Nucleotide sequence of the coat protein gene of pelargonium leaf curl virus and comparison of the deduced coat protein amino acid sequence with those of other tombusviruses

Abstract

The sequence of 1,787 nucleotides (nts) in the genomic RNA of pelargonium leaf curl virus (PLCV) was determined. It included the entire coat protein (cp) gene (nts 585 to 1,754), 558 nts of the 3' end of the putative RNA polymerase gene, 26 nts of an intercistronic region between the two genes and 33 nts downstream of the stop codon of the cp gene. The cp gene was cloned into the expression vector pET8c and expressed in E. coli. The deduced cp amino acid sequence of PLCV was compared with those of five other tombusviruses. The closer the degree of serological relatedness between two viruses, the more similarity was found in their cp amino acid sequences not only in the protruding domains, but also in their random and shell domains and in the arm regions. Nucleic acid hybridization tests, cp amino acid comparisons and serological tests all suggest the same order of sequence for the relationships in the tombusvirus group.

The complete nucleotide sequence of pelargonium leaf curl virus

“…The largest sequence (1787 nt) covers c. 550 nt from the 3' end of the P95 replicase gene and the entire CP gene (Li et al, 1993). This sequence is 97.4 % identical to that of our new, complete sequence.…”

“…We were able to amplify a full-length (c. 4.7 kb) clone of this virus by RT-PCR using 5'-and 3'-terminal primers derived from TBSV (5' terminus, ? strand, GAAATTCTCCAGGA TTTCTCGACC; 3' terminus, -strand, GGGCTGCATTTC TGCAATG); however, during the sequencing of this clone, we found that the CP sequence was identical to that described for PLCV [1] rather than for TBSV. To obtain the authentic terminal sequences of the PLCV genome, we isolated total RNA from virus-infected N. benthamiana leaves using a QIAGEN RNeasy Plant Mini Kit according to the manufacturer's instructions.…”

“…A serological study showed that PLCV isolates formed a homogeneous group that differed in reactivity to isolates of the tombusviruses tomato bushy stunt virus (TBSV), petunia asteroid mosaic virus (PAMV) and artichoke mottled crinkle virus (AMCV) [3]. Another study by Li et al [1] reported the nucleotide sequence of the coat protein (CP) gene of PLCV and described the CP amino acid sequence relationship between PLCV and five other tombusviruses. Most recently, PLCV was isolated from soil and geranium plants in Iran, and partial CP gene sequences were reported [7].…”

First Report on Occurrence of Pelargonium leaf curl virus (PLCV) in Shimla Hills, India

“…For precise identification, primers were designed for the amplification of coat protein gene on the basis of available sequences of PLCV in the NCBI database. Use of reverse transcriptionpolymerase chain reaction (RT-PCR) with specific primer for PLCV coat protein gene resulting in a product of 1,100 bp has already been reported [4]. To our knowledge, this is the first report of natural infection of PLCV on Pelargonium in India.…”