Identification of random amplified polymorphic DNA (RAPD) marker for differentiating male from female and sporophytic thalli of <Emphasis Type="Italic">Gracilaria changii</Emphasis> (Rhodophyta) |
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Authors: | M C Sim P E Lim S Y Gan S M Phang |
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Institution: | (1) Institute of Biological Sciences, University of Malaya, 50603 Kuala Lumpur, Malaysia;(2) International Medical University, Plaza Komanwel Bukit Jalil, 57000 Kuala Lumpur, Malaysia |
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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.
Presented at the 6th Meeting of the Asian Pacific Society of Applied Phycology, Manila, Philippines. |
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Keywords: | Gracilaria changii Molecular sex markers Random amplified polymorphic DNA (RAPD) |
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