PMID- 16584486
OWN - NLM
STAT- MEDLINE
DCOM- 20060505
LR  - 20171116
IS  - 1462-2912 (Print)
IS  - 1462-2912 (Linking)
VI  - 8
IP  - 4
DP  - 2006 Apr
TI  - In situ probing of Xylella fastidiosa in honeydew of a xylem sap-feeding insect 
      using 16S rRNA-targeted fluorescent oligonucleotides.
PG  - 747-54
AB  - Xylella fastidiosa is a plant pathogen that threatens a US$ 4.6 billion worldwide 
      wine and citrus industry. Monitoring its presence and distribution in plants and 
      vectors is crucial for designing control strategies, as well as for understanding 
      its ecological role and fate. We developed two fluorescent oligonucleotide probes 
      complementary to different regions of the 16S rRNA gene of X. fastidiosa. The 
      specificity of the newly designed probes S-S-X.fas-0067-a-A-18 and 
      S-S-X.fas-1439-a-A-18 was demonstrated using fluorescence in situ hybridization 
      (FISH) for 12 Xylella isolates, 15 closely related microorganisms and three plant 
      endophytes. These probes were used to detect and quantify X. fastidiosa in plant 
      sap (average value of 2.9 +/- 0.3 x 10(6) cells ml(-1)) from three different 
      citrus orchards. In a second experiment, cells were quantified in honeydew (2.2 
      +/- 0.2 x 10(4) cells ml(-1)) collected from the insect vector Bucephalogonia 
      xanthophis during the acquisition access period on an infected plant. The number 
      of pathogen cells retained or digested by the insect is 10,000 times greater than 
      the estimated minimum value to ensure an efficient transmission. Polymerase chain 
      reaction (PCR) amplification using specific primers with plant sap and honeydew 
      samples, followed by sequencing, confirmed the presence of the plant pathogen. 
      This is the first demonstration of FISH being used for environmental samples, 
      such as plant sap and insect honeydew, to estimate the abundance of a plant 
      pathogen during infection.
FAU - Rodrigues, Jorge L M
AU  - Rodrigues JL
AD  - Centro de Energia Nuclear na Agricultura, ESALQ, Universidade de Sao Paulo, 
      Piracicaba, SP, 13400-970, Brazil. rodrig76@msu.edu
FAU - Silva-Stenico, Maria E
AU  - Silva-Stenico ME
FAU - de Souza, Adriane N
AU  - de Souza AN
FAU - Lopes, Joao R S
AU  - Lopes JR
FAU - Tsai, Siu M
AU  - Tsai SM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Environ Microbiol
JT  - Environmental microbiology
JID - 100883692
RN  - 0 (Oligonucleotide Probes)
RN  - 0 (RNA, Bacterial)
RN  - 0 (RNA, Ribosomal, 16S)
SB  - IM
MH  - Animals
MH  - Cucumis melo/*microbiology
MH  - In Situ Hybridization, Fluorescence
MH  - Insecta/*microbiology
MH  - Oligonucleotide Probes
MH  - Phylogeny
MH  - Plant Diseases/*microbiology
MH  - RNA, Bacterial/*analysis
MH  - RNA, Ribosomal, 16S/*analysis
MH  - Xylella/*genetics/isolation & purification
EDAT- 2006/04/06 09:00
MHDA- 2006/05/06 09:00
CRDT- 2006/04/06 09:00
PHST- 2006/04/06 09:00 [pubmed]
PHST- 2006/05/06 09:00 [medline]
PHST- 2006/04/06 09:00 [entrez]
AID - EMI958 [pii]
AID - 10.1111/j.1462-2920.2005.00958.x [doi]
PST - ppublish
SO  - Environ Microbiol. 2006 Apr;8(4):747-54. doi: 10.1111/j.1462-2920.2005.00958.x.