Abstract

Pseudokirchneriella subcapitata is a sickle-shaped freshwater green microalga that is normally found in unicellular form. Currently, it is the best known and most frequently used species of ecotoxicological bioindicator because of its high growth rate and sensitivity to toxicants. However, despite this organism’s, our knowledge of its cell biology—for example, the patterns of nuclear and cytoplasmic division in the mitotic stage—is limited. Although it has been reported that P. subcapitata proliferates by popularity forming four daughter cells (autospores) through multiple fission after two nuclear divisions, here, we report two additional reproductive patterns by which two autospores are formed by binary fission (“two-autospore type”) and eight autospores are formed by multiple fission (“eight-autospore type”). Moreover, we found that cell reproductive patterns differed markedly with the culture conditions or with exposure to either of two typical toxicants, potassium dichromate (K2Cr2O7) and 3,5-dichlorophenol (3,5-DCP). The eight-autospore type occurred at the highest frequency in the early phase of culture, but it disappeared under 3,5-DCP at 2.0 mg/L. Under 0.3 mg/L K2CrO7 (Cr(VI)) the eight-autospore type took substantially longer to appear than in control culture. The two-autospore type occurred only in the late phase of culture. To our knowledge, this is the first detailed evaluation of the reproductive patterns of P. subcapitata, which changed dramatically in the presence of toxicants. These findings suggest that observation of the reproductive patterns of P. subcapitata will help to elucidate different cell reactions to toxicants.

Highlights

  • In water-quality monitoring and ecological risk assessment, bioassays are deployed as valuable tools to reveal the biological effects of chemicals or wastewater

  • In order to ensure identity of National Institute of Environmental Studies (NIES)-35 with phylogenetic analysis, we chose the sequence of plastidencoded Rubisco large subunit because of many genes, the only rbcL originated from S. capricornutum (UTEX 1648) has been sequenced in high quality and available in public database

  • We report two additional reproductive patterns in P. subcapitata that form two or eight autospores, apart from the already known reproductive pattern in which four autospores are formed by multiple fission after two nuclear divisions [4] (Fig 6)

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Summary

Introduction

In water-quality monitoring and ecological risk assessment, bioassays are deployed as valuable tools to reveal the biological effects of chemicals or wastewater. Skulberg, is a sickle-shaped, freshwater green microalga that is normally found in unicellular form [1,2,3] It is the best known and most frequently used ecotoxicological bioindicator species because of its high growth rate, sensitivity to toxicants, and good reproducibility compared with those of other algae. Despite this organism’s popularity, our knowledge of its cell biology—such as the patterns of nuclear and cytoplasmic division in the mitotic stage—is limited. The four autospores are typically released through rupture of the parental cell wall without forming colonies It is unclear how cytokinesis is achieved in the multinucleated cells containing four nuclei before autospore release. Similar cell-size changes have been reported in other algae [6,7,8,9,10], their mechanism is unclear

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