Ploidy determination of three Kappaphycus alvarezii strains (Rhodophyta, Gigartinales) by confocal fluorescence microscopy

Abstract

Kappaphycus alvarezii is a red alga that is commercially important as a source of carrageenan. Since K. alvarezii presents large phenotypic plasticity and rarely develops reproductive structures in culture, identification of gametophytic and tetrasporophytic phases in cultivation systems are difficult. The aim of this study was to determine the ploidy of three K. alvarezii strains previously identified as brown “tetrasporophyte”, brown “gametophyte” and “Edison de Paula” (EP). Nuclei from these strains were stained with DAPI, and analyzed using confocal fluorescence microscopy and ImageJ software. The brown “tetrasporophyte” had the highest nuclear fluorescence intensity, consistent with a diploid tetrasporophyte (2N). The brown “gametophyte” and “EP” strains had nuclear fluorescence intensities of 55.78% and 57.10% in relation to the tetrasporophyte, respectively, consistent with haploid gametophytes (N). The present study demonstrated that this technique can be used as a rapid and effective tool to distinguish between haploid (gametophytic) and diploid (tetrasporophytic) plants of K. alvarezii, in addition to help identify new strains developed through alterations of ploidy level.