The soil insect Bradysia odoriphaga (Diptera: Sciaridae) causes substantial damage to Chinese chive. Suitable reference genes in B. odoriphaga (Bradysia odoriphaga) have yet to be identified for normalizing target gene expression among samples by quantitative real-time PCR (qRT-PCR). This study was focused on identifying the expression stability of 12 candidate housekeeping genes in B. odoriphaga under various experiment conditions. The final stability ranking of 12 housekeeping genes was obtained with RefFinder, and the most suitable number of reference genes was analyzed by GeNorm. The results revealed that the most appropriate sets of internal controls were RPS15, RPL18, and RPS18 across developmental phases; RPS15, RPL28, and GAPDH across temperatures; RPS15 and RPL18 across pesticide treatments; RSP5, RPS18, and SDHA across photoperiods; ACTb, RPS18, and RPS15 across diets; RPS13 and RPL28 across populations; and RPS15, ACTb, and RPS18 across all samples. The use of the most suitable reference genes versus an arbitrarily selected reference gene resulted in significant differences in the analysis of a target gene expression. HSP23 in B. odoriphaga was found to be up-regulated under low temperatures. These results will contribute to the standardization of qRT-PCR and will also be valuable for further research on gene function in B. odoriphaga.

The effect of various flavonoids, lectins and phenyl β‐D‐glucoside on larval survival, weights and the activities of digestive (total serine protease and trypsin) and detoxifying (esterase and glutathione‐S‐transferase) enzymes of Spodoptera litura larvae at 7 days after treatment was studied through diet incorporation assay. Flavonoids (rutin, chlorogenic acid, quinic acid, caffeic acid, naringenin, quercitin, kaempferol, myricetin, catechin, and ferulic acid) were incorporated in artificial diet at 100, 500 and 1000 ppm, lectins: groundnut leaf lectin (GLL), concavalin A (ConA) and phenyl β‐D‐glucoside at 1, 2 and 5 μg/mL. Flavonoids such as rutin, quercitin and kaempferol at 1000 ppm were more toxic to S. litura larvae than quinic acid, caffeic acid, naringenin, myricetin, catechin, and ferulic acid. Larval growth and development were significantly reduced in S. litura larvae fed on a diet with GLL and ConA at 5 μg/mL compared to the larvae fed at 2 and 1 μg/mL concentrations. The larvae fed on flavonoid‐treated diets showed significant reduction in serine protease, trypsin and esterase activities. The flavonoids such as rutin, chlorogenic acid, quinic acid, naringenin, quercitin, kaempferol and myricetin, and lectins, GLL and ConA can be utilized in insect control programs.

Bradysia odoriphaga is frequently subjected to heat shock during the summer in China. Although the effects of heat shock on insect ecology and physiology have been widely explored, the effects of heat shock on the life history parameters of Bradysia odoriphaga are largely unknown. In the present study, we investigated the effects of heat shock on B. odoriphaga survival and reproduction as well as on offspring development and sex ratio. We exposed adult B. odoriphaga to 31, 33, 35, or 37 °C for different durations (from 0 to 120 min). The results showed that the survival of both sexes declined with the increase in temperature and exposure time, especially at 33, 35, and 37 °C. Longevity was markedly greater for males than females across all treatments. Fecundity generally declined as temperature and exposure time increased, and no eggs hatched when females were exposed to 37 °C for >75 min. The development of offspring larvae was significantly delayed when the parent female and male had been exposed to ≥31 °C for ≥30 min. In addition, the sex ratio of F1 progeny derived from heat-shocked parental adults was increasingly skewed to female as exposure time and temperature treatment increased. Overall, the results indicate that heat shock negatively influences B. odoriphaga.

Gut bacteria play a significant role in host insect. This study evaluated detail difference of midgut bacteria in deltamethrin-resistant, deltamethrin-susceptible and field-caught populations of diamondback moth, and studied phenomics of the predominant midgut bacterium Enterococcus mundtii. Cultivable bacteria revealed that E. mundtii and Carnobacterium maltaromaticum dominated the bacterial populations from deltamethrin-resistant and deltamethrin-susceptible larval midguts, whereas E. mundtii was predominant in field-caught population. Illumina sequencing analysis indicated that 97% of the midgut bacteria were from the phyla Firmicutes, Proteobacteria and Cyanobacteria. Both resistant and susceptible populations had more Enterococcus and Carnobacterium. Enterococcus, Carnobacterium, Bacillus, and Pseudomonas were predominant in the field-caught population. A phenomics analysis revealed that E. mundtii was able to metabolize 25.26% of the tested carbon sources, 100% of the nitrogen sources, 100% of the phosphorus sources and 97.14% of the sulfur sources, had a wide range of osmolytes and pH conditions, and showed active deaminase activity but no decarboxylase activity. This is the first report regarding different populations of DBM midgut bacteria analyzed using both high-throughput DNA sequencing and cultivation methods, and also first report concerning the phenomics of E. mundtii. The phenomics of E. mundtii provide a basis for the future study of gut bacteria functions.

Bradysia odoriphaga Yang et Zhang is a serious belowground pest of Chinese chives (Allium tuberosum). Our previous studies have indicated that B. odoriphaga females prefer to oviposit near the roots of Chinese chives rather than the roots of other plants, and that the performance (longevity and fecundity) of B. odoriphaga offspring was better on Chinese chives than on Lettuce (var. ramosa Hort.), Onion (Allium cepa) and Potato (Solanum tuberosum) but little is known about how the volatiles released by Chinese chives affect the host-finding and oviposition behaviors of B. odoriphaga. Here, we used gas chromatography-mass spectrometry and determined that Chinese chives releases the following volatiles: methyl allyl disulfide, β-myrcene, cis-ocimene, diallyl disulfide, nonane, n-dodecane, n-tetradecane, and n-hexadecane; quantities released were highest for methyl allyl disulfide and diallyl disulfide. In addition to eliciting strong responses in females in electroantennography assays, the latter two sulfur compounds and their mixtures attracted females in Y-tube olfactometer assays. The addition of methyl allyl disulfide, diallyl disulfide, or a mixture of the two compounds at a 1:5 ratio to chive plants increased oviposition when compared to control plants. These results indicate that methyl allyl disulfide and diallyl disulfide, either alone or in combination, influence the host-seeking behavior of B. odoriphaga.