Mechanical Egg Activation and Rearing of First Instar Larvae of Sirex noctilio (Hymenoptera: Siricidae).

Egg activation is a cellular transition of an arrested mature oocyte into a developing embryo through a coordinated series of events. Previous studies in Hymenoptera have indicated that mechanical pressure can induce egg activation. In this study, we developed the first egg activation protocol for the haplodiploid insect pest, Sirex noctilio (Hymenoptera: Siricidae), from two climatically different regions in South Africa to demonstrate the broad applicability of the method. In addition, activated eggs were exposed to three treatments involving water, pine sawdust, and the fungal symbiont of S. noctilio, Amylostereum areolatum (Russulales: Amylostereaceae), to determine if the symbiotic fungus is a requirement for egg development in an artificial laboratory environment, as the symbiotic fungus has been hypothesised to be necessary for egg and early larval development in a natural environment. A rearing protocol was developed for the first instar larvae using a modified Anoplophora glabripennis (Coleoptera: Cerambycidae) artificial diet. A significant difference between the mean survival rates of activated eggs from the two different regions was observed. Amylostereum areolatum was shown to be unnecessary for egg survival and adversely affected egg eclosion in an artificial laboratory environment. The maximum larval survival duration on the artificial diet was 92 days. The egg activation and rearing protocol developed in this study enables opportunities for research on the physiology, ecology, symbioses, and genetics of S. noctilio, which can be exploited for new genetic pest management strategies.

DNA extraction techniques for DNA barcoding of minute gall-inhabiting wasps.

DNA extraction from minute hymenopterans and their larvae is difficult and challenging because of their small size indicating a low amount of starting material. Hence, 11 DNA extraction methods were compared to determine their efficacy in isolating DNA. Success of each method was scored on a 2% agarose gel after PCR of the cox 1 mitochondrial locus. A silica-membrane-based approach was the most successful, followed by a method using a combination of incubation buffers and a method using magnetic beads. The method using buffers was the most cost- and time effective. Using this method, larvae from Eucalyptus seed capsule galls could be assigned a role (parasitoid, gall former or inquiline) in the gall-inhabiting complex.