Tissue-specific alternative splicing and the functional differentiation of LmLPMO15-1 in Locusta migratoria.

Abstract

Insect lytic polysaccharide monooxygenases (LPMO15s) are newly discovered copper-dependent enzymes that promote chitin degradation in insect through oxidative cleavage of glycosidic bonds. They are potential pesticide targets due to their critical role for chitin turnover in the integument, trachea, and peritrophic matrix of the midgut during insect molting. However, the knowledge about whether and how LPMO15s participate in chitin turnover in other tissues is still insufficient. Here, using the orthopteran pest Locusta migratoria as a model, a novel alternative splicing site of LmLPMO15-1 was discovered and it produces 2 variants, LmLPMO15-1a and LmLPMO15-1b. The transcripts of LmLPMO15-1a and LmLPMO15-1b were specifically expressed in the trachea and foregut, respectively. RNA interference targeting LmLPMO15-1 (a common fragment shared by both LmLPMO15-1a and LmLPMO15-1b), a specific region of LmLPMO15-1a or LmLPMO15-1b all significantly reduced survival rate of nymphs and induced lethal phenotypes with developmental stasis or molt failure. Ultrastructure analysis demonstrated that LmLPMO15-1b was specifically involved in foregut old cuticle degradation, while LmLPMO15-1a was exclusively responsible for the degradation of the tracheal old cuticle. This study revealed LmLPMO15-1 achieved tissue-specific functional differentiation through alternative splicing, and proved the significance of the spliced variants during insect growth and development. It provides new strategies for pest control targeting LPMO15-1.