The Cabinet minister said the first £25 million of investment in zero-emission buses worth up to £129 million will be focused on rural communities.
All local authorities in England outside London can apply for funding through the Zero Emission Bus Regional Areas (Zebra) programme.
Today's funding for more zero-emission buses will help decarbonise public transport— Transport Secretary Mark Harper
Applications from those that missed out in the previous stage of the scheme will also be prioritised.
Mr Harper also announced that a new research hub to develop ideas for low-carbon and resilient transport infrastructure will be created, backed by £10 million of Department for Transport funding.
The Net Zero Transport for a Resilient Future Hub will be established by Newcastle University, Edinburgh’s Heriot-Watt University, the University of Cambridge and the University of Glasgow.
Mr Harper said: “Today’s funding for more zero-emission buses will help decarbonise public transport and grow the economy by keeping our communities connected.
“We have already reached our initial target of funding at least 4,000 zero-emission buses and this additional funding will improve journeys for even more passengers, reaching those in the most remote areas.
“The UK is also cementing its position as a world leader in net zero tech with this new investment into climate resilience.
“Our Net Zero transport hub will be a centre of academic excellence, helping us keep our transport network resilient into the future.”