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FIGURES 23–28. Nitella flagellifera. 23 in Taxonomy and phylogeny of Nitella (Charophyceae, Characeae) from Brazil with emphasis on the midwest and southeast regions

FIGURES 23–28. Nitella flagellifera. 23. Primary node of a branchlet. The red arrow indicates a central secondary ray. 24. Fertile branchlet with gametangia at second (black arrow) and third (red arrow) nodes. 25. Dactyls 2-celled, elongated. Penultimate cell rounded at distal end. End cell conical and mucronate. 26. Oospore membrane with reticulate aspect under LM. 27. Overview of an oospore under SEM. 28. Oospore membrane with beaded imperfect reticulate to reticulate aspect under SEM.

FIGURES 17–22. Nitella elegans. 17. Upper whorls small and somewhat compacted. 18 in Taxonomy and phylogeny of Nitella (Charophyceae, Characeae) from Brazil with emphasis on the midwest and southeast regions

FIGURES 17–22. Nitella elegans. 17. Upper whorls small and somewhat compacted. 18. Primary node of a branchlet. The red arrow indicates a central secondary ray. 19. Dactyls 2-celled. Penultimate cell cylindrical tapering distally towards the base of end cell. End cell conical, acuminate to mucronate. 20. Oospore membrane with granulate aspect under LM. 21. Overview of an oospore under SEM. 22. Oospore membrane with finely granulate aspect under SEM.

FIGURES 5–10. Nitella acuminata. 5 in Taxonomy and phylogeny of Nitella (Charophyceae, Characeae) from Brazil with emphasis on the midwest and southeast regions

FIGURES 5–10. Nitella acuminata. 5. Upper whorls with loosely compacted heads (a diagnostic feature of N. subglomerata). 6. Compacted heads (a diagnostic feature of N. gollmeriana). 7. Dactyls 1-celled, abbreviated with acuminate apex (a diagnostic feature of N. gollmeriana). 8. Oospore membrane with finely granulate aspect under LM. 9. Overview of an oospore under SEM. 10. Oospore membrane with scabrous aspect under SEM.

FIGURES 11–16. Nitella axillaris. 11. Whorl with a in Taxonomy and phylogeny of Nitella (Charophyceae, Characeae) from Brazil with emphasis on the midwest and southeast regions

FIGURES 11–16. Nitella axillaris. 11. Whorl with a small, axillary, sessile, compacted head, at a node of the thallus. 12. Whorl with a long, axillary, stipitate and compacted head. 13. 2-furcated fertile branchlet of a head. The black and red arrows indicate, respectively, the primary and the secondary nodes. 14. Dactyls 2-celled, abbreviated, forming a crown. Penultimate cell tapering gradually towards the base of end cell. End cell conical, acute or acuminate. 15. Oospore membrane with reticulate aspect under LM. 16. Oospore membrane with beaded imperfect reticulate aspect under SEM.

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FIGURE 4 in Taxonomy and phylogeny of Nitella (Charophyceae, Characeae) from Brazil with emphasis on the midwest and southeast regions

FIGURE 4. Bayesian Phylogenetic tree based on ITS2 sequences. The numbers associated with the nodes indicate the bootstrap values (BS) for maximum likelihood and posterior probability (PP) for Bayesian analysis; nodes without values indicate BS <70% and PP <0.70; nodes with * indicate BS> 95% and PP>0.95 Sequences in bold are the new sequences generated in this study. Samples marked with "1" can be morphologically identified as N. acuminata (Wood & Imahori 1965); "2" with N. subglomerata (Wood & Imahori 1965).

FIGURES 29–34. Nitella microcarpa. 29. Upper whorls somewhat compacted. 30. Dactyls 2 in Taxonomy and phylogeny of Nitella (Charophyceae, Characeae) from Brazil with emphasis on the midwest and southeast regions

FIGURES 29–34. Nitella microcarpa. 29. Upper whorls somewhat compacted. 30. Dactyls 2-celled, abbreviated, forming a crown. Penultimate cell rounded at distal end. End cell conical and acuminate. 31. Fertile branchlet with gametangia at second (black arrow) and third (red arrow) nodes. 32. Oospore membrane with reticulate aspect under LM. 33. Overview of an oospore under SEM. 34. Oospore membrane with beaded imperfect reticulate aspect under SEM.

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FIGURE 2 in Taxonomy and phylogeny of Nitella (Charophyceae, Characeae) from Brazil with emphasis on the midwest and southeast regions

FIGURE 2. Maximum likelihood Phylogenetic tree based on rbcL sequences. The numbers associated with the nodes indicate the bootstrap values (BS) for maximum likelihood and posterior probability (PP) for Bayesian analysis; nodes without values indicate BS <70% and PP <0.70; nodes with * indicate BS> 95% and PP>0.95 Sequences in bold are the new sequences generated in this study. Samples marked with "1" can be morphologically identified as N. acuminata (Wood & Imahori 1965); "2" with N. subglomerata (Wood & Imahori 1965).

FIGURE 1. A in Taxonomy and phylogeny of Nitella (Charophyceae, Characeae) from Brazil with emphasis on the midwest and southeast regions

FIGURE 1. A—Map of Brazil. The area highlighted in gray corresponds to the study region. B—Area of study with the sampling sites. The table indicates sampling sites for each species collected.

