The deal will create 50 jobs and extend Waitrose's reach to nine million new customers.
The first shop will open next month at Oxford Services on the M40, one of Europe's busiest motorway service stations, followed by a second opening at South Mimms on the M25.
The two 2,500-square-feet shops will be Waitrose's smallest and will offer more than 850 product lines.
The development follows the introduction of new Waitrose formats in market towns and smaller convenience stores.
The deal follows the retailer's announcement in January that up to 4,000 new partners were to join Waitrose with the opening of 22 new stores in 2009.
Waitrose's 200th store is about to open in Islington.
Waitrose is also launching a new oriental range of cooks' ingredients in stores.
As the nation's appetite for pan-Asian cuisine continues to grow, the supermarket is introducing more than 30 Japanese, Chinese, and Thai specialist foods.
New ingredients include Miso Paste, Organic Coconut Milk and Shaoxing rice wine - cited by Ching-He Huang on BBC 2's Chinese Food Made Easy.
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