant Growth Regulators Auxin and Cytokinin to Callus Induction
of Some Pepper (Capsicum annuum L.) Genotypes For Genetic
Transformation (Renfiyeni, Yusniwati, J. Trisno dan Jamsari)...... ...................
Development Protocol of Agrobacterium- Mediated Genetic Transformation
of Eucalyptus pel/ita F. Muell Using Xyloglucanase Gene (N . Sri Hartati,
Andriyani Puspitaningrum, Nita Etikawati, Enny Sudarmonowati)..................
Potency Test Of Several Chitinolitic Bacteria Isolates As An Agent Of Anti
Anthracnose And Its Identification Based On 16S rRNA Sequence Gene {Irina
Sari, lr. Sutoyo, MS, Prof. Dr. sc. Agr. lr. Jamsari, MP)........................... ............
In Silico Analysis on Structure and Prediction Function Changes of Tyrosine
Kinase Domain Insulin Receptor of Diabetes Mellitus Patient (Miftakhun,
N.Y.P., Fatchiyah, Widodo)....................................................................................
Phylogeny of Mangosteen Varieties Based on ITS-nrDNA Gene
(Panca Jarot Santoso).................................... ............ ........................ .....................
Diversity of Gambier (Uncaria gambir (Hunter) Roxb.) based on RAPD
Marker in West Sumatra (Hamda Fauza, Jamsari, lstino Ferita, Azmi Dhalimi,
Ahmad Denian, and Murdaningsih H. Karmana )...............................................
Selection of Antagonist Bacteria from Green Mustard (Brassica juncea L.)
as Biofungicide for Co/letotrichum gloeosporioides, a Causal of Anthracnose
in Chili Pepper (Capsicum sp.) (Rahmi Henda Yani, Aswaldi Anwar,
Jamsari) ..ｾ Ｎ Ｎ Ｎ Ｎ Ｎ Ｎ
Ｎ Ｎ Ｎ Ｎ Ｎ Ｎ ＮＮ Ｎ Ｎ Ｎ Ｎ Ｎ
ＮＮ
ＮＮ Ｎ Ｎ ＮＮＮ Ｎ
ＮＮ
Ｎ Ｎ Ｎ ＮＮ Ｎ Ｎ
Ｎ Ｎ
Ｎ＠
A Preliminary Study: The Effect Of Progesterone To Dopamine Signaling on
Zebrafish (Danio rerio) Embryos (Christianto, A, Kishida, M)...........................
Diversity and Activities of Bacterial Isolates From Coastal Area of BandealitJerrtber, East Java - Indonesia (Dina Fitriyah, Herawati, Sattya Arimurti, and
Kartika Senjarini) ............................................................................. :...... .. :..............
Protein Profiles of Salivary Glands from Aedes aegypti (Dwi Esti F, Syubbanul
Wathon, Rike Oktarianti, Kartika Senjarini)..........................................................
Parasitemic Rates, IL-4 and IFN-y Profile on Mice Model Vaccinated by
Salivary Glands from Anopheles maculatusfor Developing TBV Against
Malaria {Imam Hanafy, lka Agus Rini, Yunita Armiyanti,
and Kartika Senjarini)..............................................................................................
Isolation and Identification of Lipase-Producing Bacteria for
the ｄ･ｧｲ｡､ｴｩｯｾ＠
of Vegetable Oil Wastes (Hishamudin Rahmat,
Teh Chin Een and Sharlina Mat Saad)...................................................................
46
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Page
Effect of Chemical Compounds to Quinine Alkaloids Content of Cinchona
ledgeriana in Cell Suspension Culture (Dedi Satriawan, Diah Ratnadewi,
Ｍ ｾ＠
Sumaryono) ................................................... ......................................................... .
63
Molecular Studies of Genes Controlling Cambial Cell Differentiation and
P21
Secondary Cell Wall Formation (Widi Sunaryo, Andrea Po lie and
Urs Fischer)............... ................................. ............ ... ...............................................
64
Cell-Specific Analysis of Symbiosis-Induced Medicago truncatula Genes
P22
via Laser Microdissection (Anna-Lena Hurter, Claudia Hogekamp, Natalija
65
Hohnjec and Helge Kuster).. ................. ............. ........................... ..................
The Use of Sulfate Reducing Bacteria (SRB) to Reduce Sulfate and Heavy
P23
66
Metals in Acid Mining Water (AMW) (Fahruddin).................. ...... .....................
Tissue Culture of Sago Palm (Metroxylon saguRottb.), a Perennial
P24
Food Crops, with Different Explants Origins in Indonesia (Sumaryono,
I. Riyadi and M.M. Sinta)... ....................................... ............................................
67
Free Energy Analysis for Naphthoquinone Derivatives Binding into HBx Protein
P25
Using Molecular Docking (Firdayani, Harry Noviardi, Pertamawati, Susi
Kusumaningrum) .... ............. .. .........................................................................
