Periodic Reporting for period 1 - NECROFUNGI (The role of necrotrophic effectors in the ability of Botrytis and Monilinia species to infect host plants)
Reporting period: 2018-03-01 to 2020-02-29
Genome sequence and annotation of the stone fruit pathogen M. fructicola using PacBio technology. The annotation was manually curated in order to discard pseudogenes and repetitive regions. Moreover, we also obtained the group of proteins that encompass the secretome and the effectorome.
Development of a reproducible infection assay for M. fructicola in stone fruit leaves to avoid heterogenous results due to fruit ripening.
Discovery of two M. fructicola effector genes that induce cell death in host and non-host plants using different methodologies.
Discovery of three B. cinerea effectors that are able to suppress the plant cell death induced by Avr4 protein. These results support the hypothesis that B. cinerea has a brief biotrophic phase prior to switching to the induction of necrosis but we still do not know the implications in the infection process.
The use of CRISPR-Cas9 technique to generate transformants in B. cinerea and M. fructicola.
The use of TRV virus to overexpress proteins in plants that possess difficulties to be infiltrated as a high throughput methodology to screen cell death inducing effectors.
The first main contribution to the state of the art was to sequence the genome of M. fructicola and to describe the transcriptome from M. fructicola-infected stone fruit leaves. Using both data we found the group of proteins that are part of the secretome and effectorome. To our knowledge there are no studies about this for M. fructicola neither from other postharvest pathogens (except B. cinerea). This data offers new research opportunities to explore the activity of those effectors in virulence of M. fructicola. Moreover, we found 3 effector proteins that induce cell death in both host and non-host plants. These effectors can influence M. fructicola virulence and can be used to screen stone fruit germplasm to find resistant/susceptible varieties in early stages of development. These results are very innovative and strongly contribute to the state of the art in the study of virulence factors in M. fructicola and can be extrapolated to the use of other postharvest diseases.
We demonstrated that B. cinerea has a short early biotrophic phase before switching to necrosis. We have 3 effectors that are able to suppress plant cell death during the early infection (8-12 hpi). There are several studies about effectors that induce cell death in B. cinerea and its relation with virulence however, this is the first evidence that a necrotrophic pathogen can have a biotrophic lifestyle during the early phases of the infection. This theory will reconsider the lifestyle of B. cinerea and will open new research approaches.