The Cutie Mark Crusaders — Apple Bloom, Scootaloo, and Sweetie Belle — appear huddled together for a short moment at the end of the first episode (Apple Bloom had already been introduced earlier, but the other two wouldn't officially meet her until episode 12).
Sweetie Belle also appears briefly at the beginning of Call of the Cutie due to an animation error, where an anonymous pony based on her model should've been shown.
Adventures in Ponyville, a flash game on the Hub's website, revealed a few tidbits about the show during the long gap between the premiere and the second episode. Snips, Snails, and Cheerilee were first seen (though unnamed), Rarity's and Pinkie Pie's occupations were made explicit, and Twilight demonstrated the power to temporarily give wings to other ponies, an ability not seen until episode 16.
The premiere of the show made the series look more like a Magical Girl show. However, the Elements of Harmony are completely forgotten until Season 2, and the show turns to focus on one-shot episodes rather than an overarching plot.
It also took a while for the visual effects for unicorn magic to be standardized, and even then the Color-Coded Wizardry didn't really show up until "Lesson Zero".
Likewise, the exact nature of unicorn magic seems different early on. In "Boast Busters", unicorn magic appears to be entirely in-born. A unicorn either has an ability, or they don't. However, later episodes seem to suggest that unicorn magic is much closer to Rule Magic, thanks to the appearances of magical schools and spell books, wherein any unicorn's ability to learn a spell is limited only by aptitude at learning magic, rather than by simply being born with a given list of 'tricks'.
The third episode of the series, "The Ticket Master" was the first episode to be written, and it shows - Spike has no interest in Rarity, non-unicorn ponies can somehow levitate the tickets above their heads etc, though this could easily be explained by Rarity or Twilight holding their tickets for them or the tickets themselves being magical.
It's also the lightest episode plot-wise, as it was originally intended for a Two Shorts format.
Easily Forgiven: Many antagonists are forgiven quickly. Justified, considering the series main theme is friendship.
Downplayed with Luna. She tried to create eternal night and is immediately forgiven by her sister, the mane cast, and Ponyville. However, Luna Eclipsed shows that they are still terrified of her.
Edible Ammunition: The citizens of Appleloosa use apple pies to repel a buffalo stampede. The entire episode was a metaphor for the American Settlers encroaching on the Native's land. And the pie battle was very intentionally a tame version of a shootout, because you wouldn't get incapacitated by a pie, would you?
In "Dragonshy", Applejack demonstrated expert marksmanship with kicked apples. She didn't get to use them against the dragon, but she was ready to.
In the same episode, Angel beans Fluttershy with a half-eaten carrot.
Catapult-launched pumpkins are a popular part of Nightmare Night celebrations. Presumably, they could be re-purposed for less festive occasions.
In "Secret of my Excess", Pinkie Pie tries to fend off Spike by throwing cakes at him. He just catches them instead.
In "Magic Duel", Trixie conjures several pies and throws them at Twilight Sparkle, who defends herself by summoning a parasprite.
Edible Theme Naming: Applejack. Her entire family qualifies too, given that they're named after brands of, types of, or products made from said fruit.
Distant relatives seem to be named after different fruits, such as Aunt and Uncle Orange.
In "Bridle Gossip", when Applejack is shrunk, Spike dubs her "Appletini". (Both names are kinds of alcoholic drinks.)
Seems to be a pattern with most (though not quite all) Earth Ponies; Pinkie hails from the Pie family, though makes her living with the Cakes; two of Equestria's founders were Chancellor Puddinghead and her secretary Smart Cookie; even Rose and Daisy could arguably count, since ponies have been seen multiple times eating flowers.
Many of the background pony names, too.
Edited for Syndication: The Japanese dub unfortunately has to cut out a few scenes in order to make room for the longer intro and outro. It remains to be seen if any future DVD releases will restore these scenes, not having to worry about ad blocks.
During the first season, the show was broadcast with an E/I logo in the upper left corner, but the only real concession it makes to being educational and informative is to have And Knowing Is Half the Battle dialogue at the end of every episode. (These days, simply claiming to encourage positive social values is often enough to allow the use of the logo.) Cable channels aren't required to air E/I programming, but there may be conditions to the partnership between Hasbro and Discovery Channel to form the Hub that the audience doesn't know about. Still, a cartoon that manages to be Edutainment, Merchandise-Driven and genuinely entertaining even for adults all at the same time is a truly astonishing achievement.
The E/I mark goes back to the Discovery Kids days; the network adds E/I tags in electronic program listings to make it easier for parents to find actual educational children's programming. Since no other cable network (like Disney Channel, Nickelodeon, Cartoon Network) uses the E/I tag, it's a really big advantage for The Hub to tag their shows E/I.
The show could be a Trope Codifier for Stealth Education. The writers are very talented at sneaking in educational tidbits without disrupting the flow of the show — going all the way back to the pilot, where Twilight explains that being "on the precipice (or threshold, or brink) of disaster" means that "something really bad is about to happen", and that "imperative" means "important".
A particularly stealthy tidbit they snuck in was actual physics equations for time dilation, like Gamma Correction, in an episode dealing with time travel. Who knew you could learn college-level physics in a TV-Y program?