68
Molecular Docking Studies on Series Of Potent Quinone Compounds
P26
as Polo-Like Kinase 11nhibitors (Susi Kusumaningrum,Harry Noviardi,
Firdayani )............... ...... ........................ ......................................................... ........ :.
69
P27
Home Composting Using the grEEn Technology (Trina Tallei, Julius Iskandar,
70
Marian Kamagi, Abdul Basith)..................... ................................................... .......
P28
Cytotoxic Activity of Graptophyllum pictum Methanolic Extracts Against Lung
Cancer Cell Line (A549) and Simian Retrovirus Insertion (A549-SRV)
(Rilianawati, Fery Azis Wijaya, Achmad Junaidi, Tarwadi,Jin Hyub Paik).......... 71
P29
Genic Characterization of Indonesian Coelacanth Latimeria menadoensis
(Daisy Monica Makapedua, lxchel Feibie Mandagi, Marco Barucca, Mariko
Forconi,Adriana Canapa, Ettore Olmo,Masa Iwata, AlexMasengi, Fransisco
Pangalila, Florence Longdong)................................................................................. 72
P30
Polymorphism Identification of BMP-15 Gene(Bone Morphogenetic
Protein-15) lnbeangoat(Capra hircus) usingPCR-RFLP Technique (Sri Rahayu,
lrham Hayati, Estri Laras
Arumingtyas-) ............................................................................................................... 73
P31
Identification and Phylogenetic Diversity Based on 16S rRNA Gene Sequence
Analysis of Thermophilic Bacteria from Rimbo Panti Hot Spring (Armaini, Abdi
Dharma, Sumaryati Syukur, Jamsari)........................ ............................................... 74
LIST OF PART)CIPANTS ............ .................................................................................................. 75
P20
Effect of Chemical Compounds to Quinine Alkaloids Content of Cinchona ledgeriana in
Cell Suspension Culture.
Dedi Satriawan 1l, Diah Ratnadewi 1l, Sumaryono 2l
1
lDepartment of Biology, Faculty of Mathematics and Natural Sciences, Bogar
Agricultural University,Darmaga Campus, Bogar 16680, Indonesia
2
llndonesian Biotechnology Research Institute for Estate Crops, Jalan Taman Kencana
No. 1, Bogar 16151, Indonesia
ABSTRACT
Quinine is used for tonic, appetite enhancer, antipyretic, antimalarial, cosmetic
ingredients and tanning. Quinine can be obtained from the bark of seven or more years
old Cinchona tree. Cell suspension culture is an alternative means to obtain secondary
metabolites rapidly and continously. Quinine, one of alkaloid compounds, is expected to
increase with the addition of abscisic acid (ABA) and paclobutrazol (PBZ) as growth
inhibitors, tryptophan as a precursor and mannitol as an osmotic stress inducer. ABA
was used at 1 and 3 ppm, PBZ at 5 and 7 ppm combined with 5.3 g/1 mannitol,
tryptophan at 0.2 and 2 ppm combined with 5.3 g/1 mannitol. ABA at 1 and 3 ppm
increased the cell suspension growth but reduced the content of quinine. PBZ at 7 ppm
combined with 5.3 g/1 mannitol and 20 g/1 sucrose significantly reduced the cell
suspension growth but remarkably increased quinine content, even higher than that was
found in the bark of Cinchona tree in the field. Combination of mannitol and
paclobutrazol was effective to inhibit the cell growth but enhance the production of
quinine.
Keywords: cell suspension culture, Cinchona, mannitol, abscisic acid, paclobutrazol,
tryptophan,quinine.
Dedi Satriawan'>, Diah Ratnadewi 1>, Sumaryono2> •
'Department of Biology, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University, Darmaga Campus, Bogor 16680, Indonesia
e-mail: dedi_nendra@yahoo.com
2
1ndonesian Biotechnology Research Institute for Estate Crops, Jalan Taman Kencana No.1, Bogor 16151, Indonesia
ABSTRACT
Quinine has been used as a tonic, appetite enhancer, antipyretic, antimalarial agent, cosmetic ingredients and tanning. Quinine can be obtained from the bar1< of seven or more years old
Cinchona tree. Cell suspension culture is an alternative means to produce secondary ｭ･ｴ｡｢ｯｬｾｳ＠
rapidly and continuously. Quinine, one of alkaloid oompounds, is expected to increase with the
addition of abscisic acid (ABA) and paclobutrazol (PBZ) as growth inhibitors, tryptophan as a precursor and mannitol as an osmotic stress inducer. ABA was used at 1 and 3 ppm, PBZ at 5 and
7 ppm combined with 5.3 gil mannitol, tryptophan at 0.2 and 2 ppm combined with 5.3 gil ｭ｡ｮｾｯｬＮ＠
ABA at 1 and 3 ppm increased the cell suspension growth but reduced the content of
quinine. PBZ at 7 ppm combined with 5.3 gil mannitol and 20 gil sucrose significantly reduced the cell suspension growth but remarkably increased quinine content, even higher than in the bar!<
of Cinchona tree in the field. Combination of mannitol and padobutrazol was effective to inhiM the cell growth but enha1ce the production of quinine.
