In 1556, Don Aguirre leads Spanish conquistadors to South America to search for the Lágrimas de Cristal Tree, whose flowers cure illness, heal injuries, lift curses, and may save his ill daughter. After many conquistadors die, the Puka Michuna tribe heals the survivors with the Tree's flowers. The tribal chief refuses to reveal the Tree's location, so Aguirre stabs him and burns the village. The dying chief curses the conquistadors, making them immortal and unable to leave sight of the Amazon River without being dragged back by the jungle itself.

In 1916 London, McGregor Houghton presents his sister Dr. Lily Houghton's "Tears of the Moon" research to the Royal Society, which bars women members. The Houghtons hope both to revolutionize medicine and aid the British war effort. They request access to a recently acquired arrowhead artifact that Lily believes can locate the Tears of the Moon Tree but are denied. Lily steals the artifact, narrowly evading Prince Joachim, who is equally intent on finding the Tree for himself and Germany.

After arriving in Brazil, Lily and MacGregor hire skipper Frank Wolff to guide them down the Amazon. Frank, who offers jungle cruises embellished with faked dangers, initially declines, citing the dangerous river and jungle, but reconsiders upon seeing the arrowhead. The trio departs after escaping Joachim's submarine.

Lily discovers Frank's Tears of the Moon research, but he insists he stopped searching long ago. They are captured by cannibals that are actually the Puka Michuna tribe who work for Frank as part of his contrived jungle cruise adventure. Angered, Lily distrusts Frank and sets off to find the Tree herself. Tribal chief Trader Sam translates the symbols on the arrowhead, revealing the Tree's location and that it only blooms under a blood moon.

Meanwhile, Joachim locates the conquistadors petrified inside a cave and partially revives them with Amazon water. Aguirre agrees to bring Joachim the arrowhead in exchange for lifting the curse when he finds the flowers. Joachim diverts river water to flood the cave which reanimates Aguirre and his conquistadors and infuses them with rainforest elements. The conquistadors attack the tribe and Aguirre stabs Frank through the heart. Lily flees with the artifact, chased by conquistadors, but the jungle vines pull them back to the river, preventing her capture.

To the Houghtons' amazement, an alive Frank soon reappears. He reveals he is also a cursed conquistador. He helped search for the Tree to save Aguirre's daughter but sided with the tribe against Aguirre's brutality. After years of endless fighting, Frank trapped his vengeful comrades in the cave away from the river, petrifying them. Failing to find the Tree, Frank remained tied to the river, becoming a tour guide and building a village.

Lily and Frank travel to the La Luna Rota Waterfall and discover a submerged temple containing the Tree. Meanwhile, Joachim has captured McGregor and forces him to reveal Lily's location. Frank, the Houghtons, the Germans, and the conquistadors all converge at the Tree.

