“LANTPYCATION” : METODE BARU BUDIDAYA MELON (Cucumis melo L.) RAMAH LINGKUNGAN

Jamusan village is one of the areas that becomes the center of melon production at Prambanan subdistrict, Sleman, Yogyakarta. However, at this moment, the villagers of Jamusan still encountered many obstacles in melon farming. This is caused by the poor quality of the melon, crops that have not been handled well and the lack of farmer’s knowledge about the management after harvesting, melon production that is not absorbed by the market. LANTPYCATION Program (Plant, Apply, and Diversification) aims to provide knowledge about quality and good management of the melons in the form of processed product diversification melon. The method applied in the implementation of this program which begins with socialization, counseling, training in management and cultivation of melons, good governance, advocacy and marketing of melon processed product. The result of the implementation of this method showed the increase of farmer’s knowledge about good quality melon, farm management and procedures for the proper cultivation of melons in creating diversificated processed melon to improve the welfare of villagers Jamusan.It could be revealed that LANTPYCATION Program is a new method on sustainable agriculture of melon cultivation in Indonesia. Keywords:Melon, Melon cultivation, Diversification processed melon

DETECTION AND MOLECULAR CHARACTERIZATION OF ODONTOGLOSSUM RINGSPOT VIRUS (ORSV) JAVA AND BALI ISOLATES

Orchids are one of the most important ornamental plants and cultivated in tropical countries, including in Indonesia. Virus infections become an important limiting factor in orchids cultivation because it causes significant losses of the plants. Odontoglossum ringspot virus (ORSV) is one of the most reported viruses infecting orchids and spread widely in the world. The common symptoms are mosaic with line pattern and necrotic ringspot on leaf surface, and also color breaking on flowers. The purpose of this research was to find out the occurrence of ORSV infecting orchids in Java and Bali, genetic relationship among ORSV isolates based on similarities of coat protein (CP) gene, and to analyze the pathogenicity test of ORSV Java and Bali isolates. Survey and samples collection were conducted in seven locations cultivating orchids in Java and Bali. Detection with Reverse Transcription-Polymerase Chain Reaction (RT-PCR) showed that ORSV was infected three leaf samples of Phalaenopsis sp., called ORSV BOC, ORSV KRB and ORSV TNBB isolates. The results showed 474 bp-amplified DNA band as the expression of ORSV CP gene. Phylogenetic analysis based on nucleotide sequences of CP gene showed that ORSV BOC have similarity close to ORSV Germany, whereas ORSV KRB and ORSV TNBB leads to speciation that possible to be a new strain. Pathogenicity test using various healthy plants showed that ORSV BOC may infected and cause systemic symptoms on Chenopodium amaranticolor, Nicotiana tabacum, Dendrobium sp., Cymbidium sp., Chattleya sp., Phalaenopsis sp., Liparis sp., Spatthoglotis sp. and Pectelis sussanae (L.) Raf.Keywords: ORSV, orchids, coat protein, RT-PC

DEVELOPMENT OF SPECIFIC PRIMERS FOR INTER SPECIES PHYLOGENY RELATIONSHIP ON Crocodilian sp

Poaching, trafficking, and illegal product trading are classic activities which frequently faced by Crocodilian group. To overcome, laws need supporting methods for a decision of these all activities which threaten crocodile species. This will require species identification that associated to taxonomy classification. Crocodilian species are very similar in morphology. This may result to a false identification especially when working on incomplete specimen. Currently, twenty-four existing Crocodilian species are continuously revised to improve the precise placement and/or acceptance of certain species on Crocodilian classification. Herein we address this issue using Cytochrome-b. The idea was to obtain genus specific primer from Cytochrome-b and then tested the precision of the designed primers using bioinformatics tools’ Primer-BLAST and CLC sequence Viewer 6. The designed primers showed a highly specificity on species level. The phylogenetic tree constructed by is relatively precise compared to reported phylogenetic trees. These specific primers together with the genus specific primers may give valuable and important support for the effective and efficient identification of Crocodilian group.Keywords: Crocodilian, illegal trading, Cytochrome-b , specific primer, bioinformati

