Xylella fastidiosa is a xylem-dwelling, insect-transmitted, gamma-proteobacterium that causes diseases in many plants, including grapevine, citrus, periwinkle, almond, oleander, and coffee. X. fastidiosa has an unusually broad host range, has an extensive geographical distribution throughout the American continent, and induces diverse disease phenotypes. Previous molecular analyses indicated three distinct groups of X. fastidiosa isolates that were expected to be genetically divergent. Here we report the genome sequence of X. fastidiosa (Temecula strain), isolated from a naturally infected grapevine with Pierce's disease (PD) in a wine-grapegrowing region of California. Comparative analyses with a previously sequenced X. fastidiosa strain responsible for citrus variegated chlorosis (CVC) revealed that 98% of the PD X. fastidiosa Temecula genes are shared with the CVC X. fastidiosa strain 9a5c genes. Furthermore, the average amino acid identity of the open reading frames in the strains is 95.7%. Genomic differences are limited to phage-associated chromosomal rearrangements and deletions that also account for the strain-specific genes present in each genome. Genomic islands, one in each genome, were identified, and their presence in other X. fastidiosa strains was analyzed. We conclude that these two organisms have identical metabolic functions and are likely to use a common set of genes in plant colonization and pathogenesis, permitting convergence of functional genomic strategies.Different microorganisms are able to survive in and to colonize plant water-conductive vessels (xylem). The result of this association is either beneficial or detrimental to the plant host.Of the latter, an example is the association of Xylella fastidiosa (38) with diverse plant hosts. X. fastidiosa is a fastidious, insecttransmitted, xylem-inhabiting bacterium known to cause several economically important diseases of both monocotyledonous and dicotyledonous plants (14,17,29). These diseases include Pierce's disease (PD) of grapevine and citrus variegated chlorosis (CVC), which have rather distinct symptoms and geographical distributions.PD, caused by certain strains of X. fastidiosa, is characterized by wilted, shriveled, raisin-like fruit and scorched leaves that detach, leaving bare petioles attached to the canes (37). The bark of affected canes may lignify or mature irregularly, leaving

In plants, the ubiquitin/26S proteasome system (UPS) plays a central role in protein degradation and is involved in many steps of defence mechanisms, regardless of the types of pathogen targeted. In addition to its proteolytic activities, the UPS ribonuclease (RNase) activity, previously detected in 20S proteasome preparations from cauliflower and sunflower (Helianthus annuus), has been shown to specifically target plant viral RNAs in vitro. In this study, we show that recombinant Arabidopsis thaliana proteasomal α(5) subunit expressed in Escherichia coli harbours an RNase activity that degrades Tobacco mosaic virus (TMV, Tobamovirus)- and Lettuce mosaic virus (LMV, Potyvirus)-derived RNAs in vitro. The analysis of mutated forms of the α(5) subunit demonstrated that mutation of a glutamic acid at position 110 affects RNase activity. Furthermore, it was demonstrated, using a bimolecular fluorescence complement assay, that the multifunctional helper component proteinase (HcPro) of LMV, already known to interfere with the 20S proteasome RNase activity in vitro, can interact in vivo with the recombinant α(5) subunit. Further experiments demonstrated that, in LMV-infected lettuce cells, α(5) is partially relocalized to HcPro-containing infection-specific inclusions. Susceptibility analyses of Arabidopsis mutants, knocked out for each At-PAE gene encoding α(5) , showed that one (KO-pae1) of the two mutants exhibited a significantly increased susceptibility to LMV infection. Taken together, these results extend to A. thaliana α(5) the range of HcPro-interacting proteasomal subunits, and suggest that HcPro may modulate its associated RNase activity which may contribute to an antiviral response.

