MOLECULAR DIAGNOSTICSFOR THE POTATO SPINDLE TUBER VIROID IN THE REPUBLIC OF KAZAKHSTAN
Main Article Content
Authors
L.T. Nadirova
M. Aitkhozhin Institute of Molecular Biology and Biochemistry, 86 Dosmukhamedov str., Almaty, 050012, Kazakhstan
G.E. Stanbekova
M. Aitkhozhin Institute of Molecular Biology and Biochemistry, 86 Dosmukhamedov str., Almaty, 050012, Kazakhstan
D.K. Beisenov
M. Aitkhozhin Institute of Molecular Biology and Biochemistry, 86 Dosmukhamedov str., Almaty, 050012, Kazakhstan
B.K. Iskakov
M. Aitkhozhin Institute of Molecular Biology and Biochemistry, 86 Dosmukhamedov str., Almaty, 050012, Kazakhstan
Abstract
Potato is one of the most consumed crop products. It is grown inan area of 190000 hectares in Kazakhstan. The potato spindle tuber viroid (PSTVd) is the causative agent of the so-called ‘gothic’ disease, and it causesthe degradation of potato tubers anda decrease in yield of up to 65%. PSTVd has been found in every continent;in Kazakhstan, molecular diagnostics for PSTVd have not yet been performed.
The first survey of PSTVd in Kazakhstan was performed, and spindle- and pear-shaped tubers collected from private farms in the Almaty region were grown under greenhouse conditions. Total RNAwas isolated from the leaves and analysed for the presence of PSTVd. Reverse transcription-polymerase chain reaction (RT-PCR) with specific primers revealed a 360-bp DNA fragment in a number of samples. The correspondence to PSTVd was confirmed using sequencing. Subsequently, the isolated PSTVd clone was used as a labelled probe for detecting the viroid in other plants. RT-PCR and nucleic acid hybridization assay revealed the presence of PSTVd in 26% of the selected spindle tubers.
Currently, there are no effective methods to prevent the infection of potatoes by PSTVd; therefore, diagnostic methods are crucial for preventing the spread of PSTVd through seed material.
Keywords
potato spindle tuber viroid, diagnostic, reverse transcription, PCR, northern blotting
Article Details
References
Ospanova G.S., Bozshataeva G.T., Turabaeva G.K., Alikhanova A. Viral solanaceae diseases in Kazakhstan. International journal of applied and fundamental research, 2014, no. 3, pp. 62-64.
Salazar L.F. Potato spindle tuber viroid. In: Plant Protection and Quarantine, 1989, vol. 2. Selected Pests and Pathogens of Quarantine Significance (Kahn R.P., ed), pp. 155-167.
Drygin Yu.F., Musin S.M., Kondakova O.A., et al. Molecular diagnostic of the recovered potato contamination by the spindle tuber viroid. Reports of RAAS, 1996, no. 6, pp. 24-25.
Diener T., Raymer W. Potato spindle tuber virus: a plant virus with properties of a free nucleic acid. Science, 1967, vol. 158, pp. 378-381. PMID: 6061889. doi: 10.1126/science.158.3799.378.
Gross H.J., Domdey H., Lossow C., et al. Nucleotide sequence and secondary structure of potato spindle tuber viroid. Nature, 1978, vol. 273, pp. 203-208. doi: 10.1038/273203a0.
Herold T., Haas B., Singh R.P., et al. Sequence analysis of five new field isolates demonstrates that the chain length of potato spindle tuber viroid (PSTVd) is not strictly conserved but is variable as in other viroids. Plant Mol. Biol., 1992, vol. 19, pp. 329-333. PMID: 1623184. doi: 10.1007/BF00027356.
Lakshman D., Tavantzis S. Primary and secondary structure of a 360-nucleotide isolate of potato spindle tuber viroid. Arch. Virol., 1993, vol. 128, pp. 319-331. PMID: 8435045. doi: 10.1007/BF01309442.
Puchta H., Herold T., Verhoeven K., et al. A new strain of potato spindle tuber viroid (PSTVd-N) exhibits major sequence differences as compared to all other strains sequenced so far. Plant Mol. Biol., 1990, vol. 15, pp. 509-511. PMID: 2103469. doi: 10.1007/BF00019169.
Behjatnia S.A.A., Dry I.B., Krake L.R., et al. New potato spindle tuber viroid and tomato leaf curl geminivirus strains from a wild Solanum sp. Phytopathology, 1996, vol. 86, pp. 880-886.
Raymer W.B., O’Brien M.J. Transmission of potato spindle tuber virus to tomato. Am. Potato J., 1962, vol. 39, pp. 401-408. doi: 10.1007/BF02909569.
Morris T.J., Wright N.S. Detection on polyacrylamide gel of a diagnostic nucleic acid from tissue infected with potato spindle tuber viroid. Am. Potato J., 1975, vol. 52, pp. 57-63. doi: 10.1007/BF02852039.
Owens R.A., Diener T.O. Sensitive and rapid diagnosis of potato spindle tuber viroid disease by nucleic acid hybridization. Science, 1981, vol. 213, pp. 670-672. PMID: 17847478. doi: 10.1126/science.213.4508.670.
Kondakova O.A., Drygin Yu.F. Diagnostic of the potato viroid disease by the (dien)Pt DNA probe. Biotechnologiya, 1999, vol. 4, pp. 83-90.
Drygin Yu.F., Chirkov S.N., Kondakova O.A., et al. Highly sensitive technologies for the molecular diagnosis of potato viral and viroid infections. Achievements of APK science and technique, 2007, no. 7, pp. 20-27.
Boonham N., Perez L.G., Mendez M.S., et al. Development of a real-time RT PCR assay for the detection of potato spindle tuber viroid. J. Virol. Methods, 2004, vol. 116, pp. 139-146. PMID: 14738980. doi: 10.1016/j.jviromet.2003.11.005.
UNECE Standard S-1 concerning the marketing and commercial quality control of Seed Potatoes. UNECE Guide to Seed Potato Diseases, Pests and Defects. United Nations, New York and Geneva, 2014, 108 p.
Góra A., Candresse T., Zagórski W. Analysis of the population structure of three phenotypically different PSTVd isolates. Arch Virol., 1994, vol. 138, no. 3-4, pp. 233-245. PMID: 7998831. doi: 10.1007/BF01379128.