Southern Rail, which runs trains between England's south coast and London, warned of severe disruption as it cancelled all of its 2,284 services after workers launched three days of industrial action.
Around 1,000 drivers are involved in the strike which will affect around 300,000 passengers, including those travelling to London's Gatwick airport.
A 48-hour walkout began at midnight Monday with a further 24-hour strike planned for Friday and another six days of action in January.
Union leaders have raised concerns about safety and possible job losses, although the rail company has said staffing levels will not be affected.
Passengers on routes from Brighton and other key commuter towns in southern England have already faced months of disruption to services in a series of walkouts that began in April.
The latest strike comes after the train operator's owners, Govia Thameslink Railway, lost a legal bid to halt the action.
The shutdown is expected to cause the worst disruption on Britain's railways since a series of strikes by signal workers in 1994.
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