Skip to main content
SpringerLink
Log in

Identification of random amplified polymorphic DNA (RAPD) marker for differentiating male from female and sporophytic thalli of Gracilaria changii (Rhodophyta)

  • Published:
Journal of Applied Phycology Aims and scope Submit manuscript

Abstract

There have been limited reports on molecular sex markers for macroalgae. We report the use of random amplified polymorphic DNA analysis (RAPD) to identify molecular sex markers for Gracilaria changii (Xia et Abbott) Abbott, Zhang et Xia. Two DNA extraction methods were used: a modified CTAB and phenol-chloroform combination method and the DNeasy Plant Mini Kit. The CTAB and phenol-chloroform method gave the best yield of DNA in quality and quantity and is suitable for larger-sized specimens like G. changii. Sixty-nine RAPD primers were screened to search for sex-linked DNA markers for G. changii, and only one sex-linked marker (716 bp) was identified using OPA 18. RAPD was also used to investigate the molecular characteristics of the three life-stages (male, female, tetrasporophyte) of G. changii. Seven (OPA7, OPA18, S14, S61, S64, S75 and S76) out of the 69 primers showed polymorphism and were selected for interpopulation analysis for DNA isolated from 23 samples collected from Morib and Sungai Pulai in Malaysia. The combination of data produced by the seven primers generated a dendrogram that grouped the specimens into different clades according to their sex and life-stage using the unweighted pair group and arithmetic averages (UPGMA) method. It showed that RAPD was able to differentiate tetrasporophytes, females, and males.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Billot C, Boury S, Benet H, Kloareg B (1999) Development of RAPD markers for parentage analysis in Laminaria digitata. Bot Mar 42:307–314

    Article  Google Scholar 

  • Bouza N, Caujapé-Castells J, González-Pérez MA, Sosa PA (2006) Genetic structure of natural populations in the red algae Gelidium canariense (Gelidiales, Rhodophyta) investigated by random amplified polymorphic DNA (RAPD) markers. J Phycol 42:304–311

    Article  CAS  Google Scholar 

  • Byrne K, Zuccarello GC, West J, Liao ML, Kraft GT (2002) Gracilaria species (Gracilariaceae, Rhodophyta) from southeastern Australia, including a new species, Gracilaria perplexa sp. Nov.: morphology, molecular relationships and agar content. Phycol Res 50:295–311

    Article  Google Scholar 

  • Deputy JC, Ming R, Ma H, Liu Z, Fitch MMM, Wang M, Manshardt R, Stiles JI (2002) Molecular markers for sex determination in papaya (Carica papaya L.). Theor Appl Genet 106:107–111

    PubMed  CAS  Google Scholar 

  • Dutcher JA, Kapraun DF (1994) Random amplified polymorphic DNA (RAPD) identification of genetic variation in three species of Porphyra (Bangiales, Rhodophyta). J Appl Phycol 6:267–273

    Article  CAS  Google Scholar 

  • Gan SY (1999) Molecular taxonomic studies of Gracilaria changii from various locations using the Random Amplified Polymorphic DNA (RAPD) technique. M Sc Thesis, University of Malaya, Kuala Lumpur

  • Gan SY, Qin S, Othman RY, Yu DZ, Phang SM (2003) Transient expression of lacZ in particle bombarded Gracilaria changii (Gracilariales, Rhodophyta). J Appl Phycol 15:351–353

    Article  Google Scholar 

  • González M, Rolando M, Candia A, Gómez P, Cisternas M (1996) Organellar DNA restriction fragment length polymorphism (RFLP) and nuclear random amplified polymorphic DNA (RAPD) analyses of morphotypes of Gracilaria (Gracilariales, Rhodophyta) from Chile. Hydrobiologia 326/327:229–234

    Article  Google Scholar 

  • Ho CL, Phang SM, Tikki P (1995a) Application of polymerase chain reaction (PCR) using random amplified polymorphic DNA (RAPD) primers in the molecular identification of selected Sargassum species (Phaeophyta, Fucales). Eur J Phycol 30:273–280

    Article  Google Scholar 

  • Ho CL, Phang SM, Tikki P (1995b) Molecular characterization of Sargassum polycystum and S. siliquosum (Phaeophyta) by polymerase chain reaction (PCR) using random amplified polymorphic DNA (RAPD) primers. J Appl Phycol 73:33–41

    Article  Google Scholar 

  • Holloway AK, Strand MR, William CB, Antolin MF (2000) Linkage analysis of sex determination in Bracon sp. near hebetor (Hymenoptera: Braconidae). Genetics 154:205–212

    PubMed  CAS  Google Scholar 

  • Huh MK, Lee BK, Lee HY (2006) Genetic diversity and phylogenetic relationships in five Porphyra species revealed by RAPD analysis. Protistology 4:245–250

    CAS  Google Scholar 

  • Iturra P, Medrano JF, Bagley M, Lam N, Vergara N, Marin JC (1998) Identification of sex chromosome molecular markers using RAPDs and fluorescent in situ hybridization in rainbow trout. Genetica 101:209–213

    Article  Google Scholar 

  • Jiang C, Sink KC (1997) RAPD and SCAR markers linked to the sex expression locus M in asparagus. Euphytica 94:329–333

    Article  CAS  Google Scholar 

  • Lemos EGM, Silva CLSP, Zaidan HA (2002) Identification of sex in Carica papaya L. using RAPD markers. Euphytica 127:179–184

