Articles
SEQUENCE ANALYSIS OF THE NUCLEOCAPSID PROTEIN GENE OF A TOMATO SPOTTED WILT VIRUS ISOLATE FROM GEORGIA, USA
Article number
431_21
Pages
237 – 243
Language
Abstract
Tomato spotted wilt virus (TSWV) has become an economically important virus of peanuts, tobacco, and vegetables in Georgia, USA. Our objective was to characterize TSWV isolates occurring in Georgia.
The nucleocapsid (N) protein gene was isolated by reverse transcription-polymerase chain reaction (RT-PCR) from total nucleic acid extracts from TSWV-infected plants.
The primer pair specific to the N gene of the TSWV-L isolate was used.
The two primers (5′ ATGTCTAAGGTTAAGCTC 3′ and 5′ TTAAGCAAGTTCTGTGAG 3′) gave a PCR product of about 800 bp from the majority of the peanut and tobacco samples tested.
The PCR product obtained from an isolate collected from the Coastal Plain Experiment Station’s Black Shank Nursery was subsequently cloned into pUC118 vector, and the nucleotide (nt) sequence was determined.
The tobacco isolate shared about 97% identity with the TSWV-L isolate at both the nucleotide and deduced amino-acid sequence levels.
Pair-wise comparisons with N genes of TSWV-Italy, TSWV-Bulgaria, TSWV-10W, TSWV-BL, and TSWV-CPNH1 showed a high degree of similarity (94–99%) at both the nt and amino-acid levels.
However, the tobacco isolate showed divergence from the known N-gene sequences at four positions (i.e., Ser18, Gly40, Cys174, and Thr205) in its deduced amino acid sequence.
The nucleocapsid (N) protein gene was isolated by reverse transcription-polymerase chain reaction (RT-PCR) from total nucleic acid extracts from TSWV-infected plants.
The primer pair specific to the N gene of the TSWV-L isolate was used.
The two primers (5′ ATGTCTAAGGTTAAGCTC 3′ and 5′ TTAAGCAAGTTCTGTGAG 3′) gave a PCR product of about 800 bp from the majority of the peanut and tobacco samples tested.
The PCR product obtained from an isolate collected from the Coastal Plain Experiment Station’s Black Shank Nursery was subsequently cloned into pUC118 vector, and the nucleotide (nt) sequence was determined.
The tobacco isolate shared about 97% identity with the TSWV-L isolate at both the nucleotide and deduced amino-acid sequence levels.
Pair-wise comparisons with N genes of TSWV-Italy, TSWV-Bulgaria, TSWV-10W, TSWV-BL, and TSWV-CPNH1 showed a high degree of similarity (94–99%) at both the nt and amino-acid levels.
However, the tobacco isolate showed divergence from the known N-gene sequences at four positions (i.e., Ser18, Gly40, Cys174, and Thr205) in its deduced amino acid sequence.
Authors
H.R. Pappu, A.K. Culbreath, P.F. Bertrand, A.S. Csinos, C.L. Niblett
Keywords
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