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The photograph shows the young prince wearing a knitted blue jumper, shirt and blue shorts.
He is seen sitting on top of a pile of leaves and grass, holding on to the wheelbarrow as Princess Catherine, 41, laughs.
The image is one of two snapped on the Windsor Estate earlier this month by photographer Millie Pilkington.
The second is a close-up of the beaming prince, whose elder siblings are Prince George, nine, and seven-year-old Princess Charlotte.
The three siblings live with their parents at Adelaide Cottage in Windsor's Home Park and all attend the private Lambrook School near Ascot in Berkshire.
They were featured in a previously unseen photo of the late Queen released on what would have been her 97th birthday, showing her smiling with some of her grandchildren and great-grandchildren, captured by the Princess of Wales at Balmoral.
Louis did not join his older brother and sister at the state funeral of his late great-grandmother at Westminster Abbey in September - and is thought to have been considered too young to attend the service with his parents.
However he is expected to accompany George and Charlotte in the procession from the abbey which will follow the King's coronation on 6 May, reports suggest.
All three siblings joined the King and Queen Consort together with other members of the Royal Family for the first Easter Sunday service of Charles reign, at St George's Chapel at Windsor Castle on 9 April.
Prince Louis famously stole the show on the Buckingham Palace balcony at the flypast for the Queen's Platinum Jubilee in June last year.
The youngest Cambridge child pulled faces and covered his ears as the Red Arrows soared overhead.
Read more:
Touching words Prince Louis said after Queen's death revealed
Prince Louis giggles in official photo to mark his christening
Prince Louis, who is fourth in line to the throne, was born on 23 April - St George's Day.
He was christened Louis Arthur Charles at 11 weeks old by the Archbishop of Canterbury, Justin Welby, at the Chapel Royal in St James Palace, watched by friends and family.