Application of polymerase chain reaction (PCR) using random amplified polymorphic DNA (RAPD) primers in the molecular identification of selected<i>Sargassum</i>species (Phaeophyta, Fucales)

Ho, C. C.; Phang, Siew‐Moi; Pang, Tikki

doi:10.1080/09670269500651051

Cited by 15 publications

(14 citation statements)

References 17 publications

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“…Random amplified polymorphism DNA (RAPD) (Williams et al, 1990;Hadrys et al, 1992) was used successfully in species differentiation on Gracilaria (Meneses, 1996;Lim et al, 2001), Furcellaria (Valatka et al, 2000), Laminaria (Billot et al, 1999) and Sargassum (Ho et al, 1995a(Ho et al, , 1995bPark et al, 1998;Engelen et al, 2001). The objective of this study is to use a combination of morphometric and RAPD analyses to differentiate between the S. baccularia and S. polycystum, which are found in close proximity.…”

Section: Introductionmentioning

confidence: 97%

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Morphological and molecular characterisation and differentiation of Sargassum baccularia and S. polycystum (Phaeophyta)

2004

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Two Sargassum species (S. baccularia and S. polycystum) collected from Teluk Kemang and Cape Rachado, Port Dickson, Negeri Sembilan, Malaysia, which are alike in morphology except for the rhizoidal system and vesicles, were characterised using random amplified polymorphism DNA (RAPD). The genomic DNA of both species was isolated from the leaves using a modified CTAB method. Four random primers, that is, OPA2, OPA3, OPA4 and OPA13, successfully amplified the DNA. The polymorphisms generated by these four primers were analysed using the Dice Coefficient of Similarity and cluster analysis was carried out using GelCompar II Version 2.0 (Applied Maths, Kortrijk, Belgium) based on UPGMA. DNA analysis showed that three primers were able to differentiate the two species. Morphological analysis using Principal Components Analysis (PCA) and discriminant function analysis supported the molecular data. Both species are characterised by heavily muricate main branches, oblonglanceolate leaves with dentate margins and discoid holdfasts and spherical vesicles; both are dioecious. The only difference is that S. polycystum has secondary holdfasts transformed into stolons. This last characteristic is therefore a very important criterion and may contribute to the difference shown by DNA analysis.

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Section: Introductionmentioning

confidence: 97%

“…There are more than 400 species of Sargassum, described primarily using morphological characters (Ho et al, 1995a). Sargassum is highly differentiated with variable phenotypes based on several factors including environment and locality.…”

Section: Introductionmentioning

confidence: 99%

Morphological and molecular characterisation and differentiation of Sargassum baccularia and S. polycystum (Phaeophyta)

2004

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“…The DNeasy Plant Mini Kit was found to be unsuitable for Gracilaria, as it could not efficiently remove the polysaccharide. This was also observed with Sargassum, which has alginic acid (Ho 1995a;Wong et al 2004). This can be overcome by using only the tips of Gracilaria that are freshly ground using liquid nitrogen.…”

Section: Discussionmentioning

confidence: 52%

“…2). Previous work using RAPD on Gracilaria and Sargassum species in the Algae Research Laboratory, University of Malaya, showed good reproducibility (Ho et al 1995a;Gan 1999;Lim et al 2001). Major bands were observed to be constant with all the amplifications, but minor bands were more variable.…”

Section: Molecular Analysismentioning

confidence: 91%

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

et al. 2007

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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 phenolchloroform 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. Sixtynine 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 lifestage using the unweighted pair group and arithmetic averages (UPGMA) method. It showed that RAPD was able to differentiate tetrasporophytes, females, and males.

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“…Recently, RAPD was used in genotoxicity studies (Atienzar et al 2000;Atienzar and Jha 2006) and in discriminating genetically modified organisms (Cheah and Radu 2006). RAPD was used successfully in species differentiation for seaweeds, namely, Gracilaria species (Martinez et al 1999;Lim et al 2001), Sargassum species (Ho et al 1995a(Ho et al , 1995bPark et al 1998;Engelen et al 2001;, Postelsia (Coyer et al 1997), Hizikia fusiformis (Park et al 1998), Laminaria (Billot et al 1999;Yotsukura et al 2001) and Furcellaria lumbricalis (Valatka et al 2000). The objective of this study is to use RAPD to differentiate between (a) the S. binderi and S. oligocystum, and (b) S. baccularia, S. polycystum and S. stolonifolium.…”

Section: Introductionmentioning

confidence: 99%

Use of RAPD in differentiation of selected species of Sargassum (Sargassaceae, Phaeophyta)

Wong

Phang

2007

J Appl Phycol

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Sargassum C. Agardh is one of the most common but little understood genera of Phaeophyta in Malaysia. The difficulty in species delineation is due to morphological plasticity. A combination of morphology and random amplified polymorphic DNA (RAPD) studies of selected Sargassum species was carried out to have a better understanding of the taxonomy. Primer OPA13 was found to be good for discriminating between Sargassum species. Sargassum binderi was shown to be different from S. oligocystum (S D >0.5=14.11%), indicating the importance of the vesicle and receptacle in species differentiation. S. baccularia was clearly separated out from S. polycystum and S. stolonifolium using primer OPA13. RAPD analysis showed that the presence of the stolon is an important character for separating S. baccularia (no stolon) from S. polycystum (stolon) and S. stolonifolium (stolon).

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Application of polymerase chain reaction (PCR) using random amplified polymorphic DNA (RAPD) primers in the molecular identification of selectedSargassumspecies (Phaeophyta, Fucales)

Cited by 15 publications

References 17 publications

Morphological and molecular characterisation and differentiation of Sargassum baccularia and S. polycystum (Phaeophyta)

Morphological and molecular characterisation and differentiation of Sargassum baccularia and S. polycystum (Phaeophyta)

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

Use of RAPD in differentiation of selected species of Sargassum (Sargassaceae, Phaeophyta)

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