More than 120 trees are to be removed from Northbourne Avenue before Christmas as part of construction on the light rail.
Work will begin on Monday in the first of three stages of tree removal for the transport project.
The first stage is expected to be completed by December 23, while the second and third stages will be carried out in the new year.
Trees will be removed from Northbourne Avenue and sections along Flemington Road and the Federal Highway.
The starting date for the tree removal was expected to be October but was pushed back as the ACT government had not finished the "design development process".
Transport and City Services Minister Meegan Fitzharris said the tree removal would allow for the track and light rail stops to be built.
"Some trees have been removed already due to recent damage they sustained from storms and poor health as identified in our ongoing monitoring of this important transport route," she said.
"Some trees are also reaching the end of their lifespan along the Northbourne Avenue transport corridor."
More than 40 trees were removed from the major road in April after they were judged to be unsafe, while the ACT government ordered the removal of 17 trees in October after storms.
The second and third stages of the removal involves trees being cut down from the road's median strip.
It's estimated 358 trees between Alinga and Antill streets will be removed.
While the chainsaws and tree cutters will descend on Northbourne Avenue, more trees are expected to be planted in coming months.
More than 800 trees have come in from Victoria to be used for the project and are maturing at Yarralumla Nursery.
The new trees are expected to be planted in late spring or early summer next year.
"Overall, more trees will be replanted as a result of the project," Ms Fitzharris said.
"We are also working with local organisations to reuse and recycle some of the removed trees."
Lanes will be closed along Northbourne Avenue while the trees are being removed.
"Safety is also a key component for workers, pedestrians, motorists and other road users during the staged tree-removal program," Ms Fitzharris.
The safety message comes just days after a light rail worker was thrown two metres by an electric shock.
An investigation into the incident has been launched by Worksafe ACT.