Skip to main content
Log in

DNA barcoding assessment of green macroalgae in coastal zone around Qingdao, China

  • Published:
Journal of Ocean University of China Aims and scope Submit manuscript

Abstract

An assessment with assistance of DNA barcoding was conducted on green macroalgae in coastal zone around Qingdao, China, during the period of April–December, 2011. Three markers were applied in molecular discrimination, including the plastid elongation factor tufA gene, the internal transcribed spacer (ITS) region of the ribosomal cistron and rubisco large subunit gene 3′ regions (rbcL-3P). DNA barcoding discriminated 8 species, excluding species of genus Cladophora and Bryopsis due to failures in amplification. We ascertained and corrected 4 species identified by morphological methods for effectively assisting the classification. The gene tufA presented more advantages as an appropriate DNA marker with the strongest amplification success rate and species discrimination power than the other two genes. The poorest sequencing success largely handicapped the application of ITS. Samples identified by tufA and rbcL as Ulva flexuosa were clustered into the clade of U. prolifera by ITS in the neighbor-joining tree. Confusion with discrimination of the complex of U. linza, U. procera and U. prolifera (as the LPP complex) still existed for the three DNA markers. Based on our results, rbcL is recommended as a preferred marker for assisting tufA to discriminate green macroalgae. In distinguishing green-tide-forming Ulva species, the free-floating sample collected from the green tide in 2011 was proved to be identical with U. prolifera in Yellow Sea for ITS and rbcL genes. This study presents a preliminary survey of green macroalgae distributed in the coastal area around Qingdao, and proves that DNA barcoding is a powerful tool for taxonomy of green macroalgae.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Buchheim, M. A., Keller, A., Koetschan, C., Förster, F., Merget, B., and Wolf, M., 2011. Internal transcribed spacer 2 (nu ITS2 rRNA) sequence-structure phylogenetics: Towards an automated reconstruction of the green algal tree of life. Plos one, 6(2): 1–10.

    Article  Google Scholar 

  • Clarkston, B. E., and Saunders, G. W., 2010. A comparison of two DNA barcode markers for species discrimination in the red algal family Kallymeniaceae (Gigartinales) with a description of Euthora timburtonii. Botany, 88: 119–131.

    Article  Google Scholar 

  • De Clerck, O., Verbruggen, H., Huisman, J. M., Faye, E. J., Leliaert, F., Schils, T., and Coppejans, E., 2008. Systematics and biogeography of the genus Pseudocodium (Bryopsidales, Chlorophyta), including the description of P. natalense sp. nov. from South Africa. Phycologia, 47: 225–235.

    Article  Google Scholar 

  • Fama, P., Wysor, B., Kooistra, W., and Zuccarello, G. C., 2002. Molecular phylogeny of the genus Caulerpa (Caulerpales, Chlorophyta) inferred from chloroplast tufA gene. Journal of Phycology, 38: 1040–1050.

    Article  Google Scholar 

  • Hall, J. D., Fučíková, K., Lo, C., Lewis, L. A., and Karol, K. G., 2010. An assessment of proposed DNA barcodes in freshwater green algae. Cryptogamie Algologie, 31(4): 529–555.

    Google Scholar 

  • Händeler, K., Wägele, H., Wahrmund, U., Rüdinger, M., and Knoop, V., 2010. Slugs’ last meals: Molecular identification of sequestered chloroplasts from different algal origins in Sacoglossa (Opisthobranchia, Gastropoda). Molecular Ecology Resources, 10: 968–978.

    Article  Google Scholar 

  • Hanyuda, T., Arai, S., and Uedak, K., 2000. Variability in the rbcL introns of Caulerpalean algae (Chlorophyta, Ulvophyceae). Journal of Plant Research, 113: 403–413.

    Article  Google Scholar 

  • Harper, J. T., and Sauders, G. W., 2001. The application of sequences of the ribosomal cistron to the systematics and classification of the florideophyte red algae (Florideophyceae, Rhodophyta). Les cahiers de Biologie Marine, 42: 25–38.

    Google Scholar 

  • Hayden, H. S., Blomster, J., Maggs, C. A., Silva, P. C., Stanhope, M. J., and Waaland, J. R., 2003. Linnaeus was right all along: Ulva and Enteromorpha are not distinct genera. European Journal of Phycology, 38: 277–294.

    Article  Google Scholar 

  • Hayden, H. S., and Waaland, J. R., 2002. Phylogenetic systematics of the Ulvaceae (Ulvales, Ulvophyceae) using chloroplast and nuclear DNA sequences. Journal of Phycology, 38: 1200–1212.

    Article  Google Scholar 

  • Hayden, H. S., and Waaland, J. R., 2004. A molecular systematic study of Ulva (Ulvaceae, Ulvales) from the northeast Pacific. Phycologia, 43: 364–382.

