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IMPROVEMENT METHOD OF GENE TRANSFER IN KAPPAPHYCUS ALVAREZII Triana, St. Hidayah; Alimuddin, .; Widyastuti, Utut; Suharsono, .; Suryati, Emma; Parenrengi, Andi
Jurnal Ilmu dan Teknologi Kelautan Tropis Vol. 8 No. 1 (2016): Elektronik Jurnal Ilmu dan Teknologi Kelautan Tropis
Publisher : Department of Marine Science and Technology, Faculty of Fisheries and Marine Science, IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (511.747 KB) | DOI: 10.29244/jitkt.v8i1.13087

Abstract

Method of foreign gene transfer in red seaweed Kappaphycus alvarezii has been reported, however, li-mited number of transgenic F0 (broodstock) was obtained. This study was conducted to improve the method of gene transfer mediated by Agrobacterium tumefaciens in order to obtain high percentage of K. alvarezii transgenic. Superoxide dismutase gene from Melastoma malabatrichum (MmCu/Zn-SOD) was used as model towards increasing adaptability of K. alvarezii to environmental stress. The treat-ments were the culture media and recovery duration, and each treatment consisted of three replica-tions. The best method was co-cultivation using liquid media, then recovery was conducted in liquid media for 10 days. That treatment allowed higher transformation percentage (90%), regeneration effi-ciency (90%), putative bud efficiency (100%), number of buds and explants sprouted (100%) and transgenic explants (100%). The transgenic explants showed an amplification PCR product of Mm-Cu/Zn-SOD gene fragment, whereas the non-transgenic explants showed no amplification product.  All results revealed that suitable method of transgenesis for K. alvarezii has been developed. Keywords:       Agrobacterium tumefaciens, culture media, Kappaphycus alvarezii, recovery duration, transformation
PERFORMA BIBIT RUMPUT LAUT GRACILARIA VERRUCOSA HASIL KULTUR JARINGAN DENGAN BUDIDAYA METODE SEBAR (BROADCAST) DI TAMBAK KABUPATEN SINJAI Rosmiati, Rosmiati; Harlina, Harlina; Suryati, Emma; Daud, Rohama; Herlinah, Herlinah
Jurnal Riset Akuakultur Vol 14, No 3 (2019): (September, 2019)
Publisher : Pusat Riset Perikanan, Badan Riset dan Sumber Daya Manusia Kelautan dan Perikanan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (301.265 KB) | DOI: 10.15578/jra.14.3.2019.145-152

