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Methicillin-Resistant Staphylococcus aureus (MRSA) is a species of S. Aureus, which shows resistance to the methicillin class of antibiotics. MRSA infection poses significant health problems because it causes severe disease in a vulnerable population and also because effective antibiotics are limited. Therefore, the research to find the new anti-MRSA compound is a necessity. The genome mining approach through Biosynthetic Gene Clusters (BGC) analysis can detect the ability of microbes to produce antibiotics. Thus, the study aimed to screen potential anti-MRSA microbes and predict their BGC. This study used ten microorganisms isolated from mangrove areas in Segara Anakan Cilacap and clinical MRSA 2983 from the Faculty of Medicine, UNSOED. All samples microorganisms were cultivated in starch casein nitrate agar. DNA isolation used Quick-DNA Fungal/Bacterial Miniprep Kit, 16S rRNA gene amplification used 27F and 1492R primers, and DNA amplicons were sequenced by the Sanger sequencing method. BCG prediction of the associated genes used antiSMASH 5.1.2. The results showed that W-5A, P-6B, and W-5B isolates have anti-MRSA activities with inhibition indexes of 0.58; 0.53; and 0.47, respectively. Species identification revealed similarity of W-5A with Streptomyces longisporoflavus strain Moghannam M1, P-6B with Ochrobactrum intermedium LMG 3301, and W-5B with Streptomyces cellulosae strain NBRC 13027. The results of BGC analysis showed that Streptomyces longisporoflavus has one region involved in the synthesis of tetronasin. Ochrobactrum intermedium has one region involved in the synthesis ambactin. Streptomyces cellulosae has eight gene clusters for tiacumicin B, actinorhodin, ulleungmycin, albaflavenone, desferrioxamine B/E, stenothricin, auricin, and prejadomycin productions. These results indicate that the microbes isolated from Segara Anakan Cilacap serve as promising antibacterial producers.

A B S T R A C TPeanut is one of food crops commonly consumed in Indonesia. This species comprises several cultivars such as Kancil, Bison, Jerapah, Talam, and Tuban, each of which has its individual advantages and disadvantages. The vast variation among peanut cultivars leads to the need of study on genetic diversity and relationship among them using particular molecular marker. This study aims to see whether variation on DNA sequences among some peanut cultivars amplified with atpB-rbcL primers exists or not and to know the relationship among the cultivars based on the amplicon sequences. The method involves some sequential steps, i.e. genomic DNA isolation using CTAB protocol, amplification of DNA sequence using atpB-rbcL primers and sequencing of the amplification products. Data on sequences were edited manually using Bioedit version 7.0.4.1. Sequence alignment was performed using ClustalW, which is also implemented in Bioedit version 7.0.4.1. Arlequin 2.0 was used to calculate nucleotide diversity . Phylogenetic analysis was performed using Maximum Parsimony in MEGA 5.0. The results showed that considerably high variation in DNA sequences of some peanut cultivars amplified with atpB-rbcL primers are observed. On the other hands, very close genetic relationship among cultivars is found. KEY WORDS: DNA sequence variation, intraspecific diversity analysis, rbcL-atpB primers Penulis korespondensi: YANI YULIANI |