    Article  Google Scholar 

  • Hebert, P. D. N., Cywinska, A., Ball, S. L., and Deward, J. R., 2003. Biological identifications through DNA barcodes. Proceedings of the Royal Society of London Biology, 270: 313–321.

    Article  Google Scholar 

  • Hebert, P. D. N., and Gregory, T. R., 2005. The promise of DNA barcoding for taxonomy. Systematic Biology, 54: 852–859.

    Article  Google Scholar 

  • Hu, C., Li, D., Chen, C., Ge, J., Muller-Karger, F. E., Liu, J., Yu, F., and He, M. X., 2010. On the recurrent Ulva prolifera blooms in the Yellow Sea and East. Journal of Geophysical Research, 115, C05017, DOI: 10.1029/2009JC005561.

    Article  Google Scholar 

  • Leliaert, F., Zhang, X., Ye, N., Malta, E., Engelen, A. H., Mineur, F., Verbruggen, H., and Clerck, O. D., 2009. Identity of the Qingdao algal bloom. Phycological Research, 57: 147–151.

    Article  Google Scholar 

  • Liu, D. Y., Wang, Z. Y., Sun, J., Huang, Z. Y., and Qian, S. B., 1999. Study of the benthic algae in the littoral of Qingdao coast. Transactions of Oceanology and Limnology, 3: 35–40.

    Google Scholar 

  • Liu, D. Y., Keesing, J. K., Xing, Q. G., and Shi, P., 2009. World’s largest macroalgal bloom caused by expansion of seaweed aquaculture in China. Marine Pollution Bulletin, 58: 888–895.

    Article  Google Scholar 

  • Liu, F., Pang, S. J., Zhao, X. B., and Hu, C. M., 2012. Quantitative molecular and growth analyses Ulva propagules in sediment of Jiangsu initially green tides. Marine Environmental Research, 74: 56–63.

    Article  Google Scholar 

  • Loughnane, C. J., McIvor, L. M., Rindi, F., Stengel, D. B., and Guiry, M. D., 2008. Morphology, rbcL phylogeny and distribution of distromatic Ulva (Ulvophyceae, Chlorophyta) in Ireland and southern Britain. Phycologia, 47: 416–429.

    Article  Google Scholar 

  • Manhart, J. R., 1994. Phylogenetic analysis of green plant rbcL sequences. Molecular Phylogenetics and Evolution, 3: 114–127.

    Article  Google Scholar 

  • Mares, J., Leskinen, E., Sitkowska, M., Skácelová, O., and Blomster, J., 2011. True identity of the European freshwater Ulva revealed by molecular and morphological methods. Journal of Phycology, 47: 1177–1192.

    Article  Google Scholar 

  • Mcdevit, D. C., and Saunders, G. W., 2009. On the utility of DNA barcoding for species differentiation among brown macroalgae (Phaeophyceae) including a novel extraction protocol. Phycological Research, 57: 131–141.

    Article  Google Scholar 

  • Medlin, L. K., Metifies, K., John, U., and Olsen, J. L., 2007. Algal molecular systematic: A review of the past and prospects for the future. In: Unravelling the Algae-the Past, Present and Future of Algal Systematic. Brodie, J., and Lewis, J., eds., CRC Press, Boca Raton, FL, 341–353.

    Chapter  Google Scholar 

  • Meier, R., Zhang, G., and Ali, F., 2008. The use of mean instead of smallest interspecific distances exaggerates the size of the ‘Barcoding Gap’ and leads to misidentification. Systematic Biology, 57: 809–813.

    Article  Google Scholar 

  • Nakazawa, A., Yamada, T., and Nozaki, H., 2004. Taxonomic study of Asterococcus (Chlorophyceae) based on comparative morphology and rbcL gene sequences. Phycologia, 43: 711–721.

    Article  Google Scholar 

  • O’Kelly, C. J., Kurihara, A., Shipley, T. C., and Sherwood, A. R., 2010. Molecular assessment of Ulva spp. (Ulvophyceae, Chlorophyta) in the Hawaiian Islands. Journal of Phycology, 46: 728–735.

    Article  Google Scholar 

  • Pang, S. J., Liu, F., Shan, T. F., Xu, N., Zhang, Z. H., Gao, S. Q., Chopin, T., and Sun, S., 2010. Tracking the algal origin of the Ulva bloom in the Yellow Sea by a combination of molecular, morphological and physiological analyses. Marine Environmental Research, 69: 207–215.

    Article  Google Scholar 

  • Pröschold, T., and Leliaert, F., 2007. Systematics of the green algae: Conflict of classic and modern approaches. In: Unravelling the Algaethe Past, Present and Future of Algal Systematic. Brodie, J., and Lewis, J., eds., CRC Press, Boca Raton, FL, 123–153.