Hasbro has also released guidelines for some episodes for parents that want their kid to learn more.
The bug has been absent in Season 2 so far, which left certain fans wondering if Hasbro has ditched the Edutainment aspect of the show. Either that, or The Hub has since deprecated the practice all together.
Rainbow Dash can manipulate cloud-borne water to create rain (her profession), and also to create rainbows, as seen most clearly in with her stunt in "Boast Busters". Her name also contains the word "rain", despite the fact that one might think that Rainbow Dash, as a great flyer, might also be associated with the air.
This seems to be a common thing with all pegasi, considering that Derpy Hooves caused lightning by hopping on a cloud, and that weather control is the exclusive job of the pegasi.
Rarity can magically locate pockets of precious or semiprecious stones within solid rock, giving her an earth affinity. This is due to her special talent dealing (at least partially) with gems.
Pinkie Pie, despite being, paradoxically, an earth pony, has a clear affinity for the air, as noted here, and her speed and apparent teleportation abilities are also suggestive of air-based powers, as is the fact that her cutie mark is a bunch of balloons.
Twilight Sparkle could be seen with a fire affinity, given the way her mane and tail are transformed into flames at the party in "Mare in the Moon" and in her rage in "Feeling Pinkie Keen"; on the other hand, Twilight might also be seen as a fifth elemental, given her association with the stars; either designation would fit with her magical powers, especially since, in the classical schema of the elements, fire was seen as being the closest to the fifth element anyway.
Queen Chrysalis adores this trope, getting one several times in the first part of the two part Season 2 finale, and about once every five minutes, give or take, in the second part.
Evil Plan: Nightmare Moon's 'everlasting night' plan in the pilot. For a god of chaos, Discord is incredibly strategic about getting his "rulership" (AKA position to make up extremely creative and nonsensical torments for everyone) of Equestria back. Queen Chrysalis's 'first Canterlot, then all of Equestria' plan in the Season 2finale.
Bizarrely, the closing credits in the first two season never reflected the episode, listing the same set of voice actors and roles for every episode regardless of which characters were present or absent. Season 3 started using proper credits.
Applejack:If y’all reckon back, I told you that I would tell you everything at breakfast! But I didn’t come to breakfast! I couldn’t come to that breakfast! Not if it meant tellin’ y’all what happened!
Hasbro occasionally mandates product placement for new toys in the show, although they are usually written in a way that is either subtle or works well with the story of that particular episode. Well, most of the time, at least...
And Granny Smith is very similar to Madame Foster from the same show.
Parasprites are flying Tribbles. Exact same MO. Mass breeders, extremely cute, and eat everything.
Extreme Omnivore: Spike can eat fruit, grass, gemstones… and was the only one unaffected by Applejack's toxic muffins. In fact, he liked them so much that he went through the trouble of rescuing the "baked bads" from the trash so he could eat them all.
Oddly, he rejected an apple in "The Ticket Master" for having a worm. Maybe he wasn't in the mood.
Pinkie Pie is an Extreme Herbivore at least. The wormy muffins made her sick (she was the only major character to appear in the infirmary, in fact), but she has no problem eating cupcakes that are badly burnt or covered in hot sauce.
Pumpkin Cake from the episode "Baby Cakes" is just being a baby and chewing on everything in sight as they are prone to do, but it's possible for her to grow into this down the line.
The Parasprites can eat ANYTHING, especially after Twilight Sparkle magically changed their diet from food to non-food.
Rarity and Sweetie Belle's parents enjoyed the latter's horrible food, which included ashes (burnt juice) and a bowl of bubbling ooze (toast).
Fanservice: Of the non-sexual variety. After Derpy Hooves took off within the fandom, animators started including her more and more often, sometimes in a scripted event and other times as a Funny Background Event - to the point that spotting her has become a Where's Waldo?-esque game in some circles.
However, by far the largest example of this is in "the Last Roundup", when the name Derpy for Derpy Hooves was made canon, as well as giving her a voice!
And then, when the name caused considerable backlash, she returned to being unnamed.
Played straight whenever Tara Strong decides to troll the fandom.
IDW's official comics are chock full of fanon shout-outs, although they also sneak in some played-straight fanservice once in awhile. It's not really more fanservicey than one might expect from a G-rated comic, though included are some sultry character poses and subtle ship teasing (Rarity's micro series issue has some of the more obvious examples thus far).
Fantasy Counterpart Culture: To some extent. The aesthetics of pegasus society and Cloudsdale — especially architecture — seem to be based on Ancient Greece, with Disney's Hercules being a particular source of inspiration. Of course, the Pegasus is a creature from Greek mythology to begin with.
This also explains Rarity's inspiration for Rainbow Dash's custom dress.
The Appleloosan settler-ponies and the buffalo herd are quite obvious analogues to Wild West settlers and Native Americans, respectively.
Canterlot seems to be inspired by France with a dash of Britain.
The official map of Equestria reveals significant similarities to North America, albeit with as many horse puns as they can stuff in ("Manehattan" and "Fillydelphia" being the most obvious)
Fantasy World Map: There are many fan-made maps of Equestria, but an official one can be found here or here◊(though it slightly contradicts Word Of God by placing the Everfree Forest south of Ponyville instead of to the west). It appears to be loosely based on the United States and part of Canada.
