Ganghua Li

Experimental verification and molecular basis of active immunization against fungal pathogens in termites

Termites are constantly exposed to many pathogens when they nest and forage in the field, so they employ various immune strategies to defend against pathogenic infections. Here, we demonstrate that the subterranean termite Reticulitermes chinensis employs active immunization to defend against the entomopathogen Metarhizium anisopliae. Our results showed that allogrooming frequency increased significantly between fungus-treated termites and their nestmates. Through active social contact, previously healthy nestmates only received small numbers of conidia from fungus-treated individuals. These nestmates experienced low-level fungal infections, resulting in low mortality and apparently improved antifungal defences. Moreover, infected nestmates promoted the activity of two antioxidant enzymes (SOD and CAT) and upregulated the expression of three immune genes (phenoloxidase, transferrin, and termicin). We found 20 differentially expressed proteins associated with active immunization in R. chinensis through iTRAQ proteomics, including 12 stress response proteins, six immune signalling proteins, and two immune effector molecules. Subsequently, two significantly upregulated (60S ribosomal protein L23 and isocitrate dehydrogenase) and three significantly downregulated (glutathione S-transferase D1, cuticle protein 19, and ubiquitin conjugating enzyme) candidate immune proteins were validated by MRM assays. These findings suggest that active immunization in termites may be regulated by different immune proteins.

Genetic Analysis of Population Structure and Reproductive Mode of the Termite Reticulitermes chinensis Snyder

The subterranean termite Reticulitermes chinensis Snyder is an important pest of trees and buildings in China. Here, we characterized genetic structure and reproductive modes of R. chinensis from China for the first time. A total of 1,875 workers from 75 collection sites in Huanggang, Changsha and Chongqing cities were genotyped at eight microsatellite loci. Analysis of genetic clusters showed two subpopulations in Chongqing city. The Huanggang population showed a uniform genetic pattern and was separated from the other populations by the largest genetic distances (F ST: 0.17–0.20). In contrast, smaller genetic distances (F ST: 0.05–0.12) separated Changsha, Chongqing-1 and Chongqing-2 populations. Chongqing-1 was the only population showing a genetic bottleneck. Isolation by distance among colonies in the Huanggang population indicated limited alate dispersal or colony budding. Lack of isolation by distance among colonies within the populations of Changsha, Chongqing-1 and Chongqing-2, suggested long-range dispersal by alates and/or human-mediated transport. Overall, extended family colonies (73.91%) were predominant in all four populations, followed by simple (20.29%), and mixed family colonies (5.80%). Most simple families were headed by inbred related reproductive pairs in the Changsha population, while most simple families in the Chongqing-1 population were headed by outbred unrelated pairs. Simple families in the Huanggang population were a mixture of colonies headed by outbred or inbred reproductive pairs. The sample size of simple families in the Chongqing-2 population was too small to yield significant results. Extended families in all four populations were headed on the average by ≤10 neotenics. Mixed families likely originated from pleometrosis. Presence of heterozygote genotypes showed that all neotenic reproductives collected in addition from five field colonies in Wuhan city were sexually produced, suggesting that these colonies did not undergo parthenogenesis. This study contributes to better understanding of the variance of genetic structure and reproductive mode in the genus Reticulitermes.

Dynamics of sex ratio, fresh weight and nutrient contents at five developmental stages of alates in the subterranean termite Reticulitermes chinensis

Sex ratio, fresh weight and nutrient contents were examined at five developmental stages of alates in the subterranean termite Reticulitermes chinensis Snyder, including sixth-instar nymphs, callow alates, dark alates, flying alates and dealates. The results showed that the sex ratio of callow alates and dark alates was highly female biased, but there were no significant differences in the percentage of females and males at sixth-instar nymphs, flying alates, and dealates. The fresh weights of alates declined with the developmental stages, suggesting a dynamic process in which the termites continue to shrink and harden. Meanwhile, the weights of female alates were significantly higher than weights of male alates in all five stages. There were no significant differences in the protein titers of the five developmental stages of alates. Flying alates were observed to have the highest triglyceride level, but a low glucose titer. This suggested that the main energy for flight of alates is provided by the breakdown of triglyceride reserves. Alates regardless of developmental stages, workers and soldiers had no significant differences in the cholesterol titers for the two extraction methods. Interestingly, there was a significantly negative linear correlation between the titer of glucose and the titer of triglyceride in both female alates and male alates during the five developmental stages, suggesting that the indirect mutual transformation between glucose and triglyceride might occur in the energy metabolism of all alates in R. chinensis.

Factors affecting mate choice in the subterranean termite Reticulitermes chinensis (Isoptera: Rhinotermitidae)

Termites are a group of eusocial insects. Mate choice is the most important step which affects the subsequent new colony foundation and development in termites. This study investigated the effects of the four factors on mate choice in the subterranean termite Reticulitermes chinensis Snyder, including colony origin, entry time, physical damage of antennae, and fresh body weights of male dealates. The results showed that the pairing number of dealates from the different colonies was significantly higher than that from the same colonies. The male dealates with early entry time achieved a significantly higher pairing number than the male dealates with late entry time. The male dealates with complete antennae achieved a the significantly higher pairing number than the male dealates lacking the six terminal segments of antennae. Moreover, the heavier male dealates achieved a significantly higher pairing number than the lighter male dealates. However, there were no significant differences in the pairing time for all the experimental treatments. These results suggested that the male dealates, which are heterogenous, healthy, heavy, and more familiar with the environment in R. chinensis, have the obvious advantages in the mate choice. This rule of mate choice is helpful for termites to avoid inbreeding and to maintain the genetic diversity of offspring, which is very important for the environmental adaptability and development of termite colonies.

