"The Palestinians must abandon their refusal to recognise the right of the Jewish people to their national state," he said in a speech at Bar Ilan University near Tel Aviv.
Such recognition was "a condition for reaching an agreement at the end of negotiations, but not for launching them", he said.
Peace talks between Israelis and Palestinians were relaunched in July under the auspices of the United States after nearly three years of impasse.
The direct talks, which are being held in Israel and the West Bank under a US-imposed media blackout, have been set to last nine months, and have so far yielded no concrete results.
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"Are you finally ready to recognise the Jewish state, the national state of the Jewish people?" he asked, directly addressing Palestinian leaders, deploring that their response so far to that question has been "no".
"So long as the Palestinians do not recognise this right, there will be no true peace," the premier warned his audience at the Bar Ilan auditorium.
It was at the same place that Netanyahu, leader of the right-wing Likud party, first announced his support for the two-state solution in a dramatic 2009 speech.