Overall it is the fourth time Bents Garden & Home - based in Glazebury, Cheshire - has held the award with the last three titles coming in three consecutive years.
Bents were once again presented with the title by the Garden Centre Association (GCA) following the annual audit which records scores from 200 garden centres throughout the UK.
MD Matthew Bent said: "To be acknowledged as the best in our industry for the third successive year is a huge honour and fantastic acknowledgement of the hard work and commitment put in by each and every one of our 200 plus colleagues."
GCA chief executive Gillie Westwood said:
"We are delighted for Bents. It is a fantastic achievement to retain the title for a third consecutive year which really is a first within our industry and even more impressive when up against such strong competition."
The award also sees Bents being acknowledged as a Garden Centre of Excellence, one of only 13 in the country.