Surprisingly enough, the original version of The Joker. In his first appearance, he was a permanently smiling psychotic gangster with no sense of humor whatsoever. In his first fight with Batman, Bats is actually the one making puns, while Joker is screaming "I am going to kill you!"
The Joker was actually able to make Batman laugh at the end of the graphic novel The Killing Joke (even though the story as a whole wasn't funny in the least).
X-Men: Professor X and Magneto are frequently flanderized as no-nonsense extremists. Patrick Stewart's performance is the first attempt to move away from this, although one might simply assume that as he gets older, he will get more serious. Also, Ian Mc Kellen gave Magneto a deadpan, cynical sense of humor which was expanded on by Michael Fassbender.
Idol Hooves in The Changeling Of The Guard. Having been banished from The Hive by Queen Chrysalis, he travels Equestria, seeing the outside world for the first time. His first exposure to humor is met with a lot of confusion, but he eventually comes to find the concept fascinating, though he doesn't really understand Wasta's jokes.
The film Rat Race features a character who tragically was born without a personality.
In No Country for Old Men, according to Carson Wells the big problem with Anton Chighurh isn't that he is batshit insane, but that he lacks a sense of humor.
The various Terminators exhibit this behavior, being emotionless robots.
From Happy Go Lucky, Scott the driving instructor, and Poppy's older sister Helen.
In Highlander, The Kurgan accuses nuns of being this when they don't appreciate him waggling his tongue at them.
Agent K in Men In Black quips (deadpan) that "we at the FBI do not have a sense of humor we are aware of."
This, and attempts to develop a sense of humor, are the focus of Sha Wujing's character arc in The Lost Empire (a sort of unofficial sequel to Journey to the West.) He's eventually told that it's better to have no sense of humor than to have the sense of humor that the villains have, and he laughs for the first time at the realization that he was worried over nothing.
Granny Weatherwax understands humor on a conceptual level, but has absolutely no sense of humor and has no understanding of how or why jokes work. Illustrated well in Witches Abroad, where she repeatedly tries to tell the "alligator sandwich" joke but always bungles the punchline with something like "And be quick about it!"Actual punchline "And make it snappy!"
In Going Postal, Adora Belle Dearheart tells Moist Von Lipwig that she has no sense of humour due to her embarrassing name.
In Making Money, the head bank clerk, Mavolio Bent, has "no sense of humour whatsoever. It has been proven by phrenology." It becomes more apparent when one considers his background as the child of a clown, being laughed out of the arena during his first performance (which failed), fleeing from the circus and vowing never to laugh again.
Archchancellor Ridcully has a similar lack as Granny, with the result that his jokes are one of many aspects of his personality that grate on the Bursar, who also has no sense of humour, but at least understands how jokes are supposed to work. In fact, when Ridcully first appears in Moving Pictures, the Bursar (who has not yet been driven completely insane by the man) attempts a joke himself:
The Archchancellor gave him a look so old-fashioned it might have belonged to an ammonite. "That a joke?" he said, in the suspicious tones of someone who wouldn't really understand the term "sense of humour" even if you sat down for an hour and explained it to him with diagrams.
Dwarves are known for their sense of humor. In a way. People point them out and say "Those little buggers haven't got a sense of humor."
Sergeant Detritus, a troll on the A-M City Watch, "treats humor as a human abberation" to be overcome by talking slowly and patiently.
Harry Potter: Percy Weasley "wouldn't recognize a joke if it danced naked in front of him wearing Dobby's tea cozy." He does have a bit of a harshly sarcastic moment in Deathly Hallows, though, which is remarked upon with astonishment by his siblings one of whom is tragically killed seconds later.
From Mercedes Lackey's SERRAted Edge novels, Unseleighe psychics have absolutely No Sense of Humor, along with little comprehension of allegory and the imaginations of bricks. In one novel, they were driven insane trying to comprehend the lyrics of They Might Be Giants.
From Animorphs, Andalites. All of them besides Ax and Arbron.
Thomas from Malevil. He even notes that he's known for not having a sense of humor but doesn't argue the point.
In Warrior Cats, Rainflower begins scolding Crookedjaw and Oakheart for telling friendly jokes about the queens, making a (quite) boring lecture about how the queens "enjoy helping their Clan". Crookedjaw and Oakheart just roll their eyes at this.
