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Ekspresi Gen Penyandi Phenylalanine Ammonia Lyase (PAL) Cabai (Capsicum annuum) Sebagai Respons Terhadap Fusarium oxysporum Dewi, Yulita Wiwik Irana; Ferniah, Rejeki Siti; Pujiyanto, Sri
Berkala Bioteknologi Vol. 1, No. 2, November 2018
Publisher : Berkala Bioteknologi

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Tanaman cabai (Capsicum annuum) merupakan salah satu sektor pertanian terbesar di Indonesia. Tanaman cabai banyak dibudidayakan oleh masyarakat di Indonesia sampai produksinya diekspor ke luar negeri. Namun pada tahun 2015 produksi cabai di Indonesia mengalami penurunan dibandingkan tahun-tahun sebelumnya. Penurunan produksi cabai di Indonesia disebabkan karena adanya serangan organisme pengganggu tanaman salah satunya adalah Fusarium oxysporum yang menyebabkan terjadinya penyakit rebah tanaman. Fusarium oxysporum adalah jamur yang berada di tanah, dan beberapa strain F. oxysporum bersifat patogen terhadap tanaman dan bersifat sulit dikendalikan. PAL adalah enzim yang mengkatalis perubahan phenilalanin menjadi ammonia dan trans-sinamat yang merespon tekanan biotik dan abiotik seperti patogen, UV radiasi, dan suhu rendah. Tujuan dari penelitian ini yaitu, untuk mengetahui ekspresi gen penyandi Phenylalanin Ammonia Lyase (PAL) pada cabai (Capsicum annuum) sebagai respons terhadap Fusarium oxysporum. Metode dari penelitian ini meliputi penanaman dan pemeliharaan tanaman, inokulasi F. oxysporum ke tanaman cabai, isolasi RNA daun cabai, analisis ekspresi gen PAL dengan menggunakan qRT-PCR dan analisis data. Hasil dari penelitian ini adalah ekspresi gen PAL pada jam ke-6 terekspresi sedikit lebih tinggi daripada kontrol dan mengalami penurunan pada jam ke-48 dan ke-96. Penelitian ini dapat disimpulkan bahwa tidak ada peningkatan ekspresi gen penyandi Phenilalanine Ammonia Lyase pada daun tanaman cabai (Capsicum annuum) sampai dengan 96 jam setelah inokulasi.

Pertumbuhan dan Produksi Pigmen Merah oleh Serratia marcescens pada Berbagai Sumber Karbon Wicaksono, Setiawan; Kusdiyantini, Endang; Raharjo, Budi
Berkala Bioteknologi Vol. 3 No. 2 November 2020
Publisher : Berkala Bioteknologi

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Bakteri Serratia marcescens adalah salah satu bakteri penghasil pigmen merah yang banyak dimanfaatkan sebagai pewarna alami. Bakteri ini diisolasi dari sedimen sumber air panas di Gedong Songo, Bandungan, Semarang. S. marcescens memiliki potensi sebagai penghasil pigmen alami. Penelitian ini dilakukan untuk mengukur pertumbuhan dan produksi pigmen dalam medium NB yang mengandung sumber karbon yang berbeda. Sumber karbon yang digunakan adalah glukosa, fruktosa, maltosa, dan laktosa. Metode yang digunakan adalah pengukuran pertumbuhan berdasarkan nilai berat kering sel dan nilai ODλ=600nm, pengukuran gula reduksi, pengukuran pH medium pertumbuhan, dan pengukuran konsentrasi pigmen merah. Hasil analisis yang diperoleh dalam penelitian ini menunjukkan bahwa pemberian sumber karbon tidak berpengaruh signifikan terhadap pertumbuhan S. marcescens. Sumber karbon terbaik untuk produksi pigmen merah adalah laktosa dengan konsentrasi pigmen sebesar 0,451 mg/L yang dicapai pada waktu inkubasi 24 jam.

