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Emir. J. Food Agric. 2011; 23(4): 355-367


Identification and molecular characterization of
Tomato Yellow Leaf Curl Virus-EG

Asmaa F. Abd El-Monem1
, Kh. A. El-Dougdoug2
, Ibtisam A. Hamad1
,
Entsar A. Ahmed1
and Hayam S. Abd El-Kader3,4?.

Abstract
Tomato Yellow Leaf Curl virus (TYLCV-Eg) was isolated from whiteflies-infectedtomato (Lycopersicon esculentum cv.Castle rock) plants growing in Nubaria and El-BeheraGovernorate. The infected plants exhibited systemic viral symptoms in the form of sever leafcurling, leaf crinkle with marginal yellowing, stem upright, twisted and stunted. TYLCV-Egreacted positively with polyclonal antibodies specific to TYLCV using DAS-ELISA. It wastransmitted by both syringe injection and whiteflies with transmission efficiency of about 80% and100%, respectively. TYLCV-Eg isolate was transmitted to different species belonging to familiesCucurbitaceae, Fabaceae, Solanaceae and Chenopodiaceae. TYLCV had TIP of 70C, DEP of10-7 and LIV of about 6 days. Electron micrograph of the partially purified TYLCV revealed thepresence of monomer and dimmer gemini particles with dimensions of 22 nm and 20 30 nm to24 30 nm, respectively when negatively stained with uranyl acetate. Using degenerateoligonucleotide primers, the viral coat protein gene was amplified successfully by PCR, producing~ 500 bp fragment from tomato infected plants. The viral genome was detected by specific DNAprobe using dot blot hybridization technique. Comparative nucleotide sequence analysis showed asimilarity of 98% between TYLCV-Eg and other isolates.

Key words: TYLCV-EG, DAS-ELISA, PCR, Dot-blot hybridization, and Nucleotide sequence



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