Fashion Shop Fashion Show: Twilight and Fluttershy go through this, courtesy of Rarity. Though Twilight doesn't buy anything and Fluttershy was just modeling.
The Fashionista: Not only is Rarity the proprietor of the Carousel Boutique, but there's an entire fashion industry in Equestria including designer Hoity Toity, photographer Photo Finish, print advertising campaigns, and trend-following audiences at fashion shows who can't seem to form opinions of their own.
Feather Fingers: This was averted initially. Ponies use their mouths, magic, or sometimes even tails to manipulate objects as long as it can be made to work. However, later Season 2 episodes features ponies using their hooves as hands more often, or in the case of pegasus characters (Rainbow Dash, Fluttershy), a literal feather finger.
In Hurricane Fluttershy, the titular character tries to improve her flying by training, including doing wing 'push-ups'.
Felony Misdemeanor: Only Rarity understands what a "crime against fabulosity" ruining the water serpent's mustache was.
Pinkie Pie's extreme attitude towards keeping secrets, which leads her to stalk Twilight all over town reminding her that she risks losing Rarity's and Fluttershy's friendship FOREVER! In a bizarre attempt to drive the point home, she takes an apple... and EATS IT. Possibly a representation of Twilight's head, considering her reaction to Applejack breaking her Pinkie Promise in "The Last Roundup".
Pinkie Pie discovers that rainbows don't really taste like Skittles, or vice versa, in "Sonic Rainboom".
First Name Basis: Pretty much anypony who refers to Twilight Sparkle simply calls her "Twilight." Interestingly, Princess Celestia is the only character to refer to her by her full name with any regularity.
Averted with Pinkie Pie and Rainbow Dash. The former can be either Pinkie or Pinkie Pie (and in one Full Name Ultimatum, "Pinkamena Diane Pie"), while the latter can be called by first name, last name, or both, with equal familiarity.
Five-Temperament Ensemble: Applejack and Rainbow Dash (choleric); Rarity, Twilight Sparkle, and Princess Luna (melancholic); Fluttershy and Princess Celestia (leuquine); Pinkie Pie and Princess Cadence (sanguine); and Spike (phlegmatic).
Flanderization: While the show is still a relatively new one, this phenomenon is already beginning to happen to some of the characters, mostly happening in season 2.
In "Applebuck Season" and "Winter Wrap Up", Big McIntosh has actual dialogue. Since then, most of his lines have been "Eeyup" and "Nope". Although later episodes give him more varied lines, though he's still usually quiet.
Rarity, who goes from a generous and occasional melodramatic Large Ham to still being generous but much more melodramatic.
Fluttershy was initially meek (to the point of being somewhat aloof) and barely able to conjure up sentences to even her closest friends. Her later appearances, though more timid or cowardly, make her more cheerful and sociable. She was exaggerated in one area and rounded out in another.
Pinkie Pie suffers this as well, going from a character with a fondness for treats that would sometimes miss the point of situations, while still being dependable and determined to prove herself to being so consumed by the mere mention of frosting that she became lost in her own fantasy of it and needs Fluttershy to wipe the drool from her chin, not to mention going for coloring books while the rest of the cast are looking for a solution to a very real and immediate danger right outside the door.
Flash Forward: In the initial version of a recipes PDF on The Hub's website, the page for Cutie Mark Crusaders Chocolate Popcorn included Side Story Bonus Art (originally fanart drawn by layout artist Kat Stenson long before becoming part of the show's crew) that depicted Apple Bloom, Scootaloo, and Sweetie Belle all grown up with cutie marks.
Flat Character: Diamond Tiara. Lampshaded; her cutie mark matches her crown. This also applies to her BFF Silver Spoon.
Fleur de Lis: Celestia, Luna, and Nightmare Moon have this integrated into the 'boots' they wear as part of their royal attire (Nightmare Moon even having it as part of the design of her collar). A unicorn mare seen in Canterlot with Fancy Pants in "Sweet And Elite" is even named this and has three of these symbols as her cutie mark.
One of the many cutie marks Apple Bloom gets in "The Cutie Pox" is a fleur de lis which causes her to speak French.
In Feeling Pinkie Keen, when the already horribly-injured Twilight Sparkle is assuring Spike that something is not going to fall, a flowerpot then crashes on her head. Then an anvil.Then a hay wagon. Then a grand piano. They all turned out to have fallen out from a delivery truck that Derpy Hooves was working at.
In The Mysterious Mare Do Well, during The Reveal that Mare Do Well is some of the Mane 6 teaching Rainbow Dash a lesson, Pinkie Pie says she stopped the building collapse with her Pinkie Sense. She then detects a falling flowerpot with it, and dashes away to avoid it.
In It's About Time, when Twilight asks the gypsy Pinkie Pie if she can use her Pinkie Sense to detect what will happen in the future, Pinkie explains that it's only for immediate emergencies. Cue flowerpot to Twilight's head.
