RT-PCR APPLICATION FOR DETECTION OF THE STRAWBERRY CRINKLE VIRUS AND STRAWBERRY MILD YELLOW EDGE VIRUS DISEASES ON STRAWBERRY PLANTS (FRAGARIA VESCA L.) CULTURED IN VITRO

Dương Tấn Nhựt
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Authors

  • Dương Tấn Nhựt Viện sinh học Tây Nguyên

DOI:

https://doi.org/10.15625/1811-4989/7/3/12450

Keywords:

Propagation, reverse transcription-polymerase chain reaction, strawberry, strawberry crinkle virus, stra-wberry mild yellow edge virus, virus-free strawberry plantlets

Abstract

The reverse transcriptase-polymerase chain reaction (RT-PCR) was applied for detection of the strawberry crinkle vims (SCV) and strawberry mild yellow edge vims (SMYEV) diseases on three strawberry cultivars: "My Da", "My Huong" and "Phap". The in vitro leaves of these three strawberry cultivars were used for this research. Three different culture media were used. Calli were induced on the MS medium containing 1 mg/l TDZ, 0.1 mg/l 2,4-D, 30 g/l sucrose and 8 g/l agar (1); shoots were regenerated on MS medium containing 0.2 mg/l BA, 30 g/l sucrose and 8 g/l agar (2); roots were formed on MS medium containing 5 ml/1 B5 vitamin, 30 g/l sucrose and 8 g/l agar (3). The RNA extraction from leaf tissues according to the method of Mazzara and James (2000) and then, the extracted RNA transposition into RT-PCR using the StrataScript® One-Tube RTPCR Kit of ITS Vietnam. In this research, two primer pairs SCIDFW-SCIDRV and SMlDFW-SMlDRV were used for the detection of these virases by RT-PCR. The amplified products had expected sizes: 345 bp and 271 bp, respectively. We found that the RT-PCR test with these two primer pairs SCIDFW-SCIDRV and SMlDFW-SMlDRV was capable to detect the SCV and SMYEV diseases on in vitro strawberry plantlets. The infection rates of SCV and SMYEV on three strawberry cutivars were "My Da": 2.66% SCV and 3.3% SMYEV; "My Huong": 4% SCV and 2.66% SMYEV; "Phap": 4.6% SCV and 1.3% SMYEV. Vims-free strawberry plantlets were obtained, and were used as a vims-free expiant source for the strawberry propagation.

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Published

22-11-2011

How to Cite

Nhựt, D. T. (2011). RT-PCR APPLICATION FOR DETECTION OF THE STRAWBERRY CRINKLE VIRUS AND STRAWBERRY MILD YELLOW EDGE VIRUS DISEASES ON STRAWBERRY PLANTS (FRAGARIA VESCA L.) CULTURED IN VITRO. Vietnam Journal of Biotechnology, 7(3), 335–340. https://doi.org/10.15625/1811-4989/7/3/12450

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