This is based on opinion. Please don't list it on a work's trope example list.
Tear Jerker / The Lion King
Mufasa's demise, obviously. The scene where Mufasa is killed shows the body pulled about by his son.
It's especially sad for kids because if something so horrible could happen to the perfect father, who's to say it couldn't happen to their own father?
The music that plays during the scene certainly doesn't help, especially when the part when Mufasa's theme comes in (fittingly it was based on parts of Mozart's Requiem Mass). Add Simba trying in vain to wake his dad up, and then in utter despair trying to cuddle up under his paw, and you have a scene that is possibly one of the saddest in any animated film ever.
Simba's Big "NO!" moment right as Mufasa dies could also qualify, since it's pretty much a given what's going to happen at that point.
The scene also became a tearjerker for the film's creators, as they received a letter from a young boy who had lost his father, thanking the creators for the film as it helped him deal with his loss. The staff were moved to tears by it.
The scene is even more heartbreaking in theaters, in 3D...
This scene is arguably worse than the death of Bambi's mother. For one thing, there isn't any Mood Whiplash to this scene. Afterwards, Scar tells HIS OWN NEPHEW that it's his fault his father is dead, to run away and never return, before sending the Hyenas to kill him. Of course, there's a little bit of much needed comedy courtesy of Banzai falling into a bramble patch (with the much memed line "Cactus Butt!"), but that's it. Plus, unlike Bambi, where audiences don't actually see any onscreen death, this movie has the visual of Mufasa ''getting pulled off the cliff and falling all the way down into the stampede.
There's also the fact that is a character with heart and humor who the audience has come to love and sympathize with. To see him snuffed out so cruelly is heartbreaking for audiences.
Afterwards, however. we see Scar giving a none-too-sincere eulogy for Simba and Mufasa just before he takes the helm, and we see the lionesses and Zazu's reaction to this, with Zazu comforting the heartbroken and shattered Sarabi. And then the hyenas come out. And these aren't the goofy trio that we know and love; these hyenas are Nightmare Fuel.
Followed by Rafiki's reaction, and then Simba almost dead in the desert. A lot of people probably consider it a good thing Timon and Pumbaa showed up when they did.
Two Meta examples with voice actors. Jonathan Taylor Thomas was recording this scene and his mom was present so his director told him to imagine his mother falling; he proceeded to screw up the take by yelling "MOM!" instead of the scripted "DAD!" And then in the Danish dub, there was a break in the recording and the director noticed that Simba's cub voice actor was unusually quiet and asked what was wrong. Turns out the kid's father had recently died.
The size of this point proves that this scene is, and will remain, within the top 5 saddest moments ever in animation history.
Prior to that, Simba briefly cries into Scar's leg. He trusts his uncle, his father's brother, and he's so desperate for comfort and for someone to tell him it wasn't his fault...which he gets, but not in a good way.
The reactions of the lionesses when Scar takes the throne and the hyenas and eerily creeping in.
Rafiki as he watches Scar take the throne followed by him smudging out a drawing of Simba, who everyone thought was dead.
Timon and Pumbaa having "sad" eyes is both hilarious and this.
And if he falls in love tonight It can be assumed His carefree days with us are history In short, our pal is doomed
Cue Timon and Pumbaa crying hysterically, for laughs.
When Rafiki tells Simba, "You see, he lives in you."
Just before this, Simba is at his lowest point since the stampede; all of his feelings of guilt and shame having been brought back by his argument with Nala. He starts off in what seems like Calling the Old Man Out/Rage Against the Heavens, but it quickly turns once again into self-blame. It's definitely a moment where Matthew Broderick proves He Really Can Act.
Simba: You said you'd always be there for me!! But you're not... (beat) And it's because of me. It's my fault...it's my...fault... (chokes up and starts to sob)
The ending when Simba takes the throne is one of these too, partly from Tears of Joy (and the intense emotional power of Zimmer's score) but also just the sense of sorrow for everything Simba went through to get to that point and the relief that he is finally there, he's honored and avenged his father, and more than earned his rightful place as king. One word sums it up: "Remember."
After Mufasa saves Simba and Nala. Imagine son being in grave danger if you're a parent. Driven home in the following scene when Mufasa confesses to his son that the possibility of losing him is the one thing that made him genuinely scared. Even moreso during the stampede, when you can clearly see the distress on Mufasa's face as he searches desperately for Simba among the mass of wildebeests.
