INTENSITAS PENYAKIT KUNING (CRINIVIRUS), PERTUMBUHAN DAN HASIL VARIETAS TOMAT HIBRIDA

Penyakit kuning yang disebabkan oleh TICV (Tomato infectious chlorosis virus) pada tanaman tomat merupakan penyakit baru di Indonesia. Penelitian ini bertujuan untuk mengetahui karakter agronomi beberapa varietas tomat terhadap intensitas penyakit kuning (Crinivirus) di Kabupaten Garut. Penelitian dilakukan di Tarogong Kaler, Garut. Penelitian menggunakan Rancangan Acak Kelompok (RAK) dengan 13 perlakuan dan 3 ulangan. Varietas yang digunakan yaitu varietas Royal 58, TM Marvel, Marta F1, Yasmin F1, Swadesi F1, Maya 353, Larisa F1, Amala 474, Toti F1, Natama Super F1, Warani F1, Agatha F1 dan Marta 9 F1. Hasil penelitian menunjukkan bahwa varietas TM Marvel secara nyata menunjukkan intensitas serangan TICV paling rendah yaitu 14, 13% dan menunjukkan penampilan terbaik pada tinggi tanaman serta bobot buah layak jual per plot. Varietas Royal 58 menunjukkan penampilan terbaik pada luas daun dan bobot buah per tanaman. Varietas Swadesi F1 mengalami intensitas serangan TICV cukup berat (26,83%) namun mampu menghasilkan bobot buah layak jual per plot cukup banyak dan menunjukkan penampilan terbaik pada bobot kering tanaman. Varietas Amala 474 menunjukkan penampilan terbaik pada jumlah buah per tanaman. Kata kunci : Karakter agronomi, Tomat, Crinivirus

INFEKSI GANDA BEGOMOVIRUS DAN CRINIVIRUS PADA TANAMAN TOMAT DI KABUPATEN MAGELANG , JAWA TENGAH (DOUBLE INFECTIONS OF BEGOMOVIRUS AND CRINIVIRUS ON TOMATO PLANTS AT MAGELANG REGENCY, CENTRAL JAVA)

ABSTRACTSince 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.Keywords: Begomovirus, Crinivirus, double infection, PCR, tomato 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.Kata kunci: Begomovirus, Crinivirus, infeksi ganda, PCR, tomat

DETEKSI MOLEKULER PENYEBAB PENYAKIT KUNING (Tomato chlorosis virus DAN Tomato infectious chlorosis virus) PADA TANAMAN TOMAT (MOLECULAR DETECTION CAUSED YELLOWING DISEASE (Tomato chlorosis virus AND Tomato infectious chlorosis virus) ON TOMATOES)

ABSTRACTThis 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 reversetranscription-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.Keywords: molecular detection, ToCV, TICVINTISARIPenelitian 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-polymerasechain 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.Kata kunci: deteksi molekuler, ToCV, TICV

EKSPRESI GEN PROTEIN SELUBUNG TOMATO INFECTIOUS CHLOROSIS VIRUS PADA ESCHERICHIA COLI

Expression of tomato infectious chlorosis virus coat protein gene on Escherichia coli. Tomato infectious chlorosis virus (TICV) is the causal agent of chlorotic disease of tomato. Detection of TICV can be carried out by RT-PCR and serological test. Titer of TICV in plant tissue is very low because TICV is limited to phloem. Serological detection of TICV requires antiserum which is not available in Indonesia. Producing antibody through cloning and coat protein gene (TICV CP gene) expression is a promising approach in producing antiserum. The objective of this study was to express TICV CP gene as antigen for antiserum production. TICV CP gene was amplified using RT-PCR from total RNA extracted from TICV infected leaves collected from Cipanas, Cianjur, West Java. The amplified CP gene was then sequenced and sub-cloned into pET 21b expression vector, transformed into Escherichia coli strain BL21 DE3(pLysS) and induced expression using IPTG 1 mM overnight at 37 °C. CP that contains 6xhistag was purified using NiNTAspin column and then confirmed by SDS-PAGE. The size of TICV CP gene was 750 bp and the gene was expressed on pET 21 b vector and SDS-PAGE showed a 29 kDa band.

Tomato infectious chlorosis virus. [Distribution map].

A new distribution map is provided for Tomato infectious chlorosis virus. Closteroviridae: Crinivirus. Main host: tomato (Solanum lycopersicum). Information is given on the geographical distribution in Europe (Bulgaria, France, Greece, Crete, Italy, Sardinia, Sicily and Spain), Asia (Indonesia, Japan, Honshu, Jordan and Taiwan), Africa (Tunisia) and North America (Mexico, USA, California and North Carolina).

IDENTIFIKASI TOMATO INFECTIOUS CHLOROSIS VIRUS PENYEBAB PENYAKIT KLOROSIS PADA TANAMAN TOMAT DI CIPANAS JAWA BARAT MELALUI PERUNUTAN NUKLEOTIDA GEN PROTEIN SELUBUNG UTAMA

Identification of tomato infectious chlorosis virus, the causal agent of chlorosis disease on tomato in Cipanas West Java by sequencing of main coat protein gene nucleotide. Tomato infectious chlorosis virus (TICV) causes chlorosis on tomato. Tomatoes infected by this virus shows interveinal yellowing, necrotic, bronzing, brittleness, and declining in productivity. This study aims to identify the causal agent of chlorotic disease on tomato by sequencing the coat protein gene. The methods involve collecting infected plants, total RNA extraction, cDNA synthesis, DNA amplification, visualization of the results of reverse transcription polymerase chain reaction (PCR), and phylogenetic analysis using BLAST, clustal w, Bioedit v 7.0.5.3, MEGA v 6:06. RT-PCR using spesific primers (CP-F TICV Bam and TICV R-Hind) amplified a DNA band of 792 bp, which has been successfully sequenced and identified as TICV. Nucleotide sequences homology analysis showed that TICV Indonesia_TWJ isolate Cipanas is the same strain as TICV from other countries (99.4 – 100%), such as Spain, Greece, USA, France, and Italy.