Identifikasi Molekuler Virus Penyebab Penyakit Daun Keriting Isolat Bantul pada Melon

One of important problems of melon farming in Yogyakarta is a leaf curl disease that show specific symptom of Begomovirus infection. The little information about the nature of virus causal agent was constraint for the disease management. The purposes of this research were to molecularly identify the leaf curl causing virus in melon in Yogyakarta and to find the genetic relationship of this virus with other Begomovirus members which cause leaf curl disease. This research was conducted through several phases, which are: curly leaves collection on the field, virus DNA isolation, Begomovirus identification using universal primer Krusty Homer, Begomovirus DNA-A and DNA-B detection using primer Gemini full BamH1 forward and reverse for full genome DNA-A, and primer BF518 and BR1641 for DNA-B, genome sequencing and genetic relationship analysis of the sequence with other Begomovirus causing leaf curl. The result of field studies which were conducted in Sewon found some melon plant with symptom of Begomovirus infection. The molecular identification result using PCR showed that leaf curl causing virus in melon is Begomovirus, having DNA-A and DNA-B. Genetic relationship analysis of this virus with other Begomovirus causing leaf curl shows that this virus is closely related with Pepper yellow leaf curl Indonesia virus (AB267834) based on nucleotide and amino acid sequencing as coat protein of Begomovirus. The result of shows that the study is the first report of PepYLCIDV infection, a bipartite genome virus on melon, and its natural leaf curl symptom in Indonesia. Salah satu kendala budidaya melon (Cucumis melo L.) di Yogyakarta yaitu adanya penyakit daun keriting dengan gejala khas infeksi Begomovirus. Belum tersedianya informasi mengenai jenis dan ciri patogen virus penyebab penyakit tersebut merupakan salah satu kendala penting dalam menentukan strategi pengelolaan virus tersebut. Tujuan penelitian ini adalah mengidentifikasi secara molekuler virus penyebab daun keriting pada melon di Yogyakarta dan mengetahui hubungan kekerabatan virus tersebut dengan virus anggota Begomovirus lain penyebab daun keriting yang telah dipublikasi di database genebank. Penelitian dilakukan dengan beberapa tahapan, yaitu deskripsi gejala daun keriting di lapangan, isolasi DNA virus, identifikasi Begomovirus dengan primer universal Krusty Homer, deteksi DNA-A dan DNA-B Begomovirus dengan primer Gemini full BamH1 forward dan reverse untuk full genome DNA-A, serta primer BF518 dan BR1641 untuk DNA-B, sequencing genom, dan analisis hubungan kekerabatan sequence tersebut dengan Begomovirus lain penyebab daun keriting. Hasil pengamatan lapangan di Sewon Bantul diperoleh tanaman melon dengan gejala khas infeksi Begomovirus. Hasil identifikasi secara molekuler dengan PCR menunjukkan bahwa virus penyebab daun keriting pada melon adalah Begomovirus, memiliki DNA-A dan DNA-B. Analisis hubungan kekerabatan virus penyebab daun keriting pada melon dengan Begomovirus lain penyebab daun keriting menunjukkan bahwa virus tersebut berkerabat dekat dengan Pepper yellow leaf curl Indonesia virus (AB267834) berdasarkan sekuen nukleotida dan asam amino sebagian coat protein Begomovirus. Hasil penelitian ini menunjukkan bahwa penelitian ini merupakan laporan pertama infeksi PepYLCIDV dengan bipartite genome pada melon dengan gejala daun keriting secara alamiah di Indonesia

Pola Agihan dan Intensitas Penyakit Busuk Pangkal Batang Lada di Provinsi Sulawesi Tenggara