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A study on the RAPD profile of some pitcher plants (Nepenthes spp.) of Baturraden Botanical Garden collection was carried out. Seven individuals of particular concern were Mount Slamet semi-endemic and Java endemic species, i.e. N. adrianii and N. gymnampora. Both species of pitcher plants were taken from Baturraden Botanical Garden collection. Samples were then subject to genomic DNA extraction, after which RAPD markers were amplified using PCR technique with ten random primers, i.e. ES10G23, ES10A26, ES10C24, ES10G33, OPA-2, OPA-9, OPA-13, OPB-3, OPB-5 and OPB-7. The results show that the 10 primers produce 43 (67.2%) polymorphic bands and 21 (32.8%) monomorphic bands with size ranging from 130 to 1,500 bp. OPA-2 and OPB-3 produce highest polymorphism, while ES10G23 and OPA-13 produce most monomorphic bands. Dendogram constructed using UPGMA in MEGA 6.06 shows that at a genetic distance of 18% two clusters are formed. Both clusters are separated seemingly by geographic location. Key words: pitcher plants, RAPD, Baturraden Botanical Garden AbstrakTelah dilakukan penelitian tentang profil RAPD beberapa tanaman kantung semar (Nepenthes spp.) koleksi Kebun Raya Baturraden.Tujuh individu yang dipelajari merupakan spesies semiendemik Gunung Slamet dan endemik Jawa, yaitu N. adrianii dan N. gymnampora. Sampel kedua spesies kantung semar ini diambil dari koleksi Kebun Raya Baturraden. DNA genomik sampel diekstraksi untuk kemudian digunakan sebagai templat dalam amplifikasi marka RAPD menggunakan teknik PCR dengan 10 primer acak, yaitu ES10G23, ES10A26, ES10C24, ES10G33, OPA-2, OPA-9, OPA-13, OPB-3, OPB-5, dan OPB-7. Hasil penelitian menunjukkan bahwa ke-10 primer tersebut menghasilkan 43 (67,2%) pita polimorfikdan 21 (32,8%) pita monomorfikdengan ukuran berkisar dari 130 hingga 1.500 bp. OPA-2 and OPB-3 menghasilkan polimorfisme tertinggi, sedangkan ES10G23 dan OPA-13 menghasilkan pita monomorfik terbanyak. Dendogram yang dikonstruksi menggunakan UPGMAdalam MEGA 6.06 memperlihatkan bahwa dua klaster terbentuk pada jarak genetik 18%. Kedua klaster tersebut cenderung dipisahkan oleh lokasi geografi.

A B S T R A C TWinged bean [Psophocarpus tetragonolobus (L.) DC] is a tropical plant that has some benefits and is very suitable to be cultivated in Indonesia. Study about diversity of winged bean is very important to support the future development of winged bean. Increased genetic diversity can be done through mutation. Gamma ray is often used for inducing mutations. One of way to observe genetic diversity resulted by molecular mutation is using RAPD method. The purpose of this study was to identify the genetic diversity of winged bean resulted by gamma ray irradiation. The method used in this study was an experiment by using RAPD technique. Samples used were plant leaves that had been induced by gamma ray with wavelength of 20 Gy, 25 Gy, and control plants that were not induced by gamma ray. RAPD were done by ten primers that were OPA 9, OPA 10, OPA 13, OPA 18, OPB 2, 3 OPB, OPB 6, 7 OPB, OPB 10, and OPB 11. Data were analyzed using GenAlex 6.1 Program. The percentage of polymorphic loci of winged bean population control was 47,54%, while in the 20 and 25 Gy treatment were 62,30% and 54,10%, respectively. The values of genetic variation based on the calculation of allele frequencies were 0,236; 0,202 and 0,194 for treatment of 20 Gy, 25 Gy and for control plants, respectively. Meanwhile, the value of genetic distance ranged from 0.08 to 0.32.

Peanut (Arachis hypogea) is a typical plant species of tropical regions that has high economic value. The plantation is widely spread over many areas and the production is being pushed to meet the increasing demand. Peanut breeding program is aimed to improve genetic quality, mainly with resepct of production and thus information on genetic diversity is necessary as a basis for consideration in breeding, management and sustainable utilization. One approach to analyse genetic diversity of peanut is by using molecular markers. Random Amplified Polymorphic DNA (RAPD) is a widely used molecular marker for genetic diversity analysis. Therefore, the aim of this study was to assess genetic diversity of peanut cultivars, i.e. Jerapah, Kancil, and Hypoma 2, based on RAPD markers. The study was conducted in a survey method, in which three individuals of each cultivar were analyzed using PCR-RAPD technique employing twelve primers, i.e. OPA-1, OPA-2, OPA-9, OPA-13, OPB-2, OPB-3, OPB-4, OPB-5, OPB-7, OPB-11, OPB-12 and OPJ-07. Data analysis based on morphological data is also included. Molecular analysis revealed that only 7.55% polymorphic band was obtained, while most of the bands were monomorphic, indicating very low variation among the cultivars. The phenogram that constructed based on literature showed that Kancil was closer to Jerapah cultivar, while RAPD-based dendogram showed that Hypoma 2 was closer to Kancil cultivar.