Construction of novel recombinant receptor proteins to improve disease resistance in wheat

[摘要] English: The aim of the current study was to construct novel recombinant RLK genes encodingchimeric receptor proteins to broaden disease resistance in wheat. Eight different RLKs wereinitially chosen as extracellular domain donors based on their ability to bind PAMPs/MAMPsor DAMPs. The kinase domain donor was LRK I0, an R protein that renders wheat resistantagainst Puccinia triticina.These recombinant genes were first designed in silico using a bio-informatic approach wherethe gene portions encoding the different extracellular domains were linked to the LRKIOkinase domain encoding gene fragment. Translation of the different recombinant genesconfirmed that full length polypeptides will be formed. General PAMP binding capacity wastherefore combined with the highly specific defence signalling ability of LRKIO within asingle protein.Due to several problems encountered, only four recombinant genes were successfullyproduced, namely BRll-LRKlO, CERK-LRKIO, LecRKI-LRKIO and PERKI-LRKIO. Toconfirm the success of the approach, LecRKI-RLKIO was successfully transcribed andtranslated in vitro, thus producing the correct sized polypeptide. Future work will include theproduction of transgenie wheat to test the ability of these chimeric proteins to increase wheatresistance against pathogen infection.

[发布机构] University of the Free State