The success of foot rot disease control is largely dependent on information data of pepper cultivation conditions, distribution pattern of the disease, and the magnitude of the intensity of the disease. This study aimed to determine the condition of pepper cultivation, distribution pattern of pepper foot rot disease, the development of disease symptoms, and intensity of pepper foot rot disease in Southeast Sulawesi. Data cultivation conditions, distribution of the disease, progression of symptoms, and the intensity of the disease were obtained by means of surveys in pepper plantations and interviews with pepper farmers. The results showed that the pepper plantations in Southeast Sulawesi were cultivated on flat to hilly topography, and the cultivation method was very conventional but herbicide use was very intensive. The development of wilt symptoms on pepper plants was very quick in dry weather but slow in the wet. Pepper foot rot disease has patch distribution. The highest intensity of the pepper foot rot disease was in less weedy plantations with intensive use of herbicides. The intensity of the pepper foot rot disease in Southeast Sulawesi was 61,2% with the spread in each district namely South Konawe at 53,8%, Konawe at 63,7% and Kolaka by 61,2%

Infeksi Ganda Begomovirus dan Crinivirus pada Tanaman Tomat di Kabupaten Magelang, Jawa Tengah

Since 2006, a yellowing disease has been observed in tomato (Lycopersicon esculentum) fields in Central Java, Indonesia. Epidemics of the diseases were mainly associated with populations of the whitefly Trialeurodes vaporariorum and Bemisia tabaci, the major whitefly pests in vegetable crops. The main symptoms were severe yellowing on lower leaves, and curling on upper leaves of plant. Total DNA was extracted from tomato leaves using CTAB methods, while total RNA was extracted using NucleoSpin RNA Plant extraction kit (Macherey-Nagel). Because of occurring mixed symptom on an individual plant, hereby it is important to detect the causal agent to manage of the disease. Samples from symptomatic plants were analyzed by polymerase chain reaction (PCR) and shown to be infected with Tomato infectious chlorosis virus (TICV) (family Closteroviridae, genus Crinivirus) and a virus species belongs to the genus Begomovirus, family Geminiviridae. The research result is the first report of Crinivirus and Begomovirus double infection in single tomato fields in Indonesia. INTISARIPenyakit kuning pada tanaman tomat (Lycopersicon esculentum) telah ditemukan sejak tahun 2006 di Jawa Tengah, Indonesia. Epidemi penyakit tersebut terutama berkaitan dengan keberadaan dua spesies whitefly (Trialeurodes vaporariorum dan Bemisia tabaci). Gejala utama yang ditemukan adalah daun-daun pada tanaman bagian bawah berwarna kuning sedangkan daun daun pada tanaman bagian atas menunjukkan gejala keriting. Adanya gejala campuran pada satu individu tanaman maka perlu dideteksi penyebabnya untuk pengelolaannya. DNA total diekstraksi dari daun tomat yang terinfeksi menggunakan metode CTAB, sedangkan total RNA diekstraksi dengan menggunaan NucleoSpin RNA Plant extraction kit (Macherey-Nagel). Analisis sampel tanaman sakit menggunakan teknik poymerase chain reaction (PCR) menunjukkan adanya infeksi Tomato infectious chlorosis virus (TICV) (famili Closteroviridae, genus Crinivirus) dan satu spesies virus anggota genus Begomovirus, famili Geminiviridae. Hasil penelitian ini merupakan laporan pertama kali adanya infeksi ganda kelompok Crinivirus dan Begomovirus pada satu individu tanaman tomat di Indonesia

Deteksi Molekuler Penyebab Penyakit Kuning (Tomato Chlorosis Virus dan Tomato Infectious Chlorosis Virus) pada Tanaman Tomat

This research was aimed to detect the ToCV and TICV caused yellowing disease on tomatoes by molecular detection. Leaf samples of symptomatic plants were taken from Ketep (Magelang), then the leaves were identified by reverse transcription-polymerase chain reactions (RT-PCR) using specific primer ToCV-CF/ToCV-CR (360 bp) and TICVCF/TICV-CR (416 bp). The result of nucleotide sequence analysis, amino acid and PCR product phylogenetic sequences were verified as TICV, it showed that TICV from Magelang belongs to the same group with TICV from Japan, North America and Europe, France, Italy, and USA. INTISARIPenelitian ini bertujuan untuk mendeteksi keberadaan ToCV dan TICV penyebab penyakit kuning pada tanaman tomat. Daun bergejala diambil dari Desa Ketep (Magelang), selanjutnya diuji denganreverse transcription-polymerase chain reactions(RT-PCR) menggunakan primer spesifik ToCV-CF/ToCV-CR (360 bp) dan TICV-CF/TICV-CR (416 bp). Hasil analisis sekuen nukleotida, asam amino, dan filogenetik produk PCR teridentifikasi sebagai TICV yang menunjukkan bahwa TICV isolat Magelang berada dalam satu kelompok dengan isolat TICV asal Jepang, Amerika Utara dan Eropa, Perancis, Italia, dan USA