Callus Induction and Differentiation on Melon From In Vitro Culture with The Addition of Indole Acetic Acid and Benzyl Amino Purine Growth Regulator

Melon plants (Cucumis melo L.) are susceptible to disease attacks primarily caused by viruses. One of the viruses often attacking melon plants is Cucumber mosaic virus (CMV). Previous studies have shown that melon callus produced from in vitro culture is detected to have CMV resistance gene (Creb-2). However, the trait of resistance to CMV in that melon (F2) is still segregated. To know the stability of the Creb-2 gene, further research is needed to detect the gene in the differentiated callus. This study aims to  callus induce and callus differentiate melon. Firstly, melon seeds were grown on ¼ MS medium with addition of 2 mg / L 2.4-Dichlorophenoxyacetate and 1 mg / L BAP (Benzyl Amino Purine) to grow callus. Secondly, the callus was differentiated in MS medium with the addition of 2,4-D , BAP and IAA in different doses. 0 mg / L; 0.01 mg / L 2.4-D and 0.1 mg / L BAP, and MS medium with 0.01 mg / L; 0.1 mg / L; 1 mg / L IAA (Iodole Acetic Acid) and 0.1 mg / L BAP. The results showed that callus was successfully differentiated into root and it grew optimally in MS medium with 0.01 mg / L 2.4-D: 0.1 mg / L BAP and 1 mg / L IAA: 0.1 mg / L BAP . Therefore the roots can be used as isolation samples and DNA amplification to detect stability of the Creb-2 gene

Variation Symptoms and Resistance Response of Different Types on Orchids (Orchidaceae) Against Odontoglossum ringspot virus (ORSV) Infection

Odontoglossum ringspot virus (ORSV) is one of the viruses that are reported to infect the most orchids in Indonesia. Viral infections cause economic losses. This research was conducted to study the introduction of symptoms of infection that appear as the initial stages of disease control. The source of the virus is orchid leaves Phalaenopsis amabilis positively infected by ORSV collected from Magelang. Virus isolation was carried out by mechanical inoculation to indicator plants (Chenopodium amaranticolor and Nicotiana tabaccum). Furthermore, the results of inoculation were multiplied using various types of orchids (Dendrobium, Cymbidium, Chattleya, Phalaenopsis, Spatthoglotis, Liparis, and Pecteilis). Virus detection on orchid tissue was done trough the DAS-ELISA and observation of virus structure using TEM. The results showed that Pectelis sussanae (L.) Raf. was the only orchid plant that was immune against ORSV approached. Dendrobium sp., Cymbidium sp., Chattleya sp., Phalaenopsis sp., and Spathoglotis sp. showed a response of susceptible of ORSV severe infection symptoms, while Liparis sp. were resistant. The variation of resistance from several orchid plant against infection ORSV showed severe symptoms with the incubation periode was seen earlier. The observation of virus structure using TEM showed rigid road shape particle, 300 x 18 nm in size, which is general characteristic of Tobamovirus. This indicate an infection of ORSV is a dangerous disease and require serious contro

The genetic variations and relationship of Madura tobacco (Nicotiana tabacum L.) based on molecular characteristics

Madura has at least 22 genotypes of local tobaccos (Nicotiana tabacum L.). This diversity could potentially produce new genotype of tobaccos with superior characters. However, information of the genetic diversity of Madura tobaccos is still limited. The aim of this study was to determine the genetic variation and relationship of 24 genotypes of Madura tobaccos with Random Amplified Polymorphic DNA (RAPD) analysis. In this research we were used 6 single primers for amplification: (OPA-18, OPB-12, OPB-14, OPC-1, OPC-8 and OPC-19) and 2 mixture primers ((OPB-12+OPC-8) and (OPC-1+OPC-19)). Genetic similarity and clustering was analyzed with Unweighted Pair Group Method Arithmetic (UPGMA) method with Numerical Taxonomy and Multivariate Analysis System (NTSYS) version 2.10 software. From this research we found that OPA18425, OPB12450, OPC8500, (OPC19+OPC1)550 and OPC8800 can be used as specific markers. Polymorphic bands percentage with mixture primers was relatively equal with single primers (<60%). The dendogram showed that Madura tobacco genotypes consist of 2 main clusters: cluster A (22 genotypes) and cluster B (2 genotypes: Bukabu Sa’ang and Prancak-95). Madura tobaccos had high genetic similarity between genotypes ranging from 0.80-1.00

