Proteômica diferencial em plantas transgênicas de Nicotiana benthamiana e testes da resistência em Solanum tuberosum induzida pelo mesmo gene contra Pectobacterium atrosepticum

Abstract

The DomB synthetic gene isolated from potato (Solanum tuberosum) apyrase fraction has demonstrated anti-gene activity in animals and resistance of tobacco (Nicotiana benthamiana) plants against Pectobacterium atrosepticum. In this work we propose differential proteic profile analysis in Nicotiana benthamiana plants genetically modified with g2S26His-DomB synthetic gene by nanoUPLC-MSE tool. We compared the R0 hemizigotic progenie protein content of transgenic N. benthamiana with native N. benthamiana protein content, and these groups challenged with P. atrosepticum. Moreover, we proposed the evaluation of this gene activity in S. tuberosum by agroinfiltration. Trypsinized proteins initial analyses suggests that defense sinalization proteins were produced by N. benthamiana. The preliminary analysis of Solanum tuberosum cv. Asterix commercial cultivar of transiently expressing the same quimeric gene by agro-infiltration method using Agrobacterium 3101 with pk7WG2D-gfp-g2S2-6His-DomB plasmid, compared to another group where exist co-infiltration with Agrabacterium EHA101 containing TCV-CP, that works like a suppressor of silencing of post- transcriptional gene, showed promising results. After the fifth day of agro-infiltration, 5 - 8weeks S. tuberosum leaves were, tested with phytopathogenic bacterium and observed for seven days. By the end of this period, a hypersensitivity reaction was observed at application site which did not expanded for the whole leaf tissue. The dot-blot immunoassay suggest that the expression of p2S26His-DomB polypeptide, in infiltrated leaf wih plasmid and TCV-CP. To get resistant cultivars against P. atrosepticum infection, this work can add information about defensive mechanism of transgenic plants with g2S2-6His-DomB gene and get a better production of transgenic potato plants resistant to phytopathogenic with DomB synthetic gene.