Articles

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
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
GROWTH, MORPHOLOGY AND GROWTH RELATED HORMONE LEVEL IN KAPPAPHYCUS ALVAREZII PRODUCED BY MASS SELECTION IN GORONTALO WATERS, INDONESIA Fadilah, Siti; Alimuddin, .; Pong-Masak, Petrus Rani; Santoso, Joko; Parenrengi, Andi
HAYATI Journal of Biosciences Vol. 23 No. 1 (2016): January 2016
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1072.105 KB) | DOI: 10.4308/hjb.23.1.29

Abstract

The use of high quality seed can support the success of the seaweed cultivation. This study was conducted to evaluate the growth performance, morphology and growth related hormone level of brown strain seaweed Kappaphycus alvarezii seed produced by mass selection. Selection was performed in the Tomini Gulf, Gorontalo, based on mass selection of seaweed seed protocol with a slight modification in cut-off 10% of the highest daily growth rate. Selection was carried out for four generations. The selected 4thgeneration of seed was then used in cultivation performance test in the Celebes Sea, North Gorontalo, for three production cycles. The results showed that the selected K. alvarezii has higher clump weight and daily growth rate, longer thallus, more number of branches, and shorter internodes compared to the unselected control and seaweed from the farmer as external control. Furthermore, total sugar content, levels of kinetin hormone and kinetin:indole-3-acetic acid ratio were higher in selected seaweeds than that of unselected control and external control. Thus, mass selection method could be used to produce high growth of seed, and kinetin and indole-3-acetic acid play an important role in growth of K. alvarezii.
REGENERASI KALUS BERFILAMEN RUMPUT LAUT KAPPAPHYCUS ALVAREZII PADA BERBAGAI PERBANDINGAN ZAT PENGATUR TUMBUH AUKSIN (INDOLE ACETIC ACID) DAN SITOKININ (KINETIN, ZEATIN) Mulyaningrum, Sri Redjeki Hesti; Parenrengi, Andi; Risjani, Yenny; Nursyam, Happy
The Journal of Experimental Life Science Vol 2, No 1 (2012)
Publisher : Graduate School, University of Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1250.749 KB) | DOI: 10.21776/ub.jels.2012.002.01.05

Abstract

Abstrak Interaksi auksin-sitokinin dianggap penting untuk mengatur pertumbuhan dan perkembangan dalam jaringan tanaman. Penelitian ini bertujuan untuk menentukan komposisi auksin dan sitokinin yang tepat untuk regenerasi filamen kalus rumput laut K. alvarezii. Kultur filamen kalus dilakukan pada media cair dengan formulasi ZPT indole acetic acid (IAA) : kinetin : zeatin, dengan komposisi konsentrasi sebagai berikut : A (0,4:0:1) ppm; B (0,4:0,25:0,75) ppm; C (0,4:0,5:0,5) ppm; D (0,4:0,75:0,25) ppm; E (0,4:1:0) ppm; kontrol (tanpa ZPT). Penelitian menggunakan rancangan acak lengkap dengan pengulangan masing-masing perlakuan 3 kali. Parameter yang diamati adalah laju pertumbuhan harian, sintasan, kecepatan regenerasi, panjang tunas dan perkembangan morfologi. Hasil yang diperoleh menunjukkan bahwa formula ZPT terbaik adalah formula A dengan laju pertumbuhan harian 1,929%/hari, sintasan 83,33%, kecepatan regenerasi 41,67% dan rata-rata panjang tunas 44,59 µm. Tunas mulai terbentuk pada 15 hari masa kultur.Kata kunci: formula zat pengatur tumbuh, auksin-sitokinin, K. alvarezii, mikropropagasi
PENGARUH APLIKASI DSRNA VP-15 IN VITRO DAN IN VIVO TERHADAP SINTASAN DAN RESPONS IMUN UDANG WINDU PENAEUS MONODON Parenrengi, Andi; Tenriulo, Andi; Mulyaningrum, Sri Redjeki Hesti; Lante, Samuel; Nawang, Agus
Jurnal Riset Akuakultur Vol 14, No 4 (2019): (Desember, 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 (277.291 KB) | DOI: 10.15578/jra.14.4.2019.213-223

