I believed she maried the mayor.
- James Henry Trotter – The protagonist of the book, who wants nothing more than to have friends and be happy. Though something of a dreamer, James is clever and ever-resourceful throughout his adventure in the giant peach, and his intuitive plans save his friends' lives on each occasion.
- The Old Man – A friendly yet mysterious individual, who initiates James' adventure. In the 1980 re-printing of the book, with illustrations by Nancy Ekholm Burkert, he can be seen in the final illustration, amongst the New York City crowd.
- Aunt Spiker – A dominating, cruel, malicious, and thoroughly repulsive woman; possibly the older of James' aunts. Spiker is described as tall and thin – almost emaciated – with steel glasses. She is destroyed by the giant peach in its escape.
- Aunt Sponge – Spiker's sister: a greedy, selfish, and morbidly obese woman, and equally as cruel and repulsive as Spiker, and destroyed concurrently.
- The Centipede – A male centipede, depicted as a boisterous rascal with a good heart, and perhaps James' closest friend in the peach. He is generally optimistic and even brave, but outspoken and rash. His sources of pride are his multitude of legs and his ancestral status of garden pest. He often asks for help with putting on his many boots, or taking them off, or shining them. In the last chapter of the book, it is revealed that he becomes Vice-President-in-Charge-of-Sales of a high-class firm of boot and shoe manufacturers.
- The Earthworm – An earthworm who often quarrels with the Centipede, and is frequently the most pessimistic of the protagonists, though on amicable terms with nearly all. In New York City, he becomes the mascot of a skin-cream company.
- The Old Green Grasshopper – A male grasshopper, who (as the eldest) assumes an almost paternal rôle to James and the others. He is an accomplished musician; wherefore, he ultimately becomes a member of the New York Symphony Orchestra where his playing is greatly admired.
- The Ladybug – A kind, motherly female ladybug who takes care of James. In the last chapter of the book, it is revealed that the Ladybug "married the head of the New York City Fire Department and lived happily ever after".
- Miss Spider – A good-natured female spider who takes care of James. Generally friendly and decent in manner, but described by Dahl as having "a large, black and murderous-looking head, which to a stranger was probably the most terrifying of all". She has particular resentment toward Sponge, who caused the deaths of Miss Spider's father and grandmother. On the journey, Miss Spider makes hammocks for the others to sleep; and in the final chapter, becomes a tightrope manufacturer.
- The Glowworm – A female glowworm, who quietly hangs from the ceiling at the center of the giant peach and provides lighting for the interior. After the adventure, she illuminates the Statue of Liberty's torch.
- The Silkworm – A female Silkworm, who assists Miss Spider in the production of thread, both before and after the adventure. She is never seen to speak.
- Cloud Men – Are minor antagonists, who threw rocks and supplies at the peach after Centipede taunts them.
- The Peach - While not a sentient character, the peach plays a vital role in helping the characters escape an unpleasant existence.
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