The Inhibition of Tobamovirus by Using the Extract of Banana Flower

Some extract of the banana plants parts have been identified to produce a compound that has efficacy as traditional medicine and human virus inhibitor. There was no previous report on the use of the banana flower as the plants antiviral sources for plant pathogen. The objective of this study was to identify the potency of two types of the banana flower as the Tobamovirus inhibitor substance. The antiviral was prepared from the extract of Ambon banana (Musa acuminata Colla) flower and Klutuk banana (Musa balbisiana Colla) flower, as the comparison extract of Mirabilis jalapa L. leaf which is widely known to have antiviral on various plant viruses was used. This study applied the Complete Randomized Design with three replications. Collected data includes the virus incubation period and the inhibitor power upon the local necrotic symptom on indicator plant. Results of this study showed that the extract of Ambon and Klutuk banana flower was able to inhibit the Tobamovirus with inhibition levels of 86.34% and 91.22%.IntisariBeberapa ekstrak bagian tanaman pisang diketahui memiliki kandungan senyawa yang berkhasiat sebagai obat tradisional dan zat yang dapat menghambat virus manusia. Belum pernah dilaporkan penggunaan bunga pisang sebagai sumber antiviral terhadap virus tumbuhan. Penelitian ini bertujuan untuk mengetahui potensi ekstrak bunga dua jenis pisang sebagai zat penghambat Tobamovirus. Antiviral disiapkan dari ekstrak bunga pisang Ambon (Musa acuminata Colla) dan pisang Klutuk (Musa balbisiana Colla), sebagai pembanding digunakan ekstrak daun Mirabilis jalapa L. yang sudah banyak diketahui mengandung antiviral pada berbagai virus tumbuhan. Penelitian ini dirancang menggunakan Rancangan Acak Lengkap dengan 3 ulangan. Data yang dikumpulkan meliputi masa inkubasi virus dan daya hambat gejala nekrotik lokal pada tanaman indikator. Hasil penelitian menunjukkan bahwa ekstrak bunga pisang jenis Ambon dan Klutuk mampu menghambat Tobamovirus dengan tingkat penghambatan sebesar 86,34% dan 91,22%

Disease Incidence of Melon Leaf Curl in East Java and Special Province of YOGYAKARTA

Geminivirus infection has caused severe losses on various economically important crops. The disease incidence on melon has been observed since 2004, and widespread at melon production centers in East Java and Special Province of Yogyakarta. In East Java and Special Province of Yogyakarta, the Disease Incidence of leaf curl on melon reached 100% and 14.3 % respectively in 2008. Detection of the causal agent using primers of CPA5 and CPA2 resulted viral specific DNA fragment of 770 bp. Infeksi penyebab penyakit yang disebabkan oleh geminivirus telah menyebabkan kerugian secara ekonomi berbagai jenis tanaman penting yang dibudidayakan. Kejadian penyakit pada tanaman melon telah diamati sejak tahun 2004, dan tersebar secara luas di pusat penanaman melon di Jawa Timur maupun Daerah Istimewa Yogyakarta (DIY). Di Jawa Timur dan Daerah Istimewa Yogyakarta (DIY), pada tahun 2008 kejadian penyakit daun keriting melon mencapai 100% dan 14,3%. Penyebab penyakit telah dideteksi menggunakan teknik polymerase chain reaction. Amplifikasi Fragmen DNA virus dari tanaman yang terinfeksi dihasilkan dengan ukuran 770 bp menggunakan sepasang primer CPA5 dan CPA2

Preferensi Wereng Batang Cokelat Terhadap Varietas Padi Dan Ketahanan Varietas Padi Terhadap Virus Kerdil Hampa

