The definition of male and female will be based on whether people are born with eggs or sperm, rather than on their chromosomes.
Under the order, prisons and settings such as shelters for migrants and rape victims would be segregated by sex, based on this criteria.
Officials also said the order would impact federal documents including passports.
The order would also block requirements at government facilities and at workplaces that transgender people be referred to using the pronouns that align with their gender.
Mr Trump’s team says those requirements violate the First Amendment’s freedom of speech and religion.
The second “common sense” order targets diversity, equity and inclusion (DEI) initiatives, and ends their federal funding.
As part of this, officials said there would be a monthly meeting of relevant agencies to assess any DEI programs and whether they should be shut down.
Speaking in a rally on Sunday ahead of his inauguration, Mr Trump also said that he would stop the participation of trans athletes in women’s sports.
The move will undo parts of one order President Joe Biden signed on his first day in office four years ago.
Throughout his campaign, Mr Trump has been vocal in his opposition to increased support and rights for transgender and nonbinary people.
The order does not appear to issue a nationwide mandate on which bathrooms transgender people can use or which sports competitions they can join, though many states have passed laws on those areas.
Civil rights groups were already preparing to challenge Mr Trump’s restrictions in court before he set foot in the Oval Office.
“We are going to persevere, we’re going to continue in our work and we’re going to continue to protect trans rights throughout the country,” said Ash Orr, a spokesperson for Advocates for Trans Equality last week, anticipating such an order.
Donald Trump has signed a raft of executive orders within hours of returning to the White House as he reversed some of his predecessor Joe Biden’s policies.
The new president told supporters at an inauguration parade event at a sports arena in Washington DC that he would revoke “80 destructive and radical executive actions of the previous administration”.
:: Pardons for about 1,500 Trump supporters who have been charged over the January 6 2021 riot at the US Capitol.
The action also cuts short the sentences of 14 members of the far-right Proud Boys and Oath Keepers organisations, including some who were convicted of seditious conspiracy.
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Climate
:: Withdrawing the US from the Paris climate agreement. The announcement further threatens the central goal of the agreement to avoid a rise in global temperatures of 1.5C.
Mr Trump withdrew the US from the agreement during his last presidency, but President Biden reversed this decision.
::Revoking a 2021 Biden order that sought to ensure half of all new vehicles sold in the US by 2030 were electric.
Energy
:: Repealing a 2023 memo from Mr Biden that barred oil drilling in 16 million acres in the Arctic. The new president declared a national energy emergency, promising to fill up strategic oil reserves and export US energy all over the world.
“We will be a rich nation again, and it is that liquid gold under our feet that will help to do it,” the American leader said.
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Trump inauguration: Key moments
Illegal immigration
:: Declaring illegal immigration at the US-Mexico border a national emergency as he plans to send US troops to help support immigration agents and restrict refugees and asylum.
The president has also designated criminal cartels as terrorist organisations, and has taken steps to block citizenship for children of immigrants who are in the US illegally.
Mr Trump reversed several immigration orders from the Biden presidency, including one that narrowed deportation priorities to people who commit serious crimes, are deemed national security threats, or were stopped at the border.
Foreign affairs
:: Temporarily suspending all US foreign assistance programmes for 90 days pending reviews to determine whether they are aligned with his policy goals.
It was not immediately clear how much assistance would initially be affected by the order as funding for many programmes has already been appropriated by Congress and is obligated to be spent, if not already spent.
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1:33
Who was at Trump’s inauguration?
Free speech
:: Restoring freedom of speech and preventing censorship of free speech.
:: The president signed a document “ending weaponisation” of government against political opponents.
The order directs the attorney general to investigate the activities of the federal government over the last four years, including at the Department of Justice, the Securities and Exchange Commission, and the Federal Trade Commission during the previous administration.
It said the government will “identify and take appropriate action to correct past misconduct by the federal government related to the weaponisation of law enforcement and the weaponisation of the intelligence community”.
Death penalty
:: Mr Trump signed a sweeping order on the death penalty, directing the attorney general to “take all necessary and lawful action” to ensure that states have enough lethal injection drugs to carry out executions.
Mr Trump said in the order that “politicians and judges who oppose capital punishment have defied and subverted the laws of our country”. A moratorium on federal executions has been in place since 2021, and only three defendants remain on federal death row after Democratic President Joe Biden converted 37 of their sentences to life in prison.
TikTok
:: Mr Trump signed an executive order to keep TikTok operating for 75 days, a boost for the social media platform’s users even as national security questions remain.
TikTok’s China-based parent company ByteDance was supposed to find a US buyer or be banned on 19 January. Mr Trump’s order could give ByteDance more time to find a buyer.
“I guess I have a warm spot for TikTok,” Mr Trump said.
Return to office
:: Ordering federal workers to return to the office five days a week.
:: The return-to-office order comes alongside a federal hiring freeze, except for the military, and the creation of an advisory body – the Department of Government Efficiency (DOGE) – aimed at reducing the federal government’s size.
Cost of living
:: There was a directive to every department and agency to address the cost of living crisis.
Gender and equality
:: Ending “wasteful” government programmes promoting diversity, equity, and inclusion, defend women from “gender ideology extremism” and restore biological truth to the federal government ( two sexes male and female).
Drug cartels
:: Designating cartels and other organisations as foreign terrorist organisations and specially designated global terrorists. “International cartels constitute a national-security threat beyond that posed by traditional organized crime.”
Donald Trump has said he will declare an emergency at the US-Mexico border as one of a host of executive orders the incoming president will sign on Monday and in the coming days.
Incoming White House officials have said the first wave of executive orders will cover a mix of campaign trail promises, reversals of outgoing President Joe Biden’s policies and restructuring of the federal government workforce.
One of the most high-profile orders will be Mr Trump declaring a national emergency on the US-Mexico border as part of a wider crackdown on immigration and other cross-border crimes.
Sky News’ US partner NBC News has reported Mr Trump is expected to sign more than 50 executive orders on Monday, and possibly more than 100, citing a person in his transition operation.
Border and immigration
On the border, Mr Trump said he would declare a national emergency in his inaugural speech.
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During the inauguration ceremony, Mr Trump said: “All illegal entry will immediately be halted and we will begin the process of returning millions and millions of criminal aliens back to the places they came.
“I will end the practice of catch and release and I will send troops to the southern border to repel the disastrous invasion of our country.
“Under the orders I sign today we will also be designating the cartels as foreign terrorist organisations.”
However, the border emergency is not the only one Mr Trump intends to declare.
During his speech, the president said he would also declare “a national energy emergency”, adding: “We will drill baby drill.”
He continued: “We will be a rich nation again, and it is that liquid gold under our feet that will help to do it.”
NBC News reported this is aimed at expanding the administration’s ability to improve domestic energy production.
Alongside that, he reportedly will: allow more drilling both offshore and on federal lands, end a freeze on liquid natural gas exports and cut off climate-related funding that was approved under the Inflation Reduction Act signed by President Biden.
In another executive order, Mr Trump reportedly will implement Schedule F.
This eliminates job protections for potentially thousands of federal workforce positions, which could make it easier to fire them.
Gender and diversity
Matching talking points from the campaign trail, Mr Trump said it would be government policy, through an executive order, that there would only be two genders recognised by the US government – male and female.
Speaking on Martin Luther King Jr Day he also promised to slash diversity, equity and inclusion (DEI) efforts, saying: “This week, I will also end the government policy of trying to socially engineer race and gender into every aspect of public and private life.
“We will forge a society that is colour-blind and merit-based.
“As of today, it will henceforth be the official policy of the US government that there are only two genders, male and female.”
Mr Trump also intends to reverse an order by President Biden that allows transgender people to serve openly in the military in their self-identified gender.
The order also provided greater access to medical care and even support for gender transition.
The Pentagon and military
Alongside that, NBC News said that Mr Trump would reverse two other decisions by the Democrat who beat him in the 2020 election.
One would change the location of the US Space Command headquarters, moving it from Colorado to Alabama.
The other would reverse a 2022 decision that provided paid leave and reimbursed transportation costs for troops and dependents who travelled out of state for abortion and other reproductive care.
TikTok and Hunter Biden
Mr Trump has said he would sign an extension for TikTok to find a buyer, to comply with the law that went into effect on Sunday.
Mr Trump would also reportedly suspend security clearance for 51 national security officials who signed a letter saying that Hunter Biden’s emails and laptop had the hallmarks of a Russian operation.
“Hats off! No bags! No drinks, no food, no water, no nothing. Everything out of your pockets. Come on – let’s go, let’s go let’s go!”
Security’s tough at the front of the line. There is a real sense of urgency.
It is bitterly cold outside Washington’s Capital One Arena and people have been queuing since 3.30am. Everyone’s keen to get in.
The queue snakes all the way around the back of the Martin Luther King Jr Memorial Library and down the block.
Inauguration Day happens to coincide with MLK Day (Martin Luther King Jr Day) this year. There was meant to be an outside match to commemorate that too, but it’s been cancelled because of the cold.
It’s now 7am and the queue starts moving. There’s a huge cheer at the front.
Despite the freezing temperatures and wind chill, people are excited. They came here for Trump, and it would have been colder on the National Mall in any case. They’re psyched.
“I spent so much money on my bills – they’re gonna go down, everything’s gonna go down and my pay rate is gonna go up,” says Dan French, who’s dressed as Uncle Sam.
Tabitha agrees. “New York, where I’m from, is a hot mess,” she says. “Everything is expensive, housing, food. Trump is going to come in and clean this up.”
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