Antibacterial Activity Tests of Endophytic Bacteria Isolates From Tea Plant (Camellia sinensis) Againts Staphylococcus aureus and Staphylococcus epidermidis Sari, Suli Arum; Pujiyanto, Sri; Suprihadi, Agung
Berkala Bioteknologi Vol. 5, No. 2, November 2022
Publisher : Berkala Bioteknologi

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Staphylococcus is one of the most common types of bacteria in Asia that causes local infectious diseases of the skin, nose, urethra, vagina, digestive tract, pneumonia, endocarditis, septic arthritis, and septicemia. Staphylococcus aureus and Staphylococcus epidermidis are the most common types of Staphylococcus in Asia. Tea plants contain bioactive compounds and endophytic bacteria which are widely used as antimicrobial agents. Endophytic bacteria are bacteria that exist in plant tissues, not pathogenic, and have the ability as the host plant. The purpose of this study was to determine the antibacterial activity of endophytic bacterial isolates of tea plants (Camellia sinensis) against the growth of Staphylococcus aureus and Staphylococcus epidermidis bacteria. The antibacterial activity test of endophytic bacteria of tea plants includes a series of processes such as sample selection, surface sterilization of samples, isolation of endophytic bacteria in agar medium, purification, screening, suspension of endophytic bacteria in 0.9 % NaCl and standardized with 0.5 McFarland, making endophytic bacterial culture in nutrient borth, making endophytic bacterial supernatant and antibacterial activity test with paper disc diffusion method. The results obtained are the antibacterial activity of the endophytic bacterial supernatant isolates B23, B14, and A2 on the growth of Staphylococcus aureus and Staphylococcus epidermidis. The best antibacterial activity was found in endophytic bacterial B14 isolates with inhibition zones of 7.75 mm and 12.5 mm followed by B23 isolates with 7.5 mm and 8.25 mm inhibition zones and A2 isolates with large inhibition zones of 7.42 mm and 8.16 mm. Endophytic bacteria isolates of tea plant show better antibacterial activity against the growth of Staphylococcus aureus than Staphylococcus epidermidis.

Identifikasi Jenis Pigmen dan Uji Potensi Antioksidan Ekstrak Pigmen Bakteri Serratia marcescens Hasil Isolasi dari Sedimen Sumber Air Panas Gedong Songo Naufal, Adhitya; Kusdiyantini, Endang; Raharjo, Budi
Berkala Bioteknologi Vol. 4, No. 1, April 2021
Publisher : Berkala Bioteknologi

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Pigments can be produced by plants, animals, and microbes, including the bacteria Serratia marcescens. Pigments are believed to be used as antibacterial, anticancer, antibiotic and antioxidant. The quality of the pigment is greatly determined by its type. The aim of this study was to identify the type of pigment produced by S. marcescens resulting from the sediment of Gedong Songo hot spring, and testing its antioxidant activity. S. marcescens was grown on Nutrient Broth (NB) medium for 96 hours at room temperature and sampled once every 6 hours for growth measurement and pigment measurement. Cell dry weight was used to measure the growth of S. marcescens, while the measurement of pigment production was done using spectrophotometer with 535 nm wavelength for red pigment and identification of pigment with Thin Layer Chromatography and UV-Vis and testing of antioxidant activity using carotene bleaching method. The results showed the measurement of pigment production has optimal point at 48 hours at 1,319 mg/L. Identification of pigment type S. marcescens using TLC obtained value of 0,8 with spectrophotometer wavelength at 536 nm. The value of antioxidant activity Pigment S. marcescens obtained at 13%.

Isolasi Khamir Fermentatif dari Batang Tanaman Tebu (Saccharum officinarum. L) dan Hasil Identifikasinya Berdasarkan Sekuens Internal Transcribed Spacer Anggraini, Ika; Ferniah, Rejeki Siti; Kusdiyantini, Endang
Berkala Bioteknologi Vol. 5, No. 2, November 2022
Publisher : Berkala Bioteknologi

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Yeast is a single-celled fungus that acts as epiphytes, endophytes or parasites. Yeast is divided into fermentative yeast and oxidative yeast. Fermentative yeast can produce primary and secondary metabolites. The role of fermentative yeast is widely used in the food industry, health and energy, so necessary to be explore fermentative yeast from sugarcane stems. The purpose of this study was to isolate fermentative yeast from sugarcane stems and identify molecular yeast based on the Internal Transcribed Spacer sequences (ITS). Isolation of epiphytic and endophytic yeast was carried out by spread plate of water soak sugarcane and sugar cane juice. Yeast isolation using 2 media, PDA and YGP with chloramphenicol. Morphological characterization was carried out by observing macroscopic and microscopic characteristics. Biochemical characterization was carried out by carbohydrate fermentation test and 50% glucose media growth test. Selected isolates were molecularly identified using Internal Transcribed Spacer (ITS) sequences. Primers used are ITS 1 and ITS 4. Phylogenetic analysis using Neighbor Joining from MEGA-6 program. The results of isolation obtained 7 yeast isolates characterized morphologically and biochemically. The based result of morphology and biochemical characterization were found 1 selected isolate with name Ed 1B. Selected yeast isolate have characteristics are round colonies, creamy white, shiny surface, raised elevation, wavy edges, ovoid cell shape, cell diameter 4,74µm, budding, glucose fermentation and sucrose fermentation, but not for lactose and grow well of 50% glucose media. The results of the Basic Alignment Search Tools (BLAST) are Ed 1B isolates had 99% homology with Kodamaea ohmeri species.

