RESUMO
Lytic polysaccharide monooxygenases (LPMOs) are important enzymes that boost the hydrolysis of recalcitrant polysaccharides, such as chitin. They are found extensively in different insect species and are classified as auxiliary activities family 15 (AA15) LPMOs (LPMO15). Some of them were identified from the insect midgut and proven to act on chitin. However, knowledge about their physiological roles during insect growth and development remains limited. Here, we found that midgut-specific LPMO15s are widely distributed in different insect orders, such as the orthopteran Locusta migratoria and the lepidopteran Bombyx mori. Using L. migratoria as a model insect, the function of midgut-specific LmLPMO15-3 during development was investigated. Double-stranded RNA-mediated downregulation of LmLPMO15-3 expression at the 4th or 5th instar nymph stage severely decreased the survival rate and resulted in lethal phenotypes. Hematoxylin and eosin staining results indicated that the deficient individuals exhibited incompletely digested peritrophic matrix (PM), which suggested that LmLPMO15-3 is essential for the deconstruction of the PM during molting. This study provides direct evidence of the physiological importance of a midgut-specific LPMO15 during insect development. As L. migratoria is one of the most destructive agricultural pests, LmLPMO15-3 is a potential target for pest management.
Assuntos
Locusta migratoria , Animais , Quitina/metabolismo , Amarelo de Eosina-(YS)/metabolismo , Hematoxilina/metabolismo , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Locusta migratoria/metabolismo , Oxigenases de Função Mista/metabolismo , RNA de Cadeia Dupla/metabolismoRESUMO
A novel fluorescent probe, phenolphthaleindialdehyde(2pyridyl) hydrazone (L), for sequentially detecting Al3+ and F- in almost 100% aqueous medium was successfully designed and synthesized. The probe offers two binding pockets for Al3+ to form a 1: 2 ligand/metal complex, leading to a significant fluorescence enhancement at 465â¯nm. Further, the in-situ formed L-Al complex acts as a secondary fluorescent chemosensor for F- by quenching the fluorescence of the complex with high selectivity. The detection limit for Al3+ and F- sensing is 2.28â¯nM and 0.13⯵M, respectively, which are far below the World Health Organization (WHO) acceptable limits (7.41⯵M for Al3+ ion and 79⯵M for F-) in drinking water. The probe L was successfully applied to the detection of Al3+ and F- in cells using fluorescence microscopy.