Keywords: abscisic acid, cell suspension culture, Cinchona, mannitol, padobutrazol, quinine, tryptophan
Introduction
• Quinine has many uses especially as an antimalarial drug and as a biltering agent of
soft drinks.
• Quinine is conventionally harvested from the bar1< of Cinchona trees after 7 to 10 years
of planting.
• Cell culture can be used to produce secondary metabolites, including alkaloids, in very
short and continuous cycles.
ABA is known to enhance as well as to inhibit the growth (Finkelstein and Rock 2002).
this study ABA increased cell growth of Cinchona in A, and A, treatments.
ABA can cause impertection in the process of cell wall formation during cell division due to
impaired synthesis of cellulose. Imperfection in the cell wall increases the rate of required
nutrients to move in and out of cells make the cells to multiply more quickly. ABA induces
the accumulation of protein reserves in somatic embryos of white spruce. The availabiley
of protein reserves increases the process of multiplication and division of plant cells.
• The objective was to determine the effect of different chemical substances on the
growth and level of quinine in a cell suspension culture of Cinchona.
Materials and Methods
2
• Culture conditions: under lamps at light intensity 20 !Jmol photon/m /sec for 12 h
photoperiod and temperature 26 °C.
• Explant: young leaves of C. ledgeriana clone QRC 315.
• Callus initiation: leaf cuts were cultured on WP medium with 3.5 gil gelrite, 30 gil
s• 1crose, 151JM pikloram, 2 !JM BA, 1 !JM phloroglucinol (Sumaryono & Riyadi 2005).
• Callus proliferation: on media similar to the initiation of callus.
30 gil sucrose, 15 !JM picloram, 0.5 !JM BA and 1
• Cell culture: WP liquid medium ｷｾ＠
!JM ploroglucinol in baffle flasks. The flasks were placed on a shaker at 90 rpm
• Treatment: The cells were cultured into WP liquid medium ｷｾｨ＠
151JM picloram, 2 !JM
BA. 1 11M phloroglucinol and the treatments were: 1) 30 gil sucrose (control), 2) 30 gil
sucrose and 1 ppm ABA (A 1), 3) 30 gil sucrose and 3 ppm ABA (A3 ), 4) 20 gil suaose,
5.3 gil mannitol and 0.2 ppm tryptophan (To. 2 ) , 5) 20 gil sucrose, 5.3 gil mannitol and 2
ppm tryptophan (T2) , 6) 20 gil sucrose, ｭ｡ｮｾｯｬ＠
gll5.3 and 5 ppm paclobutrazol (P5 ), 7)
20 gil sucrose, 5.3 gil mannitol and 7 ppm paclobutrazol (P7).
Results and Discussion
• Callus Culture
Callus was formed in the second week after planting. It produced a white callus slightly
browned crumbs mixed with brown compact callus (Figure 1 ).
Figure 3. The growth of C. ledgeriana cell suspension observed by CVS method.
14
12
10
ｾ＠
""
"2
ｾ＠
•
•
6
4
6"'weeks
7"'weeks
2
0
Figure 1. Callus of Cinchona ledgeriana
K
• Cell Culture
AI
A3
11l.2
12
P5
P7
Treatments
Suspension cultures were able to produce enough new cells in just one week on the
homogenization phase.
Figure 4. Quinine level in C. ledgeriana cell suspension
• Quinine Level
• Cell Growth
treatment (up to 15 times from the initial cell
The best cell growth was obtained in ｾ＠
volume) and lowest was in P7 treatment (only Slimes the initial cell volume). Cell growth in
A, and A 1 treatments were higher than the control treatment while in T02 T 2, P5 and P1
treatments were lower. Generally, the peak growth was reached at the 42;\,l day, except in
the treatment of T 2 which was still growing (Figure 2). The cell growth of Cinchona
observed by CVS method can be seen in Figure 2.
Quinine levels of all treatments at 7 weeks of culture were higher than at 6 weeks. The
highest quinine level at 10.9% was in P7 treatment at 7 weeks of cUlture and the lowest
was in treatment To.z at 6 weeks of Ct_Jiture (Figure 4).
