Targeted Inactivation Of The Ago1 Homeologues Of Nicotiana Benthamiana Reveals Their Distinct Roles In Development And Antiviral Defence

The Solanaceae family includes numerous highly valuable crops. Understanding the viral diseases that affect them is of great importance. Nicotiana benthamiana has contributed greatly to unravelling antiviral RNA interference, and can also be regarded as an adequate model for studying viral diseases of solanaceous crops. This species, however, as with many of its relatives, possesses an allopolyploid genome, in which homeologous gene pairs frequently occur.AGO1 is a pivotal component of most plant RNA silencing pathways. The Nicotiana benthamiana genome encodes two highly similar AGO1 homeologues: AGO1A and AGO1B. To understand their roles in planta, their genes were selectively inactivated. Given the inherent limitations of RNA interference-based techniques, we used genome editing to achieve this goal.We found that AGO1A was not required for normal development, while AGO1B was indispensable for that. By contrast, the two homeologues both contributed to antiviral defence. Additionally, we observed that AGO1B utilised miR168 poorly, which may help to retain a significant level of antiviral RNA interference during viral infection.Our results have important implications for the better understanding of viral diseases of economically important solanaceous crops.