Israel's foreign ministry protested to Jordan on December 30 after a minister was pictured stepping on an image of the Israeli flag while entering a meeting.
Jordanian minister of state for information Jumana Ghneimat was pictured last week stepping on the flag design when entering a meeting at a trade union complex.
The image of the flag has been affixed to the floor just inside the entrance to the complex for several years to protest Israel's occupation of the Palestinian territories and Jordan's normalised ties with the Jewish state, union officials say.
Israel called Jordan's ambassador in for clarifications on Sunday, the foreign ministry said.
"The foreign affairs ministry considers the incident in which a Jordanian minister insulted the Israeli flag in Amman as serious," a statement said.
Jordan's foreign ministry later responded, reaffirming "the (Jordanian) government's respect for the peace treaty with Israel".
Jordan had told Israeli officials "that the building was private property and that the minister entered via the main entrance to attend an official meeting", said spokesman Majed al-Qatarneh.
Jordanian Prime Minister Omar al-Razzaz was reported to have entered the building through a side door.
Jordan and Egypt are the only two Arab countries to have diplomatic relations with Israel.
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