Quinine level in all treatments (on average at 5.6%) was the higher than the yields
obtained by Robins eta/. (1986) at 0.01%, Rhodes et af. (1986) at 0.003%, and Ratnadewi
& Sumaryono (2010) at 0.12%.
The highest level of total quinoline (quinine, quinidine, cinchonine and cinchonidine) was
obtained in P7 treatment at 6 weeks of culture followed by P7 treatment at 7 weeks of
culture and the lowest was in T2 treatment at 6 weeks of culture.
20
II
16
ｾ＠
II
0
12
ｾ＠
10
tK
Conclusions
I AJ
• WP liquid medium added with 3 ppm ABA and 30 gil sucrose was the best medium for
the growth of cell suspension of C. /edgeriana .
I AJ
t 11))
ｾ＠ "
•n
&Pl
• PI
II
21
1J
ll
12
19
• Paclobutrazol at 7 ppm, 5.3 gil ｭ｡ｮｾｯｬ＠
and 20 gil sucrose at 7 weeks of culture was
the best for quinine production in Chincona cell culture.
References
Ratnadewi o. Somaryooo. 2010. Quinoline alklMoids in suspension cuftures of Cinchona ledgeriana treated
with vanous substances. HAYAn J. Biosa. 17(4): 179-182.
Rhodes MJC. Payne J, Robins RJ. 1986. cell suspension cultures of Cinchona ledgeriana; II. the effed of a
range of auxins and cytokinin on the production of quinoline alkaloids. Planta Medica: 52: 226-229.
Robins RJ, Payne J, Rhodes MJC. 1986. CeU suspension wltures at Cinchona Jedgeriana; I. growth and ...
quinoline alkaloid production. Planta Medica: 52: 226-225.
Culture Age
SumafYOOO. I. Riyadi 2005. Pertumbuhan kullur kalus dan suspensi sel kina (Ciochooa ledgeriana Moens).
Figure 2. Cell growth curve of C. ledgeriana
MenataPerlcebunan 73(1): 1-11 .
Presented at the IGN TTRC International Conference, 17-18 July 2012, Bogar, Indonesia
THE 1st iiC
I GN-TTRC
I NTERNATIONAL
C ONFERENCE
"BIOTECHNOLOGY FOR HUMAN LIFE"
/
PT. ELO KARSA UTAMA
PREFACE
It is always a great time to meet with scientists to talk about science and
technology since what they say is only the facts of phenomena . This first international
conference held by the IGN-TIRC, the Indonesia-German Network in Teaching, Training
and Research Collaboration, is expected to have pictures of research progresses on
biomolecule and biotechnology in the world, in particular in the Asia region as well as in
Indonesia. The conference also invites students to involve scientifically as they will be
the next developer of this world. That is the reason of having the theme of the
conference that covers large scope of biotechnology researches.
In this two day-conference, 25 research results will be presented orally and 31
are in form of posters. They were sent by scientists/academicians/students from
Germany, Malaysia, Riau, West Sumatra, East Kalimantan, South Sulawesi and North
Sulawesi, East Java, Yogyakarta, West Java, Banten and Jakarta. Six senior scientists have
been invited to give their talk about recent development of their focus of research, five
come from Germany and one from Indonesia.
Although this book contains only abstracts, we hope that the presentations
would be the main focus of the conference that enlighten the discussion among all the
participants. A list of participants along with their email address will be useful to start,
maintain or develop interaction among the participants.
Have a fruitful conference.
Organizing Committee Chairwoman
Diah Ratnadewi
FOREWORD
Since March 2011, seven Indonesian universities have engaged to collaborate
with Kassel University. This collaboration aims at developing academic and scientific
matters, especially for post-garduate studies. Directorate General of Higher Education of
the Ministry of National Education of Republic Indonesia hasbeen promoting as well as
supporting any Indonesian university to increase its quality of education and researches
to international level. Despite much efforts have been carried out at university level,
collaboration with other universities having international reputation in some areas of
study is believed to speed up the target.
Collaboration among the in-country universities is also encouraged since
scientific communication among the scientists would make the researches going faster
to rearch outputs which are needed to help developing this country, i.e. for industries,
medical or agriculture purposes. Fragmented researches in every research laboratoria
are wasting in term of time, money, as well as resources. Therefore the ign-ttrc was
designed to pioneer such collaboration.
Biotechnology is a vast field of study. The mondial progresses and achievement
of this kind of research has proven solving many human problems. Although controversy
on the application of biotechnology results remains strong, the use of biotech products
in some extent has given benefit to human being particularly in the _third world.
