Molecular Biology platform of Coniphy
CONIPHY Skills
on phytopathogen’s detection
CONIPHY has developed an expertise in phytopathogenic fungi and more particularly on the study of the phenomena of resistance to fungicides and the detection of certain pathogens
Resistance of phytopathogens
The laboratory works of the main pathogens responsible for diseases of the vineyard (downy mildew, powdery mildew and gray rot), cereals (Septoria leaf blotch on Wheat, Net Blotch and Rhychosporium on Barley), rapeseed (Sclerotinia) and potatoes (Potato Late blight and Alternarioses). CONIPHY carries out in-vitro (agar medium in Petri dish, liquid medium in microplate) and in-vivo (leaf discs) tests in order to identify resistant phenotypes to different modes of action and on the other hand to characterize the sensitivity of the POPULATIONS (study of a very large number of individuals for each population).
In addition and for several years, CONIPHY has developed new tools and acquired expertise recognized by the scientific community (publications) on molecular biology methods (qPCR, pyrosequencing, NGS). These tools are useful to determine the mechanisms of resistance to various modes of action (respiratory inhibitors, sterol biosynthesis inhibitors, etc.). These methods make it possible to genetically characterize strains and quantify the frequencies of resistant individuals in natural populations.
What are the pathogens for which CONIPHY are studying the different modes of action?
Downy mildew (Plasmopara viticola)
1 – Test on leaf discs or test «in vivo» is a qualitative Test (presence or absence of resistant phenotypes).
From the symptoms present in a plot sample, a population is reconstituted. It is used to infest leaf discs in the presence of an active molecule. After incubation, a visual scoring of sporulation (in comparison with the control) is used to calculate the effectiveness of each concentration of active ingredient tested.
2 – Test microplates or test « in vitro » is a Quantitative Test (percentage of each phenotype in a population)
From a reconstituted population picking from the symptoms present in a plot sample, a study based on the observation of the number of zoospores released or germinated, in contact with an active ingredient, is carried out to quantify the frequency of each phenotype in a population
3 – Molecular test is a quantitative test (percentage of genetic alleles in a population)
From the DNA extract of a population, a screening for genetic markers specific to the resistance sought is carried out by qPCR or by pyrosequencing.
Powdery mildew (Erysiphe necator)
1 – Test on leaf discs or test «in vivo» is a qualitative Test (presence or absence of resistant phenotypes)
From the symptoms present in a plot sample, a population is reconstituted. It is used to infest leaf discs in the presence of an active molecule. After incubation, a binocular magnifying glass (in comparison with the control) observation is performed to calculate the effectiveness of each concentration of active ingredient tested.
2 – Adhesive tape test «in vivo» is a Quantitative Test (percentage of each phenotype in a population) to design the structure of each population with respect to the active ingredient).
From leaf discs, containing an active material and infested with a population from symptoms present in a sample, an imprint of the conidia and mycelial filaments is made with an adhesive tape. Observation under a microscope makes it possible to calculate the percentage of spores that are sensitive and tolerant to the dose of active substance studied.
3 – Molecular test is a quantitative test (percentage of genetic alleles in a population)
From the DNA extract of a population, a screening for genetic markers specific to the resistance sought is carried out by qPCR or by pyrosequencing
Gray mold (Botrytis cinerea)
1 – Test microplates or test « in vitro » is a Quantitative Test (percentage of each phenotype in a population)
From a reconstituted population from the symptoms present in a plot sample, a study based on the germination and the length of the germ tube of each spore is carried out using an in-house image software, Fungi Mesure. This tool helps to quantify the frequency of each phenotype in a population.
2 – Molecular test is a quantitative test (percentage of genetic alleles in a population)
From the DNA extract of a population, a screening for genetic markers specific to the resistance sought is carried out by qPCR or by pyrosequencing.
Potato Late blight (Phytophtora infestans)
1 – Test on leaf discs or test «in vivo» is a qualitative Test (presence or absence of resistant phenotypes).
From the symptoms present in a plot sample, a population is reconstituted. It is used to infest leaf discs in the presence of an active molecule. After incubation, a visual scoring of sporulation (in comparison with the control) is used to calculate the effectiveness of each concentration of active ingredient tested.
Leaf blotch on wheat (Zymoseptoria tritici)
1 – Test microplates or test « in vitro » is a Quantitative Test (percentage of each phenotype in a population)
From a reconstituted population from the symptoms present in a plot sample, a study based on the germination and the length of the germ tube of each spore is carried out using an in-house image software, Fungi Mesure. This tool helps to quantify the frequency of each phenotype in a population.
Net blotch of barley (Helminthosporium teres)
1 – Test microplate or test « in vitro » is a Quantitative Test (percentage of each phenotype in a population)
A reconstituted population originating from the reactivation of symptoms present in the sample from a plot is brought into contact with an active molecule. An observation of the germination and the length of the germ tube of each spore is carried out using a binocular magnifying glass. It with this tool, it is possible to quantify the frequency of each phenotype in a population.
2 – Molecular test is a quantitative test (percentage of genetic alleles in a population)
From the DNA extract of a population, a screening for genetic markers specific to the resistance sought is carried out by qPCR or by pyrosequencing.
Rhynchosporium on barley (Rhynchosporium commune)
1 – Test on Petri dish or test « in vitro » is a Quantitative Test (percentage of each phenotype in a population))
A reconstituted population from lesions present in the sample from a plot is brought into contact with an active molecule in an agar dish. After incubation, a study based on the germination and germ tube length of each spore is performed using in-house image software, Fungi Mesure. It allows the quantification of the frequency of each phenotype in a population.
Detection of phytopathogens
Today, the detection of phytopathogenic elements is also a matter of molecular biology. It is possible with these reliable and robust methods not only to characterize the presence of these microorganisms but also to detect them when they can’t be detected with naked eye. An asset for adapting control treatments and validating field observations.
Detection of powdery mildew on vineyards
In partnership with Bayer Company, Coniphy has been working since 2014 on the development of a kit for the rapid detection of powdery mildew on vine leaves.
The objective of this method is to be able to detect the presence of the disease before the onset of symptoms and therefore to optimize ways of control.
The method detects the presence of contamination from two powdery mildew spores per cm2. This sensitivity could be achieved through the use of qPCR.
This technique detects the presence of powdery mildew on leaves 10 to 20 days before symptoms apparition.
After harvesting 30 leaves and shipping to Coniphy laboratory, the answer is available 48 hours later. This data allows the best possible use of preventive treatments against powdery mildew
Detection of flavescence dorée and black wood in grapevines
Since 2020 Coniphy is marketing the detection of two phytoplasma: Candidatus phytoplasma vitis responsible for Flavescence dorée and Candidatus phytoplasma solani responsible for Black wood.
Coniphy uses the triplex real-time PCR method, derived from the official MOA006 version 2a method, which allows you to have the answer within 5 days.
Coniphy is not a reference laboratory and does not have accreditation for the detection of these two phytoplasma. On the other hand, our laboratories have the equipment and skills for the application of detection methods in molecular biology.
Double-blind studies have officially confirmed the results obtained for our customers in 2020.