The awards covered 15 categories, including Large Visitor Attraction, and had 45 national finalists.
RHS Garden Wisley head of site James Rudoni said: "We are delighted to receive this award and proud that it reflects the hard work that all our staff has put in. Last year we saw record numbers of visitors to Wisley, attracted by the depth and diversity of our plant collection and varied programme of events. This year looks even more promising and we are continuing to make changes and improvements to our garden.
"Winning the Large Visitor Attraction of the Year' in the 2010 Enjoy England Awards for Excellence is really important to us because we feel it will raise the profile of RHS Garden Wisley and help the RHS in our key charitable purpose of introducing more people to the wonders of gardening. But also that everyone will see that we have so much to offer in one of the world's finest settings and come and experience a great day out at Wisley."
RHS Garden Wisley has planted a sunflower maze this year and staff are building a bird hide. Work continues on the new rose garden, due for completion next year.