Ropalidia marginata is characterized as a primitively eusocial wasp due to the absence of morphological differentiation between the queen and worker castes. Unlike other primitively eusocial wasps, however, the queen in this species is a docile individual, who does not use aggression to regulate worker reproduction, and does not act as the central pacemaker of her colony. However, if the queen dies or is experimentally removed, one of the workers steps up her aggression immensely within minutes, and if the queen is not replaced, she develops her ovaries, reduces aggression and takes over as the new queen of the colony. We call her the potential queen (PQ). When I started my work on R. marginata, two very intriguing questions were demanding to be answered, which had developed from work done by my immediate seniors in the lab. I decided to pursue both of these for my thesis. My work has been enriched by inputs from several collaborators and colleagues - I couldn’t have done all of it by myself. So, henceforth, I will be using the word “we”, instead of the first person singular to describe the work that has gone into this thesis. Question 1: Is there a designated successor to the queen in R. marginata? My senior Sujata P. Kardile has shown in her thesis, that in R. cyathiformis, a primitively eusocial wasp very closely related to R. marginata, the queen is always succeeded by the next most aggressive individual in the colony, and so the PQ is easily predictable in the presence of the queen. However, in R. marginata, the PQ appears to be an unspecialized individual, who cannot be predicted in the presence of the queen by using age, ovarian profile or behaviour as the yardsticks. However, the PQ becomes evident within minutes after queen removal. The swiftness with which the PQ is established led us to believe that perhaps the successor to the queen in R. marginata is known to the wasps, though we cannot identify her in the presence of the queen. We designed an experiment to check for the presence of such a ”cryptic successor” in R. marginata. Our experiments involved splitting a normal, queen-right nest into two halves separated by a wire mesh partition, so that the wasps could not move across the mesh. Earlier we had used this set-up to demonstrate that a PQ always establishes herself on the queen-less fragment of the nest. So, to test if there is a cryptic successor, we allowed a PQ to establish herself on the queen-less fragment, and then exchanged the queen and the PQ (designated as PQ1) between the two sides. There is a 50% probability that the cryptic successor, if present, would be on the queen-less side in the beginning. Then, upon exchange, she should be able to hold her position on the other side easily. On the other hand, if the cryptic successor is first on the queen-right side, then, upon exchange, she should take over as the PQ (PQ2), and PQ1 should not be able to hold her status. The cryptic successor hypothesis had two predictions: (i) the PQ1 would lose to a PQ2 in about half the cases, and (ii) there would never be a PQ3. We obtained a PQ2 in 5 out of 8 cases, and we never had a PQ3. So we could conclude that there is indeed one individual who is the designated successor to the queen in R. marginata. Since we could not identify her in the presence of the queen, we call her the cryptic successor. The cryptic successor did not receive even a single act of aggression from the PQ1, or from any other individual in the colony. Thus we conclude that she is acceptable to all the wasps in the colony. We next used the more sophisticated and rigorous method of network analysis to check if the PQ could be predicted due to some unique position she might be holding in the social network on her colony. Since this was a first study in a primitively eusocial insect using network tools, we began by characterizing the social networks of queen-right and queen-less colonies of R. marginata, and compared them with the R. cyathiformis networks to see how different the R. marginata society is from a typical primitively eusocial one. The R. marginata social networks based on dominant-subordinate interactions were low in their centrality measure as compared to the R. cyathiformis networks. However, in both the species, the queen-less networks were highly centralized, star-shaped networks with the PQs at the centre. Neither the queens, nor the PQs were key individuals in the queen-right colonies, but it is interesting to note that the removal of an insignificant node, the queen, resulted in a major change in the network architecture, converting the de-centralized queen-right network into a highly centralized one. Such centralized star-shaped networks are unique, and to our knowledge, the first ever described, in any social system. When we removed the queen from the data set (in silico removal), the resulting network was similar in centrality to the queen-right networks. We then did a comparative analysis of the positional importance of the PQs of the two species, and tried to see if we could use this as a tool