Identification and Characterization Analysis of Transient Receptor Potential Mucolipin Protein of Laodelphax striatellus Fallén.

Abstract

Simple SummaryThe small brown planthopper Laodelphax striatellus is a destructive pest of rice, maize and wheat crops in Asia, causing damage by directly sucking phloem sap and transmitting plant viruses, and thus seriously impacting crops yields. Transient receptor potential mucolipin (TRPML) protein plays a vital role in Ca2+ ions release, resulting in membrane trafficking, autophagy and ion homeostasis; however, till now, we have learned little about the TRPML protein of agricultural pests. In this study, we first identified the TRPML of L. striatellus. We analyzed not only the gene evolutionary features and expression profiles but also clarified the protein subcellular localization and lipid-binding characteristics of Ls-TRPML. We found that TRPML is evolutionarily conserved among agricultural pests. Ls-TRPML is predominately expressed in L. striatellus ovary. Moreover, we found that Ls-TRPML localizes in the nuclear membrane in Spodoptera frugiperda cells and the intestine and ovary of L. striatellus. The binding of Ls-TRPML with lipids was detected by lipid-binding assay, indicating the potential role of Ls-TRPML in lipid interaction. Thus, our findings first helped us analyze the gene characterization of Ls-TRPML and then identify the binding of Ls-TRPML with lipids; our findings will broaden our understanding of TRPML’s roles in agricultural pests.Transient receptor potential mucolipin (TRPML) protein in flies plays a pivotal role in Ca2+ ions release, resulting in membrane trafficking, autophagy and ion homeostasis. However, to date, the characterization of TRPML in agricultural pests remains unknown. Here, we firstly reported the TRPML of a destructive pest of gramineous crops, Laodelphax striatellus. The L. striatellus TRPML (Ls-TRPML) has a 1818 bp open reading frame, encoding 605 amino acid. TRPML in agricultural pests is evolutionarily conserved, and the expression of Ls-TRPML is predominately higher in the ovary than in other organs of L. striatellus at the transcript and protein level. The Bac–Bac system showed that Ls-TRPML localized in the plasma membrane, nuclear membrane and nucleus and co-localized with lysosome in Spodoptera frugiperda cells. The immunofluorescence microscopy analysis showed that Ls-TRPML localized in the cytoplasm and around the nuclei of the intestine cells or ovary follicular cells of L. striatellus. The results from the lipid-binding assay revealed that Ls-TRPML strongly bound to phosphatidylinositol-3,5-bisphosphate, as compared with other phosphoinositides. Overall, our results helped is identify and characterize the TRPML protein of L. striatellus, shedding light on the function of TRPML in multiple cellular processes in agricultural pests.