The Duchess of Cornwall met Pope Benedict XVI when the pontiff offered the couple a private audience at the Vatican.
The royal couple arrived Sunday and are staying with President Napolitano at the Quirinal Palace, before travelling to Venice today and then to Germany.
Under Vatican protocol the Prince was expected to first hold talks with the Pope before being joined by the Duchess.
However, the royal couple were received together for the 15-minute audience, with the Duchess wearing a black silk dress designed by Anna Valentine and a matching veil or mantilla. The prince also held talks with Cardinal Tarcisio Bertone, the Secretary of State, for over half an hour.
A Vatican communique said there had been "cordial discussions" and an "exchange of views" on "the human promotion and development of peoples, environmental protection and the importance of inter-cultural and inter-religious dialogue for furthering peace and justice in the world".
There was no immediate indication of whether the Prince had reinforced an invitation to Pope Benedict to visit Britain, extended in February by Gordon Brown, the Prime Minister.
The prince, who was wearing a dark blue suit with a black tie, gave the Pope 12 dessert plates decorated with paintings of flowers from his Highgrove estate, adding: "I don't know whether these will be of any help to you," as he handed them over.
He also gave him a framed and signed photograph of himself and his wife.
The Pope, who thanked his guests in English, gave the royal couple an etching depicting the pre-sixteenth century St Peter's Basilica, before it was redesigned by Michelangelo and other Renaissance masters, and a set of papal medals.
Prince Charles told the Pope he was "most touched" by the gifts. As he was leaving he was heard to say - in an apparent reference to John Paul II, Benedict's predecessor - "he was such a wonderful man, we miss him terribly".
The prince met Pope John Paul II in 1985 when he visited Rome with Diana, Princess of Wales. His meeting with Pope Benedict was his first audience at the Vatican since his divorce from in 1996 and her death a year later.
The prince and the duchess, formerly Camilla Parker-Bowles, were both divorcees before they married in a civil ceremony in 2005 followed by a service of blessing at Windsor. Their wedding was delayed by a day so that Prince Charles could attend the funeral of Pope John Paul II. He has made many visits to Italy, but his last official visit was five years ago.
At one point the prince presented Arthur Edwards, the royal photographer of the Sun newspaper, to the Pope, saying: "He's a marvellous man. He has been following me for 35 years, and he's a Catholic".
Before the meeting at the Vatican, the prince delivered a keynote speech on climate change to an invited audience at the Chamber of Deputies of the Italian Parliament, and later joined a meeting of Italian business leaders to discuss environmental issues.
He also held a reception to promote "Slow Food" culture of local sourcing and organic production. He has brought with him beef from his Highgrove estate and lamb from Wales, served at the reception by two top Italian chefs, Andrea Berton and Carlo Cracco.
In a front page article in La Repubblica today. the prince called for a "new Renaissance", saying that tackling climate change and "sustainable agriculture" were vital "if we want to protect our planet for future generations". He said scientific predictions on global warming were "ever more alarming" and called for "collaboration between communities, nations and continents" to provide a "global response".
The prince said: “I believe strongly that the response we make to the challenge before us will define our era. Just as the Italian Renaissance defines for so many of us in Europe a reawakening of cultural and intellectual identity at the end of the medieval period, so our actions today could be remembered as a renaissance in how we live, and the beginning of a new era in the effective stewardship of our planet.”
He added: “While in Italy, I will attempt to set out some of the forms that such co-ordinated action might take and outline some of the principles which might help us in our quest for greater environmental sustainability. But whatever the specifics, the crucial point I want to emphasise is that we need to act now - with real urgency. I wish it wasn’t the case and it was possible to continue with business as usual. But, unfortunately, it is the case and, if we continue as usual, then we will bequeath a terrible, poisoned legacy to our children and grandchildren."
He said he sincerely hoped "that our two countries, whose histories have been intertwined for over 2,000 years, will once again join with common purpose to lead the world away from disaster and towards a fair, safe and sustainable future.”
The prince also spoke of his sorrow over the earthquake which struck the Abruzzo region on April 6 and which claimed nearly 300 lives.
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