Discovering …

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• Dwayne Johnson as Frank Wolff / Francisco Lopez de Heredia: A shrewd and cynical but ultimately noble steamboat skipper who reluctantly agrees to guide two explorers on their quest for the mythical Tree. He eventually reveals his true name and identity as Aguirre's adopted brother and one of the cursed conquistadors, forever trapped by the Amazon River. He is a trained cartographer who has spent centuries searching for the Tree to break the curse that made him immortal so he can die peacefully. Meanwhile, he has built a town, and a boat he christened "La Quila" for his business under "Jungle Navigation Company" while owning a number of tamed exotic cats as a pet, each named "Proxima".
• Emily Blunt as Dr. Lily Houghton: An eccentric, adventurous and virtuous botanist working in a male-dominated field, who hopes to find the Tree in order to harness its power for modern medicine. Possessing an ancient arrowhead and one of Frank's old maps of the Amazon, she proves she is resourceful and capable in martial arts and lock picking skills, despite the fact that she cannot swim until Frank helps her out with it at La Luna Rota. She wants to prove herself equal to her chauvinistic peers but gains some notoriety for wearing trousers.
• Édgar Ramírez as Aguirre: A Spanish conquistador who once sought the Tree's power to save his ill daughter, only to be cursed with immortality for his cruelty towards its guardians. Temporarily freed by German explorers, he sets out to take revenge on his adopted brother, Frank, for turning on him. His partially-decomposed body is now composed of an infestation of snakes and as such, he possesses the ability to convert part or all of his body into snakes.
• Jack Whitehall as MacGregor Houghton: Lily's younger brother who works as her assistant. He later confesses to Frank that the rest of their family disowned him after he was outed as being gay when he refused to marry a woman, prompting Lily to hire him so he could support himself and he has been loyal to her ever since. As a somewhat foppish snob who adheres to proper etiquette and prefers wearing three-piece suits, he seems ill-suited to jungle life, but gradually develops into a more confident, rugged man by the end of the expedition.
• Jesse Plemons as Prince Joachim: A deranged and ambitious German royal who finances and leads a military expedition with mercenaries to claim the Tree of Life, both to aid Germany's war effort and achieve immortality. He receives aid from the cursed conquistadors after he frees them for his own interest, and communicates with them through their bees and snakes.
• Paul Giamatti as Nilo Nemolato: The harbormaster at Nilo's River Adventure, Porto Velho, where Frank moors his boat. Nilo owns a Moluccan cockatoo named "Rosita". He is also a business rival, and confiscates Frank's boat engine when Frank is unable to repay him a loan with Rosita constantly quoting "Frank owes me money". This leads Frank to wreck Nilo's prize boats in return.
Additionally, Verónica Falcón plays Trader Sam, the female chief of the Puka Michuna tribe in 1916, who is a longtime friend of Frank's and knows about the curse. The Puka Michuna tribe has a centuries-long history of guarding the Tree, but Sam does not know the exact location herself until Lily shows her the arrowhead. Dan Dargan Carter, Dani Rovira, and Quim Gutiérrez play Aguirre's fellow conquistadors, as, respectively: Gonzalo, whose partially-decomposed …

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In December 2004, it was announced that Jungle Cruise would be developed for Mandeville Films, with a script by Josh Goldstein and John Norville. Following the success of Pirates of the Caribbean, the film was to take place sometime in the twentieth century and was loosely inspired by the theme park attraction of the same name which featured prominently in Disneyland's grand opening in 1955. In 2006, Alfred Gough and Miles Millar were in talks to write the film. In February 2011, it was announced that Tom Hanks and Tim Allen, who had previously worked together in the Toy Story franchise, would star in the long-gestating film, with a script to be written by Roger S. H. Schulman.
In August 2015, it was announced that Walt Disney Pictures was revamping the film adaptation to star Dwayne Johnson. The previous script originally written by Goldstein and Norville would be rewritten by John Requa and Glenn Ficarra with the intention to harken back to its period roots. John Davis and John Fox signed on as producers.

Johnson, who did a lot of research before getting into the role, announced in April 2017 that he would co-produce the film under his Seven Bucks Productions, and he expressed his interest in having Patty Jenkins helm the project. However, in July 2017, Jaume Collet-Serra was announced as the director of the film. In January 2018, Michael Green was reported to have rewritten the script, previously worked on by J. D. Payne and Patrick McKay. Also, Emily Blunt signed on, as Johnson wanted her to be his co-star. Blunt was paid between $8–10 million for her involvement.
In March 2018, an open casting call was made for the other characters in the film, including men and women of all ethnicities between ages 17 to 90 and children between 6 and 14 years old.

In the same month, Jack Whitehall was cast as the brother of Blunt's character. Six months later, it was reported that he would have a coming out scene in the film with Johnson. This would be the second incidence of a gay character in a live-action Disney film, the first being Le Fou, portrayed by Josh Gad, in the 2017 adaptation of Beauty and the Beast. There was some backlash over the report, with some people online expressing anger over a straight man being cast as a "camp" gay character.

In April 2018, Édgar Ramírez and Jesse Plemons were added to the cast as villains with the former being "a man with a conquistador background". In May, Paul Giamatti was cast to portray a "crusty harbormaster". In June, Quim Gutiérrez joined the cast to portray one of the villains. In July, it was announced that Veronica Falcón, Dani Rovira, and Andy Nyman had join…

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