Dot is the younger daughter of the Queen of the Ant Colony and is a princess. Her older sister, Atta, is the heir to the throne. Dot is initially Flik's only friend and supporter. She is a member of the Blueberries, a group similar to a scout troop. Like her sister and mother, she has wings, but they are not fully grown in due to her age. By the end of the film, however, she is capable of flight.
Dot idolizes Flik and hates being small. She compliments Flik's ideas and helps convince him a number of times during the film. Dot fears the demented grasshopper Thumper, whom Hopper uses as a punishment. However, she eventually conquers her fear of Thumper when she smacks him on the face and has Dim scare him away.
Dot struggles with trying to fly at the beginning of the film and is further dismayed when her mother chastises her for trying to fly before her wings have grown in. After Flik is dismissed by Atta for holding up the harvest with his inventions, Dot chases after him and tells him she likes his inventions. Flik is disappointed that he can't make a difference in the colony and Dot feels the same way, being so little. Flik lifts her spirits by showing her how a seed becomes an enormous tree eventually (although he hands her a rock) and she just needs more time to grow. When the grasshoppers arrive and find their food gone, after being knocked into the river by Flik, the ants all take refuge in the anthill.
While Hopper is lecturing the ants over the importance of ants serving grasshoppers, Dot becomes frightened at the sight of Hopper's dog-like pet grasshopper, Thumper. She tries to slip away but Hopper grabs her and teasingly brings her closer to Thumper, presumably to feed her to him. Flik intervenes and saves her, but backs away in fear when Hopper advances on him.
When Flik is to leave Ant Island to go to the city to enlist the help of warrior bugs to help them ward off the grasshoppers, Dot wishes him good luck, hoping that he will come back in one piece. She is later seen sitting on a cloverleaf, using Flik's telescope to keep on the lookout for him, despite two of her friends poking fun at her. She spots him returning with the newly-recruited "warrior bugs," who are actually a troupe of circus bugs recently fired by their ringmaster P.T. Flea. She is the first to rush over and welcome him back. Flik playfully lifts her up.
When the circus bugs realize later on they have been scammed, they go to leave. Flik desperately tries to get them to stay, watched from afar by Dot. Although she's not sure what is going on, Dot tries to follow them across the dried up riverbed but becomes stranded in the air on a dandelion puff after trying to use her wings again. At the same time, Flik and the circus bugs have aroused a hungry bird, who spots the helpless Dot in midair and charges her, prompting her to let go of the dandelion puff. Dot starts free-falling, but Francis catches sight of her and flies under her to catch her, but the force of impact causes the two to fall into a gap in the riverbed, where Francis' leg gets smashed by a rock and he is knocked out. The bird pecks at the gap, trying to get at them but Dot pulls the unconscious Francis out of reach. As Heimlich lures the bird away, Flik uses Rosie's thread to scoop up Dot and Francis. After Flik, Dot, and the circus bugs fly to safety, the ants applaud the circus bugs for their bravery. She and the Blueberries decide to make Francis their "den mother" and have a wonderful time with him, unaware of his chagrin.
Much later on, P.T. Flea arrives at the anthill looking for his troupe and blows Flik's cover. Flik and the circus bugs are banished by the Queen and Atta for the deceit. Dot tries to go after Flik; however, her mother kindly but boldly stops her. She then calls out to Flik sadly as he trudges off.
Soon after, Hopper's gang usurp Ant Island. Dot gathers the Blueberries and leads them to take refuge in their hidden clubhouse. From inside, she overhears the grasshoppers' plan to squish the Queen after pillaging the island of food. She decides to go and get help. Unfortunately, she accidentally bumps into Thumper, who viciously chases her to the cliffside where she falls into the foggy abyss below. However, her wings have finally grown and she flies off to catch up with Flik and the others.
When she finds them, she urges Flik and the circus bugs to come back, telling them what's going on and what she overheard. Flik refuses several times, ashamed of all the troubles he has caused for the colony. Dot finally retrieves a stone, shows it to him and echoes his own previous words; "pretend it's a seed, okay?" His depression turning to determination, Flik thanks Dot and embraces her tenderly.
Once they return, Flik and Dot gather the Blueberries, and they all climb aboard his makeshift bird and commandeer it, with Dot giving instructions and making bird sounds. However, it is set on fire by P.T. Flea who mistook it for a real bird, accidentally aiding Hopper in the process, and Hopper discovers that he has been tricked. Dot is captured by him again, but Flik orders him to release her, confessing to him that the bird is his idea and Hopper did release her and beats Flik. When the ants rebel against the grasshoppers, Thumper confronts Dot again, but she plucks up courage, slaps him and says, "No! Bad grasshopper, bad grasshopper! Go home!" giving an assumption that she has overcome her fear of him. Just then, Dim lands behind her and gives out a monstrous roar to scare away Thumper, and he and Dot high-five each other.
At the end of the film, Dot and the Blueberries are seen sobbing and grieving as their "den mother," Francis, must leave with the circus troupe. Dot receives Atta's tiara when she is made the new Queen. As Flik waves goodbye to the departing circus troupe, Dot and Atta take him to a tree root for a better view.
Andrew Stanton has said that he was the creator of Dot and that he was inspired to want a little girl in the film because he just had a newborn daughter at the time, which he said was something that John Lasseter as a father of five sons "couldn't claim to have."
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