International Journal of Marine Science 2016, Vol.6, No.33, 1-7
1
Research Report Open Access
Morphological and Molecular Studies of Undescribed
Kappaphycus
Species
V. Y. Thien, W. T. L. Yong, G. J. W. L. Chin
Biotechnology Research Institute, Universiti Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah, Malaysia
Corresponding author email
International Journal of Marine Science 2016, Vol.6, No.33 doi
Received: 09 Jul., 2016
Accepted: 12 Sep., 2016
Published: 18 Sep., 2016
This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article
:
Thien V.Y., Yong W.T.L. and Chin G.J.W.L., 2016, Morphological and Molecular Studies of Undescribed
Kappaphycus
Species, International Journal of Marine
Science, 6(33): 1-7 (doi
Abstract
Morphological and molecular studies were carried on an undescribed species of
Kappaphycus
(AS 12) from Sabah,
Malaysia. The species displays unique and different physical characters from
Kappaphycus alvarezii
,
K. striatus
,
K. malesianus
and
Eucheuma denticulatum
in terms of branching patterns, thalli texture and colors. The phylogenetic position of this plant was inferred
by
cox
2-3 spacer, intergenic transcribed spacer (ITS) region and RuBisCo spacer. Based on the results from this study, it is clear that
the specimen AS 12 pointed to significant differences from
K. alvarezii
,
K. striatus
,
K. malesianus
and
E. denticulatum
has supported
by both morphological and molecular analyses.
Keywords
cox
2-3 spacer;
Eucheuma
; ITS region;
Kappaphycus
; RuBisCo spacer
1 Introduction
This study is a morphological report of an undescribed species of
Kappaphycus
encountered during our sampling
trips to Semporna, Sabah, Malaysia. The morphological characters exhibited in this plant are unique and different
as compared to the related
Kappaphycus
and
Eucheuma
species. Its phylogenetic position is proposed to be
examined using three molecular markers including mitochondrial-encoded
cox
2-3 spacer, nuclear-encoded
ribosomal internal transcribed spacer (ITS) and plastid-encoded RuBisCo spacer, to clarify the taxonomical
position of this entity at the species level.
The physical characteristic of
Kappaphycus
and
Eucheuma
tend to be variable where it is manipulated by both
genetic make-up and environmental influences and apparently from spontaneous mutations (Neish, 2008). They
are different by coloration, branch structure and other morphologies based on environments where they grow.
They tend to be highly variable, enabling them to colonize better in variety of habitats and thrive in different
environment regimes. The differences in their morphology maybe due to the interaction between light, water
currents, water depth and nutrient availability (Santelices, 1999; Munoz et al., 2004; Thirumaran and
Anantharaman, 2009; Góes and Reis, 2011).
2 Materials and Methods
Specimens for this study (AS 12) were obtained from Sebangkat Island with latitude of 4°33' 18.8994" and
longitude of 118°39' 18.7806" in Semporna, Sabah on 16 October 2012. Gross external morphology was
examined and described. Genomic DNA was extracted from approximately 100 mg of thalli ground in liquid
nitrogen by using CTAB DNA extraction procedure outlined by Zuccarello et al. (2006). PCR amplifications of
the
cox
2-3 spacer (Zuccarello et al., 2006), ITS region (White et al., 1990) and RuBisCo spacer (Tan et al., 2013)
were carried out using primers shown in Table 1. PCR amplification was conducted using TopTaq DNA
Polymerase (QIAGEN, Inc, USA) according to manufacturer’s instruction. Cycling condition of the amplification
was carried out as follow: initial denaturation at 94 °C for 3 min, followed by 30 cycles of each consisting
denaturation at 94 °C for 30 sec, annealing at 50 - 56 °C according to primers used for 30 sec and extension at
72 °C for 1 min, and lastly final extension at 72 °C for 10 min. Annealing temperature was manipulated to obtain
optimal PCR products. PCR products were purified by using the QIAquick Gel Extraction Kit (QIAGEN, Inc,
