Orlando is a fictional biography of a person called Orlando who lives over three hundred years from Queen Elizabeth's reign in the sixteenth century through to King Edwards reign in 1928, the year Virginia Woolf wrote the novel. In the beginning of the book Orlando is a nobleman who has literary ambitions. As a man he writes plays and poems every day of his life while courting some of his generation's most beautiful women. Everything changes when he turns into woman, and for the remainder, Woolf draws comparisons between the thought processes of men and women across the different eras.

In the first chapter, Orlando is a fifteen-year-old boy living with his parents. He dreams of joining his father and grandfather in the war against the Moors, but Queen Elizabeth takes to his exceptional beauty and refuses to allow him to fight. However, he falls out of favor with the Queen when she catches him kissing another girl. Shortly afterwards, King James takes over the throne and welcomes Orlando back to the royal court. By this time, the Great Frost has taken hold of the country. To increase his popularity, the King builds upon the ice, creating a carnival type atmosphere. While skating, Orlando meets a Russian girl called Sasha. Their relationship causes scandal, but Orlando is deeply in love with her. Unfortunately, during one of Orlando's melancholy moods, Sasha cheats on him. He forgives her, but soon after, she sets sail back to Russia.

In the second chapter, grief overcomes Orlando and he goes back to his country house and sleeps for seven days. When he wakes, he remembers little of Sasha, though he sinks into depression if anyone mentions Russia. To get over his mood, he begins to read and write again. After a long period of seclusion, he arranges a meeting with a famous poet named Nick Greene. Greene comes to his house and entertains everyone with stories about Shakespeare and Marlowe, and Orlando shows him some of his own work. Greene promises to read it once he gets home. When he arrives home, Greene decides to write a satirical piece about Orlando, using some of the extract of Orlando's poem. Orlando is so upset when he reads the book he denounces all men, replacing them with dogs. Eventually Orlando comes out of his mood and decides he should write about whatever takes his fancy. He decides the best thing to write about is his surroundings and his house. It is while writing he realizes his estate needs redecorating and he sets about making it all beautiful again. People come afar to see the new developments and Orlando becomes part of high society. One day he meets an Archduchess and falls in love. The feeling shocks him and he asks King Charles to assign him as Ambassador of Constantinople in Turkey.

Chapter three follows Orlando's time as Ambassador of Constantinople. His good looks mean he is popular with everyone and particularly women. Orlando, however, does not form any close relationships and focuses on his work. Such dedication impresses the King and he sends over a friend to award Orlando with the highest ranked position in his peerage. Celebrations follow the ceremony, but Orlando disappears into his room. In the morning, he does not wake up and stays asleep for seven days. During this time, the Turks rise against the Sultan and enter Orlando's room, intent on killing him. Seeing Orlando fast asleep, they think he is already dead. When Orlando awakes, he has changed into a woman. She leaves with a Gypsy and joins their way of life. Unfortunately, the difference in lifestyle is too great and she pays for a ship back to England.

Chapter four details Orlando's time aboard the ship and his struggles at accepting his new identity as a woman. Immediately, he see the power women can hold over men when the Captain gives her her own awning. The relationship with the captain develops and he invites her ashore when they anchor in Italy. On her return to the boat, she finally feels like a real woman. Soon they arrive back in England. Orlando goes back to her house in Blackfriars and discovers she has legal problems due to her change in sex and three men claiming to be her sons, therefore having a legal right to her property. She decides to go back to the country. Here she meets the Archduchess again. Strangely, when she invites the Archduchess back to her house, it turns out he is a man. The Archduke wants to marry Orlando, but she has no feeling for him whatsoever. She manages to get rid of him by disappearing into society life and finding many male admirers, including the poet Mr. Pope. She soon gets bored of their company and starts dressing up as a man and walking through London. At the end of the chapter, the city clouds over and the eighteenth century becomes the nineteenth century.

In the fifth chapter, Orlando struggles to cope with Victorian times. Everywhere she looks, people are married and women have numerous children. She knows society expects the same from her and she feels torn about wanting her independence and the need to conform. One day she takes a walk in the park and meets the seamen Shelverdine. Immediately they decide they should get married. Orlando loves him and even more so when she discovers he used to be a woman. They spend every day together until Shelverdine heads back to sea. Before he goes, one Orlando's servant marry them.

In the sixth chapter, Orlando wants to become a writer again. She has a chance meeting with Nick Greene. Greene looks completely different, but Orlando soon discovers his views on life are similar as before as he claims to hate the modern Victorian poets, but loves the Elizabethan poets he once despised. Orlando shows him her poem, the Oak Tree, and Greene promises to find a publisher and get it good reviews. Orlando then goes to a bookshop and buys all the Victorian literature available. When she has finishes reading, she has a child, and the period changes to 1928. Now in the present, she reflects on old times and even thinks she sees her old Russian flame Sasha. Such thoughts frighten her and she goes back to wait for her husband to return. The book finishes with the reuniting of Shelverdine and Orlando.