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Garbelotto, M.

Publications and source records attributed to Garbelotto, M..

2 recordsLinked to original sources

Assembly and analysis of the genome of Notholithocarpus densiflorus

Tanoak (Notholithocarpus densiflorus) is an evergreen tree in the Fagaceae family found in California and southern Oregon. Historically, tanoak acorns were an important food source for Native American tribes and the bark was used extensively in the leather tanning process. Long considered a disjunct relictual element of the Asian stone oaks (Lithocarpus spp.), phylogenetic analysis has determined that the tanoak is an example of convergent evolution. Tanoaks are deeply divergent from oaks (Quercus) of the Pacific Northwest and comprise a new genus with a single species. These trees are highly susceptible to sudden oak death (SOD), a plant pathogen (Phytophthora ramorum) that has caused widespread mortality of tanoaks. Here, we set out to assemble the genome and perform comparative studies among a number of individuals that demonstrated varying levels of susceptibility to SOD. First, we sequenced and de novo assembled a draft reference genome of N. densiflorus using co-barcoded library processing methods and an MGI DNBSEQ-G400 sequencer. To increase the contiguity of the final assembly, we also sequenced Oxford Nanopore (ONT) long reads to 30X coverage. To our knowledge, the draft genome reported here is one of the more contiguous and complete genomes of a tree species published until now, with a contig N50 of [~]1.2 Mb and a scaffold N50 of [~]2.1 Mb. In addition, we sequenced 11 genetically distinct individuals and mapped these onto the draft reference genome enabling the discovery of almost 25 million single nucleotide polymorphisms and [~]4.4 million small insertions and deletions. Finally, using co-barcoded data we were able to generate complete haplotype coverage of all 11 genomes.

genomics↗

Recombinase Polymerase Amplification Assay for the Field Detection of Mal Secco Disease by Plenodomus tracheiphilus

In this study, we developed a new diagnostic assay based on the recombinase polymerase amplification (RPA) technology to detect Plenodomus tracheiphilus, the anamorphic fungus responsible for the destructive vascular disease of lemon named mal secco, in infected tissues of host plants. A 142 bp RPA-compatible barcode was sought within the 544 bp Internal Transcriber Spacer (ITS) fragment identified in a previous study and its P. tracheiphilus-specificity was confirmed by BLAST in the NCBI database. This was the premise to design an RPA probe (RPA_Ptrach_Probe). The specificity and inclusivity of the RPA assay were tested on gDNA isolated from tissues of C. limon, isolates of P. tracheiphilus of various origins and axenic cultures of non-target organisms, including fungal and oomycete pathogens typically associated to citrus trees, such as Alternaria spp., Colletotrichum spp., Phyllosticta spp., Penicillium spp., Phytophthora spp. With a detection threshold of 1.0 pg of gDNA the RPA assay proved to be as sensitive as the SYBR(R) Green I Real Time-PCR test included in the diagnostic protocol for P. tracheiphilus of the European and Mediterranean Plant Protection Organization. RPA assay was even more sensitive than Real Time-PCR in tests on DNA samples obtained through a rapid extraction method. In tests, on naturally infected lemon twigs, molecular approaches were comparable to each other and performed better than conventional isolation method. Overall, results of this study demonstrate the potential of RPA for rapid, easy to handle and cost effective in-field diagnosis of mal secco.

molecular biology↗