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MOLECULAR PHYLOGENY OF MAIDENHAIR FERN GENUS ADIANTUM (PTERIDACEAE) FROM LESSER SUNDA ISLANDS INDONESIA BASED ON RBCL AND TRNL-F Lestari, Wenni Setyo; Adjie, Bayu; Jaruwatanaphan, Tassanai; Watano, Yasuyuki; Pharmawati, Made
REINWARDTIA Vol 14, No 1 (2014): Vol. 14 No. 1
Publisher : Research Center for Biology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (3344.96 KB) | DOI: 10.14203/reinwardtia.v14i1.409

Abstract

The Lesser Sunda Islands of Indonesia are composed of small islands scattered from Bali to Timor Island. We analyzed a molecular phylogeny of Adiantum collected from Lesser Sunda Islands to reveal its phylogenetic relationships. A total of 12 species of Adiantum from this region and seven species from Java Island were collected and used in this study. Two cpDNA regions (rbcL and trnL-F) were chosen as markers and phylogenetic analyses were conducted using Neighbour-Joining (NJ) and Maximum Parsimony (MP) methods. The tree topologies reconstructed by NJ and MP from specimens used in this study and other species downloaded from GenBank are congruent in which trees are divided into five major clades. Adiantum species of Lesser Sunda Islands are not monophyletic and comprises three clades, i. e. Clade I composed of A. hispidulum group, Clade III composed of A. peruvianum group and Clade IV or A. caudatum group, each together with extra-Lesser Sunda samples. No sample from Lesser Sunda Islands examined is located in Clade II (A. tenerum group) and V (A. capillus-veneris group).
PCR-RFLP and Sequencing of trnS/trnfM Fragment of Enhalus acoriodes from Sanur Coastal Waters, Bali, Indonesia: A Preliminary Study Pharmawati, Made; Imaniar, Eka Fibayani
Journal of Tropical Life Science Vol 6, No 2 (2016)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.06.02.10

Abstract

This study aimed to evaluate the resolution of PCR-RFLP and sequencing of trnS/trnfM in detecting genetic diversity of seagrass Enhalus acoroides at Sanur Beach, Sindhu Beach and Semawang Beach. This research used six samples E. acoroides from each location. DNA extraction method followed Doyle and Doyle. PCR amplifications were done using primers P672/P673, P674/P675 and ITS4/ITS5 followed by digestion using restriction enzymes EcoRI, PstI, HindIII, BamHI, Rsa, Mva and HinfI. Sequencing analysis used PCR products of primers trnS/trnfM. Testing of DNA sequences of E. acoroides were conducted using BLAST (Basic Local Alignment Search Tool). DNA sequences were further analyzed using MEGA 5.2 software (Molecular Evolutionary Genetic Analysis) to evaluate variations of DNA. The sequence alignments were done using ClustalW software to determine the homology between the DNA sequences. The results showed that 18 samples of E. acoroides from Sanur, Sindhu and Semawang Beach have no polymorphism based on restriction enzyme analyses. Furthermore, sequencing of trnS/trnfM region of 18 samples E. acoroides showed that the sequences were identical.   
Aksi Ethyl Methane Sulphonate terhadap Munculnya Bibit dan Pertumbuhan Cabai Rawit(Capsicum frutescens L.) (Ethyl Methane Sulphonate Action on Seed Emergence and Growth of (Capsicum frutecens L.)) Rustini, Ni Kadek Dewi; Pharmawati, Made
JURNAL BIOS LOGOS Vol 4, No 1 (2014): JURNAL BIOSLOGOS
Publisher : Universitas Sam Ratulangi

