Bettys at Harlow Carr will be closed for four weeks while it undergoes a refurbishment.
The café and shop will be closed from September 7 until October 5 for the refresh.
Devoted Bettys fans can visit branches in nearby Harrogate, Ilkley, York and Northallerton, or order favourites online.
When the tea room reopens in October, visitors can expect a 'relaxed sense of elegance', the full menu, and scenic views across the gardens.
A new 'Bettys To Go Window' will launch at the garden entrance, offering an easy way for visitors to pick up picnic essentials.
Table bookings will also be introduced to help visitors, especially families planning later afternoon visits.
Nicky Sellers, branch director at Bettys, says: 'We’re excited to be investing in Bettys Harlow Carr and creating an even more welcoming experience for our customers.
'The café tea rooms are a very special part of a visit to RHS Garden Harlow Carr, and we look forward to welcoming everyone back in early October.'
Liz Thwaite, head of site at RHS Garden Harlow Carr, adds: 'We’re delighted to see the refurbishment of Bettys Harlow Carr and to continue our long-standing partnership with such an iconic Yorkshire brand.
'We know Bettys is a much-loved part of a visit to the garden, and we’re excited to welcome guests back to a refreshed café this autumn.
'While the work takes place, RHS Garden Harlow Carr remains open as usual, with plenty for visitors to enjoy, including a range of additional catering and refreshment options available across the garden throughout the closure period.'
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