Nucleic Acids Research, 1992, Vol. 20, No. 24 6733 © 1992 Oxford University Press Nucleotide sequence of a cucumber mosaic virus satellite RNA associated with a tomato top stunting Francesco Grieco, Fabrizio Cillo1, Loredana Barbarossa and Donato Gallitelli* Dipartimento di Protezione delle Piante dalle Malattie, Universita di Bari and Centra di Studio sui virus e le virosi delle colture mediterranee del CNR, 70126 Bari, and 1Tecnogen SCpA, 81015 Piana di Monte Verna (CE), Italy EMBL accession no. X69136 Received November 23, 1992; Accepted November 25, 1992 A strain of cucumber mosaic cucumovirus (CMV) belonging to the WT subgroup has been isolated in Apulia (Southern Italy) from tomato plants showing a severe shortening of internodes of terminal shoots leading to a bushy appearance. The disease was clearly distinguishable from other CMV-induced disorders in tomato Italian crops like fruit necrosis, lethal necrosis and shoestring-fem leaf (1). Virus preparations purified from naturally infected plants contained CMV genomic RNAs 1 —4 and a satellite RNA named TTS-CARNA 5 (CARNA 5 for CMVassociated RNA 5). The virus, for which the name of CMVTTS was adopted, was cultured in Rutgers tomato and purified by a standardized procedure (2). We report here the nucleotide sequence of its satellite RNA (Figure 1). Nucleotide sequence of TTS-CARNA5 was determined using the dideoxy-chain termination method on both strands of three different pUC-cloned cDNA templates, and by direct sequencing on RNA template (3). When compared with other CMV satellite RNAs (1, Bl, B2, D, E, G, I17N, R, S, WL1, WL2, F, K8) (4, 5, 6) the following nucleotides were found to be uniques for TTS-CARNA5: T38 (vs C), C 3 9 (vs. T), A269 (vs G). The 3' half of TTS-CARNA 5 does not contain the 'necrogenic consensus' reported for other CMV satellite RNAs (4). Preliminary results strongly suggest that TTS-CARNA5 is involved in causing tomato top stunting by a modulation of symptoms induced by CMV helper strain. ACKNOWLEDGEMENTS This work was supported by a grant of MURST (art. 65 D.P.R.382/80 quota 40%) 'Genetica del virus del mosaico del cetriolo e del suo RNA satellite in isolati da pomodoro'. Grateful * To whom correspondence should be addressed thanks are due to Dr P.Iosco, Metapontum Agrobios, Metaponto (MT), Italy, for preparing the synthetic oligonucleotides used in this study. REFERENCES 1. 2. 3. 4. 5. 6. Crescenzi.A. et al. (1992) Plant Disease 76, (in press). Lot,H. et al. (1972) Ann. PhytopathoL 4, 2 5 - 3 8 . Meshi.T. et al. (1983) FEBS Lett. 162, 282285. Devic.M. etal., (1990)7. Gen. Virol. 71, 1443-1449. Hidaka.S. et al. (1990) J. Gen. Virol. 71, 439-442. Fraile A. et al. (1990) Nucleic Adds Res. 18, 4593. 10 20 30 40 50 60 GUUUUGUUUC UUAGAGAAUU GCGUGGAGCG GUUAUAUUCA CGUGAGGAUC UAUCACUCGG 110 120 70 80 90 100 CGGUGUGGGU UACCUCCCUG CUACGGCGGG UUGAGUUGAC GCACCUGCGA CUGGGGACCG 170 180 130 140 150 160 CUGGCCUGCG AGCUAUGUCC GCUACIKUCA GCACUACGCA CUCAUUUGAG CCCCCGCUCA 230 240 190 200 210 220 GUUUGCUAGC AAAACCCGGC ACGUGGUUUG CCGUUACCAC GGAACUUCGA AAGAAACACU 290 300 250 260 270 280 CUGUUAGGUG GUAUCGUGGA UGACGCACAC AGGGAGAGGC UAAAACCUAU ACGGUCAUGC 310 320 330 UGAUCUCCGU GAAUGUCUAC ACAUUCCUCU ACAGGACCC Figure 1. Complete nucleotide sequence of TTS-CARNA5.