Molecular characterisation of Sargassum polycystum and S. siliquosum (Phaeophyta) by polymerase chain reaction (PCR) using random amplified polymorphic DNA (RAPD) primers

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

doi:10.1007/bf00003547

Cited by 31 publications

(18 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Although the CTAB method (Shivji et al, 1992;Ho et al, 1995) and the combination of alginic lyase and CTAB (Hu & Zhou, 2001) have been reported successfully to extract DNA from brown algae, they are not suitable for DNA extraction from sporophyte Laminaria japonica. DNA isolated by these two methods was contaminated with large amounts of polysaccharides and RNA, and DNA solution was so viscous that it was difficult to peppete DNA into the sample wells.…”

Section: Discussionmentioning

confidence: 99%

“…To reduce the effects of viscous polysaccharides, Ho et al (1995) found that DNA extraction from Sargassum polycystum and S. siliquosum using CTAB (cetyltrimethyl ammonium bromide) were amplified successfully by random amplified polymorphic DNA (RAPD-PCR). Hu and Zhou (2001) reported a new extraction method to extract high quality DNA from Laminaria japonica, which involves enzymatic dissociation of sporophyte tissues and subsequent elimination of the remaining polysaccharides using CTAB.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

A simple method for DNA extraction from sporophyte in the brown alga Laminaria japonica

Wang

Xia

et al. 2005

J Appl Phycol

View full text Add to dashboard Cite

A simple method was developed for extracting DNA from brown algae Laminaria japonica, which possess large amounts of acidic polysaccharides. Firstly, the sporophyte were washed by eliminating polysaccaride buffer to remove the polysaccharides and then ground in liquid nitrogen. Secondly, the powders were treated with lysing buffer. Thirdly, KAc was used to eliminate the remaining acidic polysaccharides. The extracted DNA was purified using a chloroform-isoamyl alcohol (24:1 v/v), and precipitated in cold isopropanol. The yield was from 18.7 to 37.5 µg g −1 (wet weight) and the purity of total DNA was determined spectrophotometrically as the ratio of A 260 /A 280 , which was about 1.7-1.9. The extracted DNA was of high quality and suitable for molecular analyses, such as PCR, restriction enzyme digestion. This method is a reproducible, simple, and rapid technique for routine DNA extraction from sporophyte in Laminaria japonica. Furthermore, the low cost of this method makes it attractive for large-scale studies.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A simple method for DNA extraction from sporophyte in the brown alga Laminaria japonica

Wang

Xia

et al. 2005

J Appl Phycol

View full text Add to dashboard Cite

show abstract

“…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: 99%

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

2004

Self Cite

View full text Add to dashboard Cite

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.

show abstract

“…For a general review of this technique, see Weising et al (1995). In seaweed studies, RAPDs have been used for species identification (Patwary et al 1993, Dutcher & Kapraun 1994, Patwary & van der Meer 1994, Ho et al 1995, Gonzalez et al 1996, Meneses 1996, biogeographic studies (van Oppen et al 1994, Pakker et al 1996, and the determination of genetic relatedness among and within populations (Coyer et al 1997). Van Oppen et al (1996) assessed the limits of RAPDs for seaweed biogeography.…”

Section: Introductionmentioning

confidence: 99%

Assessing patterns of geographic dispersal of Gelidium sesquipedale (Rhodophyta) through RAPD differentiation of populations

Alberto¹,

Santos²,

Leitão³

1999

Mar. Ecol. Prog. Ser.

View full text Add to dashboard Cite

Randomly ampldied polymorphic DNAs (RAPDs) of bulked genomic DNA samples were used to analyse the genetlc differentiation of Gelidium sesquipedale populations. They reflect the patterns of gene flow, which in turn depend on the dispersal mechanisms of the species and on near-shore ocean currents. Fourteen populations were sampled from northern France to Morocco, covering the geographical distribution of the species. A single bulk DNA sample (from 15 individuals) was used in each population, under the assumption that the resulting patterns represent the populations' most common genetic features. To test this, we investigated the genetic variability among 5 bulk samples within a single population. Genetic distances anlong bulks were very low (average = 0.065) and were significantly lower than those observed between geographically separated populations (average = 0.241). Neighbour-joining analysis of the distance matrix of populations separated a well-supported group including populations of northern Spain and of northern France, and a less-supported group containing populations of northern Portugal. Multidimensional scaling of the genetic distance matrix revealed 2 isolated populations. Sao Rafael in southern Portugal and Algeciras in southern Spain. These patterns of genetic differentiation are discussed under the available data on the near-shore ocean currents. Results suggest that the genetic differentiatlon of G. sesqujpedale populations may be used as a biological tracer of prevailing flows and barriers of the near-shore currents. A positive correlation between geographical and genet~c distances of G. sesquipedale populations along the specles geographical distribution was found, suggesting that a continuous transport of detached fronds and their reattachment to new substrate must be an effective dispersal mechanism of the species, sustaining the gene flow among populations.

show abstract

Molecular characterisation of Sargassum polycystum and S. siliquosum (Phaeophyta) by polymerase chain reaction (PCR) using random amplified polymorphic DNA (RAPD) primers

Cited by 31 publications

References 20 publications

A simple method for DNA extraction from sporophyte in the brown alga Laminaria japonica

A simple method for DNA extraction from sporophyte in the brown alga Laminaria japonica

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

Assessing patterns of geographic dispersal of Gelidium sesquipedale (Rhodophyta) through RAPD differentiation of populations

Contact Info

Product

Resources

About