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35799/jbl.4.1.2014.4836

Abstract

Abstrak Penelitian ini bertujuan mengamati pengaruh lama perendaman biji dengan 1% EMS terhadap persentase munculnya bibit dan karakter pertumbuhan tanaman cabai rawit pada umur 4 minggu setelah tanam (MST). Benih cabai rawit direndam dalam air selama 6 jam, selanjutnya direndam 1% EMS dalam buffer fosfat pH 7, selama 6 jam, 9 jam dan 12 jam. Tiap biji disemai dalam bumbungan kertas. Munculnya bibit diamati setiap hari. Setelah berumur 3 minggu, bibit dipindahkan ke bedengan. Hasil menunjukkan perlakuan 1% EMS memperlambat munculnya bibit. Pada 10 hari setelah semai (HSS) munculnya bibit pada perlakuan dapat mencapai 100%. Perlakuan 1% EMS berpengaruh nyata terhadap tinggi tanaman dan jumlah daun, sedangkan panjang, dan lebar daun dengan perendaman 6 jam tidak berbeda dengan kontrol, namun berbeda dengan perendaman 9 jam dan 12 jam. Kata kunci : EMS, bibit, pertumbuhan, Capsicum frustescens L.   Abstract This research aimed to evaluate the influence of different exposure durations of 1% EMS on seedling emergence and growth characters of C, fustescent at 4 week after planting (WAT). Seeds were soaked in water for 6 hours, then soaked in 1% EMS in phosphate buffer pH 7, for 6 hours, 9 hours and 12 hours. Each seed was then sowed in a single paper tube. Seedling emergence was observed every day. At 3 weeks after sowing, seedlings were transferred to field. Results showed that soaking seeds in 1% EMS inhibited seedling emergence. At 10 days after sowing, the percentages of seedling emergence at control and treated seed were 100%. Treatments of 1% EMS have a significant effect on plant height, and the number of leaf, while length, and width of leaf with 6 hour exposure were not different with control, but differ from exposure of 9 hours and 12 hours. Keywords: EMS, seed emergence, growth, Capsicum frutescens L.
Induksi Mutasi Kromosom dengan Kolkisin Pada Bawang Putih (Allium sativum L.) Kultivar ‘Kesuna Bali’ (Induced Chromosome Mutation Using Colchicine in Garlic (Allium sativum Linn.) Cultivar ‘Kesuna Bali’) Pharmawati, Made; Wistiani, Ni Luh Ayu Jami
JURNAL BIOS LOGOS Vol 5, No 1 (2015): JURNAL BIOSLOGOS
Publisher : Universitas Sam Ratulangi

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35799/jbl.5.1.2015.9317

Abstract

Abstrak Tanaman bawang putih (Allium sativum L.) adalah tanaman holtikultura yang memiliki banyak manfaat terutama umbinya yang umumnya digunakan sebgai bumbu dan obat. Salah satu kultivar bawang putih yang ditanam di Bali adalah ‘Kesuna Bali’ yang hanya memiliki satu siung.  Salah satu cara untuk memperbaiki karakter tanaman adalah dengan cara induksi mutasi kromosom dengan kolkisin. Penelitian ini bertujuan menganalisis pengaruh perlakuan kolkisin terhadap indeks stomata dan jumlah kromosom dari tanaman ‘Kesuna Bali’. Penelitian menggunakan Rancangan Acak Kelompok dengan enam ulangan. Perlakuan kolkisin yang digunakan adalah 5%, 10% dan 20%. Hasil penelitian menunjukkan bahwa perlakuan kolkisin menurunkan indeks stomata dan meningkatkan jumlah kromosom. Kromosom triploid (2n=3x=24) dihasilkan pada perlakuan kolkisin 20%. Kata kunci : Allium sativum L., ’Kesuna Bali”, kolkisin, mutasi, sitologi Abstract Garlic (Allium sativum L.) is a horticultural crop that has many benefits, especially as spice and traditional medicine.  One of garlic cultivars planted in Bali is ‘Kesuna Bali’ which only has one clove. To improve characters of ‘Kesuna Bali’, modification of ‘Kesuna Bali’ properties can be done by means of induced mutation using colchicine.  This research aims to analyse the effect of colchicine on stomata index and the number of chromosomes of 'Kesuna Bali'. This research used randomized block design with six replicates. In this experiment the concentration of colchicine used were 5%, 10% and 20%. The results of this study showed that colchicine treatment of 20% resulted in the lowest stomata index and there was an increase in chromosome number. Colchicine at concentration of 20% resulted in triploid chromosome set (2n = 3x = 24). Keywords: Allium sativum L., ’Kesuna Bali”, cholchicine, mutation, cytology
Respon Morfologis dan Ekspresi Gen Aquaporin pada Padi IR 64 yang Mengalami Cekaman Kekeringan pada Fase Reproduktif (Morphological Responses and Aquaporin Gene Expression in Rice IR64 under Drought Stress at the Reproductive Stage) Pharmawati, Made; Wirasiti, Ni Nyoman; Wrasiati, Luh Putu
JURNAL BIOS LOGOS Vol 7, No 2 (2017): JURNAL BIOSLOGOS
Publisher : Universitas Sam Ratulangi

