Detection of the bacterial endosymbiont Wolbachia and determination of super groups present within New Zealand invertebrates : a thesis presented in partial fulfilment of the requirements for the degree of Masters of Science in Zoology at Massey University, Palmerston North, New Zealand

Abstract

𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 is one of the most widespread intracellular bacterium on earth, estimated to infect between 40 and 66% of arthropod species. Where significantly screened for, there is virtually no ecosystem that 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 has not managed to invade. Their impact does not come solely from their vast distribution but in their ability to modify their hosts reproductive biology. 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 is a maternally inherited endosymbiont that can induce a range of host phenotypic responses, including cytoplasmic incompatibility, male death, feminization, and parthenogenesis. This holds high potential for influencing genetic diversity and speciation of its host. 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 has yet to be formally identified in New Zealand native invertebrates and therefore a gap remains in the global understanding of 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 distribution and diversity. The first aim of this thesis (Chapter 2) was to determine if the use of high throughput sequencing (HTS) of invertebrates could be used to identify 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 sequences and establish a 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 infection. seven HTS produced a positive indication for 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 sequences, with six samples originating from native New Zealand invertebrate hosts. Once 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 was detected the second aim (Chapter 2) was to determine which super group the 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 strains detected fall into. Comparing New Zealand 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 sequences to sequences obtained from GenBank, it was determined that there were two distinct strains of 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢in New Zealand hosts. One strain was related to 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 super group A and the other matched 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 super group B. 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 had now been detected in New Zealand, the next aim (Chapter 3) was to determine the presence and distribution of 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 across New Zealand and across a number of native invertebrates. 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 was detected in both of the main islands of New Zealand across a number of species or putative species of ground and cave weta. The final aim of this thesis was to determine the diversity of the 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 detected and compare that to the diversity of the ground weta complex 𝘏𝘦𝘮𝘪𝘢𝘯𝘥𝘳𝘶𝘴 𝘮𝘢𝘤𝘶𝘭𝘪𝘧𝘳𝘰𝘯𝘴 to determine if there was any evidence for 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 affecting the genetic diversity of its host (Chapter 4). It was observed that 𝘏. 𝘮𝘢𝘤𝘶𝘭𝘪𝘧𝘳𝘰𝘯𝘴 complex was infected with both strains of 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 detected in New Zealand invertebrates. There was however not enough resolution to determine if 𝘞𝘰𝘭𝘣𝘢𝘤𝘩𝘪𝘢 has had a significant effect on the diversity and speciation of the ground weta host.