Rice ragged stunt virus (RRSV) is transmitted by brown planthopper in the persistent manner. This disease in recent years has become a serious problem in Indonesia and some other countries, such as China, Vietnam, Philippines, and Thailand. Disease control is usually conducted by the vector control using insecticides, so often causes an environmental pollution. An alternative control method is using the environmentally friendly of resistant varieties. The purpose of this study was to determine the level of preference of brown planthopper (BPH) to some varieties and rice germplasms, the resistance of several varieties against rice ragged stunt virus (RRSV) disease and its effectiveness as an inoculum source of virus (RRSV). A total of 15 varieties of rice were used in preference test of BPH and resistance test to RRSV. The test for resistance varieties to RRSV was conducted by transmission of 2nd instar of BPH for 3 days of acquisition feeding period, 10 days incubation period and 24 hours inoculation period with population density of 3 BPH/plant. Disease index was calculated and used to determine the level of plant resistance. Varieties that showed resistant, moderately resistant and susceptible responses were selected and were used as a source of inoculum to be transmitted on to TN1 variety susceptible check variety to know the effectiveness of varieties as source of virus inoculum. The results showed that of the 15 varieties tested, Situ Bagendit, Utri Merah, Mentik Wangi, Mahsuri, and Inpari 1 each was less favored by BPH to settle and to multiply. Mentik Wangi, Tetep, Utri Merah, and Swarnalata each showed resistant response to RRSV. Transmission test to susceptible variety (TN1) showed that the variety of Situ Bagendit, Inpari 13, Mentik Wangi, and Tetep each has a fairly low effectiveness as a source of inoculum as indicated by the lower percentage of infection and disease index on the transmited test plants

Survei Odontoglossum Ringspot Virus (ORSV) yang Menginfeksi Anggrek Alam Tropis di Indonesia

Natural orchids are one of the important ornamental plants that were cultivated in tropical countries, including Indonesia. Virus infections has been important limiting factor in orchids cultivation because it decreases the orchids quality. Odontoglossum ringspot virus (ORSV) is one of the most reported virus infecting orchids which spread widely in the world. During 2010–2014 surveys of viral infections were conducted in Indonesia. The orchids were found infected by virus, showed symptoms of mosaic, mottle, chlorotic, necrotic, streak, wilting leaf, and ringspot on leaf surface. Detection with serological test DAS-ELISA showed only 11 from 125 samples were infected by ORSV with total incidence of 8,8%. Nine leaf samples of Phalaenopsis sp. were infected, respectively. Thus, it was concluded that Phalaenopsis is orchids genus which is the most abundantly and susceptibly infected by ORSV. The results proved that ORSV have entered and spread widely by infected orchids in orchids landscape (nursery), semi-natural forests (botanical gardens), and natural forest (national park) throughout Indonesia. This is the first report of ORSV infecting natural tropical orchids in Indonesia. INTISARI Anggrek alam merupakan salah satu kekayaan flora asli negara tropis, termasuk Indonesia. Keberadaan di habitat aslinya sudah sangat berkurang yang disebabkan kerusakan hutan dan adanya penyakit. Infeksi virus masih menjadi faktor pembatas terpenting dalam budidaya dan pengembangan potensi anggrek alam. Odontoglossum ringspot virus (ORSV) adalah salah satu jenis virus yang dilaporkan paling banyak menginfeksi anggrek serta memiliki penyebaran yang luas di dunia. Selama 2010-2014 telah dilakukan survei lapangan terhadap infeksi virus di Indonesia. Beberapa anggrek yang ditemukan terinfeksi oleh virus menunjukkan gejala berupa mosaik, belang, klorosis, nekrosis, streak, daun layu, dan bercak cincin pada permukaan daun. Deteksi dengan uji serologis DAS-ELISA menunjukkan bahwa 11 dari 125 sampel terinfeksi oleh ORSV dengan total kejadian 8,8%. Masing-masing sembilan dari total sampel daun terinfeksi merupakan Phalaenopsis sp. Hal ini menjelaskan bahwa Phalaenopsis adalah genus anggrek yang paling cocok dan rentan terhadap infeksi ORSV. Hasil penelitian survei kejadian infeksi Odontoglossum ringspot virus (ORSV) membuktikan bahwa virus ini telah masuk dan menyebar secara luas oleh anggrek-anggrek alam tropis yang terinfeksi di pertamanan anggrek (nurseri), hutan semi-alami (kebun raya), dan hutan alam (taman nasional) di seluruh wilayah Indonesia. Ini merupakan laporan pertama mengenai infeksi ORSV terhadap anggrek-anggrek alam tropis di Indonesia