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35799/jbl.7.2.2017.18579

Abstract

Abstrak Cekaman kekeringan merupakan faktor pembatas penting bagi pertumbuhan dan produktivitas tanaman termasuk padi.      Penelitian ini bertujuan menganalisis respon padi IR64 terhadap cekaman kekeringan dengan pemberian polietilen glikol (PEG) pada fase reproduktif.  Penelitian juga bertujuan menganalisis ekspresi gen aquaporin akibat cekaman kekeringan.  Bibit padi ditanam dalam pot dan perlakuan PEG dengan konsentrasi 108g/L (-0.25MPa) dan 178g/L (-0.52 MPa) diberikan saat munculnya panikula. Perlakuan diberikan selama 2 minggu, kemudian tanaman disiram kembali.  Ekspresi gen diamati pada akhir perlakuan dengan semi kuantitatif real time PCR.  Ekstraksi RNA menggunakan RNeasy plant mini kit, sedangkan sintesis cDNA menggunakan Transcriptor First Strand cDNA Kit.  Hasil penelitian menunjukkan bahwa jumlah malai dan berat total malai berkurang akibat cekaman kekeringan.  Persentase gabah kosong mencapai 84,6% pada perlakuan PEG-0,52 MPa, sedangkan pada perlakuan PEG -0,25 MPa persentase gabah kosong sebesar 67,8%.  Pada kontrol persentase gabah kosong adalah 10,3%.  Ekspresi gen OsPIP2;7 sedikit menurun pada perlakuan PEG -0,52 MPa.Kata kunci: ekspresi gen, IR64, kekeringan, padi, PEG  Abstract Drought stress is one of the limiting factors of plant growth and productivity including rice.  The aim of this study was to analyze responses of IR64 rice to polyethylene glycol (PEG)-induced-drought stress at the reproductive stage.  This study also aimed to analyze the expression of aquaporin under drought stress.  Rice seedlings were grown in pot system and PEG treatment at concentration of -0.25MPa (108g/L) and -0.52 MPa (178g/L) were given when the panicles arose.  Treatments were conducted for 2 weeks, after that the plants were rewatered.  Gene expression was evaluated at the end of PEG treatment using semi quantitative real time PCR. RNA was extracted using RNeasy plant mini kit, while cDNA synthesis was done using Transcriptor First Strand cDNA Kit.  The results showed that the number and weight of rice ear were less in plant treated with PEG than in control.  The percentage of empty rice grain reached 84.6% at PEG -0.52 MPa, while at PEG -0.25 MPa the percentage of empty grain was 67.8%.  In control plant, the percentage of empty grain was 10.3%.  Drought stress did not alter the expression of OsPIP2;7.  Keywords: drought, gene expression, IR64, PEG, rice
Optimasi Konsentrasi DNA dan MgCl2 pada Reaksi Polymerase Chain Reaction-Random Amplified Polymorphic DNA untuk Analisis Keragaman Genetik Tanaman Faloak (Sterculia quadrifida R.Br) (Optimization of DNA and MgCl2 Concentrations in Polymerase Chain Reacti Uslan, Uslan; Pharmawati, Made
JURNAL BIOS LOGOS Vol 5, No 1 (2015): JURNAL BIOSLOGOS
Publisher : Universitas Sam Ratulangi

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35799/jbl.5.1.2015.9316

Abstract

Abstrak Faloak  merupakan tanaman yang tumbuh di lahan kritis. Sebagai upaya mendukung pemuliaan dan konservasi tanaman faloak diperlukan informasi keragaman genetiknya. Salah satu metode analisis keragaman genetik adalah menggunakan penanda DNA yang berbasis PCR. Untuk itu diperlukan kondisi PCR (Polymerase Chain Reaction) yang tepat sehingga diperoleh hasil yang dapat dianalisis lebih lanjut. Penelitian ini bertujuan menentukan kondisi optimum PCR-RAPD (Polymerase Chain Reaction-Random Amplified Polymorphic DNA) tanaman faloak. Ekstraksi DNA dilakukan dengan metode CTAB. Optimasi dilakukan dengan menggunakan beberapa konsentrasi DNA cetakan dan MgCl2. Kondisi optimum PCR-RAPD tanaman faloak yang menghasilkan pita produk PCR yang jelas diperoleh  menggunakan 50 ng/ul DNA, 3 mM MgCl2 serta jumlah siklus termal 45 x. Kata kunci : PCR-RAPD, optimasi, tanaman faloak Abstract Faloak is a plant that grows on critical lands. In an effort to support breeding and conservation of faloak, information about its genetic diversity is required. One of the methods of genetic diversity analysis is using PCR-based DNA markers. For that purpose, proper PCR conditions is needed in order to obtain results that can be further analyzed. This study aimed to determine the optimum conditions for PCR-RAPD of faloak plants. DNA extraction was conducted using CTAB. Optimization was done by using several concentrations of DNA templates and MgCl2. The optimum conditions of PCR-RAPD of faloak plants that produce clear band of PCR products were obtained using 50 ng/ ul DNA, 3 mM MgCl2 and 45x thermal cycles Keywords : PCR-RAPD, optimization, faloak plant
Chloroplast DNA Copy Number May Link to Sex Determination in Leucadendron (Proteaceae) MADE PHARMAWATI; GUIJUN YAN; PATRICK FINNEGAN
HAYATI Journal of Biosciences Vol. 16 No. 1 (2009): March 2009
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (70.649 KB) | DOI: 10.4308/hjb.16.1.21

