• ALGAE
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• v.28 no.1
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• pp.31-43
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• 2013

This review provides a comprehensive summary of the PCR primers and profiles currently in use in our laboratory for red algal DNA barcoding and phylogenetic research. While work focuses on florideophyte taxa, many of the markers have been applied successfully to the Bangiales, as well as other lineages previously assigned to the Bangiophyceae sensu lato. All of the primers currently in use with their respective amplification profiles and strategies are provided, which can include full fragment, overlapping fragments and what might best be called "informed overlapping fragments", i.e., a fragment for a marker is amplified and sequenced for a taxon and those sequence data are then used to identify the best primers to amplify the remaining fragment(s) for that marker. We extend this strategy for the more variable markers with sequence from the external PCR primers used to "inform" the selection of internal sequencing primers. This summary will hopefully serve as a useful resource to systematists in the red algal community.

• Molecular & Cellular Toxicology
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• v.4 no.3
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• pp.199-204
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• 2008

The bio-molecules involved in defense mechanisms can be used as efficient biomarkers for physiological changes in organisms caused by both of internal and external stress. Thus, the expression level of genes which encoding such molecules serve as critical 'early warning system' for environmental assessment as well as health diagnosis of biological organisms. In this study, Cytochrome P450, Heat shock protein 90, Ubiquitin and ${\beta}$-actin gene were isolated for the first time from a red alga Gracilaria textorii. The quantitative differential gene expression analyses of three genes, GteCYP1A, GteHsp90 and Gte-UB, were carried out in G. textorii sporophytes collected from two different localities, polluted Sujeong (Masan, Korea) and potentially unpolluted Danggeum (Daemaemuldo Is., Korea). The transcripts of all three tested genes were highly expressed in the Sujeong population. The results suggest: 1) the Sujeong site was more polluted than the Danggeum site; 2) G. textorii could be applicable to marine environment monitoring in coastal regions.

• ALGAE
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• v.34 no.2
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• pp.91-97
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• 2019

Chilean species of marine macroalgae with amphitropical distributions oftentimes result from introductions out of the Northern Hemisphere. This possibility was investigated using haplotype data in an amphitropical red macroalgae present in Chile, Callophyllis variegata. Published sequence records from Canada and the United States were supplemented with new collections from Chile (April 2014-November 2015). Specimens of C. variegata were amplified for the 5′ end of the cytochrome c oxidase subunit I gene (COI-5P) and the full length nuclear internal transcribed spacer region. Haplotype networks and biogeographic distributions were used to infer whether C. variegata was introduced between hemispheres, and several population parameters were estimated using IMa2 analyses. C. variegata displayed a natural amphitropical distribution, with an isolation time of approximately 938 ka between hemispheres. It is hypothesized that contemporary populations of C. variegata were established from a refugial population during the late Pleistocene, and may have crossed the tropics via rafting on buoyant species of kelp or along deep-water refugia coincident with global cooling, representing a rare case of a non-human mediated amphitropical distribution.

• ALGAE
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• v.31 no.2
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• pp.167-174
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• 2016

Biological response of cells to variable conditions should affect the expression level of certain genes. Quantification of these changes in target genes needs stable internal controls. Real-time quantitative polymerase chain reaction (PCR) has traditionally used reference or ‘housekeeping’ genes, that are considered to maintain equal expression in different conditions, to evaluate changes in target genes between samples and experimental conditions. Recent studies showed that some housekeeping genes may vary considerably in certain biological samples. This has not been evaluated in red algae. In order to identify the optimal internal controls for real-time PCR, we studied the expression of eleven commonly used housekeeping genes; elongation factor 1-alpha, glyceraldehyde-3-phosphate dehydrogenase, β-actin, polyubiquitin, 30S ribosomal gene, 60S ribosomal gene, beta-tubulin, alpha-tubulin, translation initiation factor, ubiquitin-conjugating enzyme, and isocitrate dehydrogenase in different life-history stages of Bostrychia moritziana. Our results suggest that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and 30S ribosomal gene, have the most stable gene expression levels between the different life history stages (male, female, carposporophyte, and tetrasporophyte), while the other genes are not satisfactory as internal controls. These results suggest that the combinations of GAPDH and 30S would be useful as internal controls to assess expression level changes in genes that may control different physiological processes in this organism or that may change in different life history stages. These results may also be useful in other red algal systems.

