Cloning of three epsilon-class glutathione S-transferase genes from Micromelalopha troglodyta (Graeser) (Lepidoptera: Notodontidae) and their response to tannic acid.

Micromelalopha troglodyta (Graeser) is an important pest of poplar in China, and glutathione S-transferase (GST) is an important detoxifying enzyme in M. troglodyta. In this paper, three full-length GST genes from M. troglodyta were cloned and identified. These GST genes all belonged to the epsilon class (MtGSTe1, MtGSTe2, and MtGSTe3). Furthermore, the expression of these three MtGSTe genes in different tissues, including midguts and fat bodies, and the MtGSTe expression in association with different concentrations of tannic acid, including 0.001, 0.01, 0.1, 1, and 10 mg ml-1, were analysed in detail. The results showed that the expression levels of MtGSTe1, MtGSTe2, and MtGSTe3 were all the highest in the fourth instar larvae; the expression levels of MtGSTe1 and MtGSTe3 were the highest in fat bodies, while the expression level of MtGSTe2 was the highest in midguts. Furthermore, the expression of MtGSTe mRNA was induced by tannic acid in M. troglodyta. These studies were helpful to clarify the interaction between plant secondary substances and herbivorous insects at a deep level and provided a theoretical foundation for controlling M. troglodyta.

Cap 'n' Collar C and Aryl Hydrocarbon Receptor Nuclear Translocator Facilitate the Expression of Glutathione S-Transferases Conferring Adaptation to Tannic Acid and Quercetin in Micromelalopha troglodyta (Graeser) (Lepidoptera: Notodontidae).

Micromelalopha troglodyta (Graeser) (Lepidoptera: Notodontidae) is a notorious pest of poplar. Coevolution with poplars rich in plant secondary metabolites prompts M. troglodyta to expand effective detoxification mechanisms against toxic plant secondary metabolites. Although glutathione S-transferases (GSTs) play an important role in xenobiotic detoxification in M. troglodyta, it is unclear how GSTs act in response to toxic secondary metabolites in poplar. In this study, five GST gene core promoters were accurately identified by a 5' loss luciferase reporter assay, and the core promoters were significantly induced by two plant secondary metabolites in vitro. Two transcription factors, cap 'n' collar C (CncC) and aryl hydrocarbon receptor nuclear translocator (ARNT), were cloned in M. troglodyta. MtCncC and MtARNT clustered well with other insect CncCs and ARNTs, respectively. In addition, MtCncC and MtARNT could bind the MtGSTt1 promoter and strongly improve transcriptional activity, respectively. However, MtCncC and MtARNT had no regulatory function on the MtGSTz1 promoter. Our findings revealed the molecular mechanisms of the transcription factors MtCncC and MtARNT in regulating the GST genes of M. troglodyta. These results provide useful information for the control of M. troglodyta.