Colonel Lieutenant after promotion Brendig in David Edding's Belgariad. Also Ortzel from the Elenium. The two never smile. Ortzel does later, albeit it looking unnatural on him. Brendig does show some humour, but since this is a work of Edding's, compared to the others, his humour is close to, if not, nil.
Ce'Nedra:Don't you ever smile?
Brendig:(perfectly straight face) I am smiling.
In a series full of deadpan snarkers, Baron Vengeous from Skulduggery Pleasant has no sense of humour and an aversion to humour in general. Skulduggery's constant wisecracking is one of the things he hates about him. Interestingly though, in the second book Vengeous does crack a joke, saying "Forgive the expression but you and and what army" and at one point almost laughs at one of Skulduggery's jokes. Generally though, he's dour and humourless.
In 1066 and All That, Queen Victoria "remained obdurately plural and not amused" throughout her reign despite the best efforts of her subjects to amuse her.
A Song of Ice and Fire: Stannis Baratheon is perceived as this by the people of Westeros. The reality is that he has an extremely dry and subtle sense of humor which is lost on the average Westerosi who is accustomed to bawdy and/or toilet humor.
Victarion and Aeron Greyjoy both seem to have this lack. Victarion isn't very bright and dislikes jokes because he's always worried that they'll be at his expense and he won't be able to tell. Aeron used to have a sense of humor until he found religion, at which point he lost it.
Roland from The Dark Tower freely admits that his sense of humor was "shot off in some war somewhere." He gets somewhat better the more time he spends with his True Companions.
The narrator of The Curious Incident of the Dog in the Night-time has an Ambiguous Disorder (the blurb says Asperger's, the book doesn't), and explains there will be no jokes in the book because he doesn't understand them. He can understand the concept of a pun, if it's explained to him, but if he tries to keep multiple meanings in his head at the same time, it's like trying to follow multiple conversations.
Vulcans are renowned for having No Sense of Humor, though many of them are Deadpan Snarkers instead. The Romulans are much the same in this regard.
Data from Star Trek: The Next Generation repeatedly attempts to understand humor as part of his quest to become more human. He doesn't succeed until he gets his emotion chip, which is one of the plot threads in the above-quoted movie.
Worf from the same series has almost no sense of humor either. He's always serious. At one point he shows disdain for people who rely too heavily on humor, saying they "talk much and say little." Martok eventually works out that he has a very dry and sarcastic sense of humor, he just almost never laughs.
Lampshaded on Criminal Minds: Reid (falsely) calls Aaron Hotchner a narcissist as clue to where he is taken hostage. While figuring this out, Hotch asks the team what they consider his greatest folly. Spontanous answer by Prentiss: "You have no sense of humor!" Actually something of a subversion. Hotch does have a sense of humour; it's just so dry the Sahara looks like a swamp by comparison, and at this point Prentiss hasn't caught on to it yet.
"Now, nobody likes a good laugh more than I do... except perhaps my wife and some of her friends... oh yes and Captain Johnston. Come to think of it most people likes a good laugh more than I do."
Sheldon from The Big Bang Theory. He understands the concept of humor, he just doesn't get humor. When he's told that something is a joke, he forces the most fake laugh ever heard on television. And the things he does find humorous are usually beyond the scope of "mere mortals" like the remainder of the cast. Not until the end of the fourth season does he tell any jokes that get laughs (although that's partly due to Tough Room) and when he did so, he wasn't even aware he was doing it.
In early episodes of Mork and Mindy, Mork identifies people making In-Universe jokes by saying "Oh, humor — Ar-Ar!" And almost as often he does that when people aren't joking.
A common trait for the villains on Doctor Who. And some non-villains, like the Ood.
Played with in Stargate SG-1, Colonel O'Neill has very specifically stated that his name is spelled with two Ls on several occasions; this is because there's another Colonel Jack O'Neil in the fictional version of the USAF, one with No Sense Of Humor. This is an in-joke and lampshading of the fact that Kurt Russell's version of the character in the movie (spelled with one L in the credits) was significantly less funny.
Teal'c almost never tells a joke or laughs (unless you count his occasional plays on earth idioms), with one exception. It turns out his sense of humor just doesn't translate cultures well.
