Chinese Journal of Oceanology and Limnology

, Volume 31, Issue 1, pp 97–105 | Cite as

Phylogenetic analyses of four species of Ulva and Monostroma grevillei using ITS, rbc L and 18S rDNA sequence data

  • Zhongheng Lin (林中姮)
  • Songdong Shen (沈颂东)Email author
  • Weizhou Chen (陈伟洲)
  • Huihui Li (李慧慧)


Chlorophyta species are common in the southern and northern coastal areas of China. In recent years, frequent green tide incidents in Chinese coastal waters have raised concerns and attracted the attention of scientists. In this paper, we sequenced the 18S rDNA genes, the internal transcribed spacer (ITS) regions and the rbc L genes in seven organisms and obtained 536–566 bp long ITS sequences, 1 377–1 407 bp long rbc L sequences and 1 718–1 761 bp long partial 18S rDNA sequences. The GC base pair content was highest in the ITS regions and lowest in the rbc L genes. The sequencing results showed that the three Ulva prolifera (or U. pertusa) gene sequences from Qingdao and Nan’ao Island were identical. The ITS, 18S rDNA and rbc L genes in U. prolifera and U. pertusa from different sea areas in China were unchanged by geographic distance. U. flexuosa had the least evolutionary distance from U. californica in both the ITS regions (0.009) and the 18S rDNA (0.002). These data verified that Ulva and Enteromorpha are not separate genera.


