The painting, owned by the De Morgan Foundation, which has recently lost its home in London, shows Flora the Roman goddess of flowers set against a Florentine landscape enhanced with fruit, flowers and birds.
It was created by renowned artist Evelyn de Morgan, an English painter who died in 1919 and inspired by Botticelli's Primavera.
Flora is reflected in a grand Venetian mirror and, curiously, her expression changes when viewed through the ancient glass.
The De Morgan Foundation was set up in 1967 to celebrate the work of the husband and wife team of Evelyn and William De Morgan.
After Wandsworth Council declined to renew the lease on the foundations home, its collection is being dispersed to a number of temporary exhibitions across the country and the National Trust thought Wightwick Manor would be a perfect location for the painting of Flora until a new home is secured.
Conservation and engagement manager at Wightwick Manor, John Wood, said the opportunity was too good to turn down considering the buildings history.
He said: "There are so many links here, it was an obvious thing for us to do.
"Geoffrey Mander, who owned Wightwick Manor, offered to house the De Morgan Collection during the War to protect it from the danger of the Blitz, but it went elsewhere.
"Geoffrey's mother was called Flora and the painting was created in 1894, the year after the Great Parlour was constructed.
"We just had to say yes when offered to look after the painting."
The painting will be available to view until August and is a worthy addition to the collection of fascinating works by Pre-Raphaelite artists at Wightwick Manor.
Lynn Hawthorne, learning and visitor experience co-ordinator at the Victorian manor house, said: "This is a really exciting art loan for us and for the City of Wolverhampton.
"Visitors will be able to spend time admiring the painting and the ceramics from the De Morgan Foundation, as well as our other treasures, so we look forward to sharing our enthusiasm with them.
"For a few months at least, Wightwick has its very own Mona Lisa."