Stargate villains in general tend to be this way. Exceptions include the System Lord Ba'al and Todd the Wraith who, unlike the rest of their respective races, are Deadpan Snarkers.
Admiral Brovo: Don't worry, Teddy. Junior officers get thrown to the sharks first. And lieutenant Rabb is the one who's sailing into harm's way.
Commander Lindsey: Yes, sir.
Admiral Brovo: Harm's way? Lieutenant Rabb's first name is Harm.
Commander Lindsey: Oh! Oh, yes, sir.
Admiral Brovo: You need a sense of humor, Teddy, otherwise people are gonna think you're a lawyer.
Commander Lindsey: I am a lawyer, sir.
People who don't "get" Mystery Science Theater 3000 - a show that plays bad movies and heckles them as they play - sometimes are dealing with this. They find the riffing gets in the way of their enjoyment of the movie being played. Not everyone has this issue though - some people have a good sense of humor but it's hard for them to follow two things - the movie and the riffing.
At the 2005 Academy Awards, host Chris Rock joked that Jude Law was in "every movie" released that year, remarking "Who is this guy?" and implying that Law was a low-rent Tom Cruise. Sean Penn later responded with dead seriousness, "In answer to our host's question, Jude Law is one of our finest young actors." After this, it became a bit of a Running Gag for people to poke fun at Penn's apparent lack of humor. Saturday Night Live did an entire sketch about this called "Sean Penn's Celebrity Roast", in which Penn completely misses the point of the roast and acts annoyed at the comedians making fun of their fellow celebrities.
In one of the Muppet's "at the ball" sketches, a female dancer asked her partner, "Do you know who is the most humorless man in the world?" When he said no, she replied, "You are, Fathead!" He, of course, responded, "I don't get it."
By the third game, she's now a full on aversion of the trope, and is, for the most part, able to keep up with Joker, although her taste in humor leans more towards messing with people's heads.
The Mechari of WildStar suffer from this. Ironic, as real-life manipulators frequently use humor to disarm targets and throw suspicion off themselves.
Bladewolf from Metal Gear Rising: Revengeance analyzes Raiden's jokes. When Raiden says he needs to teach him sarcasm, Bladewolf confirms that he does get the jokes. He just doesn't find them funny.
Some Champions in League of Legends just refuse to joke when you order them to. If you read their backstories, it was implied that a traumatic event from their past prevent them to take jokes and take things in utter seriousness.
Poppy: Jokes? I don't know any jokes.
Vayne: I have no time for nonsense. / Joke? What do you mean?
The warship A.I. Tag from Schlock Mercenary was originally created with no sense of humor, but has tried to learn it to better understand his opponents.
Third Character of Jayden and Crusader is a particularly violent atheist with no sense of humour at all.
The female warrior in Oglaf has absolutely no sense of humor, only getting pissed off by anyone who attempts to be funny. Of course, eventually she has to end up fighting cultists of the God of Fun.
Asia Ellis from morphE. She's socially oblivious Literal-Minded and requires jokes to be explained. She often finds them stupid when broken into their base components. Billy in particular despises the fact she exhibits this trait.
Adult Ben in the Ben 10 episode "Ben 10,000". He gets better though.
Optimus Prime in Transformers Prime. It's stated that all Primes are like this — the Matrix of Leadership, which contains the accumulated wisdom of all past Primes, gives them a sense of responsibility, dignity and gravitas that leaves little room for goofing around.
Optimus notably wasn't like this in the original series. While usually serious he was not above playing a game of basketball with his men, or cracking a joke or two, or mocking Megatron. However as the years and series went on his characterisation became more and more about being a no-nonsense messiah type figure.
Lemongrab of Adventure Time. He tries, though. He really does try... but he fails horribly.
Lemongrab: "Ha. Ha ha ha! HA HA HA HA HA HA HA HA HA! Twelve years Dungeon! All of you! Dungeon! Seven years, no trial!
Zane in Ninjago is noted at trying to understand humor but seems to fail.The reason for this is that he is a Ridiculously Human Robot that normally has all of his emotion switched off, including humor.
The Simpsons: Rex Banner never laughs. When he tries to do so, in "Homer vs. the Eighth Amendment" (due to him finding the idea of a beer baron operating under his nose without getting caught "laughable"), he fails miserably.