Application of Molecular Biology for Identification of Virus Resistance Gene in Melon

Source of resistance to an Indonesia isolate of Cucumber mosaic virus (CMV-B2) in melon cultivar Yamatouri has been reported. Moreover, Creb-2, a locus that confers resistance to CMV-B2 in Yamatouri has been determined as a single dominant gene. To elucidate the resistance mechanism conferred by Creb-2 in more detail, it is necessary to clone the Creb-2 gene and determine its molecular structure. One approach is by amplification and cloning of melon resistance gene analogs (MRGAs) based on degenerated PCR primers designed from conserved amino acids in the NBS-LRR motifs (P-loop, Kinase-2, and the GLPL) and Toll/ Interleukin-1 receptor-like region (TIR). This study was aimed to identify and characterize the resistance gene analogs from Cucumis melo L. cv. Yamatouri by employing polymerase chain reactions (PCR) as a molecular biology tools with degenerate primers based on conserved motifs of cloned R genes. The application of molecular biology such as DNA isolation, degenerate primers and PCR condition, cloning, sequencing, linkage analysis and mapping of resistance gene analogs to Creb-2 gene in melon will be widely discussed in this pape

Keseragaman dan Kestabilan Karakter Tanaman Melon (Cucumis Melo L. ‘Tacapa Gold’) Berdasarkan Karakter Fenotip dan Inter-Simple Sequence Repeat

Melon ‘Tacapa Gold’ merupakan varietas melon hasil pemuliaan yang dilakukan oleh Laboratorium Genetika dan Pemuliaan yang memiliki karakter berupa bentuk globular, kulit muda hingga tua berwarna dasar kuning dengan net pada kulit buahnya serta warna daging buah hijau. Tujuan penelitian ini adalah untuk mengetahui keseragaman dan kestabilan karakter fenotip berdasarkan karakter morfologis dan molekuler. Metode yang yaitu pengamatan karakter fenotip dan amplifikasi DNA dengan PCR-ISSR. Analisis karakter kuantitatif menggunakan analisis sidik ragam menggunakan PKBT-STAT 3.1. dengan metode Turkey (5%). Pengambilan data keseragaman secara molekuler dilakukan dengan menggunakan metode PCR-ISSR dengan primer UBC-807, UBC-808, UBC- 810, dan UBC-824. Hasil analisis fenotip kualitatif menunjukkan karakter melon ‘Tacapa Gold’ seragam. Karakter fenotip secara kuantitatif cenderung masih bervariasi dan belum seragam apabila dibandingkan dengan melon 'Tacapa Gold’ generasi sebelumnya. Hasil keseragaman fenotip melon ‘Tacapa Gold’ berdasarkan penanda ISSR memiliki tingkat keseragaman yang tinggi (indeks similaritas ≥70%). Rata-rata polimorfisme pada melon ‘Tacapa Gold’ rendah yaitu 24,25%