Deteksi dan Diferensiasi Virus Kerdil Pisang dengan Teknik PCR-RFLP

Banana bunchy top disease (BBTD) can be caused by the infection of two different viruses, Banana bunchy top virus (BBTV) or Abaca bunchy top virus (ABTV). Both viruses can be transmitted persistently by aphid Pentalonia nigronervosa Coq. The research was conducted to detect and to differentiate the virus bypolymerase chain reaction-restriction fragment length polymorphism (PCRRFLP) techniques. Infected plants were collected from Yogyakarta (Sleman, Yogyakarta city, Bantul, Gunung Kidul, and KulonProgo). Nucleon Phytopure DNA Extraction Kit method was used to extract the total DNA of infected plants. Universal primers of Common DNA region (S-CRF and S-CRR) and specific primers DNA-R (C1-CRF and CI-CRR) were used for PCR amplification. PCR products were analyzed by RFLP technique using the restriction enzyme of DraI. The results reconfirm previous reports that bunchy top disease of banana in Yogyakarta is caused by BBTV. The ABTV was not detected in this present study. Based on the RFLP analysis it was concluded that BBTV collected in this study could be divided into three groups. Group 1 consisted of BBTV isolate from Sleman and Yogyakarta city with two fragments DNA of 400 and 388 bp. Group 2 consisted of isolate BBTV from Kulon Progo and Gunung Kidul with three fragments DNA of 400, 388, and 323 bp. Group 3 consisted of isolate from Bantul with two fragments DNA of 723 and 376 bp. Further study on the complete characteristics of these groups is still needed. Penyakit kerdil pisang dapat disebabkan oleh infeksi virus yang berbeda yaitu Banana bunchy top virus(BBTV) atau Abaca bunchy top virus (ABTV). Kedua virus tersebut ditularkan secara persisten oleh kutu daun Pentalonia nigronervosa Coq. Penelitian ini bertujuan untuk mendeteksi dan membedakan virus dengan polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).Tanaman terinfeksi dikoleksi dari Yogyakarta (Sleman,Kota Yogyakarta, Bantul, Gunung Kidul, dan Kulon Progo). Metode Phytopure DNA Extraction Kit dan CTAB digunakan untuk mengisolasi DNA total. Primer universal Common DNA region (S-CRF/S-CRR) dan primer spesifik DNA-R BBTV(C1-CRF danCI-CRR) digunakan dalam amplifikasi dengan PCR. Produk PCRBBTV dianalisis dengan teknik RFLP menggunakan enzim restriksi Dra I. Hasil penelitian memperkuat laporan sebelumnya bahwa penyakit kerdil pisang di Yogyakarta disebabkan oleh BBTV, dan belum ditemukan ABTV. Berdasarkan analisis RFLP, BBTV dapat dibedakan menjadi tiga kelompok. Kelompok 1 yaitu BBTV isolat Sleman dan Kota Yogyakarta dengan dua fragmen DNA berukuran sekitar 400 dan 388 bp. Kelompok 2 yaituBBTV isolat Kulon Progo dan Gunung Kidul dengan tiga fragmen DNA berukuran sekitar 400, 388, dan 323 bp. Kelompok 3 yaitu BBTV isolat Bantul dengan dua fragmen DNA berukuran sekitar 723 dan 376 bp. Penelitian tentang karakterisasi dari ketiga kelompok tersebut masih diperlukan

