Determination of Local Tobacco Cultivars Using ISSR Molecular Marker
Jessada Denduangboripant, Sornsuda Setaphan, Wilasinee Suwanprasart, and Somsak Panha* Author for corresponding; e-mail address: jessada.d@chula.ac.th
Volume: Vol.37 No.2 (MAY 2010)
Research Article
DOI:
Received: 19 October 2009, Revised: -, Accepted: 11 December 2009, Published: -
Citation: Denduangboripant J., Setaphan S., Suwanprasart W. and Panha S., Determination of Local Tobacco Cultivars Using ISSR Molecular Marker , Chiang Mai Journal of Science, 2010; 37(2): 293-303.
Abstract
In many tobacco-growing countries, imported and local tobacco cultivars are subject to different regulations in tariff collection. However, there is a major technical problem how to distinguish between the imported and local cultivars. No standard method has been established to judge whether the plant has been grown for long time, enough to be legally called “local cultivar”. In this study, an Inter-Simple Sequence Repeat (ISSR) method was introduced to study genetic relationships between imported and local tobacco cultivars grown in Thailand. Of 20 screened-primers, five ISSR primers showed polymorphic PCR amplified products suitable for further phylogenetic analysis. Among them, two primers generated specific PCR bands that could separate local tobacco cultivars (Chorlare 1 and Chorlare 2) from other imported cultivars. Genetic relationship trees revealed four pairs of tobacco cultivars having bootstrap supporting-values higher than 50%. Among all 13 local tobacco cultivars collected and examined, only the couple of Chorlare 1 and 2 was distantly separated from the imported cultivars, suggesting their long-history of being grown in Thailand. The other 11 local cultivars were found to be morphologically and genetically similar to some imported cultivars and could have originated from them. Therefore, the ISSR technique could be developed and implemented as a simple, effective molecular marker to recognise early-introduced local tobacco cultivars.