Comparison of the trapping effect and antioxidant enzymatic activities using three different light sources in cockchafers

Light traps have been widely used for controlling underground pests. However, very little is known regarding the relationship between trapping effect and antioxidant enzymatic activities using light irradiation in underground pests. Thus, we determined the trapping effect of three light sources of the frequoscillation pest-killing lamp on two species of cockchafers, Serica orientalis Motschulsky (Coleoptera: Melolonthidae) and Anomala corpulenta Motschulsky (Coleoptera: Rutelidae), and evaluated the effect of the same three light sources on the activities of their antioxidant enzymes. The catches of S. orientalis were significantly higher compared to A. corpulenta using light source A in peanut fields in China. After irradiation by light source A, the malondialdehyde (MDA) contents and activities of superoxide dismutase (SOD) and glutathione S-transferases (GST) in S. orientalis were significantly and marginally significantly lower compared to A. corpulenta. Taken together, these results indicated a weaker antioxidant enzyme activity response to light stress and a larger quantity of trapping catches using light irradiation in cockchafers. Thus, we proposed a potential negative relationship between trapping effect and antioxidant enzymatic activities in response to light irradiation in cockchafers.

A trade-off between antipredatory behavior and pairing competition produced by male-male tandem running in three Reticulitermes species

Due to the omnipresent risk of predation, termites have evolved many antipredatory behaviors. The two related species Reticulitermes speratus and R. chinensis have been demonstrated to use homosexual tandem running to decrease individual predation risk after shedding their wings. In this study, we tested risk of predation in the termite R. flaviceps, which is distantly related to the above two species. We determined that homosexual tandem running also led to low individual predation risk in dealates of R. flaviceps. Moreover, by combining a predation model with a competition model, we observed a typical trade‐off phenomenon between antipredatory behavior and pairing competition produced by male‑male tandem running in the above three Reticulitermes species. Our results indicated that male‑male tandem running could effectively protect disadvantaged individuals from being caught, but disadvantaged individuals would be easily eliminated in pairing competition after male‑male tandem running, suggesting that male‑male tandem running can promote population evolution in termites by repeatedly removing the relatively inferior male individuals.

Physiological reason for ceasing growth of unfertilized eggs produced by unmated queens in the subterranean termite Reticulitermes chinensis

In Reticulitermes chinensis, a close relative of R. speratus with asexual queen succession, unfertilized eggs can be produced but are not incubated. To explain this phenomenon, we analysed the physiological differences between unfertilized eggs/unmated queens and fertilized eggs/mated queens. Fertilized eggs consumed significantly larger quantities of five amino acids (Cys, Met, Ile, Leu and Tyr), Ca, protein and cholesterol during incubation. The higher levels of four trace elements (Na, K, Zn and Fe) in fertilized eggs and their lower levels in mated queens indicated that mated queens might transfer these trace elements to fertilized eggs to complete incubation. The higher levels of Mn, triglycerides and serotonin in mated queens and higher levels of Mn and glucose in fertilized eggs suggested that these substances are very important for normal ovarian and embryonic growth. The different expression of three reproductive genes (vtgl, rabil and JHE1) suggested that they might be involved in the regulation of ovarian and embryonic growth. Overall, changes in these physiological indices may substantially affect ovarian and embryonic growth and prohibit the incubation of unfertilized eggs in R. chinensis.

Physiological profiles associated with ceasing growth of unfertilized eggs produced by unmated queens in the subterranean termite Reticulitermes chinensis

ABSTRACT In Reticulitermes chinensis, a close relative of R. speratus with asexual queen succession, unfertilized eggs can be produced but do not hatch as larvae. To explain this phenomenon, we analyzed the physiological differences between unfertilized eggs/unmated queens and fertilized eggs/mated queens. Fertilized eggs had significantly lower quantities of five amino acids (Cys, Met, Ile, Leu and Tyr), Ca, protein and cholesterol during development. The higher levels of four trace elements (Na, K, Zn and Fe) in fertilized eggs and their lower levels in mated queens indicated that mated queens might transfer these trace elements to fertilized eggs to aid development. The higher levels of Mn, triglycerides and serotonin in mated queens and higher levels of Mn and glucose in fertilized eggs suggested that these substances are very important for normal ovarian and embryonic growth. The different expression of three reproductive genes (vtg 1, rab 11 and JHE 1) suggested that they might be involved in the regulation of ovarian and embryonic growth. Overall, changes in these physiological indices may substantially affect ovarian and embryonic growth and inhibit development of unfertilized eggs in R. chinensis. Summary: Changes in amino acids, trace elements, nutrient content, serotonin and reproductive genes may substantially affect ovarian and embryonic growth and inhibit development of unfertilized eggs in Reticulitermes chinensis.