RT journal article
T1 Improved Protoplast Production Protocol for Fungal Transformations Mediated by CRISPR/Cas9 in Botrytis cinerea Non-Sporulating Isolates
A1 Coca Ruiz, Víctor
A1 Cabrera Gómez, Nuria
A1 González Collado, Isidro
A1 Aleu Casatejada, Josefina
A2 Química Orgánica
K1 Botrytis cinerea
K1 protoplast
K1 CRISPR/Cas9
K1 mycelium
K1 transformation
AB Botrytis cinerea is a necrotrophic fungus that causes considerable economic losses in commercial crops. Fungi of the genus Botrytis exhibit great morphological and genetic variability, rangingfrom non-sporogenic and non-infective isolates to highly virulent sporogenic ones. There is growinginterest in the different isolates in terms of their methodological applications aimed at gaining adeeper understanding of the biology of these fungal species for more efficient control of the infections they cause. This article describes an improvement in the protoplast production protocol fromnon-sporogenic isolates, resulting in viable protoplasts with regenerating capacity. The methodimprovements consist of a two-day incubation period with mycelium plugs and orbital shaking.Special mention is made of our preference for the VinoTaste Pro enzyme in the KC buffer as a replacement for Glucanex, as it enhances the efficacy of protoplast isolation in B459 and B371 isolates. Themethodology described here has proven to be very useful for biotechnological applications such asgenetic transformations mediated by the CRISPR/Cas9 tool.
PB MDPI
SN 2223-7747
YR 2024
FD 2024
LK http://hdl.handle.net/10498/35598
UL http://hdl.handle.net/10498/35598
LA eng
DS Repositorio Institucional de la Universidad de Cádiz
RD 05-nov-2026