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Wei J.-C.,Mudanjiang Teachers College | Qiu E.-J.,Mudanjiang Tobacco Research Institute | Guo H.-Y.,Mudanjiang Teachers College | Hao A.-P.,Mudanjiang Teachers College | Chen R.-P.,Mudanjiang Tobacco Research Institute
Biotechnology and Biotechnological Equipment | Year: 2014

A pair of primers was designed to amplify the propylene alcohol dehydrogenase gene sequence based on the cDNA sequence of the tobacco allyl-alcohol dehydrogenase gene. All introns were sequenced using traditional polymerase chain reaction (PCR) methods and T-A cloning. The sequences from common tobacco (Nicotiana tabaccum L.) and rustica tobacco (Nicotiana rustica L.) were analysed between the third intron and the fourth intron of the propylene alcohol dehydrogenase gene. The results showed that the alcohol dehydrogenase gene is a low-copy nuclear gene. The intron sequences have a combination of single nucleotide polymorphisms and length polymorphisms between common tobacco and rustica tobacco, which are suitable to identify the different germplasms. Furthermore, there are some single nucleotide polymorphism sites in the target sequence within common tobacco that can be used to distinguish intraspecific varieties. © 2014 The Author(s). Published by Taylor & Francis.

Wan X.,Mudanjiang Tobacco Research Institute | Qiao C.,Mudanjiang Tobacco Research Institute | Zhao S.,Mudanjiang Tobacco Research Institute | Li R.,Mudanjiang Tobacco Research Institute | And 2 more authors.
Tobacco Science and Technology | Year: 2015

To clarify the kinds and distribution states of tobacco Potato virus Y (PVY) strains in Heilongjiang tobacco planting areas, 107 suspected PVY isolates collected from Heilongjiang tobacco planting areas were identified with multiplex polymerase chain reaction (Multiplex-PCR) and sequencing techniques. The results indicated that the full length of genomes in four representative isolates (HLJ-SH8, HLJ-SC2, HLJ-SC6 and HLJ-MDJ4) were 9 698, 9 702, 9 702 and 9 698 bp, respectively. All of the four isolates contained an open reading frame (ORF) of 9 186 bp in length and encoded a polyprotein of 3 061 amino acids. Phylogenetic analysis indicated that the four isolates were PVYNTN, PVYN, PVYNW and PVYNTN-NW strains, respectively; among which, PVYNTN-NW strain was the dominant strain in Heilongjiang tobacco planting areas. © 2015, Editorial Office of Tobacco Science & Technology. All right reserved.

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