Experimental Verification And Bioinformatic Analysis Of The Key Bacterial Wilt Resistance Genes From Solanum Tuberosum

All over the world, potato (Solanum tuberosum) is an importantfood crop in the agricultural production, and it plays a significantrole in the structure of food safety. Bacterial wilt caused byRalstonia solanacearum is one of the most widely spread and verydestructive plant disease, causing enormous economic losses.Under the background of the human genome project (HGP),Bioinformatics is appeared. Its main function is the study of DNAand protein sequences, the genetic information and evolutioninformation for the analysis and prediction. Therefore, it is veryimportant to accelerate the disease control and disease-resistantbreeding, and for elucidating the molecular mechanism of potatodefense the bacterial wilt. Bioinormatical analysis of candidategenes could accelerate mining and using the resistance genes forpotato breeding.This study selected5kinds of ESTs which showed up-expressedin the EST librarys builded with from tthe laboratory of suppressionsubtractive hybridization (SSH) technology to build after thepathogen induced differentially expressed in the EST library, brushselected green blight fungus induced five key defense related genes, respectively isin our lab. These genes included: yippeeyippee-likeprotein gene, proteinase proteinase inhibitor inhibitor gene, WRKYtranscription factor gene, remorin gene, and four metallothioneingenes. Serveral bioinformatical analysis were carried for theirsnucleotide sequences, promoters, and amino acids, predictedsecondary structures and, the tertiary structures and their possiblefunctions in potato and the pathogen interactionsof conventionalanalysis and forecast.For the induced expression patterns of these genes, RT-PCR andqRT-PCR test were used. The results showed that the five genes hadsignificantly increased, and had expressed at the early stage ofdisease. These genes blong defence related genes categories.Potato stem and leaf tissues at the stage of7-8leaves wereused for digoxin in situ hybridization experiments to explore theproteinase inhibiting gene expression profile of organizationlocalization. The results showed that their site mainly expressed inthe stem and leaf of phloem. It showed that the proteinaseinhibiting gene could play its functions in vascular system againstthe bacterial wilt caused by R. solanacearum.These results could provide theoretical basis indisease-resistance breeding of potato.