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Xanthomonas euvesicatoria Effector AvrRxo1 Interacts with AIN1 to EnhanceBacterial Growth on Pepper and Tobacco Plants

Zhibo Wang: Virginia Tech

<div>Xanthomonas euvesicatoria Effector AvrRxo1 Interacts with AIN1 to Enhance Bacterial Growth on Pepper and Tobacco Plants Bacterial leaf spot disease caused by Xanthomonas euvesicatoria (Xe) is one of most important pepper diseases. We previously cloned a type III effector (T3E) gene AvrRxo1 from Xanthomonas oryzae pv. oryzicola, which encodes a NAD kinase. The kinase activity is required for the virulence function of AvrRxo1. However, the virulence mechanism of AvrRxo1 is still elusive. AvrRxo1 homologue, Xe-AvrRxo1, was identified from Xe. However, deletion of Xe-AvrRxo1 in Xe strain did not compromise the bacterial virulence on pepper. The virulence function of Xe-AvrRxo1 was masked by other Xe effectors. We developed Xe mutants with deletion of multiple virulence effectors in addition of Xe-AvrRxo1, which were used to inoculate pepper and tobacco plants. The Xe mutants carrying Xe-AvrRxo1 but with the deletion of T3E XopQ or a putative glycerol-transporter (GT) would proliferate better on pepper and tobacco than the isogenic strains without Xe-AvrRxo1. Therefore, Xe- AvrRxo1 indeed possesses virulence function that is redundant with XopQ and GT. To identify host proteins required for the virulence function of Xe-AvrRxo1, we performed a yeast two-hybrid (Y2H) screening by using Xe-AvrRxo1 as bait. Plant protein AIN1 was identified as a putative Xe-AvrRox1 interactor. Silencing or knockout Ain1 by Virus Induced Gene Silence (VIGS) or CRISPR/Cas9 approach compromised the virulence function of Xe-AvrRxo1. Therefore, we confirmed that Xe- AvrRxo1 could interact with AIN1 to enhance bacterial growth on pepper and tobacco plants. The interplays between Xe-AvrRxo1, XopQ, GT and plant protein AIN1 will be discussed in the poster.</div>

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