Shop owners who sell spray paint to young people under the age of 16 could now be fined as much as £1,000.
Shopkeepers could be fined up to £1,000 in the new anti-graffiti drive
The fine threat is the latest measure to come into force as a result of the Scottish Executive's flagship Anti-social Behaviour Bill.
Retailers will also have to put up a notice in a "prominent" position in their shops, setting out the new law.
Deputy Justice Minister Hugh Henry called the battle against graffiti a "key" part of the executive's campaign.
He said: "These new measures recognise the vital role that responsible retailers play in regulating access to potentially damaging materials.
"The ban on the sale of spray paint to under-16s by itself will not prevent graffiti, but it is a practical measure to control the misuse of spray paints by children.
"Education and crime prevention initiatives will continue to have a major role to play in building respect for the places we live and work in.
'No place for vandalism'
"But these new measures will make sure that when opportunity knocks, the shop door is closed."
"Preventing vandalism and graffiti is a key part of the Scottish Executive's strategy to promote safe, secure and attractive communities.
"The message should be clear - there is no place for vandalism in Scotland.
The most controversial provisions of the Anti-social Behaviour act came into force last month.
The measures aim to make communities safer
The act of parliament gives the police powers to disperse groups of youths in designated trouble spots.
It also extends anti-social behaviour orders to younger children aged 12 to 15 and gives the courts powers to close down drinking dens or clubs where drugs have been discovered.
The anti-social behaviour measures were demanded by backbench Labour MSPs, under pressure from their constituents to put order and respect back into their communities.
They were pushed through parliament, despite criticism from opposition parties and some senior police officers and social workers.