Molecular characterization of prophenoloxidase-1 (PPO1) and the inhibitory effect of kojic acid on phenoloxidase (PO) activity and on the development ofZeugodacus tau(Walker) (Diptera: Tephritidae)

Abstract

Phenoloxidase (PO) plays a key role in melanin biosynthesis during insect development. Here, we isolated the 2310-bp full-length cDNA of PPO1 from Zeugodacus tau, a destructive horticultural pest. qRT-polymerase chain reaction showed that the ZtPPO1 transcripts were highly expressed during larval-prepupal transition and in the haemolymph. When the larvae were fed a 1.66% kojic acid (KA)-containing diet, the levels of the ZtPPO1 transcripts significantly increased by 2.79- and 3.39-fold in the whole larvae and cuticles, respectively, while the corresponding PO activity was significantly reduced; in addition, the larval and pupal durations were significantly prolonged; pupal weights were lowered; and abnormal phenotypes were observed. An in vitro inhibition experiment indicated that KA was an effective competitive inhibitor of PO in Z. tau. Additionally, the functional analysis showed that 20E could significantly up-regulate the expression of ZtPPO1, induce lower pupal weight, and advance pupation. Knockdown of the ZtPPO1 gene by RNAi significantly decreased mRNA levels after 24 h and led to low pupation rates and incomplete pupae with abnormal phenotypes during the larval-pupal interim period. These results proved that PO is important for the normal growth of Z. tau and that KA can disrupt the development of this pest insect.

Chemical repertoire and biosynthetic machinery of the Aspergillus flavus secondary metabolome: A review

“…They have been implicated in several processes of insect physiology such as melanization, cuticular hardening, pathogen encapsulation, wound healing, and overall immunity (Binggeli, Neyen, Poidevin, & Lemaitre, 2014). Several studies have demonstrated that kojic acid‐based inhibition of PPOs is associated with a significant decrease in insect fitness and development (Alverson, 2003; Bai et al., 2014; Zhang, Luo, et al., 2019). Because PPOs play key roles in the defensive system of insects, they are an attractive target to implement insect control strategies.…”

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“…Enzymes and genes associated to pigmentations of insects were searched and identificated based on studies reported. There were 9 families of enzymes or genes as follow: Dopachrome converting enzyme family DEC(yellow) Futahashi et al, 2008;Ferguson et al, 2011;Futahashi et al, 2012), N-acetyltransferase(AANAT) (Hintermann et al, 1996;Tsugehara et al, 2007;Zhang et al, 2018b), Phenoloxidase(laccase2) (Futahashi et al, 2010;Yatsu et al, 2009;Futahashi et al, 2010), NBAD synthetase (ebony) (Hovemann et al, 1998;Richardt et al, 2002;Wittkopp et al, 2002;Futahashi et al, 2008;Pool et al, 2010;Takahashi et al, 2011;Ferguson et al, 2011;Zhang et al, 2018b), NBAD hydrolase (tan) (Futahashi et al, 2010;Ferguson et al, 2011;Du et al, 2018), Phenoloxidase(PO) (Zhang et al, 1997;Luna et al, 2011;Sachdev et al, 2017;Zhang et al, 2018a), Aspartate decarboxylase (ADC) (Arakane et al, 2009;Dai et al, 2015), DOPA decarboxylase (DDC\AADC) (Fang et al, 2000;Hawi et al, 2001;Chen et al, 2002;Ma et al, 2005;Yu et al, 2006;Giegling et al, 2008;Paskewitz et al, 2008;Bertoldi et al, 2009;Ferguson et al, 2011;Geomela et al, 2012), Phenylalanine hydroxylase(PAH) (Kwok et al, 1985;Johnson et al, 2003;Infanger et al, 2004;…”

“…The formation of melanin was related to contents of substances including phenylalanine, tyrosine, DOPA, N-acetyl-dopamine (NADA), N-beta-alanyl-dopamine (NBAD) Sekine et al, 2011) and so on. These substances were related to some enzymes including phenylalanine hydroxylase (PAH) (Kwok et al, 1985;Zhang, 2004;Infanger et al, 2004), TH (Futahashi et al, 2008), DDC(Fang et al, 2000;Chen et al, 2002;Paskewitz et al, 2008;Giegling et al, 2008;Bertoldi et al, 2009;Ferguson et al, 2011;Hodges et al, 2012), PO (Sachdev et al, 2017;Zhang et al, 2018a), Phenoloxidase(laccase2) (Yatsu et al, 2009;Futahashi et al, 2010), NBAD synthetase (ebony) (Hovemann et al, 1998;Wittkopp et al, 2002;Takahashi et al, 2008;Takahashi et al, 2011;Ferguson et al, 2011), NBAD hydrolase (tan) (Futahashi et al, 2010;Wagner et al, 2010;Du et al, 2018), dopachrome conversion enzyme family (yellow) Futahashi et al, 2008;Ferguson et al, 2011;Futahashi et al, 2012), aspartate decarboxylase (ADC) (Arakane et al, 2009;Dai et al, 2015) and arylalkylamine-N-acetyltransferase (AANAT) (Hintermann et al, 1996;Tsugehara et al, 2007;Zhan et al, 2010;Zhang et al, 2018b). These enzymes and genes were mostly studied on the growing developments of insects (Bai et al, 2014;Sachdev et al, 2017;Zhang et al, 2018b).…”

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“…Enzymes and genes associated to pigmentations of insects were searched and identificated based on studies reported. There were 9 families of enzymes or genes as follow: Dopachrome converting enzyme family DEC(yellow) Futahashi et al, 2008;Ferguson et al, 2011;Futahashi et al, 2012), N-acetyltransferase(AANAT) (Hintermann et al, 1996;Tsugehara et al, 2007;Zhang et al, 2018b), Phenoloxidase(laccase2) (Futahashi et al, 2010;Yatsu et al, 2009;Futahashi et al, 2010), NBAD synthetase (ebony) (Hovemann et al, 1998;Richardt et al, 2002;Wittkopp et al, 2002;Futahashi et al, 2008;Pool et al, 2010;Takahashi et al, 2011;Ferguson et al, 2011;Zhang et al, 2018b), NBAD hydrolase (tan) (Futahashi et al, 2010;Ferguson et al, 2011;Du et al, 2018), Phenoloxidase(PO) (Zhang et al, 1997;Luna et al, 2011;Sachdev et al, 2017;Zhang et al, 2018a), Aspartate decarboxylase (ADC) (Arakane et al, 2009;Dai et al, 2015), DOPA decarboxylase (DDC\AADC) (Fang et al, 2000;Hawi et al, 2001;Chen et al, 2002;Ma et al, 2005;Yu et al, 2006;Giegling et al, 2008;Paskewitz et al, 2008;Bertoldi et al, 2009;Ferguson et al, 2011;Geomela et al, 2012), Phenylalanine hydroxylase(PAH) (Kwok et al, 1985;Johnson et al, 2003;Infanger et al, 2004;Zhang et al, 2004;Wang et al, 2013)…”