A Brief Molecular Phylogeny focused on Twenty-Nine Species within the Testudinidae Family using Mitochondrial Cytochrome b Gene Christine Chessler.

A Brief Molecular Phylogeny A Brief Molecular Phylogeny focused on Twenty-Nine Species focused on Twenty-Nine Species within the Testudinidae Family within the Testudinidae Family

using Mitochondrial Cytochrome b using Mitochondrial Cytochrome b GeneGene

Christine ChesslerChristine Chessler

IntroductionIntroduction Family TestudinidaeFamily Testudinidae

Land-based tortoisesLand-based tortoises Widely distributed & Widely distributed &

recognized recognized Majority in Africa & AsiaMajority in Africa & Asia Few in America & EuropeFew in America & Europe

Evolved ~200 mya from Evolved ~200 mya from the Anapsid reptilesthe Anapsid reptiles

Only living reptile grouping Only living reptile grouping to retain Anapsid skull to retain Anapsid skull structurestructure

Earliest fossils found in Earliest fossils found in AsiaAsia

Members classified by:Members classified by: High-domed shellsHigh-domed shells

Exceptions exist. Exceptions exist. Stout scaly limbsStout scaly limbs TerrestrialTerrestrial

IntroductionIntroduction

Researchers feel Researchers feel their phylogenetic their phylogenetic relationships are relationships are controversial due to :controversial due to : Limited taxonomic Limited taxonomic

samplingsampling Studies focusing on Studies focusing on

only a subset within a only a subset within a familyfamily

IE: IE: GopherusGopherus IE:IE: Geochelone Geochelone

General ObjectiveGeneral Objective I initially wanted to know more about where I initially wanted to know more about where

exactly the gopher tortoise “fit” exactly the gopher tortoise “fit” phylogenetically among the tortoise family. phylogenetically among the tortoise family.

However, as I researched the existing However, as I researched the existing phylogeny, I also wanted to address where phylogeny, I also wanted to address where the the specific cladesspecific clades were situated among were situated among Testudinidae and also which clade the gopher Testudinidae and also which clade the gopher tortoise fit within.tortoise fit within. Identify Identify synapomorphiessynapomorphies shared within clade shared within clade

membersmembers

Lastly, I wanted to group each clade based on Lastly, I wanted to group each clade based on their their known geographical known geographical locations to see if the locations to see if the clades “made sense” given the biogeography of clades “made sense” given the biogeography of each of the species. each of the species.

Materials & MethodsMaterials & Methods

Sequence SelectionSequence Selection NCBI Nucleotide Database & BLASTNCBI Nucleotide Database & BLAST Gene chosen: Cytochrome b; partial cdsGene chosen: Cytochrome b; partial cds

29 species selected within Testudinidae (set 29 species selected within Testudinidae (set organism parameters in search) + 1 outgroup organism parameters in search) + 1 outgroup species (species (Deirochelys reticulariaDeirochelys reticularia))

Sequence AlignmentSequence Alignment Seaview v.4 with MAFFT v.6.240-2 (einsi Seaview v.4 with MAFFT v.6.240-2 (einsi

setting)setting) Construction Programs UtilizedConstruction Programs Utilized

RAxML v.7.0.4 (100 bootstraps, ML tree)RAxML v.7.0.4 (100 bootstraps, ML tree) FigTree v.1.3.1FigTree v.1.3.1

Results & ConclusionsResults & Conclusions

ConclusionsConclusions SynapomorphiesSynapomorphies present to link individuals into their present to link individuals into their

respective clades:respective clades: Clade #1Clade #1 (green; (green; ManouriaManouria and and GopherusGopherus ) )

No synapomorphies? Mental glands?No synapomorphies? Mental glands?

Clade #4Clade #4 (pink; (pink; Geochelone sp. + Pyxis + DipsochelysGeochelone sp. + Pyxis + Dipsochelys)) Pyxis, G.yniphora Pyxis, G.yniphora and and G.radiataG.radiata relationship has been supported by relationship has been supported by

the synapomorphy of an indistinct fenestra postotica the synapomorphy of an indistinct fenestra postotica G.radiataG.radiata and and G. yniphoraG. yniphora have been observed to both possess a have been observed to both possess a

ventral ridge on the maxilla-premaxilla suture and keels on the ventral ridge on the maxilla-premaxilla suture and keels on the supraocciptal crest. supraocciptal crest.

Clade #5Clade #5 (blue; (blue; Indotestudo + Testudo + MalacocherusIndotestudo + Testudo + Malacocherus ) ) the processus inferior parietalis meeting the quadrate and partially the processus inferior parietalis meeting the quadrate and partially

covering the prooticcovering the prootic a ventral tip of the processus interfenestralis, in addition to the a ventral tip of the processus interfenestralis, in addition to the

presence of sutures between this process and the surrounding bones. presence of sutures between this process and the surrounding bones.

What about the other clades…and those with low(er) boot What about the other clades…and those with low(er) boot strap values?strap values?

What does the biogeography What does the biogeography reveal?reveal?

Clade #1 Clade #1 (green; (green; ManouriaManouria and and Gopherus)Gopherus)

Represent the divergence between Represent the divergence between the Asian and N. American lines the Asian and N. American lines

Crossing the Bering Strait in the Crossing the Bering Strait in the EoceneEocene

Clade #2 Clade #2 (yellow; (yellow; Geochelone sp. + Homopus Geochelone sp. + Homopus + Chersine)+ Chersine)

Endemic to AfricaEndemic to Africa Clade #3 Clade #3 (purple; some species within the (purple; some species within the

Geochelone sp + Kinixys Geochelone sp + Kinixys clade)clade) ““state of taxonomic confusion for the state of taxonomic confusion for the

past 30 years”past 30 years” Westward sea currents to S.America?Westward sea currents to S.America?

Clade #4 Clade #4 (pink; (pink; Geochelone sp. + Pyxis + Geochelone sp. + Pyxis + DipsochelysDipsochelys))

Dispersed from Africa to the Indian Dispersed from Africa to the Indian Ocean area/Madagascar by way of sea Ocean area/Madagascar by way of sea currentscurrents

Possible ModificationsPossible Modifications Obtain better bootstrap values to Obtain better bootstrap values to

validate relationships?validate relationships? More sequences/more species in analysisMore sequences/more species in analysis Use different gene?Use different gene?

Some relationships (Some relationships (Geochelone sp.)Geochelone sp.) are continually muddled and skewed are continually muddled and skewed due to inadequate knowledge on which due to inadequate knowledge on which are sub-species, individual species, or are sub-species, individual species, or all the same species originating from all the same species originating from one population…one population…

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