Flying On A Cloud: While they don't fly around on them, pegasi and other creatures with wings (e.g. griffons, birds) can do this. A spell exists for non-pegasi to do it too; it gets a lot of use in Fan Fiction.
Foil: Interestingly enough, each of the mane six acts as a foil to another one. The most obvious pair in this regard is Applejack and Rarity; their scenes together emphasize Applejack's more blunt and tomboyish side while highlighting Rarity's mildly obsessive-compulsive girly-girl nature. Similarly, Fluttershy's soft-spoken and gentle personality wrapped around an iron core amplifies Rainbow Dash's personality as a loud, brash, and reckless pony with a soft center. Twilight's straight-laced no-nonsense approach to most things and love of organization contrasts particularly with Pinkie Pie's free-wheeling and scatterbrained pursuit of fun, as well as Pinkie Pie's sometimes irrational behavior against Twilight's preference for logic. These contrasts are almost certainly intentional, and used to help advance a less overt aesop about friendship; the mane Six can be such close friends and an effective team because of their differences, not in spite of them.
These are the more obvious pairings, but there are portions of each one's personality that serves to counter some part of the other in some way, shape, or form.
Food As Bribe: Twilight is faced with this several times throughout "The Ticket Master" by her friends in attempts to gain an extra ticket to the Grand Galloping Gala, but Twilight manages to turn away the bribes each time despite stating several times how hungry she is. Pinkie Pie also uses this against Spike in "Party of One" as part of her Perp Sweating sequence.
As the camera follows Twilight Sparkle when she's dictating her letter to Celestia in the pilot episode, the hourglass distorts everything behind it like you'd expect, but replaces the unicorn head sculpture outside with Nightmare Moon's head.
In the same episode, Rarity is choosing decorative ribbons. Upon selecting a particularly glittery one, she declares "Sparkle always does the trick!".
Also in the same episode, Twilight exclaims, "the fate of Equestria does not depend on me making friends." By the end of episode 2, it's pretty clear that it does.
Listen closely to the music playing at Pinkie's surprise party in episode one. It's almost identical to the tune of her song in episode 2.
When Nightmare Moon shows up at the festival, the Cutie Mark Crusaders are briefly shown cowering in fear next to each other, despite the fact that this episode was before "Call of the Cutie."
The camera also pans across each of Twilight's friends as she lists each of the Elements of Harmony as they correspond to each pony.
In "Call of the Cutie", Rainbow Dash tells Apple Bloom that she got her cutie mark after her first race. In "Sonic Rainboom", she says that she's only done a sonic rainboom once before, when she was "just a filly." These are, in fact, the same event, which is shown in detail in "The Cutie Mark Chronicles."
"The Show Stoppers" hinted what each of the Cutie Mark Crusaders' special talents are. Apple Bloom's is designing, Sweetie Belle's is singing/song-writing, and Scootaloo's is dancing.
Pinkie: I can't tell you that, silly—then it wouldn't be a surprise!
In the first episode, Twilight proclaims that "all the ponies in this town are CRAZY!". By the time the first season ended, all the mane characters had had a mental breakdown at least once.
In "Lesson Zero", Spike mentions that Twilight has a cape, and that he and Twilight dropped it off at the cleaners. In the next episode, "Luna Eclipsed", Twilight wears a cape as part of her Nightmare Night costume.
In "Return of Harmony Part 1", after Rainbow Dash says that she'll always be loyal to the princess, Discord ominously says "We'll see about that...". He later gives her a Sadistic Choice and inverts her sense of loyalty.
In the same episode, when Discord is taunting Celestia about being imprisoned in stone, he is leaning on one of the pictures in a window, and then knocks on the forehead of the pony whose image he's leaning on. The pony in question is Fluttershy the only one he had to magically touch instead of tempt to turn bad and the pony that would eventually reform him.
When we first see the Elements of Harmony in their new forms in the second episode, Twilight's isn't a necklace like the other five, instead taking the form of a tiara (which she comments on in "The Return of Harmony part 2"). In "Magical Mystery Cure", when Twilight becomes an Alicorn and a princess, the tiara changes shape slightly and becomes her crown.
One particularly clever bit of foreshadowing is also extremely subtle. Twilght's mane and tail are both cut in a style called "Hime Cut" and have been since day one. In Japanese, this literally means, "Princess Cut". What does she become at the end of Season 3 again?
Forgot I Could Fly: Rainbow Dash NEVER forgets this, and is rarely seen walking on the ground. Fluttershy, however, is a mediocre flyer at best, and rarely flies if she doesn't have to, and on occasion, doesn't even remember she can. The other ponies occasionally forget that two of them (Although in practice, only one of them) can fly. In one instance, this trope is inverted, as Twilight Sparkle asks if she can help with weather duty, forgetting she needs to be able to fly to do that.
Applejack forgets that her friends can fly when she attempts a Train Escape in "The Last Roundup".
Twilight Sparkle spends half of the first couple of episodes teleporting every few seconds, but totally forgets she can do it when the hydra in "Feeling Pinkie Keen"is chasing her toward the rocks she ended up jumping on.
