The project will create grassland habitat along the entire route of the John Muir Way and almost 2 miles either side, providing foraging habitat for pollinating bees, hoverflies and other insects but also homes for other wildlife.
Opportunities have been identified at school grounds, cemeteries, hospitals, golf courses and public parks and sites currently designated for conservation including Local Nature Reserves and Wildlife Reserves.
Suzanne Burgess, conservation officer with Buglife, said: "By identifying and mapping these new opportunities this will aid in the future development of projects that will provide real benefits to our declining populations of pollinating insects of bees, wasps, hoverflies and butterflies, as well as other wildlife that these habitats support."
Reports showing opportunities identified for each of the nine local authorities that the route of the John Muir Way passes through are now available on the Buglife website.
Buglife Scotland was the winner of the Central Scotland Green Network Ideas Fund 2015 and was awarded £5,000 for its John Muir Pollinator Way vision.
Keith Geddes, chair of the Central Scotland Green Network Trust, said: "The John Muir Pollinator Way is a fantastic project which will play a pivotal role in encouraging habitat connectivity for pollinators along the route. It will enhance the biodiversity and natural heritage of the area and bring environmental benefits to communities along the Way."
Ron McCraw, John Muir Way router developer for Scottish Natural Heritage, said: "We're very pleased to be supporting this project, helping to make it more attractive to visitors and local people using the route, and encouraging practical involvement by communities."
Through the project events have also been run promoting the importance of grassland meadows for pollinators, looking at how to create and manage these habitats and how to identify the pollinators that use the sites. Over John Muir's birthday week in April 2016, a total of 20 events were run engaging over 280 people.