Detection and Identification of Tomato Yellow Leaf Curl Virus-EG Using Molecular Technique | ||
| Egyptian Journal of Microbiology | ||
| Article 7, Volume 45, Issue 1, December 2010, Pages 95-111 PDF (639.1 K) | ||
| Document Type: Original Article | ||
| DOI: 10.21608/ejm.2010.279 | ||
| Abstract | ||
| TOMATO Yellow Leaf Curl virus (TYLCV-Eg) was isolated from whiteflies-infected tomato (Lycopersicon esculentum cv. Castle rock) plants growing in Nubaria and El-Behera Governorate. The infected plants exhibited systemic viral symptoms in the form of severe leaf curling, leaf crinkle with marginal yellowing, stem upright, twisted and stunted. TYLCV-Eg reacted positively with polyclonal antibodies specific to TYLCV using DAS-ELISA. It was transmitted by both syringe injection and whiteflies with the transmission efficiency of about 80% and 100%, respectively. TYLCV-Eg isolate was transmitted to different species belonging to families Cucurbitaceae, Fabaceae, Solanaceae, and Chenopodiaceae. TYLCV had TIP (Thermal Inactivation Point) of 70ºC, DEP (Dilution End Point) of 10-7 and LIV (Longevity) of about 6 days. Electron micrograph of the partially purified TYLCV revealed the presence of monomer and dimmer gemini particles with dimensions of 22 nm and 20 x 30 nm to 24 x 30 nm, respectively when negatively stained with uranyl acetate. DNA from infected plants was extracted and amplified successfully by polymerase chain reaction (PCR) using degenerate oligonucleotide primers V324(+) and C889 (-) producing ~ 500 bp fragment from infected tomato plants. The viral genome was detected by specific DNA probe using dot blot hybridization technique. Comparative nucleotide sequence analysis showed a similarity of 98% between TYLCV-EG and other isolates. | ||
| Keywords | ||
| nucleotide sequence; TYLCV-EG; DAS-ELISA; PCR; Dot-blot hybridization | ||
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