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Recovery of Four Novel Potato spindle tuber viroid Sequence Variants from Russian Seed Potatoes

First described in the early 1930s, the limited distribution of potato “gothic” disease made it of little economic significance in European Russia until the early 1970s when meristem-tip culture was widely adopted throughout the former USSR to increase production of virus-free seed potatoes. Shortly thereafter, the yield and quality of Russian seed potatoes began a dramatic decline. Symptoms of potato “gothic” resemble those of Potato spindle tuber viroid (PSTVd) (3), and initial suspicions that in vitro plantlets and seed potatoes might be viroid-infected were later proved correct when Kastalyeva et al. (2) showed that approximately 50 to 70% of in vitro plantlets and tubers collected from different regions of Russia as well as the in vitro germplasm collection maintained by the All-Russian Potato Research Institute (ARPRI) were infected with PSTVd. Measures have since been taken to reduce the incidence of PSTVd infection, and numerous PSTVd isolates were collected from territories of the former USSR; however, none of these isolates have been characterized at the molecular level. Overlapping reverse transcription (RT)-PCR products (1) were generated from four PSTVd isolates maintained in field-grown tubers at the VNIIF using two pairs of primers; PSTVd180F (5′-TCACCCTTCCTTTCTTCGGGTGTC-3′) + PSTVd179R (5′-AAACCCTGTTTCGGCGGGAATTAC-3′) and PSTVd112F (5′-ACT GGCAAAAAAGGACGGTGGGGA-3′) + PSTVd359R (5′-AGGAACC AACTGCGGTTCCAAGGG-3′). Automated sequence analysis of the resulting uncloned PCR products revealed the presence of four previously unknown PSTVd variants (GenBank Accession Nos. EF044302-EF044305). All four tubers were also infected with Potato virus M and Potato virus Y and one tuber also contained Potato virus S. ELISA tests for Potato leaf roll virus were negative. Each isolate appeared to contain only a single 358--359 nt variant differing from PSTVd-intermediate strain (GenBank Accession No. V01465) at 2--5 positions. The three closely related variants originating from Leningradskaya Province (Northwest Russia) contain two to three changes in the variable domain and central conserved region and induced intermediate symptoms in Rutgers tomato. The fourth variant originating from Samarskaya Province (Volga River Region) contains additional changes in the pathogenicity domain and induced mild symptoms. Minor differences among the Leningradskaya variants may represent sequence drift during extended (9 to 11 year) tuber passage. The presence of additional sequence changes in the variant from Samarskaya is consistent with independent origin and/or prolonged separation. Additional studies with a wider range of Russian isolates of PSTVd are currently underway to develop diagnostic methods suitable for future large-scale screening programs.

References: (1) Y. Hu et al. Virology 219:45, 1997. (2) T. B. Kastalyeva et al. Vestn. RASKHN 3:22, 1992. (3) Y. A. Leontyeva. Potato spindle tuber (‘gothic’) as one of the most important diseases in the Volga region. (In Russian.) Ph.D. thesis. Agricultural University of Leningrad, Pushkin, 1971.