For molecular research, the quality and integrity of DNA obtained will affect the reliability of subsequent results. Extracting quality DNA from scale insects, including mealybugs, can be difficult due to their small body size and waxy coating. In this study, we evaluate eight commonly used DNA extraction methods to determine their efficacy in PCR analysis across life stages and preservation times. We find that fresh samples, immediately upon collection or after 2 wks, resulted in the most effective DNA extraction. Methods using the DNeasy Blood & Tissue kit, NaCl, SDS-RNase A, and SDS isolated DNA of sufficient quality DNA. The SDS method gave high DNA yield, while the NaCl and SDS-RNase A methods gave lower yield. NaCl, SDS-RNase A, SDS, chloroform-isopentyl alcohol, and the salting-out methods all resulted in sufficient DNA for PCR, and performed equal to or better than that of the DNeasy Blood & Tissue kit. When time and cost per extraction were considered, the SDS method was most efficient, especially for later life stages of mealybug, regardless of preservation duration. DNA extracted from a single fresh sample of a female adult mealybug was adequate for more than 10,000 PCR reactions. For earlier stages, including the egg and 1st instar nymph samples, DNA was most effectively extracted by the Rapid method. Our results provide guidelines for the choice of effective DNA extraction method for mealybug or other small insects across different life stages and preservation status.

Helicoverpa zea, a well-documented and endemic pest throughout most of the Americas, affecting more than 100 species of host plants. It is a quarantine pest according to the Asia and Pacific Plant Protection Commission (APPPC) and the catalog of quarantine pests for plants imported to the People’s Republic of China. Based on 1781 global distribution records of H. zea and eight bioclimatic variables, the potential geographical distributions (PGDs) of H. zea were predicted by using a calibrated MaxEnt model. The contribution rate of bioclimatic variables and the jackknife method were integrated to assess the significant variables governing the PGDs. The response curves of bioclimatic variables were quantitatively determined to predict the PGDs of H. zea under climate change. The results showed that: (1) four out of the eight variables contributed the most to the model performance, namely, mean diurnal range (bio2), precipitation seasonality (bio15), precipitation of the driest quarter (bio17) and precipitation of the warmest quarter (bio18); (2) PGDs of H. zea under the current climate covered 418.15 × 10⁴ km², and were large in China; and (3) future climate change will facilitate the expansion of PGDs for H. zea under shared socioeconomic pathways (SSP) 1-2.6, SSP2-4.5, and SSP5-8.5 in both the 2030s and 2050s. The conversion of unsuitable to low suitability habitat and moderately to high suitability habitat increased by 8.43% and 2.35%, respectively. From the present day to the 2030s, under SSP1-2.6, SSP2-4.5 and SSP5-8.5, the centroid of the suitable habitats of H. zea showed a general tendency to move eastward; from 2030s to the 2050s, under SSP1-2.6 and SSP5-8.5, it moved southward, and it moved slightly northward under SSP2-4.5. According to bioclimatic conditions, H. zea has a high capacity for colonization by introduced individuals in China. Customs ports should pay attention to host plants and containers of H. zea and should exchange information to strengthen plant quarantine and pest monitoring, thus enhancing target management.

Microorganisms in the guts of insects enhance the adaptability of their hosts with different lifestyles, or those that live in different habitats. Tuta absoluta is an invasive pest that is a serious threat to tomato production in China. It has quickly spread and colonized Xinjiang, Yunnan and other provinces and regions. We used Illumina HiSeq next generation sequencing of the 16S rRNA gene to study and analyze the composition and diversity of the gut microbiota of three geographical populations of T. absoluta. At the phylum level, the most common bacteria in T. absoluta across all three geographical populations were Proteobacteria and Firmicutes. An uncultured bacterium in the Enterobacteriaceae was the dominant bacterial genus in the T. absoluta gut microbiotas. There were no significant differences in alpha diversity metrics among the Spanish, Yunnan and Xinjiang populations. The structures of the gut microbiota of the three populations were similar based on PCoA and NMDS results. The results confirmed that the microbial structures of T. absoluta from different regions were similar.

The frequency and amplitude of heat waves are predicted to increase under future climate change conditions. We still lack a detailed understanding of how changes in the frequency and amplitude of heat waves are linked to the life history traits and biocontrol efficiency of host-feeding parasitoids. In the present study, we simulated a series of heat waves as a function of amplitude and frequency to investigate the effects on the life history traits of the host-feeding parasitoid Eretmocerus hayati. We found that both the amplitude and frequency of heat waves significantly affected the adult phenotypes. In the low-amplitude heat wave group, the frequency of heat waves did not change the life history traits of the parasitoid; however, when the heat amplitude reached 42 °C, medium (four times/week) and high frequencies (seven times/week) of heat waves detrimentally affected these parameters. Hence, these findings suggest that to obtain optimal biological control with this parasitoid, we need to carefully monitor heat wave pattern (especially the amplitude and frequency) over the short term (usually 7–10 days) before releasing a host-feeding parasitoid.

To reveal the mechanism of temperature preference in Tuta absoluta, one of the top 20 plant pests in the world, we cloned and identified TaTRPA1, TaPain, and TaPyx genes by RACE and bioinformatic analysis, and clarified their expression profiles during different development stages using real-time PCR, and revealed their function in preference temperature by RNAi. The full-length cDNA of TaPain was 3136 bp, with a 2865-bp open reading frame encoding a 259.89-kDa protein; and the partial length cDNA of TaPyx was 2326-bp, with a 2025-bp open reading frame encoding a 193.16-kDa protein. In addition, the expression of TaTRPA1 and TaPyx was significantly lower in larvae than other stages, and it was significantly higher in pupae and newly emerging males for TaPain. After feeding target double-stranded RNA (dsRNA), the preferred temperature decreased 2 °C more than the control group. In conclusion, the results firstly indicated the molecular characterization of TRPA subfamily genes and their key role in temperature perception in T. absoluta, and the study will help us to understand the temperature-sensing mechanism in the pest, and will provide some basis for study of other Lepidoptera insects’ temperature preference. Moreover, it is of great significance in enriching the research progress of “thermos TRP”.