The CsMYB44-csi-miR0008-CsCER1 module regulates cuticular wax biosynthesis and drought tolerance in citrus

Abstract

• Cuticular wax covering aboveground organs serves as the first line of defense shielding plants from nonstomatal water loss and diverse environmental stresses. While there have been several wax-related genes identified, the molecular mechanisms responsible for the control of wax biosynthesis remain poorly understood in citrus, particularly at the posttranscriptional level.
• Here, we demonstrated that the CsMYB44-csi-miR0008-CsCER1 module is responsible for regulating drought tolerance in citrus through its control of cuticular wax biosynthesis.
• In this study, microRNA (miRNA) sequencing analyses of ‘Newhall’ navel oranges and the wax-deficient ‘Ganqi 3’ mutant variety led to the identification of a novel cuticular wax biosynthesis-related miRNA, csi-miR0008. csi-miR0008 suppresses the expression of CsCER1, an aldehyde decarbonylase-encoding gene associated with n-alkane biosynthesis. The leaves of csi-miR0008-silencing and CsCER1-overexpressing plants exhibited increases in total wax levels, with particularly pronounced increases in n-alkane levels, contributing to enhanced drought tolerance. csi-miR0008-overexpressing and CsCER1-silencing plants exhibited the opposite phenotypes. CsMYB44 was confirmed to promote wax accumulation by directly inhibiting the expression of csi-miR0008.
• Taken together, our study offers new insight into the mechanisms responsible for the posttranscriptional control of citrus cuticular wax biosynthesis, while also providing a foundation for the breeding of novel citrus varieties exhibiting enhanced drought tolerance.