"Avenge the blood of your martyr. Revolt against the new government," she said in the remarks broadcast on Tuesday.
Aisha fled to Algeria in August along with her mother and two of her brothers.
They were allowed to enter the country on humanitarian grounds.
She stressed that her father Moamer Kadhafi, who was killed on October 20 after his capture by forces loyal to the National Transitional Council, "did not abandon" Libya's people.
Her comments are thought to be in violation of an exile agreement with Algeria to refrain from any political activity.
However she already made similar comments back in September, when she denounced the NTC as "traitors" and called on the Libyan people to rise up against the new rulers.