Abstract

Leucadendron (Proteaceae) is a South African genus, the flowers of which have become a popular item in the Australian cut-flower industry. All species are dioecious. In general the female flowers are the more desirable as cut flowers. The availability of a molecular marker linked to sex determination is therefore needed both to maximize the efficiency of breeding programs and to supply markets with flowers from the preferred sex. The polymerase chain reaction-based method of suppression subtractive hybridization (SSH) combined with mirror orientation selection (MOS) were applied in an attempt to identify genome differences between male and female plants of Leucadendron discolor. Screening of 416 clones from a male-subtracted genomic DNA library and 282 clones from a female-subtracted library identified 13 candidates for male-specific genomic fragments. Sequence analyses of the 13 candidate DNA fragments showed that they were fragments of the chloroplast DNA, raising the possibility that chloroplast DNA copy number is linked to sex determination in Leucadendron. Key words: Leucadendron, sex determination, suppression subtractive hybridization (SSH)
The Genetic Relationships of Grevillea Hybrids Determined by RAPD Marker MADE PHARMAWATI; IAN JAMES MACFARLANE
HAYATI Journal of Biosciences Vol. 20 No. 4 (2013): December 2013
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (488.987 KB) | DOI: 10.4308/hjb.20.4.196-200

Abstract

Grevillea (Proteaceae) is native to Australia and is known as garden plant. The flowers have high diversity colors, which range over orange, yellow, pink, red and green. It is a large genus, consisted of 357 species. This genus is economically important and there are hundreds of Grevillea hybrids with beautiful color and shape of flower.  Information of the genetic relationships of Grevillea hybrids is not available. Ten Grevillea hybrids were used in this study to evaluate the genetic relationships between hybrids and with their parents. PCR-RAPD technique was employed in this study with seven RAPD primers. The dendrogram of Grevillea hybrids was performed using Neighbor-Joining analysis based on genetic distance. The analysis revealed that ‘Moonlight’, ‘Caloundra Gem’, and Little Pink Willie’ were clustered with Grevillea banksii. The three hybrids were hybrid of G. banksii and other Grevillea species.  ‘Superb’ was closely related to ‘Robyn Gordon’ as they are hybrid of G. banksii and G. bipinnatifida.
Genetic Diversity of Dacrycarpus imbricatus At Bukit Tapak, Tabanan, Bali Based on RAPD Marker Made Pharmawati; I Made Anom Sutrisna Wijaya
Biotropika: Journal of Tropical Biology Vol 9, No 2 (2021)
Publisher : University of Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.biotropika.2021.009.02.09

Abstract

Dacrycarpus imbricatus Blume is a member of the Podocarpaceae family. In Bali, D. imbricatus was found in Bukit Tapak, Tabanan Regency. This species is one of the dominant species in Bukit Tapak. This study aimed to determine the genetic variation of D. imbricatus in Bukit Tapak using molecular markers RAPD (Random Amplified Polymorphic DNA). The genetic diversity of D. imbricatus needs to be studied to obtain the information used for the conservation of this species. Leaf samples were taken from Bukit Tapak, Candikuning, Baturiti, Tabanan Regency, Bali. DNA was extracted using the CTAB method, followed by extraction using chloroform: isoamyl alcohol. DNA precipitation was carried out using ethanol. RAPD analysis was performed using polymerase chain reaction (PCR) using four primers. PCR products were visualized using agarose gel electrophoresis and ethidium bromide staining. The results showed that the amplified DNA bands ranged from 1 to 5 bands with DNA band sizes ranging from 230 bp - 422 bp. Only OPA4 and UBC106 primers can be used to detect D. imbricatus diversity based on the H, D, and R values. The detected genetic variation is low, as indicated by an average polymorphism of 32.5% and similarities between samples ranging from 0.51 to 1
RAPD Primers Selection for Genetic Variation Analysis of Banana Plant (Musa spp.) I Ketut Catur Wiguna; Made Pharmawati
Jurnal Biologi Universitas Andalas Vol 9, No 2 (2021)
Publisher : Universitas Andalas

