Major League Soccer – 8 Teams
The top six teams based on points in the 2008 MLS standings are automatic qualifiers (six) for the 2009 tournament. The remaining clubs will compete in play-in matches to determine the final two berths.

Automatic Qualifiers: Chicago Fire, Chivas USA, Columbus Crew, Houston Dynamo, Kansas City Wizards, New England Revolution
Eligible Teams for Qualification: Colorado Rapids, DC United, FC Dallas, Los Angeles Galaxy, New York Red Bulls, Real Salt Lake, San Jose Earthquakes, Seattle Sounders

USL First Division – 8 Teams
All eight US-based teams in the USL First Division will participate in the tournament.

USL-1 Teams: Austin Aztex, Carolina RailHawks, Charleston Battery, Cleveland City Stars, Miami FC Blues, Minnesota Thunder, Portland Timbers, Rochester Rhinos

USL Second Division – 8 Teams
All eight US-based teams in the USL Second Division will participate in the tournament.

USL-2 Teams: Charlotte Eagles, Crystal Palace Baltimore, Harrisburg City Islanders, Pittsburgh Riverhounds, Real Maryland Monarchs, Richmond Kickers, Western Mass Pioneers, Wilmington Hammerheads

Premier Development League – 8 Teams (qualifying through May 25)
Eight teams will participate from the PDL, where 61 U.S.-based teams are eligible to qualify for the U.S. Open Cup. The top team in each of the eight qualifying groups (their league divisions) will advance based on the results of four pre-designated league games will advance to the tournament.

U.S. Adult Soccer Association Regional Qualifiers – 8 Teams (qualifying through May 25)
Eight representatives from the U.S. Adult Soccer Association will also qualify for the 2009 U.S. Open Cup. Two teams qualify from each of the USASA’s four regions (I, II, III & IV), where qualifying tournaments will be staged on a statewide and then regional basis.