The region has come to a halt due to the impromptu strike, with one group igniting a burning wall of tyres on the busy B20 near Barcelona, in St Boi de Llobregat.
Other demonstrators halted traffic on motorways and other main highways with human chains or wooden pallets.
About 50 roads are believed to be affected as the wealthy region came to a standstill this morning, with thousands of people unable to get to work.
Huge banners have appeared across major highways, with one reading “Sorry for the inconvenience. Catalan Republic under construction”, and other with “Not a step back against repression. We are the republic. General strike” written across it.
Some train lines were also targeted as about 600 protestors entered the AVE railway station in Girona to interrupt traffic.
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Staff from a Catalan TV and radio station also joined the strike, called by a pro-independence union against the jailing of sacked members of the Catalan government.
They are also demonstrating the continued incarceration of the leaders of two pro-independence grassroots movements and the Spanish government’s imposition of direct rule through Article 155 after the Catalan Parliament’s unilateral declaration of independence on October 27.
Eight former members of the Catalan government have been imprisoned while the ex-Catalan leader, Carles Puigdemont, and four other ex-ministers try to force the European Union to intervene in the dispute.
Mr Puigdemont fled to Brussels shortly after the Madrid government imposed direct rule on Catalonia, but he is expected to be extradited to Spain in the coming days.
The disputed Catalan leader accused Madrid of carrying out a “brutal judicial offensive” against members of his deposed regional government.
Mr Puigdemont said: “The Spanish state must honour what was said so many times in the years of terrorism: end violence and we can talk about everything.
“We, the supporters of Catalan independence, have never opted for violence, on the contrary. But now we find it was all a lie that everything is up for discussion.”
Speaking to Catalan mayors who had flown to Brussels yesterday, Mr Puigdemont was greeted with cries of “freedom” and “president” as he declared: “We will never renounce this ideal of a country, of this notion of democracy.
“Is this the Europe you want, is this the Europe you want to build, with a democratically elected government in jail?
“We are the legitimate government of Catalonia and we are going to carry on.”
Additional reporting by Gerard Couzens