Ruth Davidson has slammed the SNP leader's property tax hikes
Tory leader Ruth Davidson challenged the First Minister to do more to help firms caught out by business rates rises.
Firms say they face "crippling" bills within weeks because of the first revaluation since 2010.
Many are also hammered by the SNP's "super-tax" on large and medium-sized businesses.
Ms Davidson cited engineering firm Precision Oil Tools, based in Kintore, Abderdeenshire, which will see its rates go up by 63 per cent in April.
Its finance director, Bev Robertson, has said the hike is "nothing short of daylight robbery" and faces having to "cut our staff numbers."
Peterhead firm Score group has also discovered it will have to pay an extra £120,000 – turning apprentices away as a result.
The Scottish Tory leader argued business confidence in Scotland is falling
In last month's draft Budget, Finance Secretary Derek Mackay rejected calls to provide "transitional rates relief" to help firms cope with the shake-up.
Unemployment in Scotland is up, employment is down
He also refused to scrap the controversial large business supplement, which last year doubled from 1.3p in the pound to 2.6p – although the threshold was lifted to £51,000.
Latest figures show the country is still lagging well behind the rest of the UK.
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Official statistics show unemployment has soared north of the Border – at the same time as it fell across the UK.
Rounding on the First Minister, Ms Davidson said: "The facts are these.. Unemployment in Scotland is up, employment is down.
"While confidence for small firms in other parts of the UK is going up, here it is falling through the floor.
"Yet we have a Finance Secretary who is hidden from companies that say that rates are pushing them to the wall. We have got a Government that taxes people and firms here more than anywhere else in the UK, and again this week they are threatening further instability with their own referendum, this time another one on independence.
"This Government is about to present the most important budget since devolution, deciding on the taxes that Scots pay.
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Nicola Sturgeon visits Glaxo Smith Kline.
"And the question is this, does the First Minister stick to her current plan of making Scotland the most highly taxed part of the UK, driving out businesses and jobs, or will she change course?"
From April, higher rate taxpayers will pay £314 more income tax per year in Scotland than in the rest of the UK because the Scottish Government will raise the threshold "in line with inflation" to £43,430 – while it will rise to £45,000 across the UK. Business leaders say the SNP's Budget plans could widen the gap in future.
But Ms Sturgeon echoed denials by Mackay that Scotland will become the highest-taxed part of the UK – because people receive more free services.
Mrs Sturgeon has been criticised for making Scotland the most taxed part of the UK
She dismissed the charge as "ridiculous" because of giveaways such as free university tuition and personal care.
"Taxpayers in Scotland get a far better deal than taxpayers in the rest of the UK," Ms Sturgeon said.
She also insisted her administration was "on the side of business" with with an expansion of its small business bonus scheme lifting 100,000 firms out of rates.
She added: "This Government has ensured we have a competitive, probably the most competitive, business rates regime anywhere in the UK because that is the importance we attach to small businesses."