The protagonists are three American brothers: Francis, Peter, and Jack Whitman, who haven't seen each other for a year, since their father's funeral. The film begins as they meet each other in a train that travels through India, which is named "The Darjeeling Limited". The oldest of the three brothers, Francis (Owen Wilson) has planned this journey. In the end, the journey should add up to a reunion with their mother, who conducts an abbey in India. Francis conceals this real reason and tells his brothers that they make the journey because of a spiritual self-discovery. Francis' brothers aren't very convinced of that and are annoyed with Francis' regimented behaviour. With his assistant's help (Brendan) he designs a fastidiously accurate daily plan. He also takes his brothers' passports to prevent them from leaving the train too early. Francis has suffered injuries from a motorcycle accident and wears bandages. The youngest Whitman, Jack (Jason Schwartzman), has written a short story which is strikingly similar to his own life. However, he denies those similarities. Jack obsessively listens to the answering machine of his ex-girlfriend at every stop the train makes. Moreover, he has an affair with the train's stewardess Rita (Amara Karan), whom Francis nicknames "Sweet Lime" for the drinks she repeatedly offers.

Peter (Adrien Brody), the second brother, says that he was his father's favorite son, by which he justifies keeping many of his late father's possessions. Francis is revolted. While Peter's wife, Alice (Camilla Rutherford), is expecting a baby, Peter fears without foundation that their relationship may end in divorce.

The three brothers consume Indian pharmaceuticals excessively at the beginning. In their trips through the Indian province, they visit temples and markets. At one market, Peter buys a cobra, which later escapes from its transport container. This escape results in the brothers being confined to their stateroom. Francis and Peter get into a fight over Peter being the "favorite" and Jack uses the pepper spray he bought in the village to mace his brothers until they stop fighting. Because of that the three of them have to leave the train with their large baggage and are left in the desert. The three brothers come closer again and even perform one of Francis' spiritual rituals. On their way back to civilization they see three boys get into trouble while attempting to cross a river. Jack and Francis rescue two of the boys, but Peter fails to save the third. This affects Peter deeply. In the village of the boys the three brothers attend the boy's funeral and befriend the people living in the village.

In between the scenery changes into a flashback: The three brothers and Alice are on the way to their father's funeral. They stop on the way to get to get their father's Porsche from the repair shop and to take it with them, hijinks ensue, but the car isn't ready to start and so the brothers leave.

Back in the present, the Whitmans reach a bus, which brings them to the airport. The brothers stop for a bathroom break, during which Francis removes his bandages to shave, which reveals a number of large, bright scars on his face, but his brothers offer marginal reserved comfort. However, before they take the plane, they revise their decision and they decide to go and visit their mother, who tells them in a letter that a visit at this time is impossible. Nevertheless, the three brothers travel to their mother's abbey. The greeting is very emotional (it is learned Francis' accident was in fact, a suicide attempt) and the family is reunited again for a moment. On the next morning the three brothers find out that the mother has again left her family and her children.

On the way back the three brothers get a train at the last minute and leave their baggage on the platform. Jack reads his new short story which tells the story of his meeting with his ex-girlfriend in the Hotel Chevalier...

In the last scene, Francis wants to give the passports back to his brothers, but they decide that he can keep them better than they can and they don't take them.

Anderson also wrote and directed the 2007 short film Hotel Chevalier, starring Jason Schwartzman and Natalie Portman. The 13-minute film acts as a prologue to The Darjeeling Limited; In it, Jack's ex-girlfriend turns up unexpectedly at his hotel room in Paris, and they spend the night together. Originally attached to festival screenings of The Darjeeling Limited, it was removed during the limited theatrical release and instead made available on Apple Inc.'s iTunes Store as a free download. On October 26, 2007, Hotel Chevalier was removed from iTunes in favor of releasing it in theaters with the wide release of The Darjeeling Limited.

The Darjeeling Limited includes many of Anderson's signature themes and styles, such as forbidden love, parental abandonment, sibling rivalries, a privileged class who rarely work and timeless fashions and props. Anderson has revealed that The River by Jean Renoir, the films of Satyajit Ray and documentaries on India by Louis Malle were his inspirations for this movie. The film was dedicated to Satyajit Ray and makes allusions to him and his work (e.g., the portrait of Ray in the compartment of the train Bengal Lancer towards the end of the film). In an homage shot, the three Whitman brothers are arranged in a row by the side of the train after it has broken down. This is a reference to an earlier Anderson film, Bottle Rocket, where characters Dignan, Anthony and Bob are arranged as such for the cover.[citation needed]

The Darjeeling Limited made its world premiere on 3 September 2007 at the Venice Film Festival, where it was in competition for the Golden Lion and won the Little Golden Lion. The film's North American premiere was on 28 September 2007 at the 45th annual New York Film Festival, where it was the opening film. It then opened in a limited commercial release in North America on 5 October 2007. The film opened across North America on 26 October 2007 and in the UK on 23 November 2007, in both territories preceded in showings by Hotel Chevalier. The film grossed $134,938 in two theaters in its opening weekend for an average of $67,469 for each theater. The film (widescreen edition) was released on DVD 26 February 2008 on Fox Searchlight, with features limited to a behind-the-scenes documentary, theatrical trailer, and the inclusion of Hotel Chevalier.