Santos limped off the field against Malta on Sunday
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Alex Santos has joined Akira Kaji on Japan's injury list ahead of the World Cup opener with Australia on 12 June.
The Brazilian-born winger picked up the injury during the 1-0 win over Malta on Sunday, in Japan's final warm-up match.
He said: "I've got the foot iced up but I should be okay for the Australia game. I'll see how it is on Tuesday."
Kaji, 26, remained a fitness doubt with a bruised ankle, and coach Zico said: "It is difficult to say when he will back in action. It may take some time."
Zico added: "He is still in a lot of pain."
Right-back Kaji said: "I'm getting better but I am not sure about the opening match."
First-choice strikers, Naohiro Takahara and Atsushi Yanagisawa were left out of the Malta game because they were "not 100 percent" fit due to muscle pains.