Abstract

Rumput laut Gracilaria verrucosa asal Kabupaten Sinjai memiliki kualitas paling rendah di antara semua sentra produksi Gracilaria sp. di Sulawesi Selatan. Hal ini salah satunya dikarenakan oleh bibit yang buruk. Penyediaan benih rumput laut yang berkualitas dapat dilakukan salah satunya dengan penggunaan bibit hasil kultur jaringan. Perbanyakan bibit Gracilaria verucosa dapat dilakukan dengan menggunakan metode tali panjang long line maupun metode sebar (broadcast) di tambak. Tujuan penelitian ini untuk mengetahui respons pertumbuhan, kandungan agar, dan kekuatan gel (gel strength) dari bibit G. verucosa hasil kultur jaringan di tambak Kabupaten Sinjai. Metode penelitian ini adalah eksperimen dengan dua perlakuan dan tiga ulangan yaitu perlakuan A (bibit kultur jaringan) dan B (bibit lokal) dengan berat awal masing-masing 10 kg. Pemeliharaan bibit dengan metode sebar dilakukan selama 30 hari. Hasil analisis ragam menunjukkan bahwa laju pertumbuhan harian (DGR), kandungan agar dan gel strength bibit kultur jaringan dan bibit lokal menunjukkan perbedaan yang nyata (P<0,05). Secara kuantitas hasil produksi bibit hasil kultur jaringan memiliki pertumbuhan yang lebih tinggi daripada bibit lokal dengan berat akhir bibit 44,3 ± 4,16 kg hasil kultur jaringan dan 33,0 ± 4,35 kg lokal dengan DGR 4,97% bobot/hari (kultur jaringan) dan 3,90% bobot/hari (lokal). Secara kualitas bibit hasil kultur jaringan lebih baik dari bibit lokal, ditunjukkan dengan persentase kandungan agar bibit hasil kultur jaringan lebih tinggi daripada bibit lokal dengan rendemen agar 22,19 ± 2,45% (kultur jaringan) dan 16,50 ± 0,96% (lokal), sementara gel strength sebesar 204,20 ± 0,45 g/cm2 (hasil kultur jaringan) dan 128,10 ± 1,55 g/cm2 (bibit lokal).Seaweed Gracilaria verrucosa from Sinjai Regency has the lowest quality among all Gracilaria sp. Production centers in South Sulawesi due to the low quality of the seed. The seed quality can be improved using seed selection, followed by tissue-culture methods. Long-line and broadcast methods in brackishwater ponds are the efficient seaweed culture techniques to multiply the number of Gracilaria verrucosa seeds. This research was aimed to determine growth performance, gel content, and gel strength of seeds produced from tissue-culture and local seaweed farming. The experiment consisted of two treatments: treatment A (cells culture seed) and B (local seed) with the initial weight of 10 kg, each has three replicates. Both seeds were stocked and reared in the ponds using the broadcast method for 30 days. The results of DGR, gel content and gel strength showed a significant difference between tissue-cultured and local seeds (P<0.05). The tissue-cultured seed had better growth than the local seed with 4.97% mass/day for tissue-cultured seed and 3.90 mass/day for local seed. The tissue-culture seed also had better quality in agar content and gel strength. The agar content of tissue-cultured was 22.19 ± 2.45% and the local was 16.50 ± 0.96%. The gel strength of tissue-culture was 204.20 ± 0.45 g/cm2, and the local was 128.10 ± 1.55 g/cm2.
PERTUMBUHAN DAN PERKEMBANGAN EKSPLAN RUMPUT LAUT GRACILARIA VERRUCOSA DAN GRACILARIA GIGAS PADA AKLIMATISASI DI TAMBAK Mulyaningrum, Sri Redjeki Hesti; Parenrengi, Andi; Suryati, Emma
ILMU KELAUTAN: Indonesian Journal of Marine Sciences Vol 20, No 3 (2015): Ilmu Kelautan
Publisher : Marine Science Department Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1750.963 KB) | DOI: 10.14710/ik.ijms.20.3.135-142