Forgotten Phlebotinum: After episode two, the ponies' Elements of Harmony powers and jewelry did not appear until the two-part Season 2 opener. They'd be overkill, anyway.
Four-Fingered Hands: Hands, as an unavoidable consequence of having four-legged animals as the dominant species, are rarely seen, but whenever they appear they have four fingers - an example comes from "Too Many Pinkie Pies", when one of the Pinkie clones causes three stubby fingers and a thumb to sprout from its hoof. Justified in the case of the more humanoid dragons (Spike, the teenage dragons seen in "Dragon Quest") as they aren't human.
Free-Range Children: The Cutie Mark Crusaders appear to have no constraints whatsoever, considering we see them doing things like practicing extreme sports deep in the forest or mountains with no supervision at all. Applejack never seems to worry about Applebloom unless the plot calls for it, and Rarity generally tends to get Sweetie Belle as far away from her as possible, due to Sweetie's tendency to cause havoc around her. Though we have seen their parents, they only appear in one episode for less than two minutes.
In "Griffon the Brush Off": When Rainbow Dash and Gilda race to a cloud, each claims to have been first, but Pinkie Pie claims Rainbow Dash won by just a little bit. To the naked eye it looks like a draw, with both arriving in a flash at the same time, but freezing the image at the right time shows Pinkie is right.
In "Sonic Rainboom": When Twilight Sparkle casts the wing-making spell, an outline of Rarity with her butterfly wings appears for a couple of frames in the explosion of light, several minutes before we get to see the results of the spell properly.
When her friends dismiss the broken table as who wants a hammer for a cutie mark, Apple Bloom looks dreadfully upset.
Just before Sweetie Belle starts singing, Scootaloo and Apple Bloom cover their ears.
In "The Cutie Mark Chronicles": During Fluttershy's flashback, at the end of her song, if you watch the upper-left corner of the screen carefully, you can see Rainbow Dash just as she hits her first Sonic Rainboom. See here.◊
The dog anatomy pictures in the Vet's room all have the outline of Winona, Applejack's dog.
When Spike recovers his normal size and Rarity says "YOU were the rampaging dragon?" Spike grins and strikes a "let me explain" pose for half a second before plummeting to the ground.
In "Baby Cakes": If you look closely during the opening, the babies in the nursery have the same or similar color schemes as other ponies, though most of them match more than one other pony (two that can be pinned down are the ones matching Rainbowshine and Medley).
This promo from the Hub riffs on the 'There's an App for That' adds from Apple. Every pony app shown has an appropriately punny name and a matching icon (e.g. Wikiponia, Hooftube, etc.), proper use of pony terminology for various app functions, and even the 'PNN' news article with tiny text which flashes by at the end is fully coherent and readable. Someone at The Hub obviously knew that that the Periphery Demographic would examine every frame for details like that, going so far as to give a Shout Out to popular Pony Blog Equestria Daily.
We rarely see Rarity and Rainbow Dash spend time together on-screen, let alone know what their relationship as friends are like. At most, they have a few snarky conversations and that's it. This is rather notable as this series mostly revolves around The Power of Friendship and True Companions.
There was in fact an episode revolved around Rarity wanting to be a good friend to Rainbow Dash, the plot barely required them to have a single conversation together.
Interestingly, the first season acted as something of a Deconstruction of this, since the six ponies were implied to be acquaintances before but only brought together due to their new found friendship with Twilight Sparkle, thus leading to many of them being put into situations together and gaining new bonds (eg. Applejack and Rarity in "Look Before You Sleep").
Furry Confusion: On top of the usual ponies, this world also boasts talking cows, mules, sheep, and donkeys, along with non-talking birds, bunnies, dogs, cats, and equines.
Furry Reminder: Many. Lauren Faust wanted it to be clear that the ponies were ponies, not pony-shaped humans. In fact, her reaction to the Bridlemaids ad was, "HUMAN POSES!!! AAARRRRGH!!!" Even the slight Anthropomorphic Shift of Season 2 doesn't stop the reminders from coming.
When faced with a threat and it looks like Violence is the Only Option, most of the horses lower their head, snort, and paw at the ground before charging. The pawing part alone is sometimes used to indicate displeasure.
Zecora's initial Character Tic of randomly pawing at the ground in her début episode is one of the reasons she's considered scary for this very reason.
"I heard Zecora eats hay." "Pinkie, I eat hay. You eat hay!"
Pinkie remembers this later when singing to the buffalo: "We all eat hay and oats—why be at each other's throats?"
When the ponies are getting ready for the Gala, Rarity won't let Spike come in because they're getting dressed. Applejack reminds her, "We don't normally wear clothes."
Rarity's song about dressmaking includes a line about cutting out the parts of the dress: "Piece by piece, snip by snip/Croup, dock, haunch, shoulders, hip".
In the same song there are the lines: "Making sure it fits forelock and crest/Don't forget some magic in the dress/Even though it rides high on the flank/Rainbow won't look like a tank"
In "The Ticket Master", Twilight picks off a flower's petals to determine which of her friends she gives the gala ticket to, gives up after one round... and then proceeds to eat the fallen petals.
At one point, Pinkie rolls around in the grass. You may think it's just her randomness, but real horses do that a lot.
