Abstract
Periplocoside P (PSP), a novel compound isolated from Periploca sepium Bunge, possesses insecticidal activity against some lepidopterans, such as Mythimna separata. In M. separata, the brush border membrane vesicles of the midgut epithelium are the initial site of action of periplocosides. We conducted two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time of flight/time of flight mass spectrometry analysis to analyze differentially expressed proteins (DEPs) from periplocoside P (PSP)-treated M. separata. We successfully isolated seven up-regulated and three down-regulated DEPs that have been previously identified, as well as a novel DEP. The DEPs are implicated in protein degradation, transporter, folding, and synthesis, and in juvenile hormone biosynthesis. DEPs involved in the oxidative phosphorylation energy metabolism pathway are enriched. Through real-time polymerase chain reaction assay, we confirmed that vma1 expression is significantly up-regulated expression levels in PSP-treated M. separata larvae. Enzymology validation further indicated that PSP can significantly inhibit V-type ATPase activity in a concentration-dependent manner. Given these results, we speculate that in M. separata, the V-type ATPase A subunit in the midgut epithelium is the putative target binding site of periplocosides. This finding provides preliminary evidence for the mode of action of periplocosides.
Highlights
Target-oriented design has become the modern mainstream model for the development of novel pesticides [1]
Periplocoside P (PSP), a novel compound isolated from P. sepium has high insecticidal activity (LC50 = 110 mg·L−1 ) against Mythimna separata [13]
Aminopeptidase N (APN) and alkaline phosphatase phosphatase (ALP) are usually used as marker enzymes in brush border membrane vesicles (BBMVs) from insect midguts [28]
Summary
Target-oriented design has become the modern mainstream model for the development of novel pesticides [1]. Periplocoside P (PSP), a novel compound isolated from P. sepium has high insecticidal activity (LC50 = 110 mg·L−1 ) against Mythimna separata [13]. Isolated from P. sepium has high insecticidal activity (LC50 = 110 mg·L−1) against Mythimna separata [13]. For the resistance of the diamondback moth (Plutella xylostella) against Cry1Ac [17]. Cry1Ac toxin targets may in the bollworm (Helicoverpa armigera) midgut [18]. The. Cry1Ac toxin targets may exist in exist the bollworm (Helicoverpa armigera) midgut [18]. In we utilized utilized two-dimensional two-dimensional gel gel electrophoresis electrophoresis (2-DE) In this this study, study, we laser desorption/ionization time of flight/time of flight mass spectrometry Desorption/ionization time of flight/time of flight mass spectrometry (MALDI-TOF/TOF MS) to to investigate theputative putativetarget target proteins of PSP in midgut the midgut.
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