Deteksi Gen tlh dan tdh pada Bakteri Vibrio parahaemolyticus dari Air Tambak Udang Vanname (Litopenaeus vannamei) di Kabupaten Rembang Hasrimi, Adila Nawan; Budiharjo, Anto; Jannah, Siti Nur
Berkala Bioteknologi Vol. 4, No. 2, November 2021
Publisher : Berkala Bioteknologi

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Vibrio parahaemolyticus merupakan bakteri halofilik gram negatif pada lingkungan aquatik. Semua strain Vibrio parahaemolyticus memiliki thermolabile hemolysin yang diregulasi oleh gen tlh. Thermostable direct hemolysin yang diregulasi oleh gen tdh adalah salah satu faktor virulensi utama pada Vibrio parhaemolyticus yaitu. Tujuan dari penelitian ini adalah untuk mendeteksi keberadaan gen tlh dan tdh pada bakteri Vibrio parahaemolyticus dari air tambak udang vanname di Kabupaten Rembang. Isolasi bakteri dari air tambak udang vanname pada media CD-VP menunjukkan pertumbuhan koloni bakteri berwarna hijau-kebiruan yang teridentifikasi spesifik sebagai bakteri Vibrio parahaemolyticus. Isolat bakteri teridentifikasi sebagai bakteri spesies Vibrio parahaemolyticus berdasarkan penyandi gen tlh. Analisis molekuler menunjukkan hasil tdh negatif, yang mengindikasikan bahwa bakteri tidak memiliki gen tdh untuk menyandi thermostable direct hemolysin yang menentukan faktor virulensi. Uji konfirmasi gen tdh pada isolat bakteri Vibrio parahaemolyticus dengan media agar wagatsuma menunjukkan hasil Kanagawa negatif yang mengindikasikan bahwa bakteri tidak menghasilkan thermostable direct hemolysin. Isolat Vibrio parahaemolyticus tidak mempunyai faktor virulensi untuk memulai kolonisasi pada organisme inang di lingkungan aquatik ini.

The Expression of Glucanase Encoding Gene (CaβGlu) in Chili (Capsicum annuum L.) As a Response to Fusarium oxysporum Infection. Rahmandanni, Yunnia; Pujiyanto, Sri; Ferniah, Rejeki Siti
Berkala Bioteknologi Vol. 5, No. 2, November 2022
Publisher : Berkala Bioteknologi

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Indonesia is a tropical country with highest level of biodiversity, especially in the agricultural sector. Chili (Capsicum annumm. L) is a very well-known and widely used agricultural product in the world, which makes chili becomes one of the most considerable national product. The chili production is oftentimes very susceptible to some diseases caused by virus, fungi, or bacteria. One of the most common diseases in chili cultivation is Fusarium wilt, which is caused by Fusarium oxysporum. This disease can cause a major loss and up to 50% crop failure. Many procedures have been done to find the best cultivar with a resistance trait to Fusarium oxysporum, including by observing and testing the chili’s genetic resistance. One of the resistance genes in chili is β -1, 3- glucanase-encoding gene, which produces an enzyme to hydrolize the cell wall of pathogenic fungi. This research aimed to determine the expression of the glucanase-encoding gene (CaβGlu) in chili as a response to Fusarium oxysporum infection. The methods including chili cultivation, F. oxysporum inoculation, isolation of chili leaves RNA, glucanase-encoding gene expression analysis using qRT-PCR, and data analysis. The result of CaβGlu gene expression is higher than the control in the first 6 hours after inoculation, and decreasing in the 48th and 96th hours. The conclusion was the infection of Fusarium oxysporum is activating the expression of CaβGlu gene which was expressed best in the first 6 hours after inoculation.