In the episode "Over the Barrel"" there's a saloon in Appleloosa called "The Salt Block" where ponies serve salt to customers. This may seem bizarre, but if you're familiar with real horses then you know that salt is an important source of nutrition for horses (usually served in a form of solid blocks of salt). It also helps them keep cool, especially in hot areas (like the desert environment where Appleloosa is) through the minerals the consumed salt provides.
You can hear Big McIntosh let out an authentic neigh towards the end of "Lesson Zero".
You can also occasionally hear neighing and other horse sounds being uttered through out the show.
In "Return Of Harmony Part 2", Rainbow Dash gives some very horsey snorts, complete with little clouds from her nostrils, when the other five restrain her (which they do in a way one would restrain a real horse).
Pinkie makes a very horse-like snort of exasperation at (unsuccessfully) trying to get the infant twins she's babysitting to eat. Twilight Sparkle and Rainbow Dash have also both been made mad enough to snort at others on at least one occasion.
In "A Friend In Deed" Rose tosses a flower at Pinkie Pie, who catches it in her mouth and eats it.
In "A Friend In Deed" Cranky Doodle Donkey occasionally brays when something surprising happens.
Ponies have been known to buck or rear as signs of excitement or annoyance.
In some adventure/quest-based fanfics, (like It's A Dangerous Business, Going Out Your Door), the ponies don't pack as much food as one would think, as they can eat the surrounding grass and leaves to conserve their supplies or in case of emergency.
"Sleepless in Ponyville" sees a panicked Scootaloo run out of the cave everypony's sleeping in because she thinks she's heard the neigh of the Headless Horse. Turns out the real source of the noises is a sleeping Rainbow Dash.
While it's a little inconsistent and they are sometimes seen clapping hooves together instead, from "The Show Stoppers" onward, ponies are most commonly seen drumming hooves against the ground to signify applause.
Used as a constant source of humor in Equestria Girls.
"Can I have that [milkshake] with some extra oats?"
"What are hands?"
Geographic Flexibility: Ponyville can gain features and buildings with no explanation or prior appearance should the plot require it. Equestria as a whole also qualifies, and although there is an officialmap◊, the map has the disclaimer: "Distance Not To Scale"; most likely not to constrain the development in later episodes.
This would make the above map more like the caricatured postcard maps of IRL national/global landmarks, more than an one used for established travel.
Genius Bonus: The This Day Aria ends on as a "deceptive cadence", which is a stealth pun considering it's a duet between Princess Cadence and the shapeshifter posing as her.
Genre Shift: The series premiere was a High FantasyMagical Girl story that just happened to star ponies. Once Nightmare Moon was defeated and the world was saved, the show instantly shifted to a Slice of Life ensemble comedy with An Aesop at the end of nearly every episode. While these make up the bulk of the series, the Magical Girl stories still return on occasion, usually in season premieres and finales. Note that this doesn't stop the Mane characters from being badass on frequent occasions, it just means the that thematic focus isn't on that part.
Sometimes individual episodes switch to another genre entirely, like Western or Mystery.
Gentle Giant: Turns out both the manticore and the sea serpent in episode 2 are friendly and easy-going beings.
Big McIntosh also qualifies for this, given his huge build, calm personality, and how he patiently puts up with Applejack's stubborn nature.
Celestia counts. She's bigger, and more powerful than most of the cast and is Large And In Charge.
Used more conventionally to omit any explanation for how the Cutie Mark Crusaders ran into Twilight Sparkle in "The Cutie Mark Chronicles":
Scootaloo: Come on, girls! We need action! We need Rainbow Dash!
Twilight Sparkle: As a young filly in Canterlot, I always wanted to go to the Summer Sun Celebration....
Girls Need Role Models: One of the biggest messages the show has to offer is that there is no wrong way to be a girl.
Giving Up On Logic: The episode "Feeling Pinkie Keen" is all about Twilight Sparkle trying to find a logical reason how Pinkie Pie has the strange ability called "Pinkie Sense". By the end of the episode, Twilight Sparkle gives up, and learns that not all things need to be completely understood to be considered real or true.
Goal in Life: Discovering your purpose in life creates your cutie mark on your flank. Given how permanent those things seem to be, one has to hope there's no pony equivalent to a mid-life crisis.
Well, considering that most of the mane six's breakdowns had something or other to do with their cutie marke.g. Pinkie Pie in "Party of One", having an insanity scene; Fluttershy not being able to make the creatures of the Canterlot Gardens love her in "The Best Night Ever"; et cetera, it seems that the fan-named Cutie Mark Failure Insanity Syndrome is said equivalent.
The cutie mark just represents that particular pony's special talent, which often goes hand in hand with their chosen occupation. For example, Rainbow Dash's cutie mark signifies that her special talent is high-speed flying. She definitely enjoys it, and plans to make a career out of it by joining the Wonderbolts, but she's also shown great skill in weather control (clearing the sky in ten seconds flat, etc.). Another good example is Pinkie Pie. Her special talent, as shown by her three-balloons cutie mark is organizing parties. However, she's also an excellent ice skater and is able to play several instruments at once, which don't have any direct relation to parties.
Good Parents: Twilight Sparkle's parents and Rarity's parents.
Gosh Hornet: Twilight Sparkle has disturbed a bee's nest and been covered with stings twice: in "Winter Wrap-Up" and "Feeling Pinkie Keen". Rainbow Dash was chased by a swarm of bees in "Fall Weather Friends", but managed to get away from them. Interestingly, there's a beehive hanging from one of the upper branches of the library where Twilight lives.
Gonna Fly Now Montage: An almost literal one for Fluttershy in Hurricane Fluttershy, where she trains herself to be a stronger flyer (with her animal friends acting as motivational trainers).
Fluttershy: But Rainbow Dash, just because you failed the Sonic Rainboom a hundred thousand times in practice doesn't mean you won't be able to do it in front of an entire stadium full of impatient, super-critical sports fan ponies!
Green Rooming: Princess Luna and Zecora both got entire episodes dedicated to introducing each early on in Season 1, but neither really got any further screentime until Season 2 (barring one quick scene for Zecora in the episode immediately after her introduction).
Half-Dressed Cartoon Animal: Specifically the male ponies; most of them wear vests. As stated elsewhere, female ponies generally just wear some form of accessory.
Hard Light: Rainbows are a liquid. It's safely edible, but painfully spicy. It's also sticky enough to be used as war paint.
The Hecate Sisters: The three alicorns in Season 2: Cadance is the Maiden, Celestia is the Mother, and Luna is the Crone (despite being younger than Celestia, thanks to her role and personality).
Heel-Face Turn: At the end of "Keep Calm and Flutter On" Discord finally fixes the chaos he caused after Fluttershy declares her friendship with him is over, realizing that he didn't want to lose the only friend he ever had in the first place.
It's worth pointing out that he promised to use his powers for good instead of evil "most of the time," and that he took special interest in hearing that Twilight would be left in charge of the Elements of Harmony.
Helping Granny Cross the Street: Rainbow Dash feels outshined by Mare Do Well, and unable to find anyone in need of rescuing, obstinately insists on "helping" Granny Smith cross the street even though she didn't want to cross it at all.
His Own Worst Enemy: Princess Luna's worst enemy was herself. While the mane cast and her own sister had to stop her as Nightmare Moon, her suffering and even her banishment were of her own doing, mainly for her inability to deal with her jealousy and her loneliness.
Hobbits: Of the pony breeds, Earth ponies seem to fit this mold best. They also don't have overt magic or flight, making them a little more relatable to the audience.
Also, in "Green Isn't Your Color" Spike gets mad at Twilight for telling someone about his crush on Rarity...while he's wearing a fanboyish t-shirt with her picture on, making his crush pretty darn obvious.
Lesson Zero has Rarity, who often performs cartoonishly overdone displays of despair at minor problems, call Twilight a drama queen. Cue the others glaring at her and subsequent Hypocrisy Nod.
In "May The Best Pet Win", Rainbow Dash tells the tortoise competing to be her pet, "This isn't a game, you know!" She immediately tells the rest of the prospective pets, "These games will determine which one of you has the most important qualities I'm looking for in a pet."
Idiosyncratic Wipes: During the Title Sequence, each of Twilight Sparkle's friends in the Mane Six gets one, related to their specialities (and cutie marks): a dashing rainbow for Rainbow Dash, a party balloon inflating to cover all the screen for Pinkie Pie, a growing eye twinkle for Rarity, a shower of apples for Applejack, and a swarm of butterflies for Fluttershy.
I Just Want to Be Special: The Cutie Mark Crusaders (Apple Bloom, Scootaloo, and Sweetie Belle) want desperately to learn their special talents so they can earn their Cutie Marks and no longer be "Blank Flanks".
There are only eight significant recurring male characters; Spike, Big McIntosh, Snips and Snails, Donut Joe, Shining Armor, Mr. Carrot Cake, and his son Pound Cake. There are more significant background ponies than that.
The show has passed the inverse Bechdel Test (two male characters who speak about something other than a female character) only in two episodes, once with Snips and Snails have an argument about getting stuck with bubble gum, and another where Donut Joe and Gustav were arguing about whose dessert was best.
In another situation, "I'm powerful late for... something."
Applejack "looking at the time" when her wrist is completely empty.
Rainbow Dash drawing a wristwatch onto her foreleg to look at the time. (Her and Fluttershy's excuse for avoiding Pinkie Pie's party, which was the most ridiculous, was the one that Pinkie Pie chose to believe.)
From Rarity: "I have to go, to do the thing, with the stuff, you know..."
Most of the mane characters have names taken from the previous My Little Pony cartoon, but that's just out of convenience. Lauren Faust primarily based the mane cast's personalities on how she used to play with her own My Little Pony toys as a kid — toys that weren't even of the same characters except for Applejack. She also used the ponies from the G1 cartoon as inspiration. See 'Composite Character' for more information.
Also goes for many background ponies, both those that have been given a name in the show and those that only have Fan Nicknames.
Interquel: The German magazine comicApple Blooms großes Abenteuer is about the first meetup of the Cutie Mark Crusaders, and is thus set after the episode "Call of the Cutie" and before all of the episodes involving the club.
In the Name of the Moon: the first preformance of the mares as magical girls involved this; it's completely improvised, and almost perfectly executed (you can see Twilight's eyes glancing down for a second, looking for the right words) because Twilight Sparkle is just that eloquent. An especially ironic example given who the villain is.
Meta Example: None of the American voice actors for the main characters are American, they're all Candian.
Ironic Echo: Played with. During "Sonic Rainboom", Fluttershy says to Rainbow Dash (who's suffering from a bad case of stage fright) that there's no reason to feel nervous about facing a crowd of ponies who will watch, criticize, and judge each and every one of her moves. Later, during "Green Isn't Your Color" Photo Finish gives Fluttershy a similar speech when the nervous pegasus is about to begin her first modeling pass.
Characters have spoken in familiar languages and accents (French, German, Italian, Brooklyn, etc.).
"I Want" Song: Played straight with Twilight's verses in "Winter Wrap-Up"; played with in "At the Gala" in an Almost There-ish way in that the ponies are singing about what they want with the anticipation that it's going to happen soon, not in some distant future.
I Warned You: Nearly every episode in which Twilight isn't the main focus has her basically lampshading the Aesop partway through the story. Plot-Induced Stupidity is essentially the only reason she lacks this kind of foresight in her own episodes.
I Wish It Were Real: Some bronies wished the show's ponies were real (either to show their fondness to the characters or they genuinely wished they exist in real life). Fan works depicting them as pets, friends or them popping out of TV screens/monitors or coming to life from paper drawings are common within the fandom (with Pinkie Pie being depicted quite often for obvious reasons).
Longing for Fictionland: The reverse is also prominent within the fandom. Fan works and discussions about what would happen if they get transported to Equestria are quite common. Often it involves turning into ponies themselves.
Pinkie Pie, Applejack, and the Cutie Mark Crusaders use atashi.
Princess Celestia uses watakushi.
Japanese Sibling Terminology: In the Japanese dub, Apple Bloom refers to Applejack as "onee-chan", as well as Sweetie Belle towards Rarity, and Applejack refers to Big Macintosh as "nii-chan" and "aniki".
Jerkass: Throughout the first two seasons, the mane six have encountered their fair share of them.
The Flim Flam Brothers from "Super Speedy Cider Squeezy 6000". They take this trope to such an extreme that, by the end of the episode, the entire populace of Ponyville runs them out of town.
The teenage dragons Spike encounters during "Dragon Quest". They bully Spike, call Celestia a "namby-pamby pony princess", and want to eat the Phoenix eggs.
Lightning Dust in season 3's "Wonderbolts Academy". She caused the other cadets to spin out of an obstacle course, and caused a tornado in order to clear the most clouds despite already being so far ahead it was redundant. The move went out of control and destroyed a portion of Cloudsdale, as well as swallowed up Rainbow's friends who just happened to fly in to visit. Rainbow Dash managed to save her friends, while Lightning Dash makes light of the situation afterward. Rainbow Dash angrily called her out on her stunts that could have potentially hurt some ponies or worse, but that she didn't seem to care because they "won".
Jerkass Ball: A large premise in most episodes (often combined with Idiot Ball). Expect the main dilemma of most stories to be caused by one of the ponies acting arrogant, self centered or even borderline insane. Every character bounces this at least once in the series (usually in their limelight episodes).
Jerk Jock: The three male pegasi who bully Rainbow Dash in Cloudsdale. Their cutie marks are a dumbbell, three basketballs, and three American footballs.
Rainbow Dash herself and Applejack are milder examples of Jerk Jocks when they laugh at Twilight for entering the Running of the Leaves.
Jerk with a Heart of Gold: Rainbow Dash is this the majority of time, being egotistical and aggressive, but loyal and altrustic to her friends. Do not bully them.
Rarity can be considered a mild example, she's vain and greedy at times, but it is heavily outweighed by her genorosity and empathetic moments.
Fluttershy's pet rabbit, Angel Bunny. He comes off as an abrasive spoiled child at times but he's shown that he really does care about his owner.
Spike also fits. He may be lazy and snarky, but he really cares about Twilight and the others, and is still a good guy.
Justified Trope: Revealing the source of a seemingly hard-to-justify trope is often used as a punchline.
Kick the Dog: The Villain Song "This Day Aria". "No I do not love the groom/In my heart there is no room/But I still want him to be all miiine!" Ouch.
Killer Rabbit: Angel is a light-hearted take on one. The Parasprites too — especially when they start trying to eat Ponyville.
Knockout Gas: In "Mmmystery on the Friendship Express", Pinkie gives explanations for how culprits could have committed a crime, all of which parody movies. One of these involves her getting knocked out by gas.
Know Your Vines: In "Bridle Gossip", the mane six walk through a patch of bright blue flowers to stop Apple Bloom following Zecora. The next morning they wake with embarrassing changes, such as Pinkie's swollen tongue, that they blame Zecora for. Eventually it is revealed that the flowers were 'poison joke' and the effects were pranks. Played by the plant.
Kubrick Stare: Pinkie Pie gives a surprisingly chilling one in "Party of One".