Abstract

Aklimatisasi eksplan rumput laut hasil kultur jaringan merupakan proses adaptasi eksplan dengan lingkungan budidaya. Penelitian ini bertujuan untuk mengevaluasi performa pertumbuhan dan perkembangan eksplan rumput laut G. verrucosa dan G. gigas yang diaklimatisasi di tambak dan mendapatkan informasi awal mengenai prospek pengembangan budidaya rumput laut G. gigas di tambak. Eksplan rumput laut G. verrucosa dan G. gigas hasil kultur jaringan dipelihara dalam hapa berukuran 50x50x50 cm dengan berat awal 15 g.hapa-1 dan dipelihara di tambak. Desain penelitian adalah rancangan acak lengkap (RAL) dengan enam unit penelitian terdiri dari tiga ulangan untuk masing-masing spesies. Pemeliharaan eksplan dilakukan selama 60 hari dan setiap 15 hari dilakukan pengukuran bobot, panjang dan perkembangan eksplan serta monitoring terhadap kualitas air. Pengamatan histologi sel rumput laut G. verrucosa dan G. gigas dilakukan dibawah mikroskop. Analisis data pertumbuhan dilakukan dengan uji komparatif independent t-test sedangkan data perkembangan eksplan dan histologi sel rumput laut dianalisis secara deskritif. Pada pemeliharaan di tambak kedua jenis rumput laut memiliki pertumbuhan yang berbeda nyata (P<0,05). Rumput laut G. verrucosa memiliki bobot mutlak lebih tinggi (221,82 g) dari G. gigas (51,94 g) dan LPH (laju pertumbuhan harian) bobot lebih tinggi (3,27%) dari G. gigas (2%). Rumput laut G. verrucosa juga memiliki pertambahan panjang yang lebih tinggi (5,28 cm) dari G. gigas (2,71 cm) dengan LPH panjang masing-masing sebesar 3,06% dan 2,18%. Perkembangan eksplan rumput laut G. verrucosa lebih cepat daripada G. gigas karena faktor fisika dan kimia lingkungan perairan tambak yang tidak sesuai untuk pertumbuhan rumput laut G. gigas yang memiliki susunan sel korteks lebih rapat. Kata kunci: pertumbuhan, perkembangan, G. verrucosa, G. gigas, eksplan, tambak Acclimatization of tissue culture seaweed explants was an adaptation procces of explants to cultivation environment. This study aims to evaluate the growth and development of G. verrucosa dan G. gigas explants on pond acclimatization as early information of pond aquaculture development of G. gigas. Explants of G. verrucosa and G. gigas were rearing on 50x50x50 cm cage net with 15 g.cage-1 of initial weight and cutured on pond. The study was a completely randomized design with six unit experiment including three replicates for each species. Acclimation was done in 60 days then explants weight, length, development, and water quality were monitored every 15 days. G. verrucosa and G. gigas cell histology was observed under microscope. Growth data was analyzed comparatively using independent t-test then explants development and cell histology were represented descriptively. The study showed that the growth of both species was significantly different (P<0.05) on pond cultivation. G. verrucosa had higher weight (221.82 g) than G. gigas (51.94 g) also higher DGR (daily growth rate) (3.27%) than G. gigas (2%). G. verrucosa also had higher elongation (5.28 cm) than G. gigas (2.71 cm) with length DGR of 3.06% and 2.18%, respectively. The development of G. verrucosa explants was better than G. gigas, because of the physical and chemical environment of pond water was not suitable for G. gigas which had dense cortical structure. Keywords: growth, development, G. verrucosa, G. gigas, explants, pond
THE EFFECTS OF STIMULANT GROWTH HORMONES ON TISSUE CULTURE OF SEAWEED KAPPAPHYCUS ALVAREZII IN VITRO Fadel, Ariyati H; Gerung, Grevo S; Suryati, Emma; Rumengan, Inneke F.M
AQUATIC SCIENCE & MANAGEMENT Edisi Khusus 1 (2013): Mei
Publisher : Graduate Program of Sam Ratulangi University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35800/jasm.0.0.2013.2282