Indonesia has developed some regulations dealing with genetically modified organisms
and their products that the acceptance has to be taken with careful and wise.
Several researches on molecular biology and biotechnology have been
developing at IPB. The focuses are to be mentioned: rice plants for resistance to
aluminum or diseases, Jatropha plant for more seed-productive, wheat plant for
adaptation to tropical area, and more others. Almost all the research groups are working
together with partners in Indonesia and in foreign countries.
In this occasion I would like to express that it is a great honor for IPB to be the
host of the first international conferenceof ign-ttrc. We welcome all of you, senior and
junior scientists to our university compound, and wish you all success.
Bogar, the 17th of July 2012
Vice Rector for Research and Collaboration,
· Anas M. Fauzi
TABLE OF CONTENTS
Page
PREFACE ................................................... ... ........................................................................
FOREWORD............................................................... ........................................................
TABLE OF CONTENTS............................................. ...........................................................
AGENDA OF THE CONFERENCE..................................................... .................................
IS 1
Epigenetics:Rapid Evolution, Biodiversity, Environmental Challenges
(Wolfgang Nellen, Benjamin Boesler, Sara Miller, Doreen Meier,
lndra Windhof, Christian Joppich,) ........................ ................... .. .......... ....
IS 2
Metagenome Analysis of Tempeh, a Unique Indonesian Fermented Food
(Cecillia A. Seumahu, Efriwati Sabron, Tati Barus, and Antonius
Suwanto)...... ............ .................................... ............... .................. ........ ............. ....
IS 3
3
4
5
9
12
13
Biomolecular Interaction Analysis in Functional Proteomics
(Prof. Dr. Friedrich W. Herberg).......................................... ..............................
Properties and Functions of dsRNA Binding Proteins in miRNA Mediated
Gene Silencing (Doreen Meier)................................. ......... ... ......... .....................
OP 2
Property of Myoglobin Protein in Rattus norvegicus- Normoxic Lung
Tissue (Rini Puspitaningrum, Guenter Lochnit)... ............................................
OP 3
Identification of New Interaction Partners of PKA (Jennifer Hermann).... ..
OP 4
Molecular Analysis of the Type 2 Diabetes Mellitus-Associated Region of Mitochondrial tRNA DNA in Gorontalonese Cases (Amien R. Ishak,
Risma Dwi Untari, Rini Puspitaningrum, Tri Murtiati, Chris Adhiyanto,
Takenori Nitta, Hattori Yukio, Yasuhiro Yamashiro).........................................
OP 5
Identification and Analysis of Single Nucleotide Polymorphism of Growth
Hormone Gene Exon 3, lntron 3 in Local Cattle Breeds in West Sumatera
Province of Indonesia (Yurnalis, Sarbaini, Arnim, Wolfgang Nellen,
Jamsari)...... ................................. ............ ..............................................................
OP 6
Polymorphisms Study on Several Specimen of Indonesian Coelacanth,
Latimeria menadoensis (Jxchel Feibie Mandagi, Yoshitaka Abe, Masamitsu
Iwata, Sinya Yamauchi, Fransisko Pangalila, Hefry Veibert Dien, Laurentius TX
Lalamentik, Kawilarang Warouw Alex Masengi, Kanako Koyanagi,Hiroki
Arimura and Hidemi Watanabe)............... ..........................................................
OP 7
Production of Double Haploid Orchids through Microspore Embryogenesis
(Ari lndrianto, Chairani Siregar, Sutikno Linuhung, Mekartinita,
Tri Sartikoningsih)...................................................................................................
OP 8
In 'Qtro Flowering of Orange Subfamily as an Alternative Way for Breeding
Techniques (Hasan Basri Jumin)... ......................................................... .............
OP 9
Effect of Aeration on Biomass and Saponin Content of Talinuin ｰ｡ｨｩ｣ｵｬｾｭ＠
Gaertn. Hairy Roots in Balloon-Type Bubble Bioreactor (ArifVachya, Alfinda
Novi Kristanti, Yosephine Sri Wulan Manuhara)............... ...................................
OP 10 The Effects of Saccharomyces cerevisiae and CuS0 4 on the Production of
Saponin in Talinum paniculatum (Jacq) Gaertn Callus (Agus Muji Santoso,
Alfinda Novi Kristanti, Yosephine Sri Wulan Manuhara)......................................
OP 11 Transformation of Aluminum Tolerant Gene Candidate in Rice
(Oryza sativa L.) (Arief Pambudi, Miftahudin, Tetty Chaidamsari)....................
OP 12 Modification of DNA Isolation Protocol from Dried-Leaf Tissues of
Threatened Species "Jernang" (Daemonorops draco (Willd.) Blume)
14
OP 1
(Revis Asra)....... ................................... ............................................. ... ... ...................
