A total of £6.9m is being invested to develop the National Cycle Network for 2018/19.
The move will fund a series of infrastructure improvements to make it easier and safer for people to walk and cycle across Scotland.
Work includes changes to the last two sections of the Caledonia Way (National Cycle Network Route 78), between Campbeltown and Inverness, and the development of a new route between Doune and Callander.
The funding will also help the completion of Soldiers Bridge in Fort William and enhance existing NCN routes across central Scotland.
Mr Matheson said: "Extra funding will support further development of the National Cycle Network and address current gaps on the route. This will help make Scotland even more attractive for our growing cycle tourism sector which is contributing to our economy, particularly in rural areas.
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"It will also help make Active Travel more appealing so people feel motivated to leave their car at home and go by bicycle or on foot for the benefit of their health and our environment, contributing towards our ambitions for an Active Nation.
"The Scottish Government has committed, in the Programme for Government, to maintain record levels of investment for the remainder of this parliamentary term for cycling and walking projects, including more and better infrastructure, behaviour change projects and other innovative ideas."
Grace Martin, Deputy Director at Sustrans Scotland added: "The National Cycle Network is vital for every day journeys, leisure trips and tourism in Scotland. The Scottish Government's additional investment in the network highlights its commitment to creating a sustainable and active Scotland.
"Sustrans will be releasing the results of its National Cycle Network Review in October 2018, which will help inform how the network is developed and improved in Scotland over the coming years.
"This additional funding will help create better connected communities and encourage people to walk and cycle more on the journeys they make every day."
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