PMID- 31647694
OWN - NLM
STAT- MEDLINE
DCOM- 20200128
LR  - 20200128
IS  - 0191-2917 (Print)
IS  - 0191-2917 (Linking)
VI  - 104
IP  - 1
DP  - 2020 Jan
TI  - Probe-Based Multiplex Real-Time PCR as a Diagnostic Tool to Distinguish Distinct 
      Fungal Symbionts Associated With Euwallacea kuroshio and Euwallacea 
      whitfordiodendrus in California.
PG  - 227-238
LID - 10.1094/PDIS-01-19-0201-RE [doi]
AB  - California has been invaded by two distinct Euwallacea spp. that vector unique 
      plant pathogenic symbiotic fungi on multiple hosts and cause Fusarium dieback. 
      The objective of this study was to develop multiplex real-time quantitative PCR 
      assays using hydrolysis probes targeting the beta-tubulin gene to detect, 
      distinguish, and quantify fungi associated with the polyphagous shot hole borer 
      (PSHB; Euwallacea whitfordiodendrus, Fusarium euwallaceae, Graphium euwallaceae, 
      and Paracremonium pembeum) as well as the Kuroshio shot hole borer (KSHB; 
      Euwallacea kuroshio, Fusarium kuroshium, and Graphium kuroshium) from various 
      sample types. Absolute quantification reaction efficiencies ranged from 88.2 to 
      104.3%, with a coefficient of determination >0.992 and a limit of detection of 
      100 copies microl(-1) for all targets across both assays. Qualitative detection using 
      the real-time assays on artificially inoculated avocado shoot extracts showed 
      more sensitivity compared with conventional fungal isolation from wood. All 
      symbiotic fungi, except P. pembeum, from PSHB and KSHB female heads were 
      detectable and quantified. Field samples from symptomatic Platanus racemosa, 
      Populus spp., and Salix spp. across 17 of 26 city parks were positively 
      identified as PSHB and KSHB through detection of their symbiotic fungi, and both 
      were found occurring together on five trees from three different park locations. 
      The molecular assays presented here can be utilized to accurately identify fungi 
      associated with these invasive pests in California.
FAU - Carrillo, Joseph D
AU  - Carrillo JD
AUID- ORCID: 0000-0003-1790-6776
AD  - Department of Microbiology and Plant Pathology, University of California, 
      Riverside, CA 92521.
FAU - Mayorquin, Joey S
AU  - Mayorquin JS
AD  - Department of Microbiology and Plant Pathology, University of California, 
      Riverside, CA 92521.
FAU - Stajich, Jason E
AU  - Stajich JE
AD  - Department of Microbiology and Plant Pathology, University of California, 
      Riverside, CA 92521.
FAU - Eskalen, Akif
AU  - Eskalen A
AUID- ORCID: 0000-0002-8829-7413
AD  - Department of Plant Pathology, University of California, Davis, CA 95616.
LA  - eng
PT  - Journal Article
DEP - 20191023
PL  - United States
TA  - Plant Dis
JT  - Plant disease
JID - 9882809
SB  - IM
MH  - Animals
MH  - *Ascomycota/classification/genetics
MH  - California
MH  - Female
MH  - *Fusarium/classification/genetics
MH  - Introduced Species
MH  - Limit of Detection
MH  - *Real-Time Polymerase Chain Reaction
MH  - *Weevils/microbiology
OTO - NOTNLM
OT  - ambrosia beetle
OT  - pathogen detection
EDAT- 2019/10/28 06:00
MHDA- 2020/01/29 06:00
CRDT- 2019/10/25 06:00
PHST- 2019/10/28 06:00 [pubmed]
PHST- 2020/01/29 06:00 [medline]
PHST- 2019/10/25 06:00 [entrez]
AID - 10.1094/PDIS-01-19-0201-RE [doi]
PST - ppublish
SO  - Plant Dis. 2020 Jan;104(1):227-238. doi: 10.1094/PDIS-01-19-0201-RE. Epub 2019 
      Oct 23.