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1* Luigi's death and Mario's reaction, which drives much of the first half of the story. Mario is so devastated about his brother's death that it indirectly leads to the entire rest of the series playing out the way it does, and the Princess feels like a part of her has died.
2* Mario successfully feeds Luigi the Life Mushroom, but nothing happens because he's been dead too long for it to work.
3* Princess Toadstool being catapulted into the lava and dying, with "Mario..." as her last words.
4* The ending, when everyone is dead, and Mario soon follows.

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