RESTRICTION FRAGMENT LENGTH POLYMORPHISM OF RIBOSOMAL DNA INTERNAL TRANSCRIBED SPACER AND 5.8S REGIONS IN JAPANESE ALEXANDRIUM SPECIES (DINOPHYCEAE)¹

Abstract

The 5.8S ribosomal RNA (rDNA) gene and flanking internal transcribed spacers (ITS1 and ITS2)from 9 isolates of Alexandrium catenella (Whedon and Kofoid) Taylor, 11 isolates of A. tamarense (Lebour) Taylor, and single isolates of A. affine (Inoue et Fukuyo) Balech, A. insuetum Balech, and A. pseudogonyaulax (Biecheler) Horiguchi ex Yuki et Fukuyo comb. nov. from various locations in Japan were amplified using the polymerase chain reaction (PCR) and subjected to restriction fragment‐length polymorphism (RFLP) analysis. PCR products from all strains were approximately 610 bp, inclusive of a limited region of the 18S and 28S rRNA coding regions. RFLP analysis using four restriction enzymes revealed six distinct classes of rDNA (“ITS types”). Restriction patterns of A. catenella were uniform at the intra‐specific level and clearly distinguishable from those of A. tamarense. The patterns associated with A. tamarense (“tamarense group”) were also uniform except for one strain, WKS‐1. Some restriction fragments from WKS‐1 were in common with those of A. catenella or A. tamarense, whereas some were distinct from all Alexandrium species tested. Alexandrium affine, A. insuetum, and A. pseudogonyaulax carry unique ITS types. The ITSs of the “tamarense group” exhibit sequence heterogeneity. In contrast, the ITSs of all other isolates (including WKS‐1) appear homogeneous. RFLP analysis of the 5.8S rDNA and flanking ITSs regions from Alexandrium species reveals useful taxonomic and genetic markers at the species and/or population levels.

Morphological and genetic study of Prorocentrum donghaiense Lu from the East China Sea, and comparison with some related Prorocentrum species

“…DNA extraction was carried out with the modified method of (Rajeshwar et al, 2001) PCR amplification. The ITS sequences were PCR-amplified from purified total DNA using ITS A primer (5 0 -CCAAGCTTCTAGATCGTAACAAG-G(ACT)TCCGTAGGT-3 0 ) and ITS B primer (5 0 -CCTGCAGTCGACA(TG)ATGCTTAA(AG)TTCA-GC(AG)GC-3 0 ) (Adachi et al, 1994). The PCR products were separated on 1% agarose gels (regular, BIOWEST, Spanish).…”

Fibrocapsa japonica (Raphidophyceae) occurrence and ecological features within the phytoplankton assemblage of a cyclonic eddy, offshore the Eastern Alboran Sea.

“…Eukaryoticspecific primers targeting the 5.8S-ITS regions (Adachi et al, 1994) were used in the first PCR. The PCR-amplified products from the PCR assay using the universal eukaryotic primers and not directly visualised on agarose gel were used as the template in the nested PCR amplification with species-specific primers for F. japonica using 1 µl of the first PCR-amplified product.…”

Growth characteristics and phylogenetic analysis of the marine dinoflagellate Dinophysis infundibulus (Dinophyceae)

“…For amplification of the SSU rDNA region, the following primers were used: S1F (5'-AACCTGGTTGATYCTGCCAG-3') + S1R (5'-CTACGAGCTTTTTAACCGCAACAA-3'), S2F (5'-CTGAGAAACGGCTACCACATC-3') + S2R (5'-TGGTAAGTTTTCCCGTGTTGAGTC-3'), and S3F (5'-AGCTTGCGGCTTAATTTGACTC-3') + S3R (5'-CTACGGAAACCTTGTTACGAC-3'). The ITS regions and LSU rDNA (D1-D2 region) were amplified according to Adachi et al (1994) and Scholin et al (1994), respectively. The PCR cycling conditions were as follows: 5 min at 94°C, 40 cycles of 30 s each at 94°C, 30 s at 56°C, 1 min at 72°C, and a final elongation for 5 min at 72°C.…”