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. 2017;56(4):469-475.
doi: 10.2216/17-3.1. Epub 2017 Apr 27.

Rediscovery of the species of 'ancestral Volvox': morphology and phylogenetic position of Pleodorina sphaerica (Volvocales, Chlorophyceae) from Thailand

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Rediscovery of the species of 'ancestral Volvox': morphology and phylogenetic position of Pleodorina sphaerica (Volvocales, Chlorophyceae) from Thailand

Hisayoshi Nozaki et al. Phycologia. 2017.

Abstract

Pleodorina sphaerica Iyengar was considered to be a phylogenetic link between Volvox and the type species Pleodorina californica Shaw because it has small somatic cells distributed from the anterior to posterior poles in 64- or 128-celled vegetative colonies. However, cultural studies and molecular and ultrastructural data are lacking in P. sphaerica, and this species has not been recorded since 1951. Here, we performed light and electron microscopy and molecular phylogeny of P. sphaerica based on newly established culture strains originating from Thailand. Morphological features of the present Thai species agreed well with those of the previous studies of the Indian material of P. sphaerica and with those of the current concept of the advanced members of the Volvocaceae. The present P. sphaerica strains exhibited homothallic sexuality; male and facultative female colonies developed within a single clonal culture. Chloroplast multigene phylogeny demonstrated that P. sphaerica was sister to two other species of Pleodorina (P. californica and Pleodorina japonica Nozaki) without posterior somatic cells, and these three species of Pleodorina formed a robust clade, which was positioned distally in the large monophyletic group including nine taxa of Volvox sect. Merrillosphaera and Volvox (sect. Janetosphaera) aureus Ehrenberg. Based on the present phylogenetic results, evolutionary losses of posterior somatic cells might have occurred in the ancestor of P. californica and P. japonica. Thus, P. sphaerica might represent an ancestral morphology of Pleodorina, rather than of Volvox.

Keywords: Molecular phylogeny; Morphology; Pleodorina; Pleodorina sphaerica; Sexuality; Somatic cell; Volvox.

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Figures

Figs 1–9
Figs 1–9
Light microscopy of Pleodorina sphaerica Iyengar strains 2015-1128-2P-4 (Figs 1–7) and 2015-1128-2P-12 (Figs 8, 9) originating from Thailand. Figs 1–5. Vegetative phase. Abbreviations: fb, flagellar base; p, pyrenoid; v, vacuole; s, stigma. Fig. 1. 128-celled colony. Scale bar = 50 μm. Fig. 2. Surface view of colony showing large reproductive cells and small somatic cells. Note that reproductive cell has two flagella and lacks stigma. Scale bar = 10 μm. Fig. 3. Optical section of somatic cell. Scale bar = 10 μm. Fig. 4. Semi-optical section of top view of reproductive cell. Scale bar = 10 μm. Fig. 5. Colony stained with methylene blue. Note that distinct individual sheaths are lacking in the gelatinous matrix. Scale bar = 20 μm. Figs 6–8. Asexually reproducing colonies. Fig. 6. Two- or four-celled stage. Scale bar = 50 μm. Fig. 7. Daughter colonies developing within the parental colony. Scale bar = 50 μm. Fig. 8. Portion of colony showing a transparent vesicle (tv) encompassing each daughter colony. Scale bar = 20 μm. Fig. 9. Portion of newly formed daughter colony showing short flagellum (sf) and long flagellum (lf) in each protoplast. Scale bar = 10 μm.
Figs 10–17
Figs 10–17
Light microscopy of sexual reproduction in Pleodorina sphaerica Iyengar strains 2015-1128-2P-4 (Figs 11, 12) and 2015-1128-2P-12 (Figs 10, 13–17) originating from Thailand. Fig. 10. Male colony with developing sperm packets (dsp). Scale bar = 50 μm. Fig. 11. Male colony with sperm packets (sp). Scale bar = 50 μm. Fig. 12. Sperm packets (sp) developing within the male colony. Scale bar = 20 μm. Fig. 13. Male gamete. Note no cytoplasmic protuberance at the same of the flagella (f). Scale bar = 10 μm. Fig. 14. Female colony with immature zygotes (z) and penetrating male gametes (m). Scale bar = 50 = μm. Fig. 15. Immature zygote with double-layered cell wall. Scale bar = 10 μm. Fig. 16. Female colony with mature zygotes (z). Scale bar = 50 μm. Fig. 17. Mature zygote. Scale bar = 10 μm.
Figs 18–21
Figs 18–21
Transmission electron microscopy of vegetative colonies of Pleodorina sphaerica Iyengar strain 2015-1128-2P-12 originating from Thailand. Each protoplast is enclosed by a dense fibrillar layer (cellular envelope; arrows) of the extracellular matrix inside a tripartite layer (colonial boundary; asterisks) encompassing whole colony. Abbreviations: c, chloroplast; G, Golgi body; m, mitochondrion; N, nucleus; p, pyrenoid; v, vacuole. Fig. 18. Part of colony showing somatic cell (left side) and reproductive cell (right side). Scale bar = 2 μm. Fig. 19. Median section of reproductive cell showing large vacuoles. Scale bar = 2 μm. Fig. 20. Peripheral portion of colony. Scale bar = 1 μm. Fig. 21. Part of cell showing pyrenoid in the chloroplast. Scale bar = 2 μm.
Fig. 22
Fig. 22
Phylogenetic position of Pleodorina sphaerica Iyengar within the advanced members of the Volvocaceae [Eudorina group (Nozaki et al. 2000)], as inferred from 6021 base pairs of five chloroplast genes. The tree was constructed by maximum likelihood (ML) method. Branch lengths are proportional to the genetic distances, which are indicated by the scale bar above the tree. Numbers on the left or right side at the branches represent bootstrap values (50% or more, based on 1000 replicates) obtained with the ML and maximum parsimony analyses, respectively. Asterisks at the branches indicate 100% bootstrap values by the two methods. For details of the methods, see the text. Clade I and clade II are essentially the same as those in Nozaki et al. (2015a) except for the addition of P. sphaerica and Volvox tertius Meyer strain 1-3k-4 in the present study. ‘sM’ and ‘sJ’ in clade I represent Volvox sect. Merrillosphaera and Volvox sect. Janetosphaera, respectively, based on Nozaki et al. (2015a).

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