Abstract

Teknologi RNA interference (RNAi) merupakan salah satu pendekatan yang digunakan untuk meningkatkan resistensi udang windu terhadap infeksi patogen termasuk WSSV. Pengembangan teknologi RNAi melalui aplikasi untai ganda RNA (dsRNA) yang berasal dari gen pengkode viral protein (VP) dari WSSV telah mulai dikembangkan pada udang. Penelitian ini bertujuan untuk mengkaji sintasan dan respons imun udang windu yang diberi VP-15 pasca uji tantang dengan WSSV. Udang windu (panjang 15,21 ± 1,19 cm dan bobot 32,5 ± 1,83 g) diinjeksi dengan 0,2 µg/ekor dsRNA in vitro (A), dsRNA in vivo (B), dan larutan garam/kontrol (C). Setelah tiga hari vaksinasi, udang windu ditantang dengan WSSV dengan dosis 50 µL/ekor. Pengamatan sintasan dilakukan setiap hari, sedangkan respons imun (THC dan aktivitas proPO) dilakukan pada awal dan hari ke-1, ke-3, dan ke-5 pasca uji tantang, serta analisis ekspresi gen antivirus dan histopatologi hepatopankreas dilakukan pada akhir penelitian. Hasil penelitian menunjukkan bahwa aplikasi dsRNA berpengaruh nyata (P&lt;0,05) terhadap sintasan, THC, dan proPO. Sintasan udang windu yang diberi dsRNA VP-15 in vitro dan in vivo memberikan sintasan yang lebih tinggi 75% dibandingkan dengan kontrol. Nilai proPO tertinggi didapatkan pada dsRNA in vivo (0,138); kemudian dsRNA in vitro (0,093); dan terendah kontrol (0,061); sedangkan THC tertinggi (5.704 x 104 sel/mL) pada dsRNA in vivo, kemudian dsRNA in vitro (3.516 x 104 sel/mL) dan terendah pada perlakuan kontrol (3.322 x 104 sel/mL). Ekspresi gen antivirus semakin meningkat dengan semakin lamanya udang windu terpapar dengan WSSV. Jaringan hepatopankreas udang windu pada perlakuan kontrol (tanpa dsRNA) menunjukkan adanya kerusakan sel akibat infeksi virus.RNA interference (RNAi) technology is one of the approaches used to improve tiger shrimp Penaeus monodon resistance against WSSV infection. The development of RNAi technology through double-stranded RNA (dsRNA) isolated from gene encoding viral protein (VP) of WSSV has been applied to shrimp. This study was aimed to assess the survival rate and immune response of injected-VP-15 WSSV tiger shrimp after a challenge with WSSV. The tiger shrimp (15.21 ± 1.19 cm in length and 32.5 ± 1.83 g in weight) were injected with 0.02 µg/shrimp of in vitro dsRNA (A), in vivo dsRNA (b) and saline solution (C). After three days of vaccination, the tiger shrimp were challenged with WSSV using a dosage of 50 µL/shrimp. The survival rate was observed daily. Analyses of immune responses (hemocyte total and PO activity) were performed in several stages: before the challenge test and day-1, day-3, and day-5 post-challenge test. The expression of the antivirus gene and hepatopancreas histophatology were was observed at the end of the experiment. The results showed that the application of dsRNA significantly influenced the shrimp survival rate, THC, and proPO. Tiger shrimp injected with dsRNA VP-15 of in vitro and in vivo exhibited a higher 75% survival rate than the control (P&lt;0.05). The highest proPO activity (0.138) was obtained at dsRNA in vivo, followed by dsRNA in vitro (0.093) and the lowest (0.061) in the control. The highest THC (5,704 x 104 cell/mL) was in vivo dsRNA, then in vitro dsRNA (3,516 x 104 cell/mL), and the lowest in the control (3,322 x 104 cell/mL). The longer the exposure with WSSV, the higher the antivirus gene expression. Histopathology analysis showed some damages to the hepatopancreas cells in the control shrimp (without dsRNA) caused by the virus infection.
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

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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.
CHARACTERISTICS OF VIRAL PROTEIN, VP-15, OF WHITESPOT SYNDROME VIRUS ISOLATED FROM INFECTED TIGER SHRIMP Penaeus monodon (Fabricius, 1798) Parenrengi, Andi; Alimuddin, Alimuddin; Tenriulo, Andi
Indonesian Aquaculture Journal Vol 12, No 2 (2017): (December, 2017)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (157.395 KB) | DOI: 10.15578/iaj.12.2.2017.67-75

