Holybourne is a village in the civil parish of Alton, in the East Hampshire district of Hampshire, England. It is 1.3 miles (2.2 km) northeast of the centre of Alton, is contiguous with it and shares its A31 bypass. The nearest railway station is also in Alton.

The village has a population of around 1,500 and is where Treloar School is located. Holybourne has a pub, The White Hart, and a small store.

Holybourne is recorded in the Domesday Book of 1086 as Haliborne and appears in 1418 as Halybourn.

The name is thought to be derived from the Old English Haligburna which means sacred stream, referring to the small stream whose spring is near Holybourne Church, whence it runs through the village.

English author Elizabeth Gaskell (1810–1865) bought a house in Holybourne in 1865. She died suddenly when visiting the house on 12 November 1865.

Being located close to the former RAF Lasham airfield, gliders are often spotted in the sky.

RAF Odiham is home to the British Chinooks, and has a flight path over the village at a low level, allowing good views of the aircraft.

Cuckoo's corner, near the end of the village, has the remains of a Roman road. Finds have been located in surrounding fields and cricket pitches.