Abstract:

To provide theoretical basis for the cultivation and identification of virus-free seedlings of P. praeruptorum Dunn. cv. Xinzhou, its tobacco mosaic virus was identified, its protein was expressed and its sequence was analyzed. The protein gene of tobacco mosaic virus of P. praeruptorum Dunn. cv. Xinzhou was identified by lncRNA sequencing and RT-PCR, and its protein of tobacco mosaic virus was expressed by E. coli recombinant technology, and its gene sequence was analyzed by bioinformatics method. The protein genes of tobacco mosaic virus of P. praeruptorum Dunn. cv. Xinzhou included replicase (ORF1), movement protein (ORF2) and coat protein (ORF3). The replicating enzyme, movement protein and coat protein of tobacco mosaic virus could be detected by RT-PCR, and could be recombined and expressed in E. coli, indicating that the sequencing result of lncRNA of tobacco mosaic virus was accurate. The total length of the cDNA of the replicase, movement protein and coat protein genes of the tobacco mosaic virus was 3351 bp, 807 bp and 480 bp, respectively. The replicase, movement protein and coat protein of tobacco mosaic virus from P. praeruptorum Dunn. cv. Xinzhou were closely related to tobacco mosaic virus, with the homology of 99.64%, 98.88% and 98.74% respectively. Peucedanum praeruptom tobacco mosaic virus (PpTMV) has occurred in seven districts or counties in Xinzhou, Guangxin, Yushan, Guangfeng, Wuyuan, Dexing and Yiyang, the main producing areas of P. praeruptorum Dunn. cv. Xinzhou in Shangrao, Jiangxi Province. This article is the first report of PpTMV detected in P. praeruptorum Dunn. cv. Xinzhou.

Key words: P. praeruptorum Dunn. cv. Xinzhou, tobacco mosaic virus, lncRNA sequencing identification, RT-PCR, prokaryotic protein expression, sequence analysis