SURVEY ON THE OCCURRENCE OF VIRUSES INFECTING CUCURBITS INYOGYAKARTAANDCENTRALJAVA

ABSTRACT Cucurbits are grown throughout the Java Island as dry season crops. Plants having mosaic, mottling, chlorosis and leaf distortion symptoms were frequently found in most of the cucurbit fields during the survey which conducted in Central Java including Sleman,Kulon Progo, and Klaten during July-September 2000 and 2001. Usingdouble antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA)Cucumber mosaic virus (CMV), Cucumber green mottle mosaic virus(CGMMV)and Kyuri green mottle mosaic virus (KGMMV)were found infecting cucurbits.CMV was widespread, infecting 48.9% of the samples tested followed by CGMMV (12.8%) and KGMMV (6.4%), while others samples (31.9%) were not tested, double infections were common with 8.5 % of the samples being infected with two viruses (CGMMV and KGMMV) and 34% with three viruses (CMV, CGMMV, and KGMMV). Severe mosaic and mottle symptomswere associatedmost often with single infectionofCGMMV andKGMMVrespectively. In addition, these are the first detections of CGMMVand KGMMV infecting cucurbit plants in Indonesia. Key words: CGMMV,CMV,Cucurbits, DAS-ELISA, KGMMV INT/SARI Tanaman labu-labuan umumnya tumbuh sepanjang musim kemarau di Pulau Jawa. Tanamanlabu-labuan dengan gejala mosaik, klorosis, mottling dan bentuk daun serta buah yang berubah banyak dijumpai selama survei yang dilakukan di Kulon Progo, Sleman dan Klaten pada bulan Juli sampai September tahun 2000 dan 2001. Deteksi menggunakan metode double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELlSA) telah berhasil mengetahui keberadaan dan infeksi Cucumber mosaic virus (CMV), Cucumber green mottle mosaic virus (CGMMV) dan Kyuri green mottle mosaic virus (KGMMV) pada tanaman labu-labuan di tiga kabupaten tersebut. CMV menginfeksi tanaman labu-labuan tinggi yaitu 48,9% dari jumlah sampel tanaman yang dikoleksi, kemudian CGMMV (J 2,8%) dan KGMMV (6,4%), sedangkan sebanyak 14 sampel tanaman (31,9%) tidak dideteksi. 1nfeksi ganda banyak ditemukan dan 8,5 %sampel tanaman terinfeksi oleh dua jenis virus (CGMMV dan KGMMV) sedangkan 34%sampel tanaman terinfeksi oleh tigajenis virus (CMV, CGMMV, dan KGMMV). Gejala mosaik dan mottling sering terjadi pada tanaman labu-labuan yang terinfeksi ganda oleh CGMMV dan KGMMV Hasil penelitian merupakan deteksi pertama CGMMV dan KGGMV pada tanaman labu-labuan di Indonesia. Kata kunei: CGMMV, CMV, Cucurbits, DAS-ELlSA, KGMM

Genetic Variation Analysis of Mold (Magnaporthe oryzae B.Couch) Using Random Amplified Polymorphic DNA

Magnaporthe oryzae B.Couch is a host-specific fungi, certain strain only infect certain host plant species. Genetic variety among M. oryzae isolates was explained by dendogram which was constructed using similarity data of Random Amplified Polymorphic DNA (RAPD). Dendogram construction was achieved by computer software, Numerical Taxonomy System (NTSYS). The aim of the research were to study the genetic variation among M. Oryzae using RAPD and to construct a dendogram of genetic similarities among the ten isolates from green foxtail (Setaria viridis L.), finger millet (Eleusine coracana L.) and rice (Oryza sativa L.).RAPD was performed in 30 cycles using 5 primers (OPA-02, OPA-03, OPA-04, OPA-05, OPA-07). Polymorphism data was used to constructed dendogram using Dice index and Unweighted Pair Group Method with Arithmetic Mean (UPGMA) in NTSYS software. There were 68 polymorphism fragments from 74 amplified fragments.Three clusters were formed in the dendrogram, based on host pathotype: foxtail millet type, finger millet type and rice type. There were two subclusters in foxtail millet type based on mating type, MAT1-1 dan MAT1-2. Thus, RAPD could be used as a method for genetic variation analysis of Magnaporthe oryzae to show host-specific specificity.Key words: Magnaporthe oryzae, RAPD, mating typ