Describing 27-year-old Prince Harry as a "high-value target", a militia spokesman said they will "make their best efforts to arrest or kill" him, the Sky news reported.
The Army captain, the third in line to the British throne, arrived in Afghanistan for his second tour of duty last week. He is deployed in southern province of Helmand, one of the bloodiest war zones in the 10-year-old conflict.
The London-based TV channel quoted Taliban spokesman Zabiullah Mujahid as saying: "Whoever is fighting in our country alongside the US is our enemy and we will attack him. Prince Harry came to Afghanistan and he is a high value target for us. We will try to arrest him. Because he is an Apache helicopter pilot, he will target us more. If we are not able to arrest him we will target him," he said.
The new kill notice on Harry by Taliban comes just two days after the militant group had dismissed his deployment as a propaganda stunt, saying it was apparently to divert attention from his escapades in US.
The royal Apache pilot hit the headlines last month after he was photographed naked at a party in Las Vegas, US.
He will spend four months based at the heavily fortified Camp Bastion at Helmand. The Prince is to complete training courses this week in first aid, shooting and improvised explosive device awareness, before starting his Apache-specific preparation.
During this phase of training, which starts today, he will climb into the cockpit and begin to familiarise himself with the way the deadly aircraft is configured for the country. Harry is not expected to be sent out to take on Taliban for at least another seven days, the TV channel reported quoting unnamed sources.
In 2008, Harry was hastily withdrawn from Afghanistan when a news blackout surrounding his deployment, on the ground directing aircraft in attacks on Taliban positions, was broken by foreign media.