Move comes amid shortage of taxis in Rome.
People travelling on Italo rail services in Rome and Milan can now reserve an Uber car to and from the railway station, at the time of booking their train ticket, thanks to a deal between the two companies.
The new intermodal travel solution was launched at a media presentation in Rome's central Termini train station on Wednesday.
The service will initially be offered to customers travelling through Roma Termini and Tiburtina in the Italian capital as well as Milano Centrale, Rogoredo and Rho Fiera in Milan.
Italo customers departing or arriving at one of these stations can book, from 90 days before the trip up to one hour beforehand, an Uber Black or Van via the Uber Reserve function.
Italo CEO Gianbattista La Rocca said the company is expanding its multimodal network "to offer complete solutions to our travellers", adding that the new agreement "simplifies travel and makes it safer, eliminating waiting times outside stations."
Lorenzo Pireddu, general manager of Uber Italia, said the partnership with Italo marks "a significant step forward in making the journey to and from the stations of Milan and Rome easier, more convenient and more reliable."
"The two platforms will communicate with each other, making booking quick and easy for users", said Dora Bonadies, head of network and market development at Italo, who said travellers can avail of the service "with a simple click."
The new alliance comes amid a severe shortage of taxis in Rome where lengthy queues of tourists and commuters are a common sight at the taxi rank outside Termini station.
The powerful lobby representing Italy's taxi drivers has staged industrial action in recent months and is currently in talks with the government over its opposition to sector deregulation.