15
16
17
18
19
20
21
22
23
24
25
26
Page
IS 1
IS 2
IS 3
Q
ｾ＠
(Jens Gebauer) ..................... ........................... ... ...... ...........................................
Genes Characterization of Non-aggressive and Aggressive Strain of
PepYLCV from Chilli in West Sumatera (Jamsari)..................... ............ .... ........
Cloning C1 Gene (Rep) of Geminivirus to pGem-T Easy Vector In Escerichia
coli BL21 (EIIy Syafriani, lrwan Darfis, Jamsari).................................................
Cloning of Geminivirus Coat Protein Gene (V1) From Capsicum annuum
(Ester Kristin Natalia, Jamsari, Sutoyo)....................................... ......................
Cloning of V2 Gene (Pre Coat Protein) Pepper Yellow Leaf Curl Virus
(Pepylcv) from Chili Pepper (Capsicum annuum L.) (Siti Nur Aisyah, Ujang
Khairul, Jamsari)............... ................................. ......................................................
Draft Genome Sequence of Streptomyces sp. strain GMR22, Isolated
from Cajupuut Plantation, Wanagama, Yogyakarta, Indonesia (Jaka Widada,
Alimuddin, and Camelia Herdini)..........................................................................
The Localization of Two Genes for Erucic Acid Content using lntervarietal
Substitution Lines in Rapeseed (Brassica napus L.) (Nurhasanah, Berisso
Kebede, Rubens Marschalek, Heiko C. Becker, and Wolfgang Ecke).............
Analysis of Genetic Diversity of Rodent Tuber Cultivars in Indonesia
Based on RAPD Marker (Nesti F. Sianipar, Danny Laurent, Chelen, Listiarini,
and Ariandana Wantho)............ ............ ............ ................................................ ...
Screening of Antifungal Product from Bacteria for Control of Anthracnose
OP8
Diseases Caused by Col/etotrichum gluosporioides In Chilli (Maizar,
I. Suliansyah, U. Khiarul, and Jamsari)...............................................................
Characterization of Glucanase Produced by Bacteria which Inhibit
OP9
the Growth of Pyricularia Oryzae and Rhizoctonia so/ani (Nurul Hikmawati,
Nisa Rachmania Mubarik, Yadi Suryadi, M. Sc)............... ....................................
OP 10 Test of Bacterial Consortium Formulation to Reduce Neck-Blast Disease on
Rice Plants (Zuhay Ratuz Zaffan, Nisa Rachmania Mubarik, Yadi Suryadi)....
OP' 11 Potency of Brevundimonas Sp. Hgp11solated from Gold Mining in Pongkor
Village, Bogor as Mercury Bioremediation Agent (Wahyu lrawati, Patricia,
Yenny Soraya, and Abyatar Hugo Baskoro).........................................................
OP 12 Isolation of Proteolytic Bacteria from GiliMeno Lake- West Nusa Tenggara
fnd Characterization of the Protease Enzyme (Susana Sagitha Raunsay, Ern in
Hidayati, Nisa Rachmania Mubarik) ........................ ............... :........ :....................
OP 13 Isolation and Characterization of Proteolytic Bacteria f rom Cricula
Trifenestrata Cocoon that Influence Quality of Goldsilk (Abdi Gunawan,
Dedy Duryadi Solihin, Nisa Rachmania Mubarik)..........................................
POSTER.................................... .......................................................................................... ...
P1
Determination of Sex of Salacca (Salacca edu/is L.) Based on
PCR-RPD (Ediwirman, Jamsari, I. Suliansyah, Gustian)......................................
P2
Effect of Pretreatmenton the Rice Straw as Substrate on Solid State
FermentatiQn of Trichoderma viride Strain T051 to the Cellulase Activity Sri
(Sugiwati, Siswati Setiasih, Nenci and Haznan Abimayu) ................................-.
P3
Starch Characterization and Yield Variation of "Gebang" Cassava
Regenerated from Irradiated In Vitro Shoots (Supatmi, Enny Sudarmonowati
@
ce
J
Molecular Diversity and Evolution of South-East Asian Ant-Plants (Genus
Macaranga, Euphorbiaceae) (Dr. Daniela Guicking)........................ ...............
Plant Genetic Resources and Their Preservation via Biotechnological and
Other Methods (Klaus J. Dehmer).................................... .................................
Mountain Oases of Northern Oman: Hotspots of Agrobiodiversity
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
Page
J
P4
P5
P6
J
P7
v' P8
P9
J PlO
J Pll
P12
J
P13
v'P14
J P15
v P16
J
P17
j P18
Pl 9
and N.Sri Hartati).. .................................................................................................
