ABSTRACT
MAIN CONCLUSION: CRISPR/Cas9-mediated Phospholipase C2 knock-out tomato plants are more resistant to Botrytis cinerea than wild-type plants, with less ROS and an increase and reduction of (JA) and (SA)-response marker genes, respectively. Genome-editing technologies allow non-transgenic site-specific mutagenesis of crops, offering a viable alternative to traditional breeding methods. In this study we used CRISPR/Cas9 to inactivate the tomato Phospholipase C2 gene (SlPLC2). Plant PLC activation is one of the earliest responses triggered by different pathogens regulating plant responses that, depending on the plant-pathogen interaction, result in plant resistance or susceptibility. The tomato (Solanum lycopersicum) PLC gene family has six members, named from SlPLC1 to SlPLC6. We previously showed that SlPLC2 transcript levels increased upon xylanase infiltration (fungal elicitor) and that SlPLC2 participates in plant susceptibility to Botrytis cinerea. An efficient strategy to control diseases caused by pathogens is to disable susceptibility genes that facilitate infection. We obtained tomato SlPLC2-knock-out lines with decreased ROS production upon B. cinerea challenge. Since this fungus requires ROS-induced cell death to proliferate, SlPLC2-knock-out plants showed an enhanced resistance with smaller necrotic areas and reduced pathogen proliferation. Thus, we obtained SlPLC2 loss-of-function tomato lines more resistant to B. cinerea by means of CRISPR/Cas9 genome editing technology.
Subject(s)
Solanum lycopersicum , Type C Phospholipases , Type C Phospholipases/metabolism , Solanum lycopersicum/genetics , CRISPR-Cas Systems , Plant Proteins/genetics , Plant Proteins/metabolism , Reactive Oxygen Species/metabolism , Oxylipins/metabolism , Plant Breeding , Botrytis/metabolism , Phospholipases/genetics , Phospholipases/metabolism , Plant Diseases/genetics , Plant Diseases/microbiology , Disease Resistance/genetics , Gene Expression Regulation, PlantABSTRACT
Introducción. La frecuencia de complicaciones postquirúrgicas de la colecistectomía realizada en la noche es un tema de controversia, siendo que se ha reportado una frecuencia mayor durante el horario nocturno. El objetivo de este estudio fue analizar la presentación de colecistectomía difícil dependiendo de la hora en que se realizó la cirugía, además de otras complicaciones, estancia intrahospitalaria postquirúrgica, reingreso a 30 días y reintervención. Métodos. Se realizó un estudio retrospectivo, observacional, analítico y transversal, comparando la presentación de colecistectomía difícil y su frecuencia en horario diurno (8:00 am a 7:59 pm) y nocturno (8:00 pm a 7:59 am), además de seroma, absceso, hematoma, fuga biliar, biloma, estancia intrahospitalaria postquirúrgica, reingreso a 30 días y reintervención. Resultados. Se incluyeron en el estudio 228 pacientes, 117 operados durante el día (52 %) y 111 durante la noche (48 %). La colecistectomía difícil se presentó 26 % vs 34 % de los casos intervenidos en el día y la noche, respectivamente. La complicación más frecuente fue seroma (14 %). La estancia hospitalaria media fue de 2,7 días en cirugías diurnas y de 2,5 en cirugías nocturnas; hubo 3 % de reintervenciones y 6 %, respectivamente. También hubo 2 % de reingresos a los 30 días entre los pacientes operados en el día y 3 % entre los operados en la noche. Conclusiones. La frecuencia de colecistectomía difícil y las complicaciones, la estancia intrahospitalaria postquirúrgica, el reingreso a 30 días y la necesidad de reintervención, no tuvieron diferencias significativas respecto al horario de la cirugía.
Introduction. The frequency of post-surgical complications of cholecystectomy performed overnight is a matter of controversy, and a higher rate has been reported during the night shift. The objective of this study was to analyze the presentation of difficult cholecystectomy depending on the time the surgery was performed, in addition to other complications, postoperative hospital stay, 30-day readmission, and reintervention. Methods. A retrospective, observational, analytical and cross-sectional study was carried out, comparing the presentation of difficult cholecystectomy and its frequency during daytime (8:00 am to 7:59 pm) and at night (8:00 pm to 7:59 am), in addition of seroma, abscess, bile leak, biloma, hematoma, post-surgical hospital stay, 30-day readmission, and reintervention.Results. A total of 228 patients were included in the study, 117 patients operated during the day (52%), and 111 at night (48%). Difficult cholecystectomy occurred in 26% vs. 34% of the cases operated on during the day and at night, respectively. The most frequent complication was seroma (14%). The mean hospital stay was 2.7 days in day surgeries and 2.5 in night surgeries; there were also 2% readmission at 30 days among patients operated during the day and 3% among those operated on at night. Conclusions. The frequency of difficult cholecystectomy and complications, postoperative hospital stay, 30-day readmission, and the need of reintervention, did not have significant differences with respect to the time of surgery.
