Articles
ELIMINATION OF APPLE STEM PITTING VIRUS FROM PEAR CULTIVARS BY IN VITRO CHEMOTHERAPY
Article number
923_15
Pages
111 – 115
Language
English
Abstract
The objective of the present study was to determine whether chemotherapy with Ribavirin can be used to eliminate Apple stem pitting virus (ASPV) from in vitro grown plants of infected pear cultivars Alexander Lucas, Bohemica, Elektra and Rote Williams. ASPV was detected in naturally infected field-grown trees by RT-PCR analysis.
In vitro cultures were successfully multiplied on MS medium with BAP (6-benzylaminopurine). Ribavirin was added in concentration of 20 mg L-1 after autoclaving to the same MS medium as for multiplication.
After a subculture period of 4 weeks, the apical part of the axis (about 3 mm in length comprising the apical meristem plus two-three primordial leaves) was dissected under a laminar flow hood and transferred to a fresh multiplication MS medium with 1.5 mg L-1 BAP for regeneration.
From the selected genotypes, at used concentration (20 mg L-1), low phytotoxicity of Ribavirin was observed in the case of cultivars Alexander Lucas and Rote Williams. The chemotherapy technique was able to eliminate ASPV in different percentage according to the cultivar, varying between 73.7% for cultivar Rote Williams and 90.0% for Elektra. The obtained results demonstrated the effectiveness of the system of virus elimination by a combination of in vitro cultures, chemotherapy with Ribavirin and subsequent removing of apical meristematic region.
Relatively high percentage of obtained virus free plants confirmed high antiviral activity of Ribavirin against ASPV. Achieved results are preliminary.
Other sanitation and repeated testing of pear cultivars used in the study will be carried out in the following years.
In vitro cultures were successfully multiplied on MS medium with BAP (6-benzylaminopurine). Ribavirin was added in concentration of 20 mg L-1 after autoclaving to the same MS medium as for multiplication.
After a subculture period of 4 weeks, the apical part of the axis (about 3 mm in length comprising the apical meristem plus two-three primordial leaves) was dissected under a laminar flow hood and transferred to a fresh multiplication MS medium with 1.5 mg L-1 BAP for regeneration.
From the selected genotypes, at used concentration (20 mg L-1), low phytotoxicity of Ribavirin was observed in the case of cultivars Alexander Lucas and Rote Williams. The chemotherapy technique was able to eliminate ASPV in different percentage according to the cultivar, varying between 73.7% for cultivar Rote Williams and 90.0% for Elektra. The obtained results demonstrated the effectiveness of the system of virus elimination by a combination of in vitro cultures, chemotherapy with Ribavirin and subsequent removing of apical meristematic region.
Relatively high percentage of obtained virus free plants confirmed high antiviral activity of Ribavirin against ASPV. Achieved results are preliminary.
Other sanitation and repeated testing of pear cultivars used in the study will be carried out in the following years.
Authors
J. Sedlak, F. Paprstein , L. Talacko
Keywords
in vitro culture, virus detection, shoots, RT-PCR, Ribavirin
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