    Google Scholar 

  • Robba, L., Russell, S., Baker, G., and Brodie, J., 2006. Assessing the use of the mitochondrial COX I marker for use in DNA barcoding of red algae (Rhodophyta). American Journal of Botany, 93(8): 1101–1108.

    Article  Google Scholar 

  • Shimada, S., Yokoyama, N., Arai, A., and Hiraoka, M., 2008. Phylogeography of the genus Ulva (Ulvophyceae, Chlorophyta), with special reference to the Japanese freshwater and brackish taxa. Journal of Applied Phycology, 20: 979–989.

    Article  Google Scholar 

  • Saunders, G. W., 2005. Applying DNA barcoding to red macroalgae a preliminary appraisal holds promise for future applications. Philosophical Transactions of the Royal Society B, 360: 1879–1888.

    Article  Google Scholar 

  • Saunders, G. W., and Kucera, H., 2010. An evaluation of rbcL, tufA, UPA, LSU and ITS as DNA barcode markers for the marine green macroalgae. Cryptogamie Algologie, 31(4): 487–528.

    Google Scholar 

  • Sun, S., Wang, F., Li, C., Qin, S., Zhou, M., Ding, L., Pang, S., Duan, D., Wang, G., Yin, B., Yu, R., Jiang, P., Liu, Z., Zhang, G., Fei, X., and Zhou, M., 2008. Emerging challenges: Massive green algae blooms in the Yellow Sea. Nature Precedings, hdl: 10101/ npre.2008.2266.1.

    Google Scholar 

  • Tamura, K., Nei, M., and Kumar, S., 2004. Prospects for inferring very large phylogenies by using the neighbor-joining method. Proceedings of the National Academy of Sciences of USA, 101: 11030–11035.

    Article  Google Scholar 

  • Tamura, K., Dudley, J., Nei, M., and Kumar, S., 2007. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software Ver. 4.0. Molecular Biology and Evolution, 24: 1596–1599.

    Article  Google Scholar 

  • Tseng, C. K., 1983. Common Seaweeds of China. Science Press, Beijing, 25–33.

    Google Scholar 

  • Tseng, C. K., Xia, B. M., and Zhou, X. T., 2009. Seaweeds in Yellow Sea and Bohai Sea of China. Science Press, Beijing, 254pp (in Chinese).

    Google Scholar 

  • Valentini, A., Pompanon, F., and Taberlet, P., 2008. DNA barcoding for ecologists. Trends in Ecology and Evolution, 24(2): 110–117.

    Article  Google Scholar 

  • Verbruggen, H., Tyberghein, L., Pauly, K., Vlaeminck, C., Van Nieuwenhuyze, K., Koositra, W., Leliaert, F., and De Clerck, O., 2009. Macroecology meets macroevolution: Evolutionary niche dynamics in the seaweed Halimeda. Global Ecology and Biogeography, 18: 393–405.

    Article  Google Scholar 

  • Wang, J. F., Li, N., Jiang, P., Boo, S. M., Lee, W. J., Cui, Y., Lin, H., Zhao, J., Liu, Z., and Qin, S., 2010a. Ulva and Enteromorpha (Ulvaceae, Chlorophyta) from two sides of the Yellow Sea: Analysis of nuclear rDNA ITS and plastid rbcL sequence data. China Journal of Oceanology and Limnology, 28: 763–768.

    Google Scholar 

  • Wang, J. F., Jiang, P., Cui, Y. L., Li, N., Wang, M. Q., Lin, H. Z., Hee, P., and Qin, S., 2010b. Molecular analysis of green-tideforming macroalgae in Yellow Sea. Aquatic Botany, 93: 25–31.

    Article  Google Scholar 

  • Yang, Z., Wang Y., Dong, K. S., Tang, X. X., and Zhao, X., 2009. The survey on the community of benthic marine macroalgae. Periodical of Ocean University of China, 39(4): 647–651.

    Google Scholar 

  • Zhao, J., Jiang, P., Liu, Z. Y., Wei, W., Lin, H. Z., Li, F. C., Wang, J. F., and Q, S., 2012. The Yellow Sea green tides were dominated by one species, Ulva (Enteromorpha) prolifera, from 2007 to 2011. Chinese Science Bulletin, DOI: 10.1007/ s11434-012-5441-3.

    Google Scholar 

  • Zuccarello, G., Price, N., Verbruggen, H., and Leliaert, F., 2009. Analysis of a plastid multigene data set and the phylogenetic position of the marine macroalga Caulerpa filiformis (Chlorophyta). Journal of Phycology, 45: 1206–1212.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yunxiang Mao.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Du, G., Wu, F., Mao, Y. et al. DNA barcoding assessment of green macroalgae in coastal zone around Qingdao, China. J. Ocean Univ. China 13, 97–103 (2014). https://doi.org/10.1007/s11802-014-2197-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11802-014-2197-1

Key words

Navigation