In The Venture Bros., The Monarch claims that a lab accident that turned Phantom Limb evil(er) did two things. First, it gave Phantom Limb the power to kill a man by touching him. Second, it turn him into a "completely humorless dick."
Agent Powers from the season two premiere of Gravity Falls claims to be "physically incapable of experiencing humor".
Temporarily happens to Cartman in the South Park episode "How to Eat With Your Butt". Cartman "blows a funny fuse" after a particularly hilarious prank leads to a couple with butts for faces showing up in town, and he becomes unable to laugh at anything.
Also a common technique among abusers, who verbally taunt, harass, and degrade their targets and then, when feelings are hurt and/or they are called to account for their behavior, defend themselves with "You can't take a joke." In other words, they take a case of Dude, Not Funny! and deliberately try to turn it into this trope.
Germans are often stereotyped as having No Sense of Humor. In reality, German Humour is a bit more complicated; Germans definitely do have a sense of humour, and enjoy comedy, but their ideas about what is funny and what isn't can vary greatly from those of English speakers. Adding to the confusion is the tendency of German people to smile at a joke instead of laugh, making foreigners think that they didn't get it and are just being polite.
And German people think the English have no humor, or at least this special "English Humour"...
This trope was parodied by UN Ambassador Hans Beinholtz whose sense of humour was...dry, to say the least.
In Greece, if someone tells a joke that completely fails to amuse, people are likely to comment 'That's an English joke'.
An executive for Looney Tunes, Eddie Selzer, once overheard a group of writers laughing at jokes being thought up for a specific Looney Tunes. He stormed in and asked what all the laughing had to do with the making of animated cartoons.
He was well-known for having the sense of humor of an underripe potato. They used to use him as a sort of barometer for the quality of their ideas; if he liked it, they were off the mark. Selzer was, in fact, the specific genesis of the cartoon "Bully for Bugs", when he barged into a brainstorming session and declared, sans all provocation, that bullfights weren't funny, and there were to be no cartoons about them. They made the cartoon anyway, the argument being that Selzer had, in Chuck Jones' words, "never been right about anything yet."
Similarly, at MGM Tex Avery was forced to work for executive producer Fred Quimby, who, according to the book "Tex Avery: King of Cartoons", never understood any cartoon Avery made. Even though everyone else, including the audience, considered them to be hilarious. He even warned Avery not to make "Blitz Wolf", in which Hitler was ridiculed, "because we don't know who will win the war in the end."
The Marx Brothers: Groucho Marx always claimed that actress Margaret Dumont, who was always the Butt Monkey of his insults, never seemed to understand his jokes. Recent research has found out that this was simply an urban legend.
A frequent side effect of Asperger's is the inability to understand humour, as well as sarcasm, due to their being Literal-Minded. They might understand the concepts of sarcasm and humor, but have trouble distinguishing them from sincerity, which leads to similar reactions. Not being so naturally keen on social interaction, aspies can often develop a rather dark sense of humour, as they do not have the same inhibitions against it. This of course varies from individual to individual.
Which is rather odd considering that there are quite a few hadiths about Muhammed himself cracking jokes.
Attila the Hun is reputed to have had no sense of humour. Allegedly an envoy from the Eastern Roman empire who met with Attila once at dinner described it as follows: “By mixing up the languages of the Italians with those of the Huns and Goths, [an entertainer] fascinated everyone and made them break out into uncontrollable laughter, all that is except Attila. He remained impassive, without any change of expression, and neither by word or gesture did he seem to share in the merriment ….” The impression left was that Attila lacked a sense of humor and cared about nothing but power.
A common misconception of Ayn Rand is that she had no comprehension of humor, despite a talent for devastatingly on-target satire. She was sometimes Late to the Punchline, perhaps as a result of early trauma, and part of it could have been that she was a native Russian-speaker operating in an English-speaking environment; plays on words and things like that may have just sailed right past her. She was prone to quips however and sometimes demonstrated a biting sardonic wit. For instance, whilst being pestered by a fly on a talk show, she commented "Perhaps it's protesting."
Isaac Newton is widely reputed to have had no sense of humour.
Legendary jazz pianist Bill Evans reportedly had this problem, much to Miles Davis's amusement. Apparently Miles had so much fun with Evans that on one occasion he told Evans that he'd have to have sex with all the members of Miles's band before he could join, and waited fifteen minutes before telling Evans it was a joke.