The mitochondrial genome of the little spotted kiwi : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Molecular Biosciences, Massey University, Palmerston North, New Zealand
The complete mitochondrial genome of the little spotted kiwi, Apteryx owenii, has been sequenced
and submitted to GenBank (acquisition number GU071052). A method was first developed to
extract pure mitochondrial DNA from one millilitre of fresh blood; as birds have nucleated
erythrocytes/red blood cells. The mitochondrial DNA was extracted from the isolated intact
mitochondria and the genome was amplified by long-range PCR as 1- 4kb overlapping fragments.
These fragments then became templates for the second, short-range, overlapping PCR
amplifications and subsequent DNA sequencing. This procedure was first trialled using two
millilitres of chicken blood before being successfully applied to the kiwi blood. The complete
mitochondrial genome of the little spotted kiwi is ~ 17,020bp long. The gene order is the standard
avian gene order first reported for chicken mitochondrial DNA. Phylogenetic relationships show
the kiwi is part of the Australasian ratite group with the emu and cassowary. This mitochondrial
sequence has been used as part of a larger study of the relationships of other ratite birds (such as
moa, emu, cassowary, rhea and ostrich) and the weakly flying tinamou of South America. The
implication of this analysis is that the ancestral paleognath was probably flying and that flight was
lost multiple times during ratite evolution.