In the 55-minute video posted online, Ayman al-Zawahiri pledged renewed loyalty to Afghan Taliban leader Mullah Omar.
Correspondents say his stated allegiance is an apparent snub to Islamic State (ISIS) militants.
ISIS is challenging al-Qaeda to lead worldwide Islamist militancy.
Announcing the formation of “al-Qaeda in the Indian Subcontinent” using a mixture of his native Arabic and Urdu widely spoken in Pakistan, Ayman al-Zawahiri appeared eager to regain some of the limelight, correspondents say.
“[Al-Qaeda] is an entity that was formed to promulgate the call of the reviving imam, Sheikh Osama Bin Laden. May Allah have mercy upon him,” Ayman al-Zawahiri said.
He urged the “umma”, or Muslim nation, to “wage jihad against its enemies, to liberate its land, to restore its sovereignty and to revive its caliphate”.
Ayman al-Zawahiri said “al-Qaeda in the Indian Subcontinent” would be good news for Muslims in Burma, Bangladesh and in the Indian states of Assam, Gujarat and Jammu and Kashmir, where they would be rescued from injustice and oppression.
A spokesperson for India’s ruling Bharatiya Janata Party (BJP) told the Associated Press that the statement was “a matter of serious concern”.
“But there is nothing to worry about. We have a strong government at the federal level,” the spokesperson said.
Counter-terrorism experts say al-Qaeda’s ageing leadership is vying with ISIS to recruit followers after the success of militants in the Middle East in attracting young followers worldwide by conquering large amounts of territory across Iraq and Syria.
ISIS leader Abu Bakr al-Baghdadi describes himself as a “caliph” – or head of state – and has called for the support of all Muslims around the world.
The two groups fell out in 2013 over the ISIS expansion into Syria, where Baghdadi’s followers have carried out decapitations, crucifixions and other forms of capital punishment.
On September 3, it emerged that Pakistani militants linked to IS have been distributing pamphlets in the north-western city of Peshawar calling on people to support their idea of creating an Islamic caliphate.
The material, published in the Pashto and Dari languages, urges people to support ISIS in its fight for a grand Islamic rule.
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