Cloning Geminivirus DNA-satellite Sequence of Pepper Plant
(Capsicum annum L.) (Dila Febria, Yusniwati, Herviyanti, Jamsari)............ ..
Spesific Primer Desain Related Potential High Catechin Content on
Gambier Plant (Uncaria gambir (Hunter) Roxb) (lstino Ferita, lrfan
Suliansyah, Gustian, Hamda Fauza, dan Jamsari)............ ........................ ........
DNA Finger print of Several Cassava Genotypes Comprising Different
Carotene Content and Drought Responses (Ahmad Fathoni, N. Sri Hartati,
Enny Sudarmonowati)..................... .....................................................................
Analysis of Aedes aegypti Microsatelite DNA in Endemic Regions of Dengue
Hemorragic Fever (DHF) Padang, Pariaman and Solok City, West Sumatra
(Hasmiwati, Djong Hon Tjong and Rita Maliza)................................................
Influence of Plant Growth Regulators Auxin and Cytokinin to Callus Induction
of Some Pepper (Capsicum annuum L.) Genotypes For Genetic
Transformation (Renfiyeni, Yusniwati, J. Trisno dan Jamsari)...... ...................
Development Protocol of Agrobacterium- Mediated Genetic Transformation
of Eucalyptus pel/ita F. Muell Using Xyloglucanase Gene (N . Sri Hartati,
Andriyani Puspitaningrum, Nita Etikawati, Enny Sudarmonowati)..................
Potency Test Of Several Chitinolitic Bacteria Isolates As An Agent Of Anti
Anthracnose And Its Identification Based On 16S rRNA Sequence Gene {Irina
Sari, lr. Sutoyo, MS, Prof. Dr. sc. Agr. lr. Jamsari, MP)........................... ............
In Silico Analysis on Structure and Prediction Function Changes of Tyrosine
Kinase Domain Insulin Receptor of Diabetes Mellitus Patient (Miftakhun,
N.Y.P., Fatchiyah, Widodo)....................................................................................
Phylogeny of Mangosteen Varieties Based on ITS-nrDNA Gene
(Panca Jarot Santoso).................................... ............ ........................ .....................
Diversity of Gambier (Uncaria gambir (Hunter) Roxb.) based on RAPD
Marker in West Sumatra (Hamda Fauza, Jamsari, lstino Ferita, Azmi Dhalimi,
Ahmad Denian, and Murdaningsih H. Karmana )...............................................
Selection of Antagonist Bacteria from Green Mustard (Brassica juncea L.)
as Biofungicide for Co/letotrichum gloeosporioides, a Causal of Anthracnose
in Chili Pepper (Capsicum sp.) (Rahmi Henda Yani, Aswaldi Anwar,
Jamsari) ..ｾ Ｎ Ｎ Ｎ Ｎ Ｎ Ｎ
Ｎ Ｎ Ｎ Ｎ Ｎ Ｎ ＮＮ Ｎ Ｎ Ｎ Ｎ Ｎ
ＮＮ
ＮＮ Ｎ Ｎ ＮＮＮ Ｎ
ＮＮ
Ｎ Ｎ Ｎ ＮＮ Ｎ Ｎ
Ｎ Ｎ
Ｎ＠
A Preliminary Study: The Effect Of Progesterone To Dopamine Signaling on
Zebrafish (Danio rerio) Embryos (Christianto, A, Kishida, M)...........................
Diversity and Activities of Bacterial Isolates From Coastal Area of BandealitJerrtber, East Java - Indonesia (Dina Fitriyah, Herawati, Sattya Arimurti, and
Kartika Senjarini) ............................................................................. :...... .. :..............
Protein Profiles of Salivary Glands from Aedes aegypti (Dwi Esti F, Syubbanul
Wathon, Rike Oktarianti, Kartika Senjarini)..........................................................
Parasitemic Rates, IL-4 and IFN-y Profile on Mice Model Vaccinated by
Salivary Glands from Anopheles maculatusfor Developing TBV Against
Malaria {Imam Hanafy, lka Agus Rini, Yunita Armiyanti,
and Kartika Senjarini)..............................................................................................
Isolation and Identification of Lipase-Producing Bacteria for
the ｄ･ｧｲ｡､ｴｩｯｾ＠
of Vegetable Oil Wastes (Hishamudin Rahmat,
Teh Chin Een and Sharlina Mat Saad)...................................................................
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
Page
Effect of Chemical Compounds to Quinine Alkaloids Content of Cinchona
ledgeriana in Cell Suspension Culture (Dedi Satriawan, Diah Ratnadewi,
Ｍ ｾ＠
Sumaryono) ................................................... ......................................................... .