The World Health Organization characterized COVID-19 as a pandemic in March 2020, the second pandemic of the twenty-first century. Expanding virus populations, such as that of SARS-CoV-2, accumulate a number of narrowly shared polymorphisms, imposing a confounding effect on traditional clustering methods. In this context, approaches that reduce the complexity of the sequence space occupied by the SARS-CoV-2 population are necessary for robust clustering. Here, we propose subdividing the global SARS-CoV-2 population into six well-defined subtypes and 10 poorly represented genotypes named tentative subtypes by focusing on the widely shared polymorphisms in nonstructural (nsp3, nsp4, nsp6, nsp12, nsp13 and nsp14) cistrons and structural (spike and nucleocapsid) and accessory (ORF8) genes. The six subtypes and the additional genotypes showed amino acid replacements that might have phenotypic implications. Notably, three mutations (one of them in the Spike protein) were responsible for the geographical segregation of subtypes. We hypothesize that the virus subtypes detected in this study are records of the early stages of SARS-CoV-2 diversification that were randomly sampled to compose the virus populations around the world. The genetic structure determined for the SARS-CoV-2 population provides substantial guidelines for maximizing the effectiveness of trials for testing candidate vaccines or drugs.

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16The World Health Organization characterized the COVID-19 as a pandemic in March 2020, the second 17 pandemic of the 21 st century. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a 18 positive-stranded RNA betacoronavirus of the family Coronaviridae. Expanding virus populations, as 19 that of SARS-CoV-2, accumulate a number of narrowly shared polymorphisms imposing a 20 confounding effect on traditional clustering methods. In this context, approaches that reduce the 21 complexity of the sequence space occupied by the SARS-CoV-2 population are necessary for a robust 22 clustering. Here, we proposed the subdivision of the global SARS-CoV-2 population into sixteen well-23 defined subtypes by focusing on the widely shared polymorphisms in nonstructural (nsp3, nsp4, nsp6, 24 nsp12, nsp13 and nsp14) cistrons, structural (spike and nucleocapsid) and accessory (ORF8) genes. 25Six virus subtypes were predominant in the population, but all sixteen showed amino acid 26 replacements which might have phenotypic implications. We hypothesize that the virus subtypes 27 detected in this study are records of the early stages of the SARS-CoV-2 diversification that were 28 randomly sampled to compose the virus populations around the world, a typical founder effect. The 29 genetic structure determined for the SARS-CoV-2 population provides substantial guidelines for 30 maximizing the effectiveness of trials for testing the candidate vaccines or drugs. Main 32In December 2019, a local pneumonia outbreak of initially unknown etiology was detected in 33 Wuhan (Hubei, China) and quickly determined to be caused by a novel coronavirus 1 , named Severe 34 acute respiratory syndrome coronavirus 2 (SARS-CoV-2) 2 and the disease as COVID-19 3 . SARS- 35CoV-2 is classified in the family Coronaviridae, genus Betacoronavirus, which comprises enveloped, 36 positive stranded RNA viruses of vertebrates 2 . Two-thirds of SARS-CoVs genome is covered by the 37 ORF1ab, that encodes a large polypeptide which is cleaved into 16 nonstructural proteins (NSPs) 38 involved in replication-transcription in vesicles from endoplasmic reticulum (ER)-derived 39 membranes 4,5 . The last third of the virus genome encodes four essential structural proteins: spike (S), 40 envelope (E), membrane (M), nucleocapsid (N) and several accessory proteins that interfere with the 41 host innate immune response 6 . 42Populations of RNA viruses evolve rapidly due to their large population sizes, short generation 43 times, and high mutation rates, this latter being a consequence of the RNA-dependent RNA 44 polymerase (RdRP) which lacks the proofreading activity 7 . In fact, virus populations are composed of 45 a broad spectrum of closely related genetic variants resembling one or more master sequences [8][9][10] . 46 Mutation rates inferred for SARS-CoVs are considered moderate 11,12 due to the independent 47 proofreading activity 13 . However, the large SARS-CoV genomes (from 27 to 31 kb) 14 provide to them 48 the ability to explore the sequence spa...