The 2x4s are simply there to spread the load out a bit and provide some support against the legs from bending outward. It's also a handy place to put a shelf, which would certainly brace the frame up a bit more. There's no reason at all to use 2x6 for the bottom frame, it's my opinion that a bigger bottom frame would actually decrease stability, since it would shift the transition higher up and move the center of gravity higher. At least until the bottom frame is wider than the length of the legs...
Somewhere in this thread (or the first part) there's a post that suggests a single 18" length of 2x4 can support up to something like 19,000 pounds or so on end before it would blowout. That doesn't account for bowing, warping, or any lateral (side to side) force, which would obviously cause premature failure, but still... if you figure there's 8 2x4's supporting these stands, that gives you 152,000 pounds of support - over 75 tons.
A safety factor of 5 (overkill) means you get over 30,000 pounds of support.
At 8.556 lb/gal sea water, 500 gallons would be 4,278 pounds - with rock and sand, you could get to (maybe) 5200 pounds. Provided you have adequate top frame support for the tanks weight 2x4's are more than enough to support that load for the legs and the bottom frame.
The bigger concern that the capacity of the 2x4 to support a downward force would be lateral forces... if someone leans on the tank, or if it's not 100% level, aggressive wavemaking, seismic activity... the taller the stand the more this is an issue. So while the 2x4 will suffice on their own under ideal circumstances, it would probably still be a good idea to sheath the entire stand in some 1/2 plywood, or add some kind of diagonal bracing. Over building the legs with 2x6 would add some lateral bracing, but nothing close to what a plywood skin would offer. (queue "plywood is all you need!" crowd)