Identifikasi Molekuler Virus Penyebab Mosaik pada Sembilan Varietas Tebu

Sugarcane (Saccharum officinarum L.) is very important commodity in Indonesia because of its economical value as a sugar producing plant. One of the disease in sugarcane cultivation which have to be treated is mosaic symptoms because of its potency in reducing sugar yield. Mosaic symptoms might be caused by some viruses, therefore virus identification is required to find the virus that has caused the sugarcane disease in all of sugarcane field which can be found in East Java as working area of Seeding and Protection Center for Estate Crops Surabaya. This laboratory had an authority to certify the shoot to be distributed. This research aimed to find out the virus that caused the mosaic symptoms from nine sugarcane varieties and to obtain information about the most suitable identification method for shoot health test in Seeding and Protection Center for Estate Crops Surabaya. The first method of this research were the sugarcane leaf with mosaic sampling from nine varieties: VMC 7616, PSJK 922, Kidang Kencana and Tolangohula 2 (Jombang); PS 881 and PS 862 (Pasuruan); PS 864 dan PSBM 901 (Kediri). The next experiments were conducted in the Laboratory of Phytopathology, Faculty of Agriculture, Universitas Gadjah Mada. RNA was isolated from the sample and continued with RT-PCR technique, sequencing and bioinformatic analysis. Primers which was used in this research were MJ1-MJ2 and SCSMV cpF – SCSMV AP3. The result showed that mosaic symptoms of eight sugarcane varieties was caused by two species of virus: SCMV (Sugarcane mosaic virus) found in PS 881 and SCSMV (Sugarcane streak mosaic virus) found in seven another varieties. Sequencing analysis showed that SCMV isolate found in PS 881 was known to have a high nucleotide homology (93.6%) and related to the isolate from Kenya (KT630805.1), while the SCSMV had a high homology (96.9%−98.5%) and related to SCSMV isolate from Indonesia (AB563503.1) and Pakistan (GQ388116.1). INTISARITebu (Saccharum officinarum L.) merupakan tanaman penting di Indonesia. Salah satu penyakit yang perlu diwaspadai pada tanaman tebu adalah penyakit mosaik karena potensinya dalam menurunkan produktivitas tebu sehingga perlu dilakukan identifikasi virus penyebab mosaik pada tanaman tebu. BBPPTP Surabaya sebagai instansi yang melaksanakan sertifikasi pada bibit tebu perlu mengetahui metode identifikasi virus penyebab mosaik sebagai dasar dalam pengujian kesehatan benih untuk mendukung tersedianya bibit tebu yang sehat. Penelitian ini bertujuan untuk mengetahui virus penyebab mosaik pada sembilan varietas tanaman tebu dan mengetahui metode identifikasi virus pada tanaman yang cepat dan akurat untuk pengujian kesehatan benih. Penelitian dilakukan di Laboratorium Fitopatologi, Fakultas Pertanian Universitas Gadjah Mada. Penelitian dilakukan dengan mengambil sampel daun tanaman tebu pada sembilan varietas yang diambil dari tiga lokasi berbeda yaitu VMC 7616, PSJK 922, Kidang Kencana dan Tolangohula 2 (Jombang); PS 881 dan PS 862 (Pasuruan); PS 864 dan PSBM 901 (Kediri). Sampel diisolasi ss-RNA dilanjutkan dengan teknik RT-PCR (Reverse-Transcription Polymerase Chain Reaction), sekuensing dan analisis bioinformatika. Primer yang digunakan dalam penelitian ada dua yaitu MJ1-MJ2 untuk target sekuen dari SCMV dan SCSMV cpF-SCSMV AP3 untuk target sekuen dari SCSMV. Hasil penelitian ditemukan bahwa gejala mosaik yang ditimbulkan pada 8 (delapan) sampel varietas tebu disebabkan oleh 2 (dua) jenis virus penyebab mosaik yaitu SCMV (Sugarcane mosaic virus) yang ditemukan pada varietas PS 881 dan SCSMV (Sugarcane streak mosaic virus) yang ditemukan pada 7 (tujuh) varietas lain. Analisis sekuensing menunjukkan bahwa SCMV yang ditemukan pada varietas PS 881 memiliki homologi nukleotida (93,6%) dan kekerabatan terdekat dengan isolat di Kenya (KT630805.1). SCSMV hasil penelitian ini dikatakan mempunyai homologi (96,9%−98,5%) dan kekerabatan tertinggi dengan isolat SCSMV asal Indonesia (AB563503.1) dan Pakistan (GQ388116.1)