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.25077/jbioua.9.2.47-53.2021

Abstract

Identification of genetic diversity using molecular markers is important for basic information for plant conservation. Banana is a fruit bearing plants that important for food sources in human life. This study aimed to determine the optimum conditions of  PCR-RAPD reaction and RAPD primers that suitable to amplify DNA fragmens. DNA isolation was done using modified of CTAB and chloroform isoamil alcohol. The samples used was young leaves of nine banana cultivar plants. Optimation was done using variety of DNA and MgCl2 concentration. Eight primers produced by Operon Primer Technology were tested. The DNA genomic concentration obtained was in the range of 23,3 ng/µl – 70 ng/µl. The optimum conditions of PCR-RAPD of banana plants that produce clear band were 50 ng/µl DNA template, 3 mM MgCl2 with the number of thermal cycles was 40 x. There were six RAPD primers that successfully amplified DNA : OPA 02, OPA 04, OPB 12, OPD 20, OPH 01, and OPH 03. The primer OPA-04 had the lowest resolving power value (4,4) , while  OPH 01 had the highest (11,3) resolving power.
Co-Authors A. A. Putri Ardyanti A.A.S. A. Sukmaningsih Agriani Dewinta Alifya Ibnu Aziz Anak Agung Ayu Putri Ardyanti Anak Agung Gde Raka Dalem Andi Bahtiar Batti Bayu Adjie Bayu Adjie Bram Setyadji Deny Suhermawan Yusup Dian Catur Prayantini Dwi Nur Rikhma Sari Eka Fibayani Imaniar Eka Fibayani Imaniar Ema Hendriyani Fransiska Nitti GUIJUN YAN Gusti Ayu Putri Arnila I Gde Suryadi I Gede Putu Ardhana I Gusti Agung Eka Saraswati I Gusti Ayu Agung Pradnya Paramitha I Gusti Ngurah Kade Mahardika I Kadek Aridena Putera I Ketut Catur Wiguna I Ketut Junitha I Ketut Suada I Komang Adi Widyastama I MADE AGUS WIARTANA I Made Anom Sutrisna Wijaya I Putu Agus Tirta Cahyana I Putu Satwika I Wayan Arnata I. Setyawati I.G.A.S. Wahyuni I.M.A.S. Wijaya IAN JAMES MACFARLANE IDA AYU ASTARINI Ida Ayu Ratih Purnama Dewi Imaniar, Eka Fibayani Indah Tria Hoky Inna Narayani Iriani Setyawati Junita Hardini Ketut Queena Fredlina Luh Putu Wrasiati Made Ria Defiani Maliza Kurnia Maya Agustina Maya Agustina Mery Wintari Ngurah Intan Wiratmini Ni Kadek Dewi Rustini NI KADEK YUNITA SARI Ni LUH ARPIWI Ni Luh Ayu Jami Wistiani, Ni Luh Ayu Jami Ni Luh Putu Agustini Ni Luh Watiniasih Ni Luh Wayan Yulia Mirayanti NI MADE DIAN PRATIWI Ni Made Gari Ni Made Sastriyani Wiendra Ni Made Sastriyani Wiendra Ni Made Suartin Ni Nyoman Wirasiti Ni Putu Adriani Astiti Ni Putu Eka Krisnayanti Ni Wayan Karolina Ni Wayan Sudatri PATRICK FINNEGAN Putu Apriliani Putu Yunita Putri Aryani Retno Kawuri Rifky Firmansyah Anwar Septarini Dian Anitasari Septarini Dian Anitasari Tassanai Jaruwatanaphan Tassanai Jaruwatanaphan, Tassanai Tiwi Wati Uslan UUL SHOVI NURKAMILA Vanesa Martida Vanesa Martida Viryanando Evan Rahardja Wawan Sujarwo Wenni Setyo Lestari Wenni Setyo Lestari, Wenni Setyo Wildan Mujahidul Basyar Yasuyuki Watano Yasuyuki Watano, Yasuyuki Yenni Ciawi