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The new bioluminescent system of the dipteran Orfelia fultonii Vadim R. Viviani*; John Woodland Hastings; Therese Wilson Molecular and Cell Biology, Harvard University, Cambridge, MA, USA      *(viviani@fas.harvard.edu) Orfelia fultonii is the only known bioluminescent dipteran in America. Their larvae have anterior and posterior lanterns, which emit the bluest bioluminescence reported among insects (max= 460 nm), whereas larvae of the related mycetophilid Arachnocampa spp. have only one caudal, histologically different lantern. We carried out experiments to characterize and compare the bioluminescent systems of Orfelia and of the Australian Arachnocampa fava. An Orfelia luciferin-luciferase reaction was demonstrated by mixing, under aerobic conditions, a cold crude extract and a hot extract treated with DTT under argon. The in vitro bioluminescence spectrum is blue-shifted compared to that of A. fava (max= 460 vs. 484 nm). Unlike A. fava, Orfelia bioluminescence is not activated by ATP, but is strongly stimulated by DTT and ascorbic acid. By gel filtration we demonstrated the presence of a 130 kD luciferase and have indications for the involvement of a second component of high molecular mass (>500 kD). In the presence of luciferase, this high MW fraction activates the bioluminescence, suggesting a luciferin binding protein. The present findings indicate that the bioluminescence of O. fultonii constitutes a novel bioluminescent system. [Talk: viviani.vadimr.54231]

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