The Bemisia tabaci Mediterranean (MED) cryptic species has been rapidly invading to most parts of the world owing to its strong ecological adaptability, which is considered as a model insect for stress tolerance studies under rapidly changing environments. Selection of a suitable reference gene for quantitative stress-responsive gene expression analysis based on qRT-PCR is critical for elaborating the molecular mechanisms of thermotolerance. To obtain accurate and reliable normalization data in MED, eight candidate reference genes (β-act, GAPDH, β-tub, EF1-α, GST, 18S, RPL13A and α-tub) were examined under various thermal stresses for varied time periods by using geNorm, NormFinder and BestKeeper algorithms, respectively. Our results revealed that β-tub and EF1-α were the best reference genes across all sample sets. On the other hand, 18S and GADPH showed the least stability for all the samples studied. β-act was proved to be highly stable only in case of short-term thermal stresses. To our knowledge this was the first comprehensive report on validation of reference genes under varying temperature stresses in MED. The study could expedite particular discovery of thermotolerance genes in MED. Further, the present results can form the basis of further research on suitable reference genes in this invasive insect and will facilitate transcript profiling in other invasive insects.

Spotted seal (Phoca largha) is a critically endangered pinniped in China and South Korea. The conventional method to protect and maintain the P. largha population is to keep them captive in artificially controlled environments. However, little is known about the physiological differences between wild and captive P. largha. To generate a preliminary protein expression profile for P. largha, whole blood from wild and captive pups were subjected to a label-free comparative proteomic analysis. According to the results, 972 proteins were identified and predicted to perform functions related to various metabolic, immune, and cellular processes. Among the identified proteins, the expression level of 51 were significantly different between wild and captive P. large pups. These differentially expressed proteins were enriched in a wide range of cellular functions, including cytoskeleton, phagocytosis, proteolysis, the regulation of gene expression, and carbohydrate metabolism. The abundances of proteins involved in phagocytosis and ubiquitin-mediated proteolysis were significantly higher in the whole blood of wild P. largha pups than in captive individuals. In addition, heat shock protein 90-beta, were determined as the key protein associated with the differences in the wild and captive P. largha pups due to the most interactions of it with various differentially expressed proteins. Moreover, wild P. largha pups could be more nutritionally stressed and have more powerful immune capacities than captive pups. This study provides the first data on the protein composition of P. largha and provides useful information on the physiological characteristics for research in this species.

Age is known to affect the gut microbiota in various animals; however, this relationship is poorly understood in seabirds. We investigated the temporal succession of gut microbiota in captive chinstrap penguins of different ages using high‐throughput sequencing. The gut microbiota exhibited a significant age succession pattern, reaching maturity in adults and then declining with increasing age. Only 15 amplicon sequence variants were shared among the gut microbiota in chinstrap penguins at all studied ages, and these contributed to most of the age‐related variations in total gut microbiota. Co‐occurrence networks found that these key bacteria belonged to the genera Acinetobacter, Clostridium sensu stricto, and Fusobacterium, and more species interactions were found within the same taxonomy. Functional prediction indicated that most of the metabolic functions were more abundant in the gut microbiota in adult chinstrap penguins, except for carbohydrate metabolism, which was significantly more abundant in older individuals.

Insects are poikilothermic animals, and temperature is one of the most important abiotic factors affecting their spread and distribution. For example, differences in thermal tolerance may underlie the significant differences in geographical distributions between the native AsiaII3 and invasive MED (Mediterranean) cryptic Bemisia tabaci species in China. Transient receptor potential (TRP) channels are key components of the insect temperature perception system and act as molecular thermometers since they can be activated by specific changes in temperature. In this study, we cloned and characterized the AsiaII3 BtTRP gene and revealed its functions in the response to thermal stress. The full-length cDNA of BtTRP was 3821 bp, with a 3501-bp open reading frame encoding a 132.05-kDa protein. Comparing the deduced amino acid sequences of AsiaII3 BtTRP and MED TRP revealed five amino acid differences. In situ hybridization indicated that BtTRP might be widely expressed throughout the AsiaII3 adult body. BtTRP mRNA expression reached the highest levels after exposure to mild thermal stimuli (12 and 35°C), showing that BtTRP expression can be induced by temperature stress. Furthermore, the thermal tolerance of AsiaII3 after BtTRP dsRNA feeding was significantly lower than that of the control. Taken together, the present study highlights the importance of TRP channels for B. tabaci thermal resistance, and allows us to infer that the differences in amino acids between AsiaII3 and MED might cause the differences in thermal tolerance of these two cryptic species. This study provides a new direction for investigating geographic distribution differences between invasive and native insects.