The mass shooting, which saw gunman Omar Mateen open fire in the Pulse nightclub in a possible Islamic State-inspired shooting, took place on Sunday and left 49 victims dead.
The local gathering was organised by equality charity Liberate and followed similar acts of solidarity held around the world.
Islanders including the Health Minister, Senator Andrew Green, and members of the gay community took part in a two-minute silence and the Rev Graeme Halls paid tribute to the victims in a prayer.
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