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The bumblebee (Bombus lantschouensis Vogt) is an important pollinator of wild plants. Sphingomyelin phosphodiesterase (SMPD) is a hydrolase that plays a major role in sphingolipid metabolism reactions. We report the preparation and characterization of a polyclonal antibody for bumblebee SMPD. We then use the polyclonal antiserum to detect the SMPD protein at different development stages and in different tissues. Our results showed that a 1228bp fragment homologous with the B. terrestris SMPD gene was successfully amplified. The molecular weight of the fusion protein was about 70 kDa by SDS-PAGE. An effective polyclonal antibody against SMPD was also obtained from mice and found to have a higher specificity for bumblebee SMPD. Western blotting detection showed that SMPD was expressed at a high level in queen ovaries, although expression was lower in the midgut and venom gland. SMPD expression decreased from the egg stage until the pdd stage. We interpret our results as showing that the development of an effective polyclonal antiserum for the SMPD protein of a bumblebee, which provides a tool for exploring the function of the SMPD gene. In addition, the work has confirmed that SMPD should be considered as an important enzyme during bumblebee egg and larval stages.

Trehalose provides the main energy source for the physiological activities of insects, especially in adverse conditions. Trehalase is the only enzyme that hydrolyzes trehalose, therefore it is important to clarify the distribution and expression of trehalase under adverse conditions such as high temperatures and starvation. Here, we have cloned the trehalase genes and investigated their expression in different tissues, at multiple development stages, and with the treatments of high temperature and starvation in Bombus lantschouensis, which is considered to be one of the most commercially viable native species in China. The results suggest that the membrane-bound (BlTre-2) cDNA has an open reading frame of 1986 nucleotides, which encodes a protein of 662 amino acids, and two putative transmembrane domains. qRT-PCR analysis indicated that BlTre-2 was expressed in 10 tissues and at nine development stages, with the highest expression in general in 30-day-old workers, and in ovarian tissue in particular. The expression of BlTre-1 for 15-day-old workers which were exposed to a pre-treatment of 45°C increased over the first 5 h and subsequently decreased over time. In contrast the expression of BlTre-2 consistently decreased over time. The highest expression levels of BlTre-1 and BlTre-2 were observed the newly emerged adult workers when starved for 16-20 h. These results indicate that BlTre-2 may be part of the carbohydrate metabolism of the bumblebee, and that BlTre-1 is a key gene regulating energy metabolism and providing trehalose when exposed to a high temperature. Both BlTre-1 and BlTre-2 may balance trehalose and provide energy when B. lantschouensis is starved.