to predict the PQ in the queen-right network. In R. cyathiformis, the PQs had consistently high ranks (mostly rank 2) in the network based on the degree index, while the PQs in R. marginata had random ranks in the hierarchy. However, since the PQs are known not to have unique ranks in the dominance hierarchies, we repeated the analysis using data on all interactions from the Q-PQ exchange experiments described above. Neither the cryptic successors nor the losers occupied any unique ranks in the all interactions networks. Thus the successors in R. marginata are truly cryptic, even in their social networks. Since R. marginata is known to be more evolved than typical primitively eusocial species, it is likely that the queen’s successor is identified by the wasps through some subtle cue like smell, and so we cannot identify her using the methods that are adequate for the identification of the PQ in a typical primitively eusocial species like R. cyathiformis. Question 2: How does the queen signal her presence and reproductive status to her workers or, how do the workers perceive the presence of their queen? The fact that in spite of her docility, the queen in R. marginata manages to maintain complete reproductive monopoly in her colony, gives rise to the obvious question of how she suppresses worker reproduction. The most attractive hypothesis is that she uses a pheromone like queens of highly eusocial species. My senior A. Sumana had shown that the queen pheromone, if present, is not a volatile substance. She also showed that the queen interacts at a very low rate with her workers, and so they cannot possibly perceive her by means of direct interactions. Since the PQ steps up her aggression within minutes of queen removal, we used her as a proxy to know how soon the queen’s absence is felt in the colony. We built a model to delineate the relationship between the decay time of the pheromone (td), the average age of the queen’s signal present with the PQ (ta), and the average realization time (tr); where tr = td − ta. Using Dijkstra’s algorithm, we showed that the queen could interact faster with the PQ by using relay interactions. Then using experimental data from 50 colonies, we obtained a ta of 102.9 minutes. The td was 340 minutes, and so we obtained a tr of 237.1 minutes; which meant that the PQ should not perceive the queen’s absence within 237 minutes of queen removal, if the queen pheromone is transmitted by a relay mechanism. However, from our experimental data, we had obtained a tr of 30 minutes. So we concluded that physical interactions, both direct and indirect were inadequate for the workers to perceive their queen. As we had ruled out physical interactions, we then wanted to check if it is possible that the queen applies her pheromone to the nest material, from where it is perceived by the workers when they walk or sit on the nest, or antennate the nest surface. The “rub abdomen behaviour (RA)” has been observed to be quite typical of R. marginata queens, and is not very common in the workers of the species. RA involves rubbing the ventral side of the tip of the abdomen or dragging it on the nest surface while walking. We thought that the queen might be using this behaviour to apply her pheromone on the nest material. So we characterized this behaviour using focal behaviour sampling, and found that the queen rubs her abdomen on the nest once in every 23 minutes. Since the observed tr is 30 minutes, it is quite likely that the queen uses the rub abdomen behaviour to apply her pheromone on the nest. The next step was to check for the source of the queen pheromone. We looked for glands that open near the base of the sting, and the Dufour’s gland was a good choice, as it is known to be involved in the recognition of egg-laying workers in the honeybees. We performed a bioassay in the blind using the crude extract of the Dufour’s gland (prepared in Ringer’s solution) from the queen. The Dufour’s gland extract of a randomly chosen worker and the solvent were used as controls. We found that the PQ responds to the queen’s Dufour’s gland extract by lowering her aggression to 65% of what she was showing on queen removal and before the application of the extract. However, the PQ did not change her behaviour significantly when the worker’s extract or Ringer’s solution was applied. The PQ’s reduction of aggression on application of the queen’s extract mimicked the reaction of PQ’s when the queen is re-introduced on the nest some time after removal. So we hypothesize that the Dufour’s gland is the source of the queen pheromone (signal) in R. marginata. This thesis has opened up newer questions pertaining to the power of the queen and the intricacies of the succession to power in R. marginata. For example, we need to pursue chemical analyses of the Dufour’s gland extract of R. marginata to have conclusive proof of it’s being the source of the queen pheromone. But that is perhaps suitable topic for my juniors in the lab, who can continue the tradition of beginning with questions opened up by their seniors!