"The Islamic umma (nation) should focus on jihad in Al-Sham (Syria) ... And unite the ranks of mujahedeen there," said 23-year-old Hamza in an audio message posted online.
"There is no longer an excuse for those who insist on division and disputes now that the whole world has mobilised against Muslims," he said.
His undated message comes after Al-Qaeda leader Ayman al-Zawahiri also urged jihadists in Syria to unite, despite his continued rejection of the rival Islamic State (IS) group and its proclaimed caliphate.
"Either you unite to live as Muslims with dignity, or you bicker and separate and so are eaten one by one," he added, according to SITE Intelligence monitoring group.
Al-Qaeda's Syria affiliate Al-Nusra Front is a rival of IS, which is an Al-Qaeda offshoot whose leader Abu Bakr al-Baghdadi in 2014 declared an Islamic caliphate across territory seized in Iraq and Syria.
Bin Laden's son said Syria is the "best battlefield" leading to "liberating Al-Quds", the Arabic name for Jerusalem.
US intelligence officials have said that Hamza was the favourite son of the 9/11 mastermind who had been grooming him to take over as Al-Qaeda's leader.
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