This is a list of airlines that are based in Ontario which have an air operator's certificate issued by Transport Canada, the country's civil aviation authority.
| Airline | Image | IATA | ICAO | Callsign | Hub airport(s) or headquarters | Notes |
|---|---|---|---|---|---|---|
| 30000 Island Air |  | Parry Sound Harbour | 2000? - ?, charter rental and leasing service air operator based in Parry Sound, Ontario using de Havilland Canada DHC-2 Beaver | |||
| Air Ontario |  | GX | ONT | ONTARIO | London | 1987 - 2001, to Air Canada Jazz |
| Air Toronto | CS | CNE | CONNECTOR | Toronto Pearson | 1984-19991 | |
| AllCanada Express |  | CNX | CANEX | Toronto Pearson | 1992 - 2005 | |
| Austin Airways |  | AAW | Timmins/Victor M. Power | 1934 - 1987, to Air Ontario | ||
| Canada 3000 |  | 2T | CMM | ELITE | Toronto Pearson | 1988 - 2001, formerly Canada 2000; Canada 3000 Cargo sold to Cargojet and 2005 revival failed |
| City Express |  | Peterborough | 1971 - 1991, founded as Air Atonabee 1971 | |||
| Great Lakes Airlines | Sarnia Chris Hadfield | 1958 - 1983, to Air Ontario | ||||
| Intair |  | ND | INT | INTAIR | Toronto Pearson | 1989 - 1991, established by City Express as a successor to Skycraft Air Transport |
| Millardair |  | Toronto Pearson | 1962 - 1990, continued as aircraft maintenance and servicing firm Millard Air Incorporated (Millardair MRO) 1990-2012 | |||
| NAC Air | HMR | HAMMER | Thunder Bay | 2000 - 2008, North American Charters was 100% First Nations owned | ||
| NorOntair |  | NOA | NORONTAIR | Sault Ste. Marie | 1971 - 1996, airline operations of Ontario Northland Transportation Commission | |
| Odyssey International |  | OL | ODY | ODYSSEY | Toronto Pearson | 1988 - 1990, name, aircraft and some employees were merged with Nationair, and operated as a separate division of Nolisair for a short period of time |
| Ontario Central Airlines | NUN | NUNASI | Kenora | 1947 - 1991?, became Nunasi-Central Airlines in 1984 and then to Nunasi-Northland Airlines in 1987 | ||
| Ontario Express | 9X | OEL | PARTNER | 1980s - 1998, to Canadian Airlines | ||
| Pem-Air | PD | OEM | PEM-AIR | Pembroke | 1970 - 2002 | |
| Roots Air | 6J | SSV | SKYTOUR | Toronto Pearson | 2000 - 2001, part of Roots Canada, ceased operations when ownership acquired by Air Canada | |
| Skycraft Air Transport |  | Oshawa | 1977 - 1989, reincarnated in 1989 and operating until 1994. | |||
| Skyservice Airlines |  | 5G | SSV | SKYTOUR | Toronto Pearson | 1986 - 2010, differs from existing Skyservice Business Aviation - same owners |
| Soundair | SDR | SOUNDAIR | Toronto Pearson | 1973-1990, owned Odyssey International and Air Toronto. Previously Owen Sound Air Services. DBA Soundair Express. | ||
| Starratt Airways | Hudson, Kenora District | 1932 - 1942, to Canadian Pacific Airlines | ||||
| Vacationair |  | Toronto | 1988 - 1990, initiated by Gray Coach Lines | |||
| Vision Airways Corporation | V6 | VSN | VISION | Timmins/Victor M. Power | ? - 1994 | |
| Vistajet | VJT | VISTA | Ottawa, Toronto, Windsor | 1997 - 1997 | ||
| Zoom Airlines |  | Z4 | OOM | ZOOM | Ottawa Macdonald–Cartier | 2002 - 2008, named acquired for new XPO Airlines 2009 |
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