In 2009, in the framework of surveying for pospiviroids, samples of various ornamental plants from the Netherlands were tested by reverse transcription (RT)-PCR with the primer pairs Pospi1-RE/FW and Vid-RE/FW (2). With primer pair Pospi1-RE/FW, amplicons of the expected size were obtained in two samples of symptomless plants of Lycianthes rantonnetii and Streptosolen jamesonii. Sequencing of the amplicons, which were expected to correspond with partial pospiviroid genomes, showed identities of 100 and 98% to the sequence of Tomato apical stunt viroid (TASVd), NCBI GenBank Accession No. AM777161 (3). For the amplification of the complete viroid genomes, RT-PCRs were performed with primer pair CEVd-FW/RE (1). Sequencing of these amplicons yielded sequences of 364 nt and identities to TASVd AM777161 of 100 and 98.1%, respectively. Therefore, both isolates were identified as TASVd. The sequence variant from S. jamesonii was submitted to the NCBI GenBank as No. GU911351. In addition, both isolates were mechanically inoculated to four tomato plants (Solanum lycopersicum) of cv. Moneymaker. All inoculated plants developed chlorosis and growth reduction after 4 weeks and TASVd infections were confirmed in a bulked sample by RT-PCR with primer pair CEVd-FW/RE after 6 weeks. Hence, two more ornamental host plant species have been identified that may act as symptomless sources of pospiviroid inoculum.
References: (1) N. Önelge. Turk. J. Agric. For. 21:419, 1997. (2) J. Th. J. Verhoeven et al. Eur. J. Plant Pathol. 110:823, 2004. (3) J. Th. J. Verhoeven et al. Plant Dis. 92:973, 2008.
