Full Length CDNA Isolation And Vector Construction Of CaNACs Of Its Genetic Transformation

Pepper(Capsicum annuum)is important but diseases susceptible solanaceae vegetable worldwide, and functional genomics of Capsicum annuum is the basal work to find and utilize disease resistance gene for the genetic improvement of its disease resistance. In this study, we isolated two full length cDNAs of CaNACs from a cDNA library of Capsicum annuum which may involved in stresses adaption. The corresponding overexpression vectors were constructed and transformed into tobacco via a agrobacterium mediated method. The main results are as follows:1) 2 pairs of specific primers were designed according to the ESTs primed by a NAC protein of Arabidopsis. With the specific primers, 2 candidated cDNAs were isolated from a cDNA library by a 96 well based PCR cDNA screening method. Their shared a high homology in sequence with NAC proteins from other plants, containing a conserved NAC functional domain. These two NACs may belong toα-NAC molecular chaperone family and named CaNAC1 and CaNAC2 respectively.2) To get the overexpression as well as RNAi transgenic plants for functional genomics study, the overexpression and RNAi vectors of the 2 candidate cDNAs were constructed via a gateway cloning technology employed gateway distinction vectors. With the vectors,transformation was carried out via a agrobacterium mediated method, and 50 and 30 T0 overexpression transgenic tobacco plants of CaNAC1 and CaNAC2 were acquired respectively.The results of this study laid a important foundation for the further functional analysis of CaNAC1 and CaNAC2.