Pengaruh Iradiasi Sinar Gamma TerhadapInduksi Kalus dan Seleksi Tingkat Toleransi Padi (Oryza sativa L.) terhadap Cekaman Salinitas secara In-Vitro Lisdyayanti, Novita Dwi; Anwar, Syaiful; Darmawati, Adriani
Berkala Bioteknologi Vol. 5, No. 1, April 2022
Publisher : Berkala Bioteknologi

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The use of saline for rice cultivation needs to be support bytolerant rice varieties in saline land. The rice variety can be obtained through plant breeding programs, one of which is by physical mutation methods using gamma rays.. Physical mutation is random mutation, so we need to do selection after that treatment. Selection to get rice mutants that are tolerant of salinity stress can be done by planting on culture media that have been modified witt addition of NaCl.The perpose of this research is to examine the effect of gamma rays irradiation doses on rice callus induction and its tolerance level in saline media. This research used 2 designs, a monofactor complete randomize design at callus induction stage with a faktor in the form of gamma rays irradiation doses (0, 100, and 200 Gy) and factorial complete randomize designat the regeneration stage with a factor in form of gamma rays irradiation doses (0, 100 and 200 Gy) and the second factor is NaCl concentration (0 and 50 mM). The steps of this research were irradiation of Ciherang variety rice seed, making media for tissue culture, sterilization of rice seed explants, initiation of explants in callus induction media, subculture to regenerated media that had been modified with the addition of NaCl according to treatment, and then observation. The results showed that gamma rays irradiation could inhibit the growth of callus diameter and reduce the grofth of callus from explants of ciherang variety. All callus subcultured into regeneration media are unable to formbuds and turn black or die.

Deteksi Gen DXS dan Penentuan Jalur Biosintesis Karotenoid pada Chlorella pyrenoidosa Monalita, Ramadhebi; Kusumaningrum, Hermin Pancasakti; Budiharjo, Anto
Berkala Bioteknologi Vol. 5, No. 1, April 2022
Publisher : Berkala Bioteknologi

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Natural carotenoid synthesis has never exceeded synthetic products on a commercial scale. Lack of understanding of the microbiological and ecophysiological aspects of carotenoid-producing isolates leads to misidentification of species. One local isolate of green algae is used as a natural food source of carotenoids in the food industry, namely Chlorella pyrenoidosa. Carotenoid accumulation of the nonMVA pathway in green algae is determined by the enzyme D-1-deoxyxylulose 5-phosphate synthase, which is encoded by the D-1-deoxyxylulose 5-phosphate synthase (DXS) gene. The main purpose of this study was to detect the DXS gene as a carotenoid biosynthetic key enzyme encoder in C. pyrenoidosa whether or not to follow the non-MVA pathway for carotenoid biosynthesis or not. Morphological and ecophysiological characterization methods are carried out based on periodic observations and DXS gene detection using the guide Kuzuyama (2000). The results of the analysis of the similarity of the C. pyrenoidosa DXS gene in sustainable areas show that it can detect partial DXS genes from Chlamydomonas reinhardtii. The absence of growth inhibition in C. pyrenoidosa with lovastatin shows a non-MVA pathway that is the pathway used in carotenoid biosynthesis.

Biofilm Inhibition Activity from Obligate Marine Fungi Against Pathogenic Vibrio Bacteria in Whiteleg Shrimp, Penaeus vannamei Kristina, Kristina; Soowanayan, Chumporn; Suetrong, Satinee; Budiharjo, Anto; Rukmi, Isworo
Berkala Bioteknologi Vol. 5, No. 2, November 2022
Publisher : Berkala Bioteknologi

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Early mortality syndrome (EMS) is caused by toxic strains of Vibrio bacteria that are produced when bacteria colonize and form biofilm in the digestive tract of cultivated shrimp. One possible control strategy for shrimp Vibriosis is biofilm inhibition. In the present study we tested cell-free culture broth (CF-CB) of 31 strains obligate marine fungi for its ability to inhibit growth and biofilm formation with 2 isolates of Vibrio bacteria, Vibrio harveyi (VH1) and Vibrio parahaemolyticus (3HP). CF-CB with the highest biofilm inhibitory activity were then prepared as feed additives and mixed with commercial feed (1 ml to 1 g ratio) to examined its potency on Vibrio-challenged shrimp. The supplemented feed were administered to post-larvae (PL) shrimps for 7 days before and after they were challenged with VH1 and 3HP which shrimp health and mortality were monitored. Overall, CF-CB from 9 out of 31 isolates examined inhibited biofilm formation by VH1 and 17 out of 31 isolates inhibited biofilm formation by 3HP. Survival rate in the un-challenged negative control was 66% while PLs fed with MCR00984 (Linocarpon appendiculatum) and challenged with VH1 and 3HP were 42% and 60% which was not significantly different (p≥0,05). This results shows some promise for possible application against Vibriosis in shrimp.

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Kota semarang,

Jawa tengah

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