PMID- 22022408
OWN - NLM
STAT- MEDLINE
DCOM- 20120217
LR  - 20211020
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 6
IP  - 10
DP  - 2011
TI  - Development of a PNA probe for fluorescence in situ hybridization detection of 
      Prorocentrum donghaiense.
PG  - e25527
LID - 10.1371/journal.pone.0025527 [doi]
LID - e25527
AB  - Prorocentrum donghaiense is a common but dominant harmful algal bloom (HAB) 
      species, which is widely distributed along the China Sea coast. Development of 
      methods for rapid and precise identification and quantification is prerequisite 
      for early-stage warning and monitoring of blooms due to P. donghaiense. In this 
      study, sequences representing the partial large subunit rDNA (D1-D2), small 
      subunit rDNA and internal transcribed spacer region (ITS-1, 5.8S rDNA and ITS-2) 
      of P. donghaiense were firstly obtained, and then seven candidate DNA probes were 
      designed for performing fluorescence in situ hybridization (FISH) tests on P. 
      donghaiense. Based on the fluorescent intensity of P. donghaiense cells labeled 
      by the DNA probes, the probe DP0443A displayed the best hybridization 
      performance. Therefore, a PNA probe (PP0443A) analogous to DP0443A was used in 
      the further study. The cells labeled with the PNA probe displayed more intensive 
      green fluorescence than that labeled with its DNA analog. The PNA probe was used 
      to hybridize with thirteen microalgae belonging to five families, i.e., 
      Dinophyceae, Prymnesiophyceae, Raphidophyceae, Chlorophyceae and 
      Bacillariophyceae, and showed no visible cross-reaction. Finally, FISH with the 
      probes PP0443A and DP0443A and light microscopy (LM) analysis aiming at 
      enumerating P. donghaiense cells were performed on the field samples. Statistical 
      comparisons of the cell densities (cells/L) of P. donghaiense in the natural 
      samples determined by FISH and LM were performed using one-way ANOVA and Duncan's 
      multiple comparisons of the means. The P. donghaiense cell densities determined 
      by LM and the PNA probe are remarkably higher than (p<0.05) that determined by 
      the DNA probe, while no significant difference is observed between LM and the PNA 
      probe. All results suggest that the PNA probe is more sensitive that its DNA 
      analog, and therefore is promising for the monitoring of harmful algal blooms of 
      P. donghaiense in the future.
FAU - Chen, Guofu
AU  - Chen G
AD  - State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of 
      Technology, Harbin, China.
FAU - Zhang, Chunyu
AU  - Zhang C
FAU - Zhang, Baoyu
AU  - Zhang B
FAU - Wang, Guangce
AU  - Wang G
FAU - Lu, Douding
AU  - Lu D
FAU - Xu, Zhong
AU  - Xu Z
FAU - Yan, Peishen
AU  - Yan P
LA  - eng
SI  - GENBANK/AY465116
SI  - GENBANK/DQ336054
SI  - GENBANK/DQ336340
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20111014
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (DNA Probes)
RN  - 0 (Peptide Nucleic Acids)
SB  - IM
MH  - Cell Count
MH  - DNA Probes
MH  - Dinoflagellida/cytology/*genetics/*isolation & purification
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Molecular Sequence Data
MH  - Peptide Nucleic Acids/*metabolism
MH  - Staining and Labeling
PMC - PMC3194811
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2011/10/25 06:00
MHDA- 2012/02/18 06:00
PMCR- 2011/10/14
CRDT- 2011/10/25 06:00
PHST- 2011/04/14 00:00 [received]
PHST- 2011/09/06 00:00 [accepted]
PHST- 2011/10/25 06:00 [entrez]
PHST- 2011/10/25 06:00 [pubmed]
PHST- 2012/02/18 06:00 [medline]
PHST- 2011/10/14 00:00 [pmc-release]
AID - PONE-D-11-06810 [pii]
AID - 10.1371/journal.pone.0025527 [doi]
PST - ppublish
SO  - PLoS One. 2011;6(10):e25527. doi: 10.1371/journal.pone.0025527. Epub 2011 Oct 14.