Coaches will replace trains on the Bendigo, Swan Hill, and Echuca lines for 24 hours.

It follows industrial action in December that saw trains on every regional line cancelled for a day.

"It's pretty disappointing for us," V/Line chief executive James Pinder said. "It's very unnecessary - we've been negotiating now in good faith with the union for quite some time.

"We believe we've got a really good package to offer them that takes into consideration the fact that V/Line is the fastest growing railway.

"We want to modernise our railway. This is not about paying people any less or taking away allowances. This is all about making sure we create the V/Line that all Victorians want for the future."

Read more: Commuters could be left stranded after V/Line workers plan strike

The RTBU and V/Line have been negotiating since January, 2019.

"We don't take any action lightly and ultimately would prefer it be avoided, but RTBU members are fed up and the public deserves a way forward," RTBU branch secretary Luba Grigorovitch said. "This campaign is all about conditions.

"Our members take great pride in their work, getting people from A to B, but when Government and the Company team up to attack their ability to deliver that service they take it very personally."

The RTBU has planned industrial action on other regional lines, including the Maryborough line, on February 18.

But Mr Pinder said he hoped an agreement could be reached before further strikes.

"We're meeting with them again next week and we'd hope they would cancel any further action on the back of those meetings," he said.

"We have got a government now that has offered mediation in this space, either independently or through the Fair Work Commission.

"We don't think the action is necessary at all. We'd like to encourage the union to get back to the negotiating table."

Public Transport Users Association regional spokesperson Paul Westcott said the ongoing industrial action was affecting regular train commuters.

"While V/Line has been given notice from the union and therefore have been able to employ buses to replace most of the train services, these buses have to run to a slower schedule than the trains," Mr Westcott said.

"This can extend peoples' journeys for sometimes up to an hour. It really is annoying for people, even if they have been given notice."

Mr Westcott called on V/Line and the union to work towards an agreement.

"We have certainly urged both parties to resolve the situation but we won't get involved," he said.

"We represent the passengers who are the meat in the sandwich. We want to be assured that both sides are continuing to talk."

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