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Amplified fragment length polymorphism (AFLP)-based differentiation of selected Chara species

Jacek Urbaniak
Jacek Urbaniak
 and 
Michał Combik
Michał Combik
   | Mar 06, 2017
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Article Category: Original research paper
Published Online: Mar 06, 2017
Page range: 74 - 84
Received: Feb 05, 2016
Accepted: May 12, 2016
DOI: https://doi.org/10.1515/ohs-2017-0008
© Faculty of Oceanography and Geography, University of Gdańsk, Poland. All rights reserved.
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Figure 1

LM images of the studied species: a. Stipulodes (upper arrow) and long spine cells (arrow in the middle of the axis) on C. aspera axis; b. Antheridia on male C. aspera; c. Upper branchlet of C. globularis axis; d. Rudimentary stipulodes below C. globularis branchlet; e. Short and reduced spine cells on C. globularis axis; f. Long spine cells on axis C. hispida; g. Spine cells in bunches on C. intermedia main axis; h. Antheridia and oogonia on dioecious C. strigosa; i. General habit of C. tenuispina; j. Long spine cells of C. tenuispina on the upper part of the main axis; k. Spine cells of C. tenuispina; l. Long stipulodes below the branchlet on C. virgata Scale bars: Figs 1, 4, 12, 100 µm; Figs 2, 3, 6, 8, 9, 10, 300 µm; Figs 5, 50 µm; Figs 7, 11, 150 µm
LM images of the studied species: a. Stipulodes (upper arrow) and long spine cells (arrow in the middle of the axis) on C. aspera axis; b. Antheridia on male C. aspera; c. Upper branchlet of C. globularis axis; d. Rudimentary stipulodes below C. globularis branchlet; e. Short and reduced spine cells on C. globularis axis; f. Long spine cells on axis C. hispida; g. Spine cells in bunches on C. intermedia main axis; h. Antheridia and oogonia on dioecious C. strigosa; i. General habit of C. tenuispina; j. Long spine cells of C. tenuispina on the upper part of the main axis; k. Spine cells of C. tenuispina; l. Long stipulodes below the branchlet on C. virgata Scale bars: Figs 1, 4, 12, 100 µm; Figs 2, 3, 6, 8, 9, 10, 300 µm; Figs 5, 50 µm; Figs 7, 11, 150 µm

Figure 2

Neighbor-joining dendrogram based on AFLP markers
Neighbor-joining dendrogram based on AFLP markers

Figure 3

Principal Component Analysis of AFLP of the analyzed species
Principal Component Analysis of AFLP of the analyzed species

Classification of the analyzed Chara species according to different authors

Krause (1997) Wood & Imahori (1965) Cirujano et al. (2008)
C. asperaDeth. ex Willd. 1809 Chara globularis var. aspera(Deth. ex Willd. 1809) R.D. Wood 1962 Chara aspera var. asperaDeth. ex Willd. 1809
C. globularisThuill. 1799 Chara globularis var. globularis f. globularisR.D. Wood 1962 Chara fragilisDesv. in Loisel. 1810
C. hispidaL. Chara hispida var. major f. major(Hartm.) R.D.Wood 1962 Chara hispida var. major(Hartm.) R.D. Wood 1962
C. intermediaA.Br. 1859 Chara hispida var. major f. intermedia(A.Br.) R.D.Wood 1962 Chara hispida var. hispida(Hartm.) R.D. Wood 1962
C. strigosaA.Br 1847 Chara globularis f. strigosa(A. Br.) R.D. Wood 1962 -
C. tenuispinaA.Br. 1835 Chara globularis var. tenuispina(Kütz.) R.D. Wood 1962 -
C. virgata (C.delicatula)Kütz. 1834 Chara globularis var. virgata(A. Br.) R.D. Wood 1962 -

Jaccard’s genetic similarity index between pairs of plants belonging to different species

C. globularis C. hispida C. intermedia C. strigosa C. tenuissima C. virgata
C. aspera 0.16 0.18 0.14 0.68 0.37 0.21
C. globularis 0.27 0.30 0.47 0.69 0.49
C. hispida 0.71 0.29 0.08 0.1
C. intermedia 0.20 0.12 0.25
C. strigosa 0.31 0.38
C. tenuissima 0.46

The studied species and populations

Species Locality with the number of individuals Date of collection
C. aspera Lake Mauda, Suwałki Lakeland, (31-34)Lake Pobłędzie, Suwałki Lakeland (35-38)Lake Wigry, Augustów Lakeland, (51-55)Lake Hańcza, Suwałki Lakeland (81-85) Jul. 2014Aug. 2013Aug. 2013Jul. 2014
C. globularis fish pond near Szczodre, Silesian Lowland, (201-205)Lake Czarne, Łęczna-Włodawa Lakeland, (206-210)pond near Niemodlin, Opolska Upland, (211-216) Jul. 2014Jul. 2014Jul. 2014
C. hispida Lake Okmin, Suwałki Lakeland, (21-24)Lake Szelment, Suwałki Lakeland, (25-28) Jul. 2014Jul. 2014
C. intermedia Lake Czarne, Łęczna-Włodawa Lakeland, (91-94)pond near Staszów, Staszów Uppland, (95-97) Jul. 2014Aug. 2013
C. strigosa S part of Lake Hańcza, Suwałki Lakeland, (71-76)SE part of Lake Hańcza, Suwałki Lakeland, (131-134) Jul. 2014Jul. 2014
C. tenuispina Lake Drążynek, Wielkopolskie Lakeland, (101-104)Lake near Wągrowiec, Wielkopolskie Lakeland, (105-108) Sept. 2013Sept. 2013
C. virgata Lake Wigry, Augustów Lakeland, (1a-4a)Lake Serwy, Augustów Lakeland, (301-305) Aug. 2013Aug. 2013

Percentage of polymorphic and non-polymorphic fragments per primer combination for the analyzed Chara species

Primer combination All bands Monomorphic Polymorphic
(n) (n) (n) (%)
ACA/CGA 185 6 179 97
ACA/CGC 177 16 161 91
ACT/CGA 184 15 169 92
ACG/CAT 201 11 190 93
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