Sally Williams, 69 and her husband Brian, 80, planted the flowerbeds near their home on Hollin House Lane, Loxley around 28 years ago, to bring colour to the quiet, rural lane and encourage wildlife to the area.
However, following a anonymous complaint to the council, the couple have now been told that both the flowers, and the stones that line the lane have to go.
Sally, who received the letter from Sheffield City Council on Saturday, says the stones have been there since the 90's, but still the council are adamant they must be removed.
She said: 'They just said it's their policy, nobody is allowed bulbs, wildflower seeds on the verges or on the roadside.
'Hollin House Lane is a quiet little backwater, there are only nine houses, it's a little lane. The residents care for the area, we've never been a problem for the council.
'We clear up dog poo, sweep the streets, encourage wildlife, we've successfully reared barn owls, everyone is very environmentally friendly.'
Sally and Brian bought the house in 1991, living there two days a week before setting up permanent residence ten years later.
The stones were originally placed to stop the erosion of the road, which had fallen into a state of disrepair before it was eventually tarmacked a few years ago.
Sally says that Amey tarmacked the stones into the road, so the couple are not prepared to try and move them.
She added: 'They've been there a long time. We also replace the dahlias every year, and maintain it all the time, as with the streets.
'It is deeper than a few flowers, in a time when things were bad we were leading wildlife and doing our bit for the environment. We increased butterflies, bees and birds in the area which was our primary aim.
'It has given people such pleasure, when they're down. It's been a talking point, we've had coffee with people. It's just sad, really sad.
'The council have categorically said no, and they are definitely getting rid.'
Neighbours of the couple have since started a petition, to keep '˜Britain in bloom' and save the roadside garden.
It has already amassed over 1,000 signatures.
Tom Finnegan-smith, head of strategic transport and infrastructure at SCC said: 'We have sympathy with the nearby home owner who has attempted to stop cars driving over the highway verge on what is a very narrow, single-track lane.
'Unfortunately the placing of flower beds, as well as stones and water barrels, on the highway outside the property, could be a danger to other road users and is a contravention of Section 149 of the Highways Act 1980. We received a complaint about this and had to take appropriate action.'
You can sign the petition to keep the garden here.