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Behavioral thermoregulation is a key strategy for insects to cope with heat stress. The rice leaf folder Cnaphalocrocis medinalis Guenée (Lepidoptera: Pyralidae) larvae usually fold one leaf to construct a leaf shelter. The larvae are vulnerable to heat stress, and the temperature in summer is often beyond the optimal range of them. Shelters confer protection against environmental stress but unclear whether larvae will alter shelter-building behavior when encountering heat stress. We observed the shelter-building behavior of larvae during and after heat shock, and then examined the shape and structure of shelters. Larvae spent more time in selecting a site and building a shelter during and after heat shock than at the optimal temperature. More than 70% of larvae folded two or three leaves to build a shelter during and after heat shock, but more than 60% of larvae only folded one leaf at the optimal temperature. Larvae built more single-leaf longitudinal shelters at the optimal temperature, but they built more multileaf overlapping shelters during and after heat stress. Larvae constructed a short leaf shelter using a small amount of silk binds when they were exposed to 40°C for 4 h. The rice leaf folder larvae can alter their shelter-building behavior and shelter structure in response to heat stress.

Under ongoing climate warming, both the degree and number of high-temperature events in summer may increase, and behavioural adaptation is an important ecological strategy employed by insects to cope with such events. The rice leaf folder, Cnaphalocrocis medinalis Güenée, is a serious insect pest of rice fields in summer. Population outbreaks have become more frequent in the last ten years. In addition to adult migration, rice leaf folders are thought to have other thermal adaptations. Therefore, the behaviours of larval and adult rice leaf folders, such as leaf folding (making shelter) and habitat selection for pupae and eggs, were observed on rice plants under heat stress. The results showed that larval shelter-making velocities significantly decreased during or after four hours of heat exposure, and shelter size decreased as the temperature increased. Larvae preferred to pupate on young rice leaves at 27°C and middle-aged leaves at 30°C, but they strongly preferred older leaves when reared at 34°C. Female moths generally preferred to oviposit on the top of young leaves, but they preferred the middle and lower leaves for egg deposition when exposed to 36 and 40°C, respectively. Furthermore, more eggs were distributed on the lower surfaces of rice leaves with an increase in heat stress. These behavioural responses of rice leaf folders to heat stress indicate that this pest has great potential to adapt to high temperatures; therefore, the possibility of a population outbreak will remain high despite global warming.

Physiological response and acclimation to thermal stress is a key strategy of insects to cope with changing climate. The underlying mechanism of heat acclimation in insects is still unclear. Here, the heat selection and transcript level response in the larvae of the rice leaf folder Cnaphalocrocis medinalis Guenee, a serious pest of rice in summer, were studied. The survival and fecundity of larvae during multigenerational heat selection at 39 °C were examined, and heat tolerance and mRNA expression of heat shock protein 70 (Hsp70) and 90 (Hsp90) were examined under heat stress. The results showed that survival and fecundity of larvae increased notably and then kept constant after two or three generations of heat selection. Heat selection improved thermal tolerance of larvae. The Hsp70 mRNA expression of the 3rd-instar larvae increased in all five generations of heat selection, but Hsp90 increased only in the first two generations. The response of Hsp70 to 39 °C heat treatment in the larvae kept at 27 °C was different from the larvae exposed to the conditioning heat treatments, but the response of Hsp 90 was similar. Moreover, the Hsp70 and Hsp90 mRNA expression levels were significantly higher in the heat-acclimated larvae than that in the unacclimated larvae at a comparable duration of exposure to 37 and 41 °C. Selection at a high temperature across multiple generations led larvae to heat acclimation, and Hsp70 and Hsp90 were involved in this acclimation process.

Single-particle electron microscopy and hydrogen–deuterium exchange mass spectrometry are used to characterize the structure and dynamics of a G-protein-coupled receptor–arrestin complex. Much has been learned about the structure of G-protein-coupled receptors (GCPRs) over the past seven years, but we still don't know what an activated GPCR looks like when it is bound to a β-arrestin. (Arrestins are cellular mediators with a broad range of functions, many of them involving GPCRs.) In this study the authors use single-particle electron microscopy and hydrogen–deuterium exchange mass spectrometry to characterize the structure and dynamics of a GPCR–arrestin complex. Their data support a 'biphasic' mechanism, in which the arrestin initially interacts with the phosphorylated carboxy terminus of the GPCR before re-arranging to more fully engage the membrane protein in a signalling-competent conformation. G-protein-coupled receptors (GPCRs) are critically regulated by β-arrestins, which not only desensitize G-protein signalling but also initiate a G-protein-independent wave of signalling1,2,3,4,5. A recent surge of structural data on a number of GPCRs, including the β2 adrenergic receptor (β2AR)–G-protein complex, has provided novel insights into the structural basis of receptor activation6,7,8,9,10,11. However, complementary information has been lacking on the recruitment of β-arrestins to activated GPCRs, primarily owing to challenges in obtaining stable receptor–β-arrestin complexes for structural studies. Here we devised a strategy for forming and purifying a functional human β2AR–β-arrestin-1 complex that allowed us to visualize its architecture by single-particle negative-stain electron microscopy and to characterize the interactions between β2AR and β-arrestin 1 using hydrogen–deuterium exchange mass spectrometry (HDX-MS) and chemical crosslinking. Electron microscopy two-dimensional averages and three-dimensional reconstructions reveal bimodal binding of β-arrestin 1 to the β2AR, involving two separate sets of interactions, one with the phosphorylated carboxy terminus of the receptor and the other with its seven-transmembrane core. Areas of reduced HDX together with identification of crosslinked residues suggest engagement of the finger loop of β-arrestin 1 with the seven-transmembrane core of the receptor. In contrast, focal areas of raised HDX levels indicate regions of increased dynamics in both the N and C domains of β-arrestin 1 when coupled to the β2AR. A molecular model of the β2AR–β-arrestin signalling complex was made by docking activated β-arrestin 1 and β2AR crystal structures into the electron microscopy map densities with constraints provided by HDX-MS and crosslinking, allowing us to obtain valuable insights into the overall architecture of a receptor–arrestin complex. The dynamic and structural information presented here provides a framework for better understanding the basis of GPCR regulation by arrestins.

The rice leaf folder, Cnaphalocrocis medinalis (Guenee), will frequently be under heat stress because of global warming. The young larvae would be liable to heat exposure owing to inability to make leaf fold for hiding themselves. Consequently, their heat tolerance might be closely related to population outbreak in summer. Therefore, the heat tolerance of the first-instar larvae was examined in this study. The results showed that the heat tolerance of young larvae was lower than that of the older ones. Survival rates of the first-instar larvae were more than 60% on exposure to 39°C for 5 h per day for 3 successive days, but it dropped to below 20% on exposure to 41°C for 5 h per day for 2 successive days. The first-instar larvae exposed to 39°C for less than 8 h per day for 1, 2, and 3 successive days exhibited similar rates of pupation and adult emergence, fecundity, and egg hatchability as the control. High temperature of 39°C influenced the first-instar larvae on a small scale, but 41°C influenced greatly. Young larvae of the rice leaf folder can tolerate short-term heat stress in the present summer, and population outbreaks might not be seriously restricted to global warming.