Analysis of Draft Genome Resources of Thirty-Three Canadian Strains of Pseudomonas syringae pv. tomato isolated between 1992 and 2008 Reveals achromobactin virulence cluster that is absent in the reference strain DC3000

Pseudomonas syringae pv. tomato is the causal agent of bacterial speck disease of field and greenhouse tomato plants. Only one Canadian whole genome sequence of this economically important pathogen is publicly available in NCBI GenBank. Here, we report 33 whole genome sequences of Canadian strains of P. syringae pv. tomato isolated in Ontario, Canada, between 1992 and 2008. The genome sequences exhibited average nucleotide identity values of 98.64-98.72 % with P. syringae pv. tomato ICMP 2844PT and DC3000, validating the taxonomic standing of these Canadian strains. The genome sizes ranged from 6.20-6.39 Mbp with G+C content of 58.6% and comprised 5,889-6,166 protein-coding sequences (CDSs). The strains had pan- and core-genomes of 6808 and 4,993 gene clusters, respectively. Genome mining of the strains for virulence factors identified typical adherence genes, proteins related to antiphagocytosis, secretion system apparatuses and effectors. Also, partial or complete achromobactin biosynthetic cluster and iron transport genes were identified in all the Canadian strains but absent in P. syringae pv. tomato DC3000 or ICMP 2844 (pathotype). These new whole genome data of Canadian strains of P. syringae pv. tomato could be useful resources in understanding the evolution of this pathogen.

A Green Nanostructured Pesticide to Control Tomato Bacterial Speck Disease

Bacterial speck disease, caused by Pseudomonas syringae pv. tomato (Pst), is one of the most pervasive biological adversities in tomato cultivation, in both industrial and in table varieties. In this work synthesis, biochemical and antibacterial properties of a novel organic nanostructured pesticide composed of chitosan hydrochloride (CH) as active ingredient, cellulose nanocrystals (CNC) as nanocarriers and starch as excipient were evaluated. In order to study the possibility of delivering CH, the effects of two different types of starches, extracted from a high amylose bread wheat (high amylose starch—HA Starch) and from a control genotype (standard starch—St Starch), were investigated. Nanostructured microparticles (NMP) were obtained through the spray-drying technique, revealing a CH loading capacity proximal to 50%, with a CH release of 30% for CH-CNC-St Starch NMP and 50% for CH-CNC-HA Starch NMP after 24 h. Both NMP were able to inhibit bacterial growth in vitro when used at 1% w/v. Moreover, no negative effects on vegetative growth were recorded when NMP were foliar applied on tomato plants. Proposed nanostructured pesticides showed the capability of diminishing Pst epiphytical survival during time, decreasing disease incidence and severity (from 45% to 49%), with results comparable to one of the most used cupric salt (hydroxide), pointing out the potential use of CH-CNC-Starch NMP as a sustainable and innovative ally in Pst control strategies.

The Effect of Pesticides on the Tomato Bacterial Speck Disease Pathogen Pseudomonas Syringae pv. Tomato

A significant part of the used pesticides does not reach the target organisms and, while remaining in the agrophytocenosis, influences all living organisms in it. Having a toxic and often mutagenic effect, pesticides induce morphological and physiological changes in the cells of microorganisms and are the cause of phenotypic heterogeneity of their populations. However, the effect of pesticides on phytopathogenic bacteria as non-target microorganisms remains out of the field of view for most researchers. However, the use of pesticides can lead to expansion of the diversity of existing phytopathogens and, as a consequence, complications of identification of the pathogens, loss of resistance by plants varieties, and increased harm from diseases caused by them. This study is focused on the effect of pesticides used in tomato plantations on the causative agent of bacterial speck of this crop—Pseudomonas syringae pv. tomato. The studies were carried out using the methods of classical microbiology. The mutagenic action of pesticides was recorded, taking into account the increase of the number of streptomycin resistance mutations in bacteria in the case of pesticide action. It is established that the fungicide aluminium phosethyl is characterised by a bacteriostatic effect on P. syringae pv. tomato. Deltamethrin insecticide does not affect the growth of P. syringae pv. tomato. However, there is an increase in the frequency of streptomycin resistance mutations in both studied strains of P. syringae pv. tomato after using deltamethrin. It is shown that the frequency of occurrence of R (rough colonies) forms of P. syringae pv. tomato IZ28 and IZ46 after using deltamethrin increased by 100 times when in comparison to the frequency of spontaneous morphological dissociation, or smooth-to-rough (S-R) mutation, of these bacteria. Therefore, aluminium phosethyl is characterised by moderate bacteriostatic action against P. syringae pv. tomato. Deltamethrin does not influence the growth of the pathogen of tomato speck but increases the frequency of formation of StrR mutants and R forms of phytopathogenic bacteria.

Gmbak1 and gmfls2, two genes response to bacterials disease of soybean

Soybean bacterial disease is a worldwide disease that can cause yield and quality decline. Although the recognition of soybean bacterial diseases has been reported in detail, but for the identification of resistant and susceptible cultivars, different researchers have different methods and standards. To detect the pathogenic effect of bacterial diseases, we first separated bacterial strains from soybean field. Finally, eight strains of soybean bacterial diseases were isolated and identified. By the screening the sequence of the bacterial 16S rDNA, seven of eight strains were identified belong to Pseudomonas, another strain belong to Xanthomonas. In order to study the relevant pathogen of soybean defense response, we used the isolated pathogen strain to inoculate the resistant cultivar Hefeng35 and sensitive cultivar Suinong14. Then the genes might involve in regulate soybean resