Buses will also replace ScotRail trains between Cowdenbeath and Leven from Saturday 24 to Tuesday 27 October.
Network Rail teams will spend 18 days carrying out some of the biggest and most complex parts of the project, including rebuilding bridges, track changes and upgrading signaling equipment.
The main road in Thornton will also be closed from Monday 19 October 2026 to 31 March 2027 to allow for bridge works.
The work is part of the Scottish Government’s £311.5 million investment to electrify 118km of railway in Fife and introduce battery-electric trains.
Some train services will be affected (Image: NQ)
The investment will reduce reliance on diesel trains, helping to deliver quieter, more sustainable journeys, while supporting Scotland’s Net Zero ambitions.
Before the overhead power lines can be installed, parts of the railway have to be changed to make room for the new equipment.
During this phase of work, bridges near Bennochy Road in Kirkcaldy, Muriespot near Coaltown by Balgonie and Main Street in Thornton will be demolished and rebuilt, while the bridge on Cardenden Road in Cardenden will be upgraded.
Crews will also continue to install foundations for the trackside pylons that support the overhead lines, and continue to make track and signal improvements.
Michael McArthur, project manager at Network Rail, said: “This is a really significant stage of the project and an opportunity to complete a substantial amount of work in a relatively short space of time.
“Teams have been preparing for this closure for months, carefully planning every activity to make the best use of the time available. By the end of these 18 days, we have completed some of the biggest and most important work to prepare the railway for the future.
“I would like to thank passengers and local communities for their patience. We do not take the need for closures lightly, but making these changes now means we can move towards a quieter, greener and more reliable railway for Fife sooner.”
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Mark Ilderton, ScotRail’s service delivery director, said: “This is the next important phase of work to electrify the railways across Fife, and it will mean significant changes to services for customers traveling through the area.
“We will keep people where possible, with trains running on alternative routes and replacement buses serving affected stations.
“Travel arrangements will change at various stages throughout the works, so we ask customers to check their journey before departure and allow some extra time to travel.
“We appreciate everyone’s patience while this important investment in Scotland’s rail continues.”