Willie Rennie has said Liberals will need to "stand up and be counted" if Nicola Sturgeon does call a second independence referendum.
But he insisted that if Scotland's future in the UK was put to the vote again, his party could "send a message of hope" and "turn back the tide of division".
The Scottish Liberal Democrat leader spoke out ahead of the start of his party's spring conference in Perth.
READ MORE: Fresh split in independence movement over referendum date
The gathering comes at a time of continued speculation that Ms Sturgeon will press for a second vote on independence to be held.
While almost two-thirds (62%) of Scots voted to keep Britain in the European Union in June 2016, the UK as a whole voted to leave, with this difference heightened further by Conservative plans for a hard Brexit.
Mr Rennie said: "If there is to be another independence referendum, the responsibility on Liberals is great.
"We must stand up and be counted for our values."
He claimed the Conservatives "have done more damage to the Union with their Brexit gamble" and said that the "complete disarray" among Labour had turned it "from the party of opposition to the party of compliance".
The Lib Dem MSP continued: "The argument of independence is a battle of ideas and values, not of identities and flags.
"We must stand up for our family whether in Britain or across Europe.
"We must make the positive, open, internationalist case.
"Send a message of hope that things can be better still.
READ MORE: Fresh split in independence movement over referendum date
"We can turn back the tide of division.
"We can celebrate both our differences and the ties that bind us.
"We can say no to independence and yes to partnership.
"We can, once and for all, put an end to the claim that if you do not believe in independence you do not believe in Scotland.
"Our job is to turn back the tide of division."
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