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Monitoring viable cells of the biological control agent Lactobacillus plantarum PM411 in aerial plant surfaces by means of a strain-specific viability quantitative PCR method
- Source :
- Applied and Environmental Microbiology, © Applied and Environmental Microbiology, 2018, vol. 84, p., Articles publicats (D-EQATA), DUGiDocs – Universitat de Girona, instname
- Publication Year :
- 2020
- Publisher :
- American Society for Microbiology, 2020.
-
Abstract
- A viability quantitative PCR (v-qPCR) assay was developed for the unambiguous detection and quantification of Lactobacillus plantarum PM411 viable cells in aerial plant surfaces. A 972-bp region of a PM411 predicted prophage with mosaic architecture enabled the identification of a PM411 strain-specific molecular marker. Three primer sets with different amplicon lengths (92, 188, and 317 bp) and one TaqMan probe were designed. All the qPCR assays showed good linearity over a 4-log range and good efficiencies but differed in sensitivity. The nucleic acid-binding dye PEMAX was used to selectively detect and enumerate viable bacteria by v-qPCR. The primer set amplifying a 188-bp DNA fragment was selected as the most suitable for v-qPCR. The performance of the method was assessed on apple blossoms, pear, strawberry, and kiwifruit leaves in potted plants under controlled environmental conditions, as well as pear and apple blossoms under field conditions, by comparing v-qPCR population estimations to those obtained by qPCR and specific plate counting on de Man-Rogosa-Sharpe (MRS)-rifampin. The population estimation did not differ significantly between methods when conditions were conducive to bacterial survival. However, under stressful conditions, differences between methods were observed due to cell death or viable-but-nonculturable state induction. While qPCR overestimated the population level, plate counting underestimated this value in comparison to v-qPCR. PM411 attained stable population levels of viable cells on the flower environment under high relative humidity. However, the unfavorable conditions on the leaf surface and the relatively dryness in the field caused an important decrease in the viable population. IMPORTANCE The v-qPCR method in combination with plate counting and qPCR is a powerful tool for studies of colonization and survival under field conditions, to improve formulations and delivery strategies of PM411, and to optimize the dose and timing of spray schedules. It is expected that PEMAX v-qPCR could also be developed for monitoring other strains on plant surfaces not only as biological control agents but also beneficial bacteria useful in the sustainable management of crop production.
- Subjects :
- 0301 basic medicine
DNA, Bacterial
030106 microbiology
Population
biological control
Biology
Real-Time Polymerase Chain Reaction
Applied Microbiology and Biotechnology
Fragaria
Pyrus
03 medical and health sciences
chemistry.chemical_compound
viability-qPCR
Plant Microbiology
Species Specificity
Molecular marker
TaqMan
Food science
education
Plants -- Diseases and pests
DNA Primers
Plant Diseases
2. Zero hunger
education.field_of_study
PEAR
Microbial Viability
Ecology
Bacterial diseases of plants
Plantes -- Malalties bacterianes
Amplicon
Plant Components, Aerial
Plantes -- Malalties i plagues
biology.organism_classification
Pests -- Control
Plagues -- Control biològic
chemistry
Biological Control Agents
Malus
Nucleic acid
Lactobacillus plantarum
Bacteria
Food Science
Biotechnology
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Applied and Environmental Microbiology, © Applied and Environmental Microbiology, 2018, vol. 84, p., Articles publicats (D-EQATA), DUGiDocs – Universitat de Girona, instname
- Accession number :
- edsair.doi.dedup.....b6bba99a9c6935149191df03183fbbb7