The separatist protesters from across the south gathered at Parade Square in the city of Aden on Saturday, that is the anniversary of the 1967 independence of former South Yemen.
The protesters waved flags of former South Yemen and carried banners with pro-independence slogans.
They also denounced the ongoing national talks aimed at drawing up a new constitution and preparing for elections.
"No to dialogue, yes to independence and liberation," and "Our demand is independence,” the protesters chanted.
Majed al-Shuwaibi, a member of the Southern Movement, said October 12 is a significant day, as the southerners "celebrate the anniversary of the October revolution ... which will continue for the re-establishment of the state.”
North and south Yemen unified in 1990 after the southern government collapsed. But four years later, the south tried to break away and this led to a civil war. The conflict ended with northern troops taking control of the south after winning the war.
The Southern Movement gained strength during mass demonstrations which forced former Yemeni president, Ali Abdullah Saleh, to step down in February 2012.
The secessionist movement calls for autonomy or the complete independence of the south. Southerner residents complain that they have been economically and politically marginalized by the central government.
HH/HH