Ulva ITS region rbc18S rDNA phylogeny sequences analysis 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Booton G C, Floyd G L, Fuerst P A. 1998. Polyphyly of tetrasporalean green algae inferred from nuclear small-subunit ribosomal DNA. J. Phycol., 34: 306–311.CrossRefGoogle Scholar
  2. Daugbjerg N, Moestrup Ø, Arctander P. 1994. Phylogeny of the genus Pyramimonas (Prasinophyceae, Chlorophyta) inferredfrom the rbcL gene. J. Phycol., 30: 991–999.CrossRefGoogle Scholar
  3. Daugbjerg N, Moestrup Ø, Arctander P. 1995. Phylogeny of genera of Prasinophyceae and Pedinophyceae (Chlorophyta) deduced from molecular analysis of the rbcL gene. Phycol. Res., 43: 203–213.CrossRefGoogle Scholar
  4. Fletcher R T. 1996. The occurrence of ‘green tide’. In: Schramm W, Nienhuis P H eds. Marine Benthic Vegetation-Recent Changes and the Effects of Eutrophication. Springer Verlag, Berlin. p.7–43.Google Scholar
  5. Friedl T. 1996. Evolution of the polyphyletic genus Pleurastrum (Chlorophyta): inferences from nuclear-encoded ribosomal DNA sequences and motile cell ultrastructure. Phycologia, 35: 456–469.CrossRefGoogle Scholar
  6. Hall T A, 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp. Ser., 41: 95–98.Google Scholar
  7. Hayden H S, Blomster J, Maggs C A, Silva P C, Stanhope M J, Waaland J R. 2003. Linnaeus was right all along: Ulva and Enteromorpha are not distinct genera. Eur. J. Phycol., 38: 277–294.CrossRefGoogle Scholar
  8. Hayden H S, Waaland J R. 2002. Phylogenetic systematics of the Ulvaceae (Ulvales, Ulvophyceae) using chloroplast and nuclear DNA sequences. J. Phycol., 38: 1 200–1 212.CrossRefGoogle Scholar
  9. Jiang P, Wang J F, Cui Y L, Li Y X, Lin H Z, Qin S. 2008. Molecular phylogenetic analysis of attached Ulvaceae species and free-floating Enteromorpha from Qingdao coasts in 2007. Chin. J. Oceanol. Limnol., 26(3): 276–279.CrossRefGoogle Scholar
  10. Kudo T, Notoya M. 1999. The free-floating Ulva species growing in the calm seas. Case of the Sea Park of Yokohama City. In: Notoya M ed. Use of Ulva and Restoration of the Environment. Seizando, Tokyo. p.55–70. (in Japanese)Google Scholar
  11. Kumar S, Tamura K, Jakobsen I B, Nei M. 2001. MEGA 2: molecular evolutionary genetics analysis software. Bioinformatics, 17: 1 244–1 245.CrossRefGoogle Scholar
  12. Leliaert F, Rousseau F, Reviers De B, Coppejans E. 2003. Phylogeny of the Cladophorophyceae (Chlorophyta) inferred from partial LSU rRNA gene sequences: is the recognition of a separate order Siphonocladales justified? Eur. J. Phycol., 38: 233–246.CrossRefGoogle Scholar
  13. Linnaeus C. 1753. Species plantarum, exhibentes plantas rite cognitas, ad genera relatas, cum differentiis specificus, nominibus trivialibus, synonymis selectis, locis natalibus, secundum systema sexuale digestas. Ed. 1. Vols. 1–2, Salvii, Stockholm. Vol. 1, pp [i–xii] 1–560. Vol. 2, pp. [i], 561–1200, [1–30 index].Google Scholar
  14. Li Y Y, Shen S D, He L H, Xu P, Wang G C. 2009. Sequence analysis of the ITS region and 5.8S rDNA of Porphyra haitanensis. C hin. J. O ceanol. L imnol., 27(3): 1–9.Google Scholar
  15. Li Y Y, Shen S D, He L H, Xu P, Lu S. 2010. Sequence analysis of rDNA intergenic spacer (IGS) of Porphyra haitanensis. J. Appl. Phycol., 22: 187–193.CrossRefGoogle Scholar
  16. Lobban C S, Harrison P J. 1997. Seaweed Ecology and Physiology. Cambridge University Press, Cambridge. 366p.Google Scholar
  17. Manhart J R. 1994. Phylogenetic analysis of green plant rbc L sequences. Mol. Phylogenet. Evol., 3: 114–127.CrossRefGoogle Scholar
  18. Malta E J, Draisma S G A, Kamermans P. 1999. Free-floating Ulva in the southwest Netherlands: species or morphotypes? A morphological, molecular and ecological comparison. Eur. J. Phycol., 34: 443–54.CrossRefGoogle Scholar
  19. Nozaki H, Ito M, Sano R, Uchida H, Watanabe M M, Takahashi H, Kuroiwa T. 1997a. Phylogenetic analysis of Yamagishiella and Platydorina (Volvocaceae, Chlorophyta) based on rbcL gene sequences. J. Phycol., 33: 272–278.CrossRefGoogle Scholar
  20. Nozaki H, Ito M, Watanabe M M, Takahashi H, Kuroiwa T. 1997b. Phylogenetic analysis of morphological species of Carteria (Volvocales, Chlorophyta) based on rbcL gene sequences. J. Phycol., 33: 864–867.CrossRefGoogle Scholar
  21. Okamura K. 1921. Illustrations of the Marine Algae of Japan, IV, No. 4. Keigyosha, Tokyo. p.79–83, l.169-70.Google Scholar
  22. Olsen J L, Stam W T, Berger S, Menzel D. 1994. 18S rDNA and evolution in the Dasycladales (Chlorophyta): modern living fossils. J. Phycol., 30: 729–744.CrossRefGoogle Scholar
  23. Shimada S, Hiraoka M, Nabata S, Iima M, Masuda M. 2003. Molecular phylogenetic analyses of the Japanese Ulva and Enteromorpha (Ulvales, Ulvophyceae), with special reference to the free-floating Ulva. Phycol. Res., 51: 99–108.CrossRefGoogle Scholar
  24. Shimada S, Yokoyama N, Arai S, Hiraoka M. 2008. Phylogeography of the genus Ulva (Ulvophyceae, Chlorophyta), with special reference to the Japanese freshwater and brackish taxa. J. Appl. Phycol., 20: 979–989.CrossRefGoogle Scholar
  25. Sun S, Wang F, Li C L, Qin S, Zhou M J, Ding L P, Pang S J, Duan D L, Wang G C, Yin B S, Yu R C, Jiang P, Liu Z L, Zhang G T, Fei X G, Zhou M. 2008. Emerging challenges: massive green algae blooms in the Yellow Sea. Nature Proceedings, hdl: 10101/npre. 2008. 2266.1.Google Scholar
  26. Thompson J D, Gibson T J, Plewniak F, Jeanmougin F, Higgins D G. 1997. The Clustal X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucl. Acids. Res., 24: 4 876–4 882.Google Scholar
  27. Tan I H, Blomster J, Hansen G, Leskinen E, Maggs C A, Mann D G, Sluiman H J, Stanhope M J. 1999. Molecular phylogenetic evidence for a reversible morphogenetic switch controlling the gross morphology of two common genera of green seaweeds, Ulva and Enteromorpha. Mol. Biol. Evol., 16: 1 011–1 018.CrossRefGoogle Scholar
  28. van Oppen M J H. 1995 Tracking Trails by Cracking Codes. Ph.D. Dissertation. University of Groningen, Groningen. 166p.Google Scholar
  29. White T J, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innes M A, Gefland D H, Sninsky J J, White T J eds. PCR Protocols: A Guide to Methods and Applications. Academic Press, New York, NY, USA. p.315–322.Google Scholar
  30. Wilcox L W, Lewis L A, Fuerst P A, Floyd G L. 1992. Assessing the relationships of autosporic and zoosporic chlorococcalean green algae with 18S rDNA sequence data. J. Phycol., 28: 381–386.CrossRefGoogle Scholar
  31. Yang J, Wang Q, Liu M H, An L J. 1999. A simple method for extracting total DNA of seaweeds. Biotechonl., 9(4): 39–42.Google Scholar
  32. Zechman F W, Theriot E G, Zimmer E A, Chapman R L. 1990. Phylogeny of the Ulvophyceae (Chlorophyta): cladistic analysis of nuclear encoded rRNA sequence data. J. Phycol., 26: 700–710.CrossRefGoogle Scholar

Copyright information

© Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Zhongheng Lin (林中姮)
    • 1
  • Songdong Shen (沈颂东)
    • 1
    Email author
  • Weizhou Chen (陈伟洲)
    • 2
  • Huihui Li (李慧慧)
    • 1
  1. 1.Department of Cell Biology, School of Biology and Basic Medical SciencesSoochow UniversitySuzhouChina
  2. 2.Institute of Marine BiologyShantou UniversityShantouChina

Personalised recommendations