Genetic diversity of an effector gene, AvrPi9, of rice blast pathogen in Thailand and characterization of its promoter

Rice blast is one of the most destructive diseases of rice and is caused by the fungus Magnaporthe oryzae. The disease causes enormous yield losses in rice production worldwide. The rice blast fungus delivers effector proteins into rice cells. The effector proteins play an essential role in fungal virulence by manipulating and controlling host cellular pathways and inhibiting host immune responses to enhance pathogenicity. An effector gene, AvrPi9, which corresponds to the resistance gene Pi9, was cloned and characterized. However, a regulatory molecular mechanism for AvrPi9 gene expression has not been determined. In this study, the genetic variation of the AvrPi9 and its promoter function were characterized. The results showed that 98% (116/118) of the samples carried the AvrPi9 gene without any sequence variation, whilst two isolates, 10576 from Kalasin and NYK56003 from Nakhon Nayok, lacked the AvrPi9 gene. A homeobox domain-containing protein (MoHOX6) was identified as a candidate transcription factor. The AvrPi9 gene expression was delayed in the MoHOX6 knockout mutant. Moreover, the AvrPi9 promoter was able to drive the expression of a luciferase gene in rice protoplasts. This study provides the first insight into the function and regulation of the AvrPi9 promoter of rice blast fungus.