Recep Tayyip Erdogan, the Turkish Prime Minister, said he was "saddened" by recent comments from Paris and ruled out the prospect of a shift of position over Cyprus before the beginning of the accession negotiations.
However, France immediately stepped up its pressure on Ankara when Philippe Douste-Blazy, the Foreign Minister, said that the Turkish position was "not acceptable".
The rift could wreck plans to open the EU accession talks on 3 October and provide a diplomatic impasse for Britain, which holds the rotating presidency of the EU. It also threatens to further damage Anglo-French political relations which are already poisonous. The UK supports Turkey's ambitions to join the EU and the start of negotiations with Ankara is a central element in its presidency plans.
However, before the talks can start, all 25 EU countries - including France and Cyprus - have to agree on a document setting out the framework for discussions, which are expected to last a decade.
Ankara recognises only the breakaway Turkish Cypriot enclave in the north of Cyprus. The island was partitioned in 1974 when Turkish troops invaded following a failed coup by supporters of a union with Greece. That position has become increasingly difficult since the Greek Cypriot government in the south joined the EU last year as the sole legitimate representative of the island off Turkey's south coast.
Britain has the backing of the European Commission in arguing that recognition of Cyprus was never identified as a precondition for starting membership talks with Turkey. It says that the status of the island should be resolved within the framework of separate United Nations discussions.
Yesterday Mr Erdogan said bluntly that it is "out of the question for us to discuss or consider any new conditions with regard to October 3." He added: "We are saddened by the statements of the French Prime Minister and of President [Jacques] Chirac."
On Tuesday, the French Prime Minister, Dominique de Villepin, said it was "inconceivable" that Turkey could start talks with the EU without recognising one of its 25 member states.
M. Chirac has not publicly commented on the issue this week, but the French daily newspaper Le Figaro, reported that the President told a cabinet meeting he agreed with his Prime Minister.
Britain has been hoping that M. de Villepin's comments do not represent the definitive French position, and remains sceptical that Paris will take the political risk of blocking the start of talks. M. Chirac has so far backed Turkey's bid to join the EU but the President faces growing internal opposition over the prospect of the large, relatively poor and mainly Muslim country joining the bloc.
The row has come to a head because of the way Ankara fulfilled a demand to extend its customs union to all EU nations including Cyprus. Turkey agreed to do this last Friday, but also issued a declaration saying this did not mean recognition of Cyprus. M. Douste-Blazy said that statement was a "new element" and a "unilateral action which raises a serious problem".
In a separate development the European Commission said it had written to warn Turkey that a draft law on religious institutions does not meet EU standards on religious freedom.