Harikrishna, 57, said he met Rajya Sabha Chairman M Hamid Ansari this morning and submitted his resignation.
Deputy Chairman P J Kurien informed the House that the Chairman has accepted his resignation.
Talking to PTI, Harikrishna alleged that the decision to bifurcate the state was taken to reap political benefit in favour of Congress in the coming Lok Sabha elections.
"By this decision, they (Congress) killed the Telugu spirit. They did it to make Rahul Gandhi next Prime Minister after the Lok Sabha polls," he said.
"My father stood firmly for the cause of united Andhra Pradesh. Now it is shattered. My conscience would not allow me to continue (as Rajya Sabha MP)," he said.
The TDP leader, who had announced his resignation few days back, had alleged that the Congress leadership announced the division without resolving the crucial issues like water sharing.