Abstract

In order to anticipate the qualified and sustainable seed requirement for seaweed culture, it is necessary to conduct tissue culture for vegetative cultivation of isolated leaves, bud, and stemin an artificial medium enriched with nutrient and growth regulator. The purpose of this study is to obtain newly grown plant in a big quantity in relatively short period of time, with physiological and morphological properties similar to the stocks. Culture media used were Grund Medium and PES with an addition of a growth regulator, IAA (Indol acetic acid) and BAP (Benzil amino purin). The buds produced were buds with similar properties as the parent. The longest bud (1,851 mm) was obtained in Grund Medium with IAA treatment, while the length of bud in PES medium was only 0.612 mm. The number of buds was highest (10,6)  in Grund media with IAA+BAP (1:1) treatment, and 6,82 with IAA treatment in PES media. The survival rate of explants was highest in media enriched with 0.5 mg/L IAA (indol acetic acid). The best media for growing seaweed Kappaphycus alvarezii was Grund Medium© Untuk mengantisipasi kebutuhan bibit yang berkualitas dan tersedia  secara kontinyu, diperlukan suatu upaya kultur jaringan untuk perbanyakan tanaman secara vegetatif dengan mengisolasi bagian tanaman seperti daun, mata tunas, serta batang dalam media buatan secara aseptik yang diperkaya dengan nutrien dan zat perangsang tumbuh. Tujuannya untuk mendapatkan tanaman baru dalam jumlah banyak dalam waktu yang relatif singkat, yang mempunyai sifat fisiologi dan morfologis sama dengan tanaman induknya. Media kultur yang digunakan adalah media Grund Medium dan PES dengan penambahan zat perangsang tumbuh yaitu IAA (Indol acetic acid) dan BAP (Benzil amino purin). Tunas yang dihasilkan merupakan anakan yang mempunyai sifat yang sama dengan induknya.  Panjang tunas tertinggi dicapai pada media Grund Medium dengan perlakuan IAA (1,851 mm) dan media PES sebesar 0,612 mm. Sedangkan jumlah tunas tertinggi dicapai perlakuan IAA+BAP (1:1) sebesar 10,6 pada media Grund dan perlakuan IAA sebesar 6,82 pada media PES. Untuk tingkat kelangsungan hidup (sintasan) eksplan yang paling baik pada media yang diberikan pupuk IAA (indol acetic acit) dengan kosentrasi 0,5 mg/L. sedangkan media yang baik untuk pertumbuhan rumput laut Kappaphycus alvarezii adalah media Grund Medium©
BINARY VECTOR CONSTRUCTION OF KAPPA(κ)-CARRAGEENASE GENE AND TRANSFORMATION TO AGROBACTERIUM TUMEFACIENS AS MEDIATOR FOR SEAWEED TRANSGENIC GENERATION Rajamuddin, Muh Alias L.; Alimuddin, .; Widyastuti, Utut; Harris, Enang; Suryati, Emma
Jurnal Ilmu dan Teknologi Kelautan Tropis Vol. 8 No. 1 (2016): Elektronik Jurnal Ilmu dan Teknologi Kelautan Tropis
Publisher : Department of Marine Science and Technology, Faculty of Fisheries and Marine Science, IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (852.637 KB) | DOI: 10.29244/jitkt.v8i1.13602

Abstract

Increasing of kappa (?)-carrageenan content in Kappaphycus alvarezii seaweed is potentially be achieved by applying transgenesis technology. This study was performed to obtain a construction of  ?-Carrageenase gene and Agrobacterium tumefaciens to carry those construction genes.  The ?-Carrageenase (?-Car) gene was involved in ?-carrageenan biosynthesis. The ?-Car gene sequence was ligated between the 35S CaMV promoter and tNos terminator sequences to generate pMSH/?-Car expression vector. Transformation of pMSH/?-Car plasmid to Escherichia coli was performed by heat-shock method, and to Agrobacterium tumefaciens by tri-parental mating method. The results showed that several colonies of E. coli and A. tumefaciens grew in the selective culture mediums containing antibiotic. PCR analysis using primers 35S-Forward and tNos-Reverse with DNA template from those bacterial colonies resulted DNA fragment of about 2,000 bp, the same as the total length of 35S CaMV promoter, ?-Car gene and tNos terminator sequences. Therefore, the construction of pMSH/?-Car gene was succeeded and a colony of A. tumefaciens transformant carrying pMSH/?-Car plasmid was successfully produced.                                                                                   Keywords:  Agrobacterium tumefaciens, kappa(?)-Carrageenase gene, transgenesis, vector
BIOSORPSI CAMPURAN LOGAM PB2+ DAN ZN2+ OLEH CHAETOCEROS CALCITRANS Hala, Yusafir; Suryati, Emma; Taba, Paulina
CHEMISTRY PROGRESS Vol 5, No 2 (2012)
Publisher : Sam Ratulangi University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35799/cp.5.2.2012.772

