Rouhani, who is seeking a deal with the West on a nuclear standoff, called in a UN speech for the abolition of all nuclear weapons and urged Israel to join the Non-Proliferation Treaty.
Yuval Steinitz, the head of the Israeli delegation at the UN General Assembly, said Rouhani was trying "to smile his way to the bomb."
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"Instead of saying that Iran will finally comply with the Security Council resolutions, it tries to shift attention to Israel," he said.
"Israel is a very responsible country, a responsible democracy, that needs to survive and defend itself in one of the most difficult and hostile neighborhoods on the face of the earth," he said.
Rouhani is hoping for a deal to end biting sanctions imposed over Iran's sensitive uranium work, which Western officials and Israel says could be used to develop a nuclear bomb.
Israeli Prime Minister Benjamin Netanyahu has called Rouhani, a self-described moderate who swept to power in June elections, a "wolf in sheep's clothing" and refused to rule out a military strike.
Israel is widely believed to have nuclear weapons but does not publicly acknowledge its arsenal and, unlike Iran, has not signed the Non-Proliferation Treaty that allows for international regulation.
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