    Article  CAS  Google Scholar 

  • Lim PE, Phang SM (2004) Gracilaria species (Gracilariales, Rhodophyta) of Malaysia including two new records. Malays J Sci 23:71–80

    Google Scholar 

  • Lim PE, Thong KL, Phang SM (2001) Molecular differentiation of two morphological variants of Gracilaria salicornia. J Appl Phycol 13:335–342

    Article  CAS  Google Scholar 

  • Liu ZJ, Li P, Argue BJ, Dunham RA (1999) Random amplified polymorphic DNA markers: usefulness for gene mapping and analysis of genetic variation of catfish. Aquaculture 174:59–68

    Article  CAS  Google Scholar 

  • Mandolino G, Carboni A, Forapani S, Faeti V, Ranalli P (1999) Identification of DNA markers linked to the male sex in dioecious hemp (Cannabis sativa L.). Theor Appl Genet 98:86–92

    Article  Google Scholar 

  • Mandolino G, Carboni A, Bagatta M, Moliterni VMC, Ranalli P (2002) Occurrence and frequency of putatively Y chromosome linked DNA markers in Cannabis sativa L. Euphytica 126:211–218

    Article  CAS  Google Scholar 

  • Martinez EA, Destombe C, Quillet MC, Valero M (1999) Identification of random amplified polymorphic DNA (RAPD) markers highly linked to sex determination in the red alga Gracilaria gracilis. Mol Ecol 8:1533–1538

    Article  PubMed  CAS  Google Scholar 

  • McHugh DJ (2003) A guide to seaweed industry. FAO Fisheries Technical Paper 441. FAO, Rome

    Google Scholar 

  • Mizukami Y, Kito H, Kunimoto M, Kobayashi M (1998) Effect of DNA preparation from laver (Porphyra yezoensis) thalli on reproducibility of RAPD (random amplified polymorphic DNA) patterns. J Appl Phycol 10:23–29

    Article  CAS  Google Scholar 

  • Park JW, Cho YC, Nam BH, Jin HY, Sohn CH, Hong YK (1998) RAPD identification of genetic variation in seaweeds Hizikia fusiformis (Fucales, Phaeophyta). J Mar Biotechnol 6:62–64

    CAS  Google Scholar 

  • Patwary MU, MacKay RM, Van der Meer JP (1993) Revealing genetic markers in Gelidium vagum (Rhodophyta) through the random amplified polymorphic DNA (RAPD) technique. J Phycol 29:216–222

    Article  CAS  Google Scholar 

  • Phang SM, Lewmanomont K (2001) Gracilaria changii (B. M. Xia & I. A. Abbott) I. A. Abbott, C. F. Chang & B. M. Xia. In: Reine P, Trono GC Jr (eds) Plant resources of South-East Asia, no. 15(1). Cryptogams: algae. Backhuys, Leiden, pp 178–180

    Google Scholar 

  • Phang SM, Shaharuddin S, Noraishah H, Sasekumar A (1996) Studies on Gracilaria changii (Gracilariales, Rhodophyta) from Malaysian mangroves. Hydrobiologia 326/327:347–352

    Article  CAS  Google Scholar 

  • Törjék O, Bucherna N, Kiss E, Homoki H (2002) Novel male-specific molecular markers (MADC5, MADC6) in hemp. Euphytica 127:209–218

    Article  Google Scholar 

  • Valatka S, Makinen A, Yli-Mattila T (2000) Analysis of genetic diversity of Furcellaria lumbricalis (Gigartinales, Rhdophyta) in Baltic Sea by RAPD-PCR technique. Phycologia 39:110–117

    Article  Google Scholar 

  • Wong CL, Gan SY, Phang SM (2004) Morphological and molecular characterization and differentiation of Sargassum baccularia and S. polycystum (Phaeophyta). J Appl Phycol 16:439–445

    Article  CAS  Google Scholar 

  • Xu WJ, Wang BW, Cui KM (2004) RAPD and SCAR markers linked to sex determination in Eucommia ulmoides Oliv. Euphytica 136:233–238

    Article  CAS  Google Scholar 

  • Yakubov B, Barazani O, Golan-Goldhirsh (2005) Combination of SCAR primers and touchdown-PCR for sex identification in Pistacia vera L. Sci Hort 103:473–478

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors gratefully acknowledge the following: Ministry of Science, Technology and Innovation, Malaysia (IRPA Grant Nos. 39-02-03-9002), and Vote F (0187/2004B), University of Malaya, for providing financial assistance and Professor Kwai Lin Thong for her advice and the use of her equipment.

Author information

Authors and Affiliations

  1. Institute of Biological Sciences, University of Malaya, 50603, Kuala Lumpur, Malaysia

    M. C. Sim & S. M. Phang

  2. International Medical University, Plaza Komanwel Bukit Jalil, 57000, Kuala Lumpur, Malaysia

    P. E. Lim & S. Y. Gan

Authors
  1. M. C. Sim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P. E. Lim
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. Y. Gan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S. M. Phang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to S. M. Phang.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sim, M.C., Lim, P.E., Gan, S.Y. et al. Identification of random amplified polymorphic DNA (RAPD) marker for differentiating male from female and sporophytic thalli of Gracilaria changii (Rhodophyta). J Appl Phycol 19, 763–769 (2007). https://doi.org/10.1007/s10811-007-9224-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10811-007-9224-1

Keywords

Search

Navigation