Abstract

Penelitian ini bertujuan untuk menentukan dampak jerapan campuran ion logam Pb(II) dan Zn(II) terhadap pertumbuhan mikroalga Chaetoceros calcitrans dan menentukan efisiensi penjerapan kedua ion logam dalam campuran. Pemaparan ion logam dengan berbagai variasi konsentrasi Pb(II) terhadap Zn(II)dilakukan setelah diperoleh pertumbuhan optimum C. calcitrans, yakni pada hari ke-9. Konsentrasi ion logam Pb(II) dan Zn(II) setelah pemaparan ditentukan dengan spektrofotometer serapan atom. Penambahan ion Pb(II) ke dalam Zn(II)membuat penjerapan ion Zn(II) oleh C. calcitrans turun dibandingkan dengan penjerapan ion tunggal Zn(II). Hal yang sama juga terjadi pada penambahan ion Zn(II)ke dalam ion Pb(II). Hasil penelitian menunjukkan bahwa ion Pb(II) lebih banyak terjerap oleh C. calcitrans dibanding ion Zn(II). Effisiensi penjerapan optimum ion Pb(II) sebesar 64,44% pada perbandingan konsentrasi Pb(II) terhadap Zn(II) 45 : 30 ppm sedangkan penjerapan ion Zn(II) yaitu 56,33% pada perbandingan konsentrasi Zn(II) terhadap Pb(II) 15 : 30 ppm.The main purpose of this research was to determine the adsorption effect of the mixture of Pb(II) and Zn(II) on the growth of Chaetoceros calcitrans and to determine the adsorption efficiency of the ions in the mixture. Exposure of Pb(II) and Zn(II)ions was conducted after the optimum growth of C. calcitrans was obtained, that is at the ninth day with the variation of the concentration ratio of Pb(II) to Zn(II). Concentration of Pb(II) and Zn(II) ions after exposure was determined by atomic absorption spectrophotometer. Addition of Pb(II) ion in solution Zn(II) ion resulted in the decrease of the Zn(II)ion adsorbed by C. calcitrans compared to the adsorption of the single ion of Zn(II). The same result was obtained when Zn(II) ion was added in Pb(II) solution. Results showed that the adsorption of Pb2+ ion by C. calcitrans was higher than that of Zn(II) ion. The maximum adsorption efficiency of Pb(II) ion was 64.44% at the Pb(II):Zn(II) ratio of 45:30, whereas that of Zn(II)ion was 56.33% at the Zn(II):Pb(II) ratio of 15:30.
TOXICITY OF Aaptos suberitoides EXTRACT ON TIGER SHRIMP Penaeous monodon POST LARVAE Rosmiati, Rosmiati; Suryati, Emma
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 9, No 2 (2014): August 2014
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v9i2.98

Abstract

Marine sponges contain bioactive compounds which can be used not only for human diseases treatment but also for animal diseases treatment. Aaptos suberitoides has been reported to have potential as antibacterial against Vibrio harveyi in vitro. This study aims to study the toxicity effect of Aaptos suberitoides extract on tiger shrimp Penaeous monodon post larvae. The toxicity study of this extract was done by using soaking method. This experiment used Completely Randomized Design with 6 treatments of A.suberitoides extract namely A). 0 ppm, B). 31.3 ppm, C). 62.5 ppm, D). 125, E). 250 ppm, and F). 500 ppm. Each treatment consisted of 3 replications. Findings showed that A.suberitoides butanol extracts at the concentration up to125 ppm were safe for tiger shrimp post larvae. On the other hand, the higher concentration of that gave a toxic effect on the post larvae. The lowest mortality (20.0%) was obtained on the post larvae treated with butanol extract at the concentration of 31.3 ppm and the highest was found at the concentration of 500 ppm (97.0%). Compared to the negative control (0 ppm), there was no any difference exhibited by the post larvae treated with butanol extract at the concentrations of 31.3, 62.5, and 125 ppm. Beside that, these concentrations gave a normal growth and no abnormal behavior. The histological observation did not also show damage on post larvae haepatopancreas tissue.
SPONGE MOLECULAR SCREENING FOR ANTIMICROBIAL GENES BY PCR Rosmiati, Rosmiati; Suryati, Emma; Parenrengi, Andi; Sulaeman, Sulaeman
Indonesian Aquaculture Journal Vol 2, No 2 (2007): (December 2007)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (160.132 KB) | DOI: 10.15578/iaj.2.2.2007.127-131

