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Detection of Latent Sphaeropsis sapinea Infections in Austrian Pine Tissues Using Nested-Polymerase Chain Reaction
Phytopathology
|October 24, 2008
Summary
Latent Sphaeropsis sapinea infections in pine shoots are common. A new nested-polymerase chain reaction (PCR) method efficiently detects these latent Sphaeropsis tip blight infections in symptomless pine.
Area of Science:
- Plant Pathology
- Mycology
- Molecular Biology
Background:
- Sphaeropsis sapinea causes Sphaeropsis tip blight in pines.
- Latent infections by S. sapinea are frequently found in symptomless pine shoots, but their role in disease development is unclear.
Purpose of the Study:
- To develop a minimally destructive sampling technique and a nested-polymerase chain reaction (PCR) protocol to detect latent S. sapinea in symptomless pine shoots.
Main Methods:
- Developed specific primers for S. sapinea DNA, avoiding amplification from other common endophytic fungi.
- Validated the nested-PCR protocol's sensitivity, detecting as little as 0.93 pg of S. sapinea DNA.
- Compared nested-PCR detection efficiency with fungal isolation methods.
Main Results:
- The nested-PCR protocol demonstrated high specificity for S. sapinea, with no amplification from 13 other common endophytic fungi.
- The method successfully detected latent S. sapinea in both bud and bark samples.
- Nested-PCR detection efficiency was statistically equivalent to traditional fungal isolation.
Conclusions:
- The developed nested-PCR protocol provides a rapid and efficient means to identify latent S. sapinea infections in symptomless pine shoots.
- This method will facilitate future research into the latency phenomenon of S. sapinea and its role in tip blight disease.