• ALGAE
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• v.29 no.2
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• pp.101-109
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• 2014

Small red algae, especially those previously referred to as 'primitive' are often overlooked, but can be quite abundant. These 'primitive' red algae are now placed in several classes distinct from the Florideophyceae, for example the Stylonematophyceae. A brownish-red filamentous alga was collected from a sandy tide pool at Cape Tribulation, Queensland, Australia. Cultured specimens were identified as Bangiopsis and conformed to the morphological characters of the genus (multicellular base, erect filaments branched or unbranched, uniseriate to multiseriate-tubular, single multilobed purple-red to red-brown plastid with central pyrenoid, vegetative cells released directly as spores). Molecular data of two plastid genes (rbcL, psbA) support placement of the Australian isolate and isolates from India in Bangiopsis. The genetic variation between these isolates and isolates from Puerto Rico previously attributed to B. subsimplex indicates that these should be considered as a separate species. As the type locality is in the Atlantic Ocean, French Guiana, and not far from Puerto Rico, and the Puerto Rican isolate has been used often in phylogenetic analyses, we propose that the Indian and Pacific Ocean isolates be designated a new species, B. franklynottii, to acknowledge Ott's many years of research on inconspicuous freshwater and marine red algae. Our research also highlights the lack of careful descriptions in many of the records of this genus and the lack of morphological characters to distinguish species. Especially within the morphologically simple red algae, morphological distinctness does not necessarily reflect evolutionary divergences.

• ALGAE
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• v.33 no.3
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• pp.269-278
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• 2018

Ocean warming can have significant negative impacts on population genetic diversity, local endemism and geographical distribution of a wide range of marine organisms. Thus, the identification of conservation units with high risk of extinction becomes an imperative task to assess, monitor, and manage marine biodiversity for policy-makers. Here, we surveyed population structure and genetic variation of the red seaweed Gracilaria vermiculophylla along the coast of China using genome-based amplified fragment length polymorphism (AFLP) scanning. Regardless of analysis methods used, AFLP consistently revealed a south to north genetic isolation. Populations at the southern coast of China showed unique genetic variation and much greater allelic richness, heterozygosity, and average genetic diversity than the northern. In particular, we identified a geographical barrier that may hinder genetic exchange between the two lineages. Consequently, the characterized genetic lineage at the southern coast of China likely resulted from the interplay of post-glacial persistence of ancestral diversity, geographical isolation and local adaptation. In particular, the southern populations are indispensable components to explore evolutionary genetics and historical biogeography of G. vermiculophylla in the northwestern Pacific, and the unique diversity also has important conservation value in terms of projected climate warming.

• ALGAE
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• v.35 no.4
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• pp.389-404
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• 2020

Red algal fertilization is unusual and offers a different model to the mechanism of intracellular transport of nuclei and polyspermy blocking. A female carpogonium (egg) undergoes plasmogamy with many spermatia (sperm) simultaneously at the receptive structure, trichogyne, which often contains numerous male nuclei. The pattern of selective transport of a male nucleus to the female nucleus, located in the cell body of the carpogonium, remain largely unknown. We tracked the movement of spermatial nuclei and cell organelles in the trichogyne after plasmogamy using time-lapse videography and fluorescent probes. The fertilization process of Bostrychia moritziana is composed of five distinctive stages: 1) gamete-gamete binding; 2) mitosis in the attached spermatia; 3) formation of a fertilization channel; 4) migration of spermatial nuclei into the trichogyne; and 5) cutting off of the trichogyne cytoplasm from the rest of the cell after karyogamy. Our results showed that actin microfilaments were involved in the above steps of fertilization, microtubules are involved only in spermatial mitosis. Time-lapse videography showed that the first ("primary") nucleus which entered to trichogyne moved quickly to the base of carpogonium and fused with the female nucleus. The transport of the primary male nucleus to the egg nucleus was complete before its second nucleus migrated into the trichogyne. Male nuclei from other spermatia stopped directional movement soon after the first one entered the carpogonial base and oscillated near where they entered trichogyne. The cytoplasm of the trichogyne was cut off at a narrow neck connecting the trichogyne and carpogonial base after gamete nuclear fusion but gamete binding and plasmogamy continued on the trichogyne. Spermatial organelles, including mitochondria, entered the trichogyne together with the nuclei but did not show any directional movement and remained close to where they entered. These results suggest that polyspermy blocking in B. moritziana is achieved by the selective and rapid transport of the first nucleus entered trichogyne and the rupture of the trichogyne after gamete karyogamy.