Abstract

Molecular biotechnology approach has been applied on sponge for preventingdiseases on fishery culture. This is important for anticipating and avoiding the using of amount of sponge in nature. The present study aims to screen the antimicrobial (oxytetracycline and chloramphenicol) genes of sponge. DNA extraction of samples was done using the DNeasy Plant mini kit, Phenol-Chloroform and modification of Phenol-Chloroform methods. The presence of oxytetracycline and chloramphenicol genes in sponge was detected using Polymerase chain reaction (PCR) technique. Result of the study showed that four species (Sylotella aurantium, Acanthella kletra, Gelliodes fibulatus and Auletta sp. were amplified for oxytetracycline and two species (Auletta sp. and Pericharax sp.) of sponge were amplified for chloramphenicol at each 226 bp.
GENETIC, COLORATION, AND GROWTH PERFORMANCE OF TWO DIFFERENT VARIETIES OF Kappaphycus alvarezii Sulaeman, Sulaeman; Parenrengi, Andi; Suryati, Emma; Rosmiati, Rosmiati
Indonesian Aquaculture Journal Vol 2, No 1 (2007): (June 2007)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (81.59 KB) | DOI: 10.15578/iaj.2.1.2007.23-26

Abstract

Two different colors (green and brown) of Kappaphycus alvarezii have been farmed in Indonesian waters for many years. This study aimed at comparing two ‘varieties’, i.e. green and brown, both genetically and morphologically. Samples for DNA analysis were collected from a farmer in Pinrang Regency, South Sulawesi. Five universal primers i.e. Ca-01, Ca-02, P-40, P-50, and DALRP were selected to obtain DNA genetic markers in differentiating the green and brown varieties. To compare coloration patterns during cultivation and the growth performance of both varieties, a field experiment was performed in a seaweed farming area in Pinrang Regency, during dry season of August-September 2004. The result of genetic assessment showed that the five selected primers revealed different RAPD banding pattern for both varieties. P-50 and DALRP primers demonstrated the greatest amplification in differentiating RAPD fragment between green and brown varieties. Fragment 900 bp and 1.300 bp were consistently generated in the green variety but were not amplified in the brown variety. The result of the field study confirmed that the coloration pattern of green and brown varieties was fixed; no interchange in color occurred during one crop cultivation.
IN VITRO GROWTH RATE OF Kappaphycus alvarezii MICROPROPAGULE AND EMBRYO BY ENRICHMENT MEDIUM WITH SEAWEED EXTRACT Suryati, Emma; Rosmiati, Rosmiati; Parenrengi, Andi; Tenriulo, Andi
Indonesian Aquaculture Journal Vol 10, No 1 (2015): (June 2015)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (416.68 KB) | DOI: 10.15578/iaj.10.1.2015.13-17

Abstract

The development of micropropagule and embryo of seaweed depend on nutrient and fertilizer used. Seaweed has been reported contain hormone regulators such as auxine, cytokinine, gibbereline, and various minerals applied in stimulating the growth ocra plant and wheat culture. The objectives of this study were to determine the potential of Kappaphycus alvarezii extract and its optimal concentration in accelerating of Kappaphycus alvarezii micropropagule and embryo growth. Micropropagule and embryo produced through callus induction were planted into PES 1/20 liquid medium supplemented with seaweed extract at the concentrations of 0 (control), 25, 50, 75, and 100 μL in 10 mL of medium. The results showed that medium enrichment with 50 μL of seaweed extract had the highest survival rate and growth of thallus. In addition, this concentration was also resulted in a good performance of K. alvarezii thallus with the lighter color. The advantage of this study for seaweed cultivation in Indonesia, among others, seaweed can be used as fertilizer, especially in the maintenance of seaweed seed, so that cultivation can be better develop.