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FIGURE 3 in Taxonomy and phylogeny of Nitella (Charophyceae, Characeae) from Brazil with emphasis on the midwest and southeast regions

FIGURE 3. Bayesian Phylogenetic tree based on ITS1 sequences. The numbers associated with the nodes indicate the bootstrap values (BS) for maximum likelihood and posterior probability (PP) for Bayesian analysis; nodes without values indicate BS <70% and PP <0.70; nodes with * indicate BS> 95% and PP>0.95. Sequences in bold are the new sequences generated in this study.

FIGURE 1–4 in Nitella singaporensis (Charophyceae, Charales): new species and implications for the taxonomy of the genus Nitella

FIGURE 1–4. General appearance and whorl and branchlet of Nitella singaporensis: 1—pressed and dried material (holotype: SING); 2—pressed and dried material (isotype: LE); 3—reconstruction of general habit of branchlet whorl (view from above) from the photo of ethanol-fixed specimen (isotype: LE) showing mean of terms used for branchlet description: 1—primary ray, 2—lateral secondary ray, 3—central secondary ray, 4—lateral tertiary ray, 5—central tertiary ray, 6—dactyls, yellow—lateral secondary segments, green—central secondary segment, blue—central tertiary segment, orange—lateral tertiary segment; 4—fertile branchlet of rewetted herbarium specimen showing oogonia arrangement (isotype: LE). Scale: 1 mm. Scale: 2—1 cm, 4—1 mm. Photo 1 by D. Liew, drawing and other photos by R. Romanov.

FIGURES 5–19 in Nitella singaporensis (Charophyceae, Charales): new species and implications for the taxonomy of the genus Nitella

FIGURES 5–19. The whorls and branchlets of Nitella singaporensis (all from isotype: LE): 5—apical part with mucilage recognised because of adhering of fine particles at its surface; 5–7—whorls of branchlets, showing long primary ray; 8–18—branchlets, showing variability in length of central secondary ray, from moderately elongated, i.e. exceeding the length of lateral secondary rays with their dactyls (8 & 9), nearly equal with them (10, 12, 14), to abbreviated, i.e. clearly shorter than lateral second segments (19), mainly spreading (8–16) or adpressed to the central one, resulting in last case in tassel-like general appearance of the branchlet (19); 8, 12, 15, 16—fine particles adhered to the surface of a branchlet, indicating presence of a mucilage at its surface; actually they are outlining the mucilage patches; 9—central secondary ray without furcation, appearing as long robust dactyl (arrowheads); 10 & 11—branchlets with central tertiary ray (arrowheads); 11—branchlet with central secondary ray with furcation (double arrowhead), producing central tertiary ray with 3 dactyls (arrowhead); 12—dactyls at the furcation at the top of central secondary ray (arrowhead); 13—central tertiary ray (arrowhead) at lateral secondary ray; 17—unfurcated lateral secondary rays (arrowheads), central secondary ray at the branchlet below is not clearly visible because of artificial deformation at its base. Scale: 5–11—2 mm, 12–17—1 mm, 18 & 19—0.5 mm. All photos by R. Romanov.

FIGURE 38 in Nitella singaporensis (Charophyceae, Charales): new species and implications for the taxonomy of the genus Nitella

FIGURE 38. ML-phylogenetic tree inferred in PAUP with GTR+I+Г nucleotide substitution model from 89 rbcL sequences of Nitella. ML BP (> 50%) and BI PP (> 0.95) are shown. Thick branches received 100% BP and 1.00 PP support. The branch leading to the outgroup was 50% reduced in length. The newly obtained sequence of Nitella singaporensis is given in bold. Clade designation is according to Sakayama (2008).

FIGURES 32–37 in Nitella singaporensis (Charophyceae, Charales): new species and implications for the taxonomy of the genus Nitella

FIGURES 32–37. Oospores of Nitella singaporensis (isotype: LE): 32—general habit, 33–37—surface ornamentation of fossa and ribs at different magnification. Scale: 32—40 μm, 33, 34—10 μm, 35—8 μm, 36—4 μm, 37—1 μm. All photos by R. Romanov.

FIGURES 20–31 in Nitella singaporensis (Charophyceae, Charales): new species and implications for the taxonomy of the genus Nitella

FIGURES 20–31. Nitella singaporensis from Singapore (isotype: LE), N. gracilis from Sicily (LE): 20 & 21—abbreviated central second segment (arrowhead), looking as an additional branchlet at first furcation; 21—rare case of 8 lateral secondary rays; 22—end parts of a branchlet with clearly visible central secondary ray (arrowhead); 23–25—2-celled dactyls, some of them appearing 1-celled after loss of end cell during storage in living state in the laboratory; 26—gametangia, [arrowhead—shortly stipitate antheridia, double arrowhead— production of antheridium in penultimate not end furcation], 27—oospore surface in LM, appearing as unclear reticulations, 28–30—stem nodes with easily visible nodal cell extruding from the base of branchlets below the whorl, resulting in nodal bulbil formation (30), 31—stem node of N. gracilis with nodal cell extruding from the base of branchlets and situated around it. Scale: 20–22—0.5 mm, 23–26, 28–31—0.2 mm, 27—20 μm. All photos by R. Romanov, except 26 by R.S.W. Yeo.