63
Molecular Studies of Genes Controlling Cambial Cell Differentiation and
P21
Secondary Cell Wall Formation (Widi Sunaryo, Andrea Po lie and
Urs Fischer)............... ................................. ............ ... ...............................................
64
Cell-Specific Analysis of Symbiosis-Induced Medicago truncatula Genes
P22
via Laser Microdissection (Anna-Lena Hurter, Claudia Hogekamp, Natalija
65
Hohnjec and Helge Kuster).. ................. ............. ........................... ..................
The Use of Sulfate Reducing Bacteria (SRB) to Reduce Sulfate and Heavy
P23
66
Metals in Acid Mining Water (AMW) (Fahruddin).................. ...... .....................
Tissue Culture of Sago Palm (Metroxylon saguRottb.), a Perennial
P24
Food Crops, with Different Explants Origins in Indonesia (Sumaryono,
I. Riyadi and M.M. Sinta)... ....................................... ............................................
67
Free Energy Analysis for Naphthoquinone Derivatives Binding into HBx Protein
P25
Using Molecular Docking (Firdayani, Harry Noviardi, Pertamawati, Susi
Kusumaningrum) .... ............. .. .........................................................................
68
Molecular Docking Studies on Series Of Potent Quinone Compounds
P26
as Polo-Like Kinase 11nhibitors (Susi Kusumaningrum,Harry Noviardi,
Firdayani )............... ...... ........................ ......................................................... ........ :.
69
P27
Home Composting Using the grEEn Technology (Trina Tallei, Julius Iskandar,
70
Marian Kamagi, Abdul Basith)..................... ................................................... .......
P28
Cytotoxic Activity of Graptophyllum pictum Methanolic Extracts Against Lung
Cancer Cell Line (A549) and Simian Retrovirus Insertion (A549-SRV)
(Rilianawati, Fery Azis Wijaya, Achmad Junaidi, Tarwadi,Jin Hyub Paik).......... 71
P29
Genic Characterization of Indonesian Coelacanth Latimeria menadoensis
(Daisy Monica Makapedua, lxchel Feibie Mandagi, Marco Barucca, Mariko
Forconi,Adriana Canapa, Ettore Olmo,Masa Iwata, AlexMasengi, Fransisco
Pangalila, Florence Longdong)................................................................................. 72
P30
Polymorphism Identification of BMP-15 Gene(Bone Morphogenetic
Protein-15) lnbeangoat(Capra hircus) usingPCR-RFLP Technique (Sri Rahayu,
lrham Hayati, Estri Laras
Arumingtyas-) ............................................................................................................... 73
P31
Identification and Phylogenetic Diversity Based on 16S rRNA Gene Sequence
Analysis of Thermophilic Bacteria from Rimbo Panti Hot Spring (Armaini, Abdi
Dharma, Sumaryati Syukur, Jamsari)........................ ............................................... 74
LIST OF PART)CIPANTS ............ .................................................................................................. 75
P20
Effect of Chemical Compounds to Quinine Alkaloids Content of Cinchona ledgeriana in
Cell Suspension Culture.
Dedi Satriawan 1l, Diah Ratnadewi 1l, Sumaryono 2l
1
lDepartment of Biology, Faculty of Mathematics and Natural Sciences, Bogar
Agricultural University,Darmaga Campus, Bogar 16680, Indonesia
2
llndonesian Biotechnology Research Institute for Estate Crops, Jalan Taman Kencana
No. 1, Bogar 16151, Indonesia
ABSTRACT
Quinine is used for tonic, appetite enhancer, antipyretic, antimalarial, cosmetic
ingredients and tanning. Quinine can be obtained from the bark of seven or more years
old Cinchona tree. Cell suspension culture is an alternative means to obtain secondary
metabolites rapidly and continously. Quinine, one of alkaloid compounds, is expected to
increase with the addition of abscisic acid (ABA) and paclobutrazol (PBZ) as growth
inhibitors, tryptophan as a precursor and mannitol as an osmotic stress inducer. ABA
was used at 1 and 3 ppm, PBZ at 5 and 7 ppm combined with 5.3 g/1 mannitol,
tryptophan at 0.2 and 2 ppm combined with 5.3 g/1 mannitol. ABA at 1 and 3 ppm
increased the cell suspension growth but reduced the content of quinine. PBZ at 7 ppm
combined with 5.3 g/1 mannitol and 20 g/1 sucrose significantly reduced the cell
suspension growth but remarkably increased quinine content, even higher than that was
found in the bark of Cinchona tree in the field. Combination of mannitol and
paclobutrazol was effective to inhibit the cell growth but enhance the production of
quinine.
Keywords: cell suspension culture, Cinchona, mannitol, abscisic acid, paclobutrazol,
tryptophan,quinine.