MOLECULAR ANALYSIS OF YEAST SPLICEOSOME COMPONENTS FOR THEIR ROLES IN AGROBACTERIUM-YEAST DNA TRANSFER

Abstract

Yeast spliceosome component BUD31 and its associated NineTeen Complex (NTC) were found to play important role in Agrobacterium-mediated transformation (AMT) in this study. Deletion of yeast BUD31 and SNT309 decreased the efficiency of AMT while deletion of SYF2 increased the transformation efficiency. Subsequent studies of BUD31 showed that the decrease of AMT efficiency of bud31? probably attributes to the decrease in the competency to receive Agrobacterium-delivered T-DNA and virulence protein VirE2. Sequence analysis showed that BUD31 gene is highly conserved across eukaryotes. BUD31 homolog of Nicotiana benthamiana (NbBUD31) was identified and NbBUD31-silencing N. benthamiana was generated by VIGS. Silencing of NbBUD31 in N. benthamiana did not affect tumor formation, transient expression and VirE2 delivery. The different effects of yeast BUD31 and its plant homolog on AMT suggested a different pathway of AMT in natural host plant from the non-natural recipient yeast.