Quantitative real-time PCR and high-resolution melting (HRM) analysis for strain-specific monitoring of fluorescent pseudomonads used as biocontrol agents against soil-borne pathogens of food crops

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

“…Monitoring the persistence and traceability of L. plantarum PM411 in plants is a key task for understanding its behavior in the crop environment and to improve formulations and delivery strategies for biological control of plant diseases. The fate and persistence of target microorganisms in the environment have been traditionally assessed with a variety of culture-dependent and independent methods ( 6 – 8 , 10 , 11 ). Among the various approaches used, PCR-based methods have been the most popular because they are very sensitive and specific to properly identify the inoculated strains, distinguishing them from the resident population.…”

“…To determine the optimal concentration of PEMAX (50, 75, and 100 μM), an appropriate volume of PEMAX stock solution (25, 37.5, or 50 μl) was added into 1 ml of viable or dead PM411 cell suspension, both adjusted to 1 × 10 6 CFU · ml −1 in apple flower washings. Dead cells were obtained by heating a cell suspension at 100°C for 15 min ( 10 , 26 ) (ThermoMixer F1.5; Eppendorf). The loss of cell viability was checked by plating on MRS agar, followed by incubation for 48 h at 30°C.…”

“…It is necessary to develop strain-specific quantitative methods for monitoring strain PM411 in the environment in order to study its ecological fitness and to optimize formulations and application strategies in the phyllosphere of plants ( 10 – 12 ). Metagenomic studies in apple trees have revealed the presence of Lactobacillus spp.…”

Application of high‐resolution melting analysis for authenticity testing of valuable Dendrobium commercial products

“…The advantages of HRM include a closed‐tube environment that greatly reduces the possibility of false positive results due to amplification product carry‐over contamination, the co‐dominant property to identify and quantify admixture samples, and time‐saving that makes a large‐scale genotyping project efficient in comparison to previously described methods . Due to the advantages of HRM analysis, it has already been used for the rapid and accurate detection of botanical origin, species composition, or pathogenic microorganisms in foodstuffs, herbal medicines and clinical specimens …”

“…21,22 Due to the advantages of HRM wileyonlinelibrary.com/jsfa analysis, it has already been used for the rapid and accurate detection of botanical origin, species composition, or pathogenic microorganisms in foodstuffs, herbal medicines and clinical specimens. [23][24][25] Herein, we report a rapid and reliable semi-quantitative HRM analysis method for identifying the botanical origin of three commonly cultivated Dendrobium species and their functional food products in terminal markets. The method was proven capable not only of distinguishing among three cultivated Dendrobium species but of detecting as low as 1% D. moniliforme adulterants in D. huoshanense or D. officinale products.…”