PMID- 30386410
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20230928
IS  - 1746-4811 (Print)
IS  - 1746-4811 (Electronic)
IS  - 1746-4811 (Linking)
VI  - 14
DP  - 2018
TI  - Visualization of the impatiens downy mildew pathogen using fluorescence in situ 
      hybridization (FISH).
PG  - 92
LID - 10.1186/s13007-018-0362-z [doi]
LID - 92
AB  - BACKGROUND: Plasmopara obducens is the biotrophic oomycete responsible for 
      impatiens downy mildew, a destructive disease of Impatiens that causes high crop 
      loss. Currently, there are no available methods for the microscopic detection of 
      P. obducens from leaves of impatiens, which may be contributing to the spread of 
      the disease. Fluorescence in situ hybridization (FISH) is a sensitive and robust 
      method that uses sequence-specific, fluorescence-labeled oligonucleotide probes 
      to detect target organisms from the environment. To study this important 
      pathogen, we developed and standardized a FISH technique for the visualization of 
      P. obducens from Impatiens walleriana tissues using a species-specific 24-mer 
      oligonucleotide probe designed to target a region of the rRNA internal 
      transcribed spacer 2 (ITS2). RESULTS: Since P. obducens cannot be propagated in 
      vitro, we developed a custom E. coli expression vector that transcribes the P. 
      obducens rRNA-ITS target sequence (clone-FISH) for use as a control and to 
      optimize hybridization conditions. The FISH assay could detect P. obducens 
      sporangiophores, sporangia and oospores, and hyphae from naturally infected I. 
      walleriana leaves and stems. Cross-reactivity was not observed from plant tissue, 
      and the assay did not react when applied to E. coli with self-ligated plasmids 
      and non-target oomycete species. CONCLUSIONS: This FISH protocol may provide a 
      valuable tool for the study of this disease and could potentially be used to 
      improve early monitoring of P. obducens, substantially reducing the persistence 
      and spread of this destructive plant pathogen.
FAU - Salgado-Salazar, Catalina
AU  - Salgado-Salazar C
AD  - 1Agriculture Research Service (ARS), Mycology and Nematology Genetic Diversity 
      and Biology Laboratory, U.S. Department of Agriculture, 10300 Baltimore Avenue, 
      Beltsville, MD 20705 USA. ISNI: 0000 0004 0478 6311. GRID: grid.417548.b
AD  - 2ARS Research Participation Program, Oak Ridge Institute for Science and 
      Education, MC-100-44, P.O. Box 117, Oak Ridge, TN 37831 USA. ISNI: 0000 0001 1013 
      9784. GRID: grid.410547.3
FAU - Bauchan, Gary R
AU  - Bauchan GR
AD  - 3Agriculture Research Service, Electron and Confocal Microscopy Unit, U.S. 
      Department of Agriculture, 10300 Baltimore Avenue, Beltsville, MD 20705 USA. 
      ISNI: 0000 0004 0478 6311. GRID: grid.417548.b
FAU - Wallace, Emma C
AU  - Wallace EC
AD  - 1Agriculture Research Service (ARS), Mycology and Nematology Genetic Diversity 
      and Biology Laboratory, U.S. Department of Agriculture, 10300 Baltimore Avenue, 
      Beltsville, MD 20705 USA. ISNI: 0000 0004 0478 6311. GRID: grid.417548.b
AD  - 2ARS Research Participation Program, Oak Ridge Institute for Science and 
      Education, MC-100-44, P.O. Box 117, Oak Ridge, TN 37831 USA. ISNI: 0000 0001 1013 
      9784. GRID: grid.410547.3
AD  - 4Present Address: Department of Plant Pathology and Environmental Microbiology, 
      The Pennsylvania State University, 120 Buckhout Lab, University Park, PA 16802 
      USA. ISNI: 0000 0001 2097 4281. GRID: grid.29857.31
FAU - Crouch, Jo Anne
AU  - Crouch JA
AD  - 1Agriculture Research Service (ARS), Mycology and Nematology Genetic Diversity 
      and Biology Laboratory, U.S. Department of Agriculture, 10300 Baltimore Avenue, 
      Beltsville, MD 20705 USA. ISNI: 0000 0004 0478 6311. GRID: grid.417548.b
LA  - eng
PT  - Journal Article
DEP - 20181024
PL  - England
TA  - Plant Methods
JT  - Plant methods
JID - 101245798
PMC - PMC6199785
OTO - NOTNLM
OT  - Downy mildew
OT  - Natural environment
OT  - Oomycota
OT  - Oospores
OT  - Peronosporales
EDAT- 2018/11/06 06:00
MHDA- 2018/11/06 06:01
PMCR- 2018/10/24
CRDT- 2018/11/03 06:00
PHST- 2018/08/22 00:00 [received]
PHST- 2018/10/19 00:00 [accepted]
PHST- 2018/11/03 06:00 [entrez]
PHST- 2018/11/06 06:00 [pubmed]
PHST- 2018/11/06 06:01 [medline]
PHST- 2018/10/24 00:00 [pmc-release]
AID - 362 [pii]
AID - 10.1186/s13007-018-0362-z [doi]
PST - epublish
SO  - Plant Methods. 2018 Oct 24;14:92. doi: 10.1186/s13007-018-0362-z. eCollection 
      2018.