Isolation of tagged INUIT gene by establishment of TAIL-PCR in Tobacco

• Main Author: Vodnala, Sharvani Munender
• Format: Master Thesis
• Language: English
• Published: Fysiologisk botanik 2007
• Subjects: inuit
• Online Access: http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-26083

Chilling injury is one of the key factors restraining plant growth in tropical and subtropical crop species. It is recognized that stability of microtubules is directly linked to systemic cold resistance of entire organism. We used a mutant approach in order to examine a possible function of microtubules in cold resistance. For this reason we characterized a mutant called inuit generated by T-DNA activation tagging (Koncz et al. 1994; Ahad et al. 2003). Inuit mutants were directly screened for long term cold resistance. The analysis of these mutants is targeted to isolate new components of so far hardly known pathway that links cold sensing to microtubular response. A mutant called ATER was screened for resistance to anti microtubular herbicide EPC are altered in microtubular dynamics and are cross resistant to chilling stress (Ahad et al. 2002; 2003). From one of these mutant lines a novel member of cytochrome-P450 oxidase superfamily was identified by a tag. This gene might represent a central element of environmental signaling towards cytoskeleton. Although plasmid rescue technique works very well in Arabidopsis, it seems in tobacco does not work in same extent. Therefore we established a technique called TAIL-PCR in tobacco which is an efficient method to amplify unknown sequences adjacent to known insertion sites was applied here in this study to isolate INUIT gene.