MtNOOT HETEROLOGOUS EXPRESSION WITH FaWRKY1 OVER- EXPRESSION CONFER STRWBERRY RESISTANCE
Abstract
Strawberry (Fragaria x ananassa) is one of the favorite fruit worldwide due to its wide health benefits, and distinct flavor and aroma. FaWRKY1 gene was suggested as a significant element intermediate defense response against various pathogens attack in strawberry. Because of the influencing role of WRKY family involving in defense network, it has become a favorable candidate for improving crops quality. WRKY can precisely recognition and binding to the down-stream promoters of transcription factors activating defense cascades. The ability of FaWRKY to enhance resistance against Macrophomina phasiolena was investigated by performing Agrobacterium-mediated transformation protocol for transient overexpression of FaWRKY1 gene in strawberry leaves (Fragaria x ananassa cv. Camarosa) to evaluate its function upon the fungal infection. However, MtNOOT gene were transiently hetero-expressed in strawberry leaves separately and in accomplishment with the FaWRKY1. We demonstrated that the existing of W-box sequences within the MtNOOT (NPR1-like gene) promoter region, which are recognized definitely by SA-induced FaWRKY DNA binding protein, increased strawberry resistance activity when the two genes are transformed in combined to each other. The severity of leaf injury was observed at three, five, and seven days post pathogen inoculation on FaWRKY1, MtNOOT, and FaWRKY1 & MtNOOT combination-transformed strawberry plants compared to the control untransformed infected plants as positive control and a healthy untransformed non-infected plants as negative control. Susceptibility to fungal infection was obviously detected and showing that the two genes combination (FaWRKY & MtNOOT) revealed the best resistance against the pathogen fungal attack followed by MtNOOT and finally by the FaWRKY1 transcription factor. Our results evidence that FaWRKY1 gene acts upstream of the heterologous MtNOOT (NPR1-like gene) and positively regulates its expression throughout plant defense activation during pathogen attack.
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