PCR-SSCP as a molecular tool for the identification of Benedeniinae (Monogenea: Capsalidae) from marine fish.
PCR-based single strand conformation polymorphism (SSCP) analysis was used to characterize monogenean specimens of the subfamily Benedeniinae of morphologically uncertain specific status from different marine fish species, using Neobenedenia melleni, N. girellae and Entobdella corona for comparison. The first internal transcribed spacer (ITS-1) and the 5' terminal variable region (D1-D3 domains) of the large subunit ribosomal DNA (lsrDNA) were amplified separately from individual monogeneans, and the amplicons were subjected to PCR-SSCP analyses, followed by direct sequencing. Both SSCP patterns and the ITS-1 sequences data allowed specimens representing Entobdella spp. from the host Taeniura melanospilos to be unequivocally distinguished from those representing Neobenedenia spp. and those representing Benedenia spp. Neobenedenia girellae, a morphologically controversial species, had identical SSCP banding pattern and ITS-1 sequence to that of N. melleni, supporting the proposal that N. girellae is a synonym of N. melleni. Neobenedenia spp. and Benedenia spp. had identical SSCP patterns and ITS-1 sequences. These findings and the PCR-SSCP approach taken should have implications for the accurate identification and assessment of taxonomic validity of other important monogenean groups of the marine fish.
Li
An-Xing
AX
Center for Parasitic Organisms, School of Life Sciences, Zhongshan (Sun Yat-Sen) University, Haizhu District, Guangzhou 510275, The People's Republic of China.
Wu
Xiang-Yun
XY
Ding
Xue-Juan
XJ
Lin
Rui-Qing
RQ
Xie
Ming-Quan
MQ
Lun
Zhao-Rong
ZR
Zhu
Xing-Quan
XQ
D000818:Animals; D001483:Base Sequence; D002965:Classification; D004247:DNA / Q000032:analysis; D005393:Fish Diseases / Q000175:diagnosis / Q000469:parasitology*; D005399:Fishes / Q000469:parasitology*; D008969:Molecular Sequence Data; D010271:Parasites / Q000235:genetics*; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational
D004247:DNA