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Sōki (蒼鬼 Sōki?) is a strong warrior who possesses the dark power of the Oni, referred as the Blue Oni. Having learned about Hideyoshi's insidious scheme, he sets out on a journey to burn the Genma trees and defeat the Genma running rampant across the land. Wielding two swords with such grace, precision and power, Sōki is soon referred to by townsfolk as "Oni of the Ashes" (灰燼の蒼鬼 Kaijin no Sōki?). He is also called the "Black Oni" due to his extreme resemblance to the Oni God. Sōki uses a variety of heavy broadswords and katanas. Soki is confirmed to appear as a playable character in the upcoming crossover fighting game, Tatsunoko vs. Capcom: Cross Generation of Heroes.
Akane Yagyū (柳生茜 Yagyū Akane?) is a young woman with exceptional skills who carries on the name of the famous Yagyū clan, possessing the rare Eye of the Oni trait. Prior to the beginning of the game itself, Akane is assigned the duty of hunting down Munenori Yagyū to restore her clan's honor. Assuming the name of Jubei (十兵 Jubei?), Akane hears of Soki's deeds and finds him at the Plains Outpost at Suruga to defeat him. But after losing to him, she joins him in his fight against the Genma. Near the end of the game, she is depicted wearing eye patch to cover her Oni Eye. Akane's swift, iaijutsu-like style of attack is similar to that of Kaede from Onimusha: Warlords. At one point in the game, Akane is confused as to whether she is romantically interested in Sōki, or he is just her elder 'brother'. Shikomizue swordsticks are her main weapons.
Ohatsu (王初 Ohatsu?) is the younger sister to Lady Yodo. Since the birth of Hideyori to her older sister, Ohatsu has been very troubled by the change she has noticed in the two. Friends since childhood, Ohatsu holds intense feelings for Hideyasu. She is an expert marksman under her mentor, Magoichi Saika of Onimusha 2, able to use variety of firearms as well as explosives and a grapple hook. In the aftermath, she married to a noble of a Kyoto house.
Roberto Frois in a Christian missionary born to a Japanese mother and Spanish father. After being betrayed by Luís Fróis, who was possessed by a Genma who tried to use him as a experiment, Roberto is now on a mission of pure revenge. Roberto's weapon of choice are his enhanced arms and fists, capable of numerous destructive boxing combinations. Roberto's gauntlets are based on the Christian archangels, Raphael, Uriel, Gabriel and Michael. Ordered from weakest gauntlet to strongest respectively. In the end, Roberto leaves for Spain to battle the Genma there so his country wouldn't suffer as Japan had.
Tenkai Nankobo (南光坊 天海 Nankobo Tenkai?) in a mysterious monk residing at the ruins of the temple on Mt. Hiei, he senses a evil starting to spread across the land and eventually discovers Hideyoshi Toyotomi's bold and insidious scheme. He then sets out on a journey to find one who possesses the power of Oni to help him bring Hideyoshi down. He uses pole arms that are named after Hindu deities for example, Varuna, Indra, and Rakshasa. Tenkai's skills are based on Buddhist and Hindu terminology, such as Enlightenment and Nirvana. During the game's end credits it is revealed that Tenkai is none other than Samanosuke Akechi.
- Minokichi (蓑吉?): A bagworm creature who aids Sōki with instant "teleported" travel, insight on the various creatures & places they traverse, and quips about the allies they eventually obtain.
- Arin: A girl with mysterious powers and a nun-like quality who travels with Tenkai.
- Toyotomi Hideyoshi (豊臣 秀吉?): Formerly known as Tokichiro Kinoshita, he succeeded Nobunaga in not only setting up the new government of Japan, but in league with the Genma to resurrect Fortinbras. However, Hideyoshi dies fighting Sōki in a suit of Genma armor.
- Mitsunari Ishida (石田 三成 Ishida Mitsunari?): Hideyoshi's retainer, who is actually Claudius of the three Genma who intend to resurrect Fortinbras. Upon being revived, Caludias reveals his true form being a Mukade Genma before slained by Tenkai.
- Luís Fróis: Roberto's guardian who betrayed him after becoming the Genma high priest/scientist Rosencrantz, one of the three Genma bend on the resurrection of Fortinbras. Once revived, he assumes his true form, a giant fly demon, before slained by Roberto.
- Lady Yodo (淀殿 Yodo-dono?): The daughter of Oyu who became a royal concubine to Hideyoshi and High Priestess Ophelia, one of the three Genma bend on the resurrection of Fortinbras. Her true form being a humanoid mantis Genma before slained by Ohatsu.
- Danemon (右衛門 Danemon?): A Genma samurai who battled Sōki and died at his hands.
- Sakon Shima (島 左近?): A human made into a Genma general after his mind is twisted and brainwashed by Ophelia, fighting Sōki and company until he was defeated at the Port of Seikai and abandoned by his masters once Tenkai Nankobo uses his purification skills to free Sakon of the Genmas' hold over him.
- Munenori Yagyū (柳生宗矩 Yagyū Munenori?): A member of the Yagyū whose heinous acts in betraying in joining the Genma and killing his kin are the reason Jubei accepts the task to capture her cousin dead or alive. However, though she defeat him, Munenori lived long enough to make a vain assassination attempt on Fortinbras that cost him his life.
- Fortinbras: The Genma Lord, also known the God of Light or the White Serpent. He is portrayed as a gargantuan serpent-like demon lord, with a pair of large wings intact on his back and a third eye on top of his head. He attempted to rule the world with aid from Oda Nobunaga, only to be slain by Samanosuke Akechi. However, Fortinbras is eventually resurrected by the Genma Triumvirate, assuming his true form, a human-like figure resembling a European noble in dress and sporting a pocket watch who has been viewing the events of the game before revealing himself. Though he was the most powerful, Fortinbras died against Sōki and his friends.