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White spot syndrome virus (WSSV) has caused mass mortality on tiger shrimp (Penaeus monodon) culture and adversely affects prawn industry worldwide including Indonesia. It is well known that the protein structure of WSSV plays an important role in the virus infection and morphogenesis process. A viral protein structure called VP-15 is located in the nucleocapsid of virion virus. The protein structure involves in the life cycle of WSSV in host cells. A gene encoding VP-15 could be involved in constructing the RNA interference (RNAi), so it is needed to isolate and characterize for RNAi technology purpose. The study was aimed to isolate and characterize the VP-15 from the infected WSSV tiger shrimp. The characterization of VP-15 was undertaken through assessment of nucleotide sequence, amino acid deduction, alignment nucleotide/protein searches using Genetyx and BLAST program, and dendrogram construction analysis. The results showed that VP-15 was successfully isolated in form of ORFDNA with a fragment size of 243 bp. The phylogenetic tree analysis revealed three clusters corresponding to the time (year) of isolates collection. The VP-15 consisted of 80 amino acids, two start codons (ATG), one stop codon (TAA), and one Kozak context (AAAATGG). Hydrophilic amino acid was the highest composition (44.2%), followed by neutral (31.2%) and hydrophobic (24.6%) amino acid groups. The VP-15 was rich in amino acid of lysine (21.3%), arginine (22.9%) and serine (24.6%). The successful isolation of VP-15 is a very important step in providing a basic yet suitable material in constructing the dsRNA vaccine to control shrimp diseases in aquaculture.
EXPRESSION OF ANTIVIRAL GENE ON TIGER SHRIMP Penaeus monodon AT DIFFERENT TISSUE AND BODY SIZE Parenrengi, Andi; Tenriulo, Andi; Lante, Samuel
Indonesian Aquaculture Journal Vol 7, No 2 (2012): (December 2012)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (484.161 KB) | DOI: 10.15578/iaj.7.2.2012.95-104

Abstract

The role of tiger shrimp defense against invading pathogen on molecular level such antiviral gene expression is limited to be reported. Gene expression is a process which codes information of genes that is converted to the protein as a phenotype. Distribution of PmAV antivirus gene, that has been reported as an important gene on non-specific response immune, is needed to be observed to several organs/tissues and size of tiger shrimp. The aim of this study is to determine the distribution of gene antiviral expression at several organ/tissue and size of shrimp. The organs/tissues observed in this study were: gill, hepatopancres, muscle tissue, eyes, heart, stomach, gonad, and intestine. While the size of shrimp consisted of three groups, those are: (A) 10-20 g/ind., (B) 30-40 g/ind., and (C) 60-70 g/ind. Analysis of antiviral gene expression was performed by RNA extraction, followed by the cDNA syntesis, and amplification of gene expression by semi-quantitative PCR. The result of PCR optimation showed the optimal concentration of cDNA and primer was 1 μL and 50 mol, respectively for PCR final volume of 25 μL. Antiviral gene was expressed on the hepatopancreas and stomach in percentage of 50.0% and 16.7%, respectively. While the highest percentage of individual expressing the antiviral gene was observed in the shrimp size of C (66.7%), followed by B (50.0%) and A (16.7%). The result of study implied that the hepatopancreas has importantly involed in tiger shrimp defense mechanism on viral infection.
ANALYSIS OF IMMUNE RESPONSES ON TRANSGENIC TIGER SHRIMP (Penaeus monodon) AGAINST PATHOGENIC BACTERIUM Vibrio harveyi Parenrengi, Andi; Tampangallo, Bunga Rante; Tenriulo, Andi
Indonesian Aquaculture Journal Vol 9, No 1 (2014): (June 2014)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (406.087 KB) | DOI: 10.15578/iaj.9.1.2014.23-32

Abstract

Vibriosis is one of main diseases of the black tiger shrimp Penaeus monodon infected by pathogenic bioluminous bacterium Vibrio harveyi that can cause mass mortalities in shrimp culture. The bacteria can also trigger the disease white spot syndrome virus (WSSV). An effort to produce shrimp disease-resistant strains has been done through transgenesis technology with antiviral gene transfection. By this technology, it is expected an increase in the immune response of shrimp in a variety of diseasecausing pathogens. This study aimed to determine the immune responses (total haemocytes, haemocyte differentiation, and phenoloxydase activity) of transgenic tiger shrimp against pathogenic bacterium V. harveyi. Research using completely randomized design, which consists of two treatments and three replications. Test animals being used were transgenic and non-transgenic shrimp with size, weight 3.93±1.25 g and a total length of 7.59±0.87 cm. Treatments being tested were the injection of bacterium V. harveyi (density of 5x106 cfu/mL) of 0.1 mL/individual on transgenic (A) and non-transgenic shrimp (B). Immune response parameters such as total haemocytes, haemocyte differentiation, and phenoloxydase activity were observed on day 1, 3, and 6 days after challenging. Data were analyzed using t-test by SPSS software. The results showed that the total haemocyte of transgenic shrimp was not significantly different (P&gt;0.05) from non-transgenic shrimp, but haemocyte differentiation and phenoloxydase activity were significantly different (P&lt;0.05) especially on sixth days after being exposed to the bioluminescent bacteria. The study results implied that transgenic shrimp has a better immune response compared than non-transgenic shrimp.