Identification and Expression Profiling of the 5-HT Receptor Gene in Harmonia axyridis.

Abstract

It has been found that 5-hydroxytryptamine (5-HT) modulates the feeding of some insects, and this phenomenon was found in Harmonia axyridis (Pallas) by our previous study. An understanding of the 5-HT system in this beetle is helpful for utilizing 5-HT to modulate its predation to improve biological control efficiency, especially in greenhouses in winter in north China. This is because 5-HT influences diapause in insects by modulating the synthesis and release of prothoracic hormone (PTTH) and, therefore, influences feeding. To elucidate the molecular basis of the H. axyridis 5-HT system, reverse-transcription polymerase chain reaction (RT-PCR), multiple sequence alignment, and phylogenetic tree construction were used to identify the 5-HT receptor in H. axyridis, and quantitative real-time PCR (qRT-PCR) was used to analyze the expression pattern of these receptor genes in different developmental stages and in the nervous system (brain + ventral nerve cord), digestive tract, pectoral muscles, and gonads of the adult ladybird. The results showed that four 5-HT receptors were identified in H. axyridis, named 5-HT1AHar, 5-HT1BHar, 5-HT2Har, and 5-HT7Har. The four receptors were expressed at high levels in the adult stage, especially in 2-day-old adults, with expression levels of 18.72-fold (male) and 14.21-fold (female) of that in eggs for 5-HT1A, 32.27-fold (male) and 83.58-fold (female) of that in eggs for 5-HT1B, 36.82-fold (male) and 119.35-fold (female) of that in eggs for 5-HT2, and 165.47-fold (male) and 115.59-fold (female) of that in eggs for 5-HT7. The level of expression decreased with the advance of day-age in adults. The levels of expression of 5-HT1BHar, 5-HT2Har, and 5-HT7Har were low at the egg, larval, and pupal stages, and 5-HT1AHar was not expressed in the larval stage. The four receptors were expressed in the nervous system, digestive tract, pectoral muscles, and male and female gonads. The 5-HT1AHar was expressed at a high level in the pectoral muscle (6.75-fold of that in the nervous system), 5-HT1BHar in male gonads (1.02-fold of that in the nervous system) and the nervous system, 5-HT2Har in male gonads (5.74-fold of that in the nervous system), and 5-HT7Har in the digestive tract (1.81-fold of that in the nervous system). The results of this study will lay a foundation for research on the function of the 5-HT receptor by RNA interference in the regulation of predation by H. axyridis.