H S Shankarlinge Gowda, who represents Chamaraja Assembly constituency, waited for more than 90 minutes for Speaker K G Bopaiah and submitted his resignation letter, official sources said.
Bopaiah immediately accepted the resignation of Gowda, who is now all set to join JDS.
Gowda's resignation comes two days after Forest Minister C P Yogeeshwar and Small Scale Industries Minister Rajugouda quit the Jagadish Shettar Cabinet. Yogeeshwar had also submitted his resignation from the assembly membership to Bopaiah in person at Madikeri yesterday.
However, the Speaker is yet to announce his decision on Yogeeshwar's resignation.
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With Gowda quitting his assembly membership, BJP's strength in the 225-member House has come down to 104, followed by Congress 71, JDS 25, Ind 7, nominated one, speaker one and 16 vacancies. If Yogeeshwar's resignation is accepted, the strength will further go down to 103, posing threat to its survival.
Of the seven independents, Varthur Prakash, who is in the ministry, supports the government and B Sreeramulu of BSR Congress has also declared his support.
The resignation letters of Vittal Katakadonda and D S Suresh of BJP, who submitted their resignation two weeks ago, are yet to be accepted.
"Let those who want to quit, let them resign. There is no threat to the government. I will complete term", he told reporters here.
Assembly polls are due in May.
Meanwhile, Opposition Congress leader in the Assembly Siddaramaiah said his party would approach Governor H R Bhardwaj "if the government loses majority".