The Cloning And Expression Analysis Of TaADF-like In Wheat Resistance To Stripe Rust Fungus

Previous studies showed that actin actively participate in the plant defense responses,in a way of rearrangement and remodeling towards the infection site of pathogens,and also indispensable to a multiple of other defense responses.The dynamic change of actin is primarily regulated by a variety of actin-binding proteins.As one of the most important ABPs,little is known about whether actin depolymerizing factor has a role in wheat resistance to stripe rust fungus.Research on this theme will definitely further our understanding in the relationship between actin and plant immune system,and elucidate the molecular mechanism of rust disease resistance in wheat from a new aspect,thus providing a novel strategy in wheat breeding program.In this study a TaADF-like gene was cloned in wheat,using in silicon cloning and RT-PCR verification.Sequence analysis showed that TaADF-like has a complete open reading frame,encoding a protein sequence composed of 138 amino acids.This protein contains a conserved ADF_cofilin_like domain,suggesting that it belongs to the ADF_gelsolin superfamily.The expression of TaADF-like in wheat-stripe rust fungus interaction was analyzed using real-time PCR.The result showed that the expression of TaADF-like in wheat was down regulated in incompatible interaction after the inoculation with stripe rust fungus,.In compatible interaction,the expression of TaADF-like maintained a relatively stable status in the first 24 hours,and then an obvious decrease occurred after 48 hours.There was a significant difference between the expressions of TaADF-like in incompatible interaction and compatible interaction.An approximate 2.5-fold decrease was detected both in incompatible interaction when 24 hours and 48 hours passed by.These results indicated that the down regulation of TaADF-like may contribute to the resistance to stripe rust disease in wheat and the depolymerization of actin may be required in the disease development.Additionally,36 ESTs representing potential ABPs were retrieved from the 4 downloaded databases consisting of numerous ESTs derived from wheat-stripe rust fungus interaction.To explore whether these ESTs were involved in the regulation of wheat resistance to stripe rust fungus,real-time PCR was performed using 8 selected sequences.Two ESTs were screened out,presenting significant differences between the expressions in incompatible interaction and compatible interaction.The GenBank accession numbers of the ESTs are EV253826.1 and GR304651.1,having high homology with myosin-Va-like protein and K+efflux antiporter 2,respectively,which will provide useful material for the oriented gene cloning of actin-binding proteins.