Last year Sheffield Council used 1,750 litres of Glyphosate on pavements, verges and parks but campaigners say it poses a risk to the public, environment and horticultural workers.
It has been banned in Holland, Denmark, Sweden and France and by some councils around the UK such as Trafford, Brighton, Bristol and Croydon.
Graham Wroe will present the petition to the full council meeting at Ponds Forge and because it has more than 5,000 signatures, it will automatically trigger a debate.
There will be a rally outside the meeting at 1pm on Wednesday, July 7, with street theatre from Act Now and speeches from Coun Alison Teal, Executive member, Mr Wroe and Extinction Rebellion.
Act Now is encouraging audience participation and inviting people to come dressed as insects.
Mr Wroe said: “At the June meeting the council declared a nature emergency so there is a big expectation for them to start putting this into practice by protecting biodiversity in the city.
“In Sheffield we spray ridiculous amounts of poisonous Glyphosate on our streets, parks and playgrounds every year.
“It is unnecessary, unsightly – leaving dead vegetation at the side of verges and walls – and dangerous, not just to insects and birds but to humans and pets too.
“There are plenty of safe alternatives, from letting the wild-flowers grow, hand weeding, mechanical weeding and spraying hot foam or acetic acid.”