Every parent who genuinely loves their kids will instantly sympathize with Mufasa on this. Imagine your son and his close friend go out to the place you explicitly tell him not to, knowing full well that the place they're going to is VERY dangerous, without any parental supervision to boot. And then strangers (who may not actually be evil, but the other person isn't going to know that) start chasing them about. Virtually any parent is suitably freaked the hell out at the idea.
There's a very understated bit during the stampede. Zazu is flying through the gorge searching for Simba and finds him desperately clinging to a tree branch over the rampaging wildebeest. Simba cries out for Zazu to help him, and all Zazu can do is reassure him that his father is coming and tell him to hold on. Simba has caused Zazu plenty of irritation in the past, but Rowan Atkinson's excellent voice acting definitely gives the impression that right now Zazu would give anything to be able to pull Simba out of harm's way. But he knows he's not strong enough to lift Simba and therefore can't save him. That probably haunted him for years.
Nasty Fridge Horror when considering Zazu for this whole thing. After Mufasa goes down into the gorge to save Simba, Zazu tries to fly back to Pride Rock and get help, only to be smacked into a wall by Scar and knocked unconscious. Only he might not realize that. He probably thought he panicked and hit the wall himself and failed to get potential help for Mufasa and Simba because of it. Until the truth comes out later, it's not unreasonable to think that he blamed himself and felt that he failed both Simba and his king. It makes him seem more woobieish than usual.
Some more fridge horror for that scene; how do we know Scar didn't pull the "it's all your fault" trick on Zazu, too?
The look of utter horror and betrayal on Mufasa's face when he realizes his own brother is about to throw him to his death. In spite of the vitriol shown between Scar and Mufasa at the beginning, Mufasa is still a good and noble character and he probably still cares about his brother and is completely unaware of Scar's hatred for him until the very end. Someone he knew as family, someone he probably loved and wanted to see happy, tricked and brutally murdered him.
As he falls to his death, he lets out an absolutely gut wrenching, completely helpless scream. The king of Pride Rock is completely helpless and can only scream in his last moments.
When Mufasa pleads for Scar to help him, he addresses him as "Brother". It demonstrates that he would've done anything to save his brother were the situation reversed, and thought Scar would do the same for him. Instead...
The hyenas looking back at Scar, appealing for food.
The scene of Mufasa and Simba's last night together, after Mufasa saved Simba from the Hyenas. First he is angry at Simba but then they are nudging and playing with each other, laughing. If you don't know what will happen on the next day this scene won't touch you that much but if you know it, then you might have some watery eyes. And the music isn't helping avoiding this.
Simba's "we'll always be together, right?" and Mufasa's answering speech about the Great Kings of the Past is a sad bit of foreshadowing, but when you think about it, the fact that he responds that way at all is very tragic. He doesn't feel he can respond with a simple "yes" or even something to the effect of them being together a good long while yet. Whether it's because nature is uncertain and dangerous or the role of being king is, Mufasa is painfully aware that he could die at any time.
Mufasa even foreshadows this earlier with his speech about the reign of kings acting like the sun's rising and setting. In both cases, Simba just can't grasp that his father one day won't be around— he innocently believes that he'll be king without anything changing. His childhood naiveté can be downright heartbreaking for anyone who's experienced a life-changing loss.
The stage musical
Although a great deal of The Lion King is deeply emotional and affecting (as is the film it was based on), there's one particular addition not in the movie: the solo number performed by Simba, "Endless Night". In particular the lines: "You promised you'd be there/Whenever I needed you/Whenever I call your name/You're not anywhere/I'm trying to hold on/Just waiting to hear your voice/One word, just a word will do/To end this nightmare." Also, the earlier lines "Sleepless, I dream of the day/When you were by my side/Guiding my path/Father, I can't find the way." By the time the end of the song comes, Simba sings such a stirring, heartfelt testimonial to hope for the future, but the way the song uses actual lines from the movie and builds upon them to reference both the deep love Simba and Mufasa shared (and how he was such a perfect father) and the guilt, despair, and suffering Simba went through as mentioned above is just so overpowering and gut-wrenching.