The University of Southern Mississippi The Aquila Digital Community Dissertations Summer 8-2007 PHYLOGEOGRAPHY OF THE INDONESIAN WATER PYTHON, LIASIS MACKLOTISSP. (SQUAMATA: BOIDAE: PYTHONINAE): A COMPARATIVE APPROACH TOWARD RESOLVING PHYLOGENY Christopher Knight Carmichael University of Southern Mississippi Follow this and additional works at: https://aquila.usm.edu/dissertations Part of the Aquaculture and Fisheries Commons, Biology Commons, and the Marine Biology Commons Recommended Citation Carmichael, Christopher Knight, "PHYLOGEOGRAPHY OF THE INDONESIAN WATER PYTHON, LIASIS MACKLOTISSP. (SQUAMATA: BOIDAE: PYTHONINAE): A COMPARATIVE APPROACH TOWARD RESOLVING PHYLOGENY" (2007). Dissertations. 1273. https://aquila.usm.edu/dissertations/1273 This Dissertation is brought to you for free and open access by The Aquila Digital Community. It has been accepted for inclusion in Dissertations by an authorized administrator of The Aquila Digital Community. For more information, please contact [email protected]. The University of Southern Mississippi PHYLOGEOGRAPHY OF THE INDONESIAN WATER PYTHON, LIASIS MACKLOTISSP. (SQUAMATA: BOIDAE: PYTHONINAE): A COMPARATIVE APPROACH TOWARD RESOLVING PHYLOGENY by Christopher Knight Carmichael A Dissertation Submitted to the Graduate Studies Office of The University of Southern Mississippi in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy Approved: August 2007 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. COPYRIGHT BY CHRISTOPHER KNIGHT CARMICHAEL 2007 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. The University of Southern Mississippi PHYLOGEOGRAPHY OF THE INDONESIAN WATER PYTHON, LIASIS MACKLOTISSP. (SQUAMATA: BOIDAE: PYTHONINAE), OF INDONESIA’S LESSER SUNDAS ARCHIPELAGO: A COMPARATIVE APPROACH TOWARD RESOLVING PHYLOGENY by Christopher Knight Carmichael Abstract of a Dissertation Submitted to the Graduate Studies Office of The University of Southern Mississippi in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy August 2007 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ABSTRACT PHYLOGEOGRAPHY OF THE INDONESIAN WATER PYTHON, LIASIS MACKLOTI SSP. (SQUAMATA: BOIDAE: PYTHONINAE), OF INDONESIA’S LESSER SUNDAS ARCHIPELAGO: A COMPARATIVE APPROACH TOWARD RESOLVING PHYLOGENY by Christopher Knight Carmichael August 2007 Liasis mackloti is currently recognized as three subspecies(L. m. savuensis, L. m. dunni and L. m. mackloti) inhabiting several islands of the Outer and Inner Banda Arc of islands of Indonesia’s Lesser Sundas Archipelago. We used partial mitochondrial cytochromeb sequences and morphological character states to examine and resolve the phylogenetic relationship of these three subspecies. Maximum likelihood and parsimony analysis showed thatLiasis fuscus is the sister lineage to theL. mackloti ssp. complex. There is strong support for the recognition of three clades that are delineated by savuensis, dunni and mackloti. The current subspecies taxonomic scheme accurately delineates the evolutionary distinctiveness of the various insular populations based on congruent topologies revealed by both the molecular and morphological data sets. The lineage is monophyletic, and each of the three subspecies differs from the other two both morphologically and genetically. Given the morphological and genetic distinctiveness of each taxon, we believe there is substantial empirical evidence and justification for the elevation of the three species to full species status. Morphologically, there are several characteristics that differentiate the three subspecies including the location of the heat ii Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. sensing pit on the labial scales L.( m. mackloti has heat sensing pits on labial scales 9, 10, 11 and 12, L. m. savuensis on labial scales 10, 11, 12 and 13, and L. m. dunni on 9, 10, 11, 12, and 13), number of loreal scales (L. m. savuensis andZ. m. dunni have 1 while L. m. mackloti have 2), midbody scale row count (Z.m. savuensis has 46-49, Z. m. dunni has 52-63, and Z. m. mackloti has 59-65), and number of postanal scales (Z. m. savuensis has 65-76, Z. m. dunni has 85-89 and Z. m. mackloti has 88-93). In terms o f cytochrome b mitochondrial DNA sequence data, there is the greatest sequence divergence between Z. m. savuensis andZ. m. mackloti (5.6%), followed by Z. m. savuensis andZ. m. dunni (5.2%) and Z. m. mackloti and Z. m. dunni (2.6%). There was no sequence divergence between the three insular poulations studied of Z. m. mackloti. Both molecular and morphological data sets reveal similar patterns of phylogeny. Their distribution and evolution appears to have been shaped by combined effects of dispersal and vicariance. We conducted pheromone trailing experiments to investigate the level of geographic variation present in this behavior in the macklot’s python,Liasis mackloti (Serpentes: Boidae). Three subspecies (Z. m. mackloti, L. m. savuensis, and Z. m. dunni) are currently recognized and are found on several of the Lesser Sundas islands of Indonesia including Sawu, Wetar, Timor, Semau and Roti. Based on prior studies, three clades have been delineated (Wetar, Sawu, and a Timor-Semau-Roti clade). The three subspecies display remarkable interpopulational (morphological, genetic and behavioral) variation but only slight intrapopulational polymorphisms. A modified Y-maze was used to test homotypic and heterotypic and male and female preferences both within and between insular populations. The results of this study of the Z.mackloti complex indicate that during the breeding season male pythons were able to discriminate between iii Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. homotypic and heterotypic odors within each of the clades (P<0.05). However, male pythons from Timor, Semau and Roti were unable to differentiate pheromone trails produced by females from these three islands (P>0.05). MaleL. m. dunni from the island of Wetar are generally longer than females and also exhibited specificity toward homotypic male trails. This homotypic male trailing behavior, in addition to several male-male combat interactions observed during the study suggests that males from Wetar may attempt to displace males for access to females. Pheromone trailing discrimination is an important pre-zygotic reproductive isolating mechanism that may have played an important role during speciation. We also present the role of dispersal and vicariance in shaping current patterns of geographic variation. Courtship behaviors were compared between three subspecies of the Indonesian water python,L. m. mackloti, L. m. savuensis, and L. m. dunni, to elucidate patterns of geographic variation in male courtship behavior in standard laboratory conditions. The three subspecies are found on a series of island that are part of the Outer Banda Arc and Inner Banda Arc of the Lesser Sundas Archapelago. The insular populations ofL. mackloti ssp. within this archipelago indicate that separation between islands during the Pleistocene played a role in determining current assemblages and variation within species. The islands o f eastern Indonesia form biogeographic subregions that have relatively high levels of endemism and evidence o f incipient speciation as a consequence of changes in sea-levels and climate during the Pleistocene. Two predominant models to explain the biogeographyL. of mackloti ssp. include vicariance and dispersion, both of which likely played an important role in shaping the distribution pattern of these pythons that we currently observe and are discussed in this chapter.L. mackloti generally adheres iv Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. to the triphasic schema including tactile-chase, tactile-alignment, and intromission and coitus. However, the use of the pelvic spurs during tactile-chase and tactile-alignment is a unique boid-typical motor pattern and likely plays an important premating reproductive isolating mechanism that prevents interbreeding between conspecific individuals. Behavioral sequence chains were derived from videotaped experimental tests and subjected to transition analysis. We observed variation in both the frequency of occurrence and the sequence of the principal courtship behaviors and, when compared statistically, most of these behaviors differed between populations. We observed geographically unique island patterns in the sequence in which male courtship behaviors are displayed. Our data yielded the following information: 1) the courtship sequence in all three subspecies is not random; 2) the sequence ofL. m. savuensis is much less complex than that ofL. m. mackloti and L. m. dunni', and 3) the tactile pelvic spurring target sites used by males on the females’ dorsum is different between the three subspecies. We also determined whether sexual isolation among selected populations existed. In the first study, we performed male-female reciprocal crosses of pythons between two different insular populations and measured mating success and whether eggs were produced. The results of this test reveal that sexual isolation occurs between the three defined subspecies. The behavioral differences in regards to courtship sufficiently delineate the subspecies and no