Anas Sarwar is to trigger a vote of no confidence against the Scottish Government next week.
Unlike the Scottish Conservative's planned vote of no confidence against Humza Yousaf, losing a vote of no confidence in the administration would have consequences, with the First Minister and his ministers forced to resign.
The Tories and the Scottish Lib Dems have said they will support the Scottish Labour leader's bid to oust the SNP leader and ultimately spark an early election.
However, the Scottish Greens and Alba have dismissed the motion, meaning it will likely fail.
News of the parliamentary push came as the First Minister insisted he would not be resigning.
Speaking to LBC, Mr Sarwar, who today was campaigning in Dunbar, said: "I think it's now a matter of when not if Humza Yousaf resigns as First Minister, but I think it'd be completely untenable for the SNP to presume they can impose another unelected First Minister on Scotland.
"And that's why Scottish Labour has said already we don't have confidence in terms of Humza Yousaf, but today we will also be laying a motion before Parliament, saying that the Scottish Parliament does not have confidence in the Scottish Government.
"Because ultimately, I think it's the people of Scotland that should set aside who leads this country not just a small group of SNP members."
The motion, tabled on Friday, simply states "that the Parliament has no confidence in the Scottish Government."
If passed, the First Minister would be required, as per the Scotland Act, to tender his resignation.
The Presiding Officer would then nominate an acting first minister but there would be a vacancy and Holyrood would have 28 days to choose a replacement.
If MSPs do not select a new first minister in that time frame, Parliament would be dissolved and an early election would be sparked.
Mr Sarwar said MSPs from across parliament should support the motion.
“Removing Humza Yousaf alone will not deliver the change we need – we need to get rid of the entire SNP government which has left almost one in six Scots on an NHS waiting list, public finances in chaos and public services at breaking point.
“The people of Scotland deserve a fresh start – this can only come with a Scottish election.
“Scottish Labour is ready and willing to deliver the change we need – from restoring our NHS, to creating jobs and making Scotland a global leader in clean energy.
“The time for change has come.”
There are 129 MSPs in Holyrood, including Presiding Officer Alison Johnstone, who would have a casting vote if the result was tied.
The SNP currently has 63 MSPs, the Scottish Conservatives 31, Scottish Labour 22, the Scottish Greens seven, the Scottish Liberal Democrats four and Alba has Ash Regan.
Ms Regan, who defected from the SNP last year to join Alex Salmond’s party, is the only opposition MSP not to state how they will vote on the motion of no confidence in Mr Yousaf.
She wrote to the First Minister this morning calling for a "reset.”
The ex-minister said her priorities were Scottish independence, “the dignity, safety and rights of women and children” and providing competent government.
Asked if they could support Labour's motion of no confidence in the government, an Alba spokesman told The Herald: "Anas Sarwar’s grandstanding is just an attempt to remain relevant. However, unionist parties should beware in their glee.
"As much as Humza Yousaf and the SNP find themselves in a tricky predicament, independence supporters will not turn back to the yoke of Tory and Labour mediocrity.
"Alba Party are now ensuring that independence supporters have a party they can turn to."
Green MSP Mark Ruskell said Labour "clearly don’t want this motion to pass."
"It was the poor judgement of Humza in ending the Bute House Agreement that is in question, not the record of the SNP/Green Gov," he tweeted.
Labour clearly don’t want this motion to pass.
— Mark Ruskell (@markruskell) April 26, 2024
It was the poor judgement of Humza in ending the Bute House Agreement that is in question, not the record of the SNP/Green Gov. https://t.co/UndPqVW0Wv
Scottish Tory chairman Craig Hoy, a former East Lothian councillor, said: “The Scottish Conservatives will always push to remove the SNP from power at every turn and will support a confidence motion in the government, but we believe the initial focus of all opposition parties needs to be solely on removing Humza Yousaf from office until that has been achieved.
“The Greens have lost confidence in Humza Yousaf but they may not vote to bring down the entire SNP Government.
"Labour must avoid an overstep that lets Humza Yousaf wriggle free when he is there for the taking. Anas Sarwar has let the SNP off the hook before on the Hate Crime Act and Gender Recognition Reform Bill. He should not be naive and hand Humza Yousaf a lifeline.
“The Scottish Conservative confidence motion has forced Humza Yousaf to the brink and we urge opposition parties to focus on delivering that first.”
Lib Dem leader Alex Cole-Hamilton said: "I want to see a government that is focused on driving down waits for healthcare, tackling the sewage in our rivers and taking action on the cost of living.
“This is clearly not that government.
“For that reason, Scottish Liberal Democrats are prepared to vote no confidence in this failed nationalist regime and will provide the remaining signatures required to bring this motion to Parliament."
Mr Sarwar was in Dunbar today, where he visited the BeGreen Energy Advice Centre on High Street, accompanied by the party's East Lothian Westminster candidate, Douglas Alexander.
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