HRM, the innovative technique for the detection of Botrytis pseudocinerea, a species recently discovered in fruit and crops

Laboratorios Diagnofruit Ltda., Is a Chilean company founded in the year 2012 by Agronomists, with the aim of developing research and provide services that respond to the needs of the country's fruit industry.

Among its clients are some of the most important exporters in the country such as Frusan, Dole and Subsole, agrochemical companies and trade associations such as the Kiwi Committee.

In addition to the diagnosis of fruit quality - tool that supports exporters in the process of segregation of orchards for extended guarding - Diagnofruit is an active developer of techniques and technologies for the detection, diagnosis of pathogens and the study of phylogenetic relationships between species , pathovars and / or genotypes.

In this last area of ​​development, Diagnofruit in cooperation with the Biomedical Research Center of the Autonomous University of Chile, generated a highly sensitive and highly efficient technique for the detection of two important species of the genus. Botrytis, as it is Botrytis cinerea and Botrytis pseudocinerea.

Why is it important to know which species of botrytis dominates within my crop?

There are some cryptic and sympatric variants of botrytis, this means that they can not be differentiated with the naked eye and that they can occupy the same ecological niche. Table grape research in the 90's years in France and more recent in Chile, characterized B. pseudocinerea as natural and intrinsically resistant to fenhexamid, isolates resistant to the molecule were detected even without having received applications of fenhexamid before.

This important characteristic conditions the design of chemical control programs based on the dominant botrytis population in the field, generating the importance of knowing the frequency of both species.

What is this new technique?

The technique is based on a variant of qPCR and is called HRM for its acronym in English (High Resolution Melting). This technique, among other characteristics, can detect minimal variations in the genome of any organism, being able to discriminate between B. cinerea and B. pseudocinerea.


What are the advantages of HRM over conventional techniques?

The conventional technique has several steps and often if the extraction of DNA is not efficient enough it generates bands difficult to visualize in a typical agarose gel.

The HRM technique does not use agarose gel, but is a direct interpretation of a small sector of the genome of the fungus, therefore, it is much more effective and efficient, reducing the analysis times in an 55% and increasing the sensitivity of detection significantly.

How the season is projected in terms of quality and conservability of fruit species such as grapes, where varieties such as Thompson S. are highly susceptible to attacks by B. cinerea and B. pseudocinerea?

Héctor García, general manager of Laboratorios Diagnofruit Ltda., Said that "Although it has been a very warm spring in the central area, above normal and with successive heat waves, which is generally positive if the risk is well managed, it has been detected that extreme heat in summer can cause problems in the maturation (precocity) accompanied by greater difficulty in the accumulation of dry matter due to thermal stress, same behavior recorded in early cherries this year".

"In terms of colonization by botrytis in flower, there are two very marked realities, in early zones where there were rains near the flower, the control was deficient, there being a high risk of infection development in harvest and post-harvest, which is why we have generated with our clients specific management plans in those units. On the other hand, the zones of later flowering presented, in general, levels of colonization by botrytis at lower frequencies, resulting in a lower risk fruit"He added.

Source: Fruit Portal


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