The main organisation representing Jewish settlers in the occupied West Bank and Gaza Strip has pledged to fight against the implementation of the roadmap.
The Yesha Council said the peace plan threatened to undermine the existence of Israel.
On Wednesday thousands of settlers and their sympathisers gathered in Zion Square in Jerusalem in protest against the plan.
More than 200,000 Israelis live on land seized in 1967
And at a news conference, a senior member of the Yesha Council, Shaul Goldstein, attacked the Israeli prime minister, saying he had lost his way and his leadership by signing up to the roadmap.
He said the demonstration would mark the beginning of a struggle against what he called Israel's surrender to terrorism.
Mr Goldstein said Jewish settlers in the West Bank and Gaza Strip should never be forced to leave so a Palestinian state could be created in these areas.
This is our homeland and we want to fight for the right that we can live here
"The Jewish people for generations dreamed about this piece of land, longed to come back and - after 2,000 years - came back to this land," he said.
"If we don't have the right to live in Bethlehem or in Hebron we don't have the right to live in Tel Aviv.
"This is our homeland and we want to fight for the right that we can live here."
Most of the protesters gathered at Zion Square were Orthodox Jews, saying no to a Palestinian state.
They do not want any kind of territorial compromise with the Palestinians and reject any dismantlement of the settlements, as spelled out by the Israeli Prime Minister Ariel Sharon in Aqaba on Wednesday.
In speeches, the settlement leaders from the West Bank and a number of right wing members of parliament have said the road map is a threat to Israel's existence.
They are also describing the roadmap as a road to destruction.
The roadmap explicitly calls for the dismantling of all settlement outposts established over the past two years.
It also demands a halt in the expansion of settlements built before then.
What ultimately will happen to the settlers is to be discussed in the final phase of the peace plan.
Around 400,000 Israelis have moved into the West Bank and Gaza Strip since the army seized the land in the war of 1967.
It is one of the most difficult issues facing the Israeli and Palestinian leaders, as they now try to push ahead with the implementation of the roadmap.