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Grevo S. Gerung
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Isolation and amplification of 16S rRNA gen of Associated Microbial isolates in Red Algae Kappaphycus alvarezii from Belang, Southeast Minahasa Regency, North Sulawesi Letha L. Wantania; Stenly Wullur; Elvi L. Ginting; Desy M. H. Mantiri; Suzanne L. Undap; Deiske A. Sumilat; Grevo S. Gerung
Jurnal Ilmiah PLATAX Vol. 7 No. 1 (2019): ISSUE JANUARY-JUNE 2019
Publisher : Sam Ratulangi University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35800/jip.7.1.2019.22808

Abstract

This study aims to obtain isolates and amplify the associative bacterial 16SrRNA gene in K. alvarezii algae. The K. alvarezii algae was collected from seaweed cultivation area in Belang, Southeast Minahasa, North Sulawesi. Associative bacteria were sampled from K. alvarezii algae, grown in Nutrient Agar and separated based on their morphological characteristics.  Each isolates were extracted their DNA genome and.the16S rRNA gene of each isolate was amplified using PCR.  Eight associative bacterial from K. alvarezii algae were successfully isolated based on morphological characteristics which were dominated by round shape, smooth edges, convex elevation, and white color of the isolates. The results of genomic DNA extraction from each of these isolates were successfully used as templates to amplify the 16s rRNA gene.Key word: Bacteria, Kappaphycus alvarezii, Taxsonomy, 16S rRNAABSTRAKPenelitian ini bertujuan untuk mendapatkan isolat dan mengamplifikasi gen 16SrRNA bakteri asosiatif pada alga K. alvarezii. Metode penelitian diawali dengan pengambilan sampel alga K. alvarezii di lokasi pembudidayaan rumput laut di desa Belang, Minahasa Tenggara, Sulawesi Utara. Selanjutnya, bakteri asosiatif pada alga K. alvarezii tersebut ditumbuhkan dalam media Nutrient Agar, isolat yang tumbuh kemudian dipisahkan berdasarkan karakteristik morfologinya. dan gen 16S rRNA masing-masing isolat diamplifikasi menggunakn PCR.  Delapan isolat bakteri asosiatif pada alga K. alvarezii berhasil diisolasi dengan karakteristik morfologi berbeda yang didominasi dengan bentuk round, tepian smooth, elevasi convex, dan warna putih. Hasil ekstraksi DNA genom dari masing-masing isolat tersebut berhasil digunakan sebagai template untuk mengamplifikasi gen 16s rRNA.Kata Kunci: Bakteri, Kappaphycus alvarezii, Taksonomi, 16S rRNA
In Vitro Culture of Seaweed Kappaphycus alvarezii under Different Formulation of Growth Stimulating Substances and Culture Media Mega D. Dalero; Grevo S. Gerung; Edwin L.A. Ngangi; Lawrence J.L. Lumingas; Markus T. Lasut
Jurnal Ilmiah PLATAX Vol. 7 No. 1 (2019): ISSUE JANUARY-JUNE 2019
Publisher : Sam Ratulangi University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35800/jip.7.1.2019.23375

Abstract

This study aims at obtaining a sustainably superior seed stock following the characteristics of the parent plant, determining the best formulation of the growth stimulating substance. In general, cytokinin and auxin combination was used, but this study also added with the combination of cytokinin and giberelin and cytokinin and abscisic acid (AA).Parameters measured were bud length, number of buds, and survival rate. Bacterial Vibrio sp test was also done as a cause of the explant mortality. Results showed that the longest bud was recorded in treatment C (S+A 1:2.5) cultured in a jar, 1.343 mm long, 38% of survival, while the highest number of buds was found in treatment B (S+A 1 : 2) 8.86. The shortest bud was recorded in treatment J (S + AA 1:2.5) cultured in a jar, 0.093 mm long, 2.64 buds, 10% of survival, while the explant cultured in the bottle had a length of 0.051 mm long, 1.50 buds, 4% of survival. As conclusion, the best growth stimulating substance was found in the treatment C for the bud length and the survival rate, while the best number of bud was recorded in the treatment B. The best culture tank was topless bottle (aerated). In vitro culture could also use S + G formulation. The explant mortality was caused by Vibrio charchariae. The use of S + AA formulation had lower growth than that of control treatment.Keywords :in vitro, growth stimulating substance, culture media, Kappaphycus alvarezii, Vibrio charchariae ABSTRAKPenelitian ini bertujuan untuk memperoleh benih unggul secara berkelanjutan yang mengikuti karakteristik dari tanaman induk, menentukan formulasi terbaik dari substansi pertumbuhan merangsang. Secara umum, kombinasi sitokinin dan auksin digunakan, tetapi penelitian ini juga menambahkankombinasi sitokinin, giberelin, sitokinin dan asam absisat (AA). Parameter yang diukur adalah panjang tunas, jumlah tunas, dan tingkat kelangsungan hidup. Bakteri Vibrio Uji sp juga dilakukan sebagai penyebab kematian eksplan . Hasil penelitian menunjukkan bahwa tunas terpanjang terdapat pada perlakuan C (S + A 1: 2,5)  kultur dalam toples, 1,343 mm, 38% hidup, sementara jumlah tertinggi tunas ditemukan pada perlakuan B (S + A 1: 2) 8.86 . Jumlah tunas paling sedikit terdapat pada perlakuan J (S + AA 1: 2,5) yang dikultur dalam toples, 0,093 mm, 2,64 tunas, 10% hidup, sedangkan eksplan yang dikultur dalam botol memiliki panjang 0.051 mm, 1. 50 tunas , 4% bertahan hidup. Sebagai kesimpulan, pertumbuhan terbaik merangsang zat ditemukan dalam perlakuan C untuk panjang tunas dan tingkat kelangsungan hidup, sementara jumlah tunas terbanyak ditemukan pada perlakuan B. Penggunaan wadah budaya terbaik adalah topless yang diaerasi. Kultur in vitro juga dapat menggunakan formulasi S + G. Kematian eksplan disebabkan oleh Vibrio charchariae . Penggunaan formulasi S + AA memiliki pertumbuhan yang lebih rendah dari pada pengobatan kontrol .Kata kunci : in vitro, zat perangsang tumbuh, media kultur, Kappaphycus alvarezii, Vibrio charchariae
DNA Isolation And Amplification of the rbcL (ribulose-1,5- bisphosphate carboxylase/oxygenase large subunit) gene of Caulerpa sp., Gracilaria sp., And Sargassum sp. Biondi Tampanguma; Grevo S. Gerung; Veibe Warouw; Billy Th Wagey; Stenly Wulllur; Deiske A. Sumilat; Hens Onibala
Jurnal Ilmiah PLATAX Vol. 8 No. 2 (2020): ISSUE JULY-DECEMBER 2020
Publisher : Sam Ratulangi University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35800/jip.8.2.2020.30003

Abstract

DNA isolation and gene amplification of algae are significantly influenced by various factors such as characteristics and components of the algae cell wall. Therefore techniques and methods of DNA isolation in certain algae, sometimes only succeed in one particular species and can not be applied to another algae species. Based on that issue, this study was conducted with the aims to determine the succeed of DNA isolation and amplify the rbcL gene as a target gene for identification. Algae DNA was isolated by using innuPrep Plant DNA commercial kit, and the second one with a modified conventional Cetyl Trimetyl Ammonium Bromide (CTAB) method,  for the amplification process was using rbcL gene (ribulose-1,5-bisphosphate carboxylase / oxygenase large subunit) with two pairs of primers : rbcL 7F-753R and rbcL 577F-rbcSR. The results showed that the DNA of Gracilaria sp was succeed isolated by using CTAB method and it was denoted by the presence of DNA bands in agarose gel. Meanwhile DNA amplification for Gracilaria sp., and Sargassum sp., were succeed amplified with the appearance of DNA bands. But in algae Caulerpa sp., was only succeed on 1 pair of primary rbcL 7F and 7.Keywords : DNA, gene rbcL, algae Caulerpa sp., Sargassum sp., Gracilaria sp;AbstrakIsolasi DNA dan amplifikasi gen pada alga sangat dipengaruhi oleh beberapa faktor seperti karakter dan komponen pada dinding sel alga. Oleh karena itu proses isolasi DNA terkadang bisa berhasil pada satu jenis alga, namun tidak berhasil pada jenis alga lainnya. Oleh karena alasan tersebut, maka penelitian ini dilakukan untuk menentukan keberhasilan Isolasi DNA dan mengamplifikasi gen rbcL sebagai gen target identifikasi. Penelitian ini dilakukan dengan tahapan awal Isolasi DNA yang menggunakan kit komersil innuPrep Plant DNA Kit, dan metode konvensional Cetyl Trimetyl Ammonium Bromide (CTAB) yang telah dimodifikasi. Sedangkan untuk proses amplifikasi, menggunakan gen rbcL (ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit) digunakan dua pasang primer yaitu rbcL 7F-753R dan rbcL 577F-rbcSR. Hasil isolasi DNA dari alga Gracilaria sp berhasil diisolasi menggunakan metode CTAB yang ditandai dengan adanya pita DNA pada gel agarose. Amplifikasi DNA pada alga Gracilaria sp., dan Sargassum sp., berhasil diamplifikasi dengan munculnya pita DNA. Namun pada alga Caulerpa sp. hanya berhasil pada 1 pasang primer rbcL 7F dan753R.Kata kunci : DNA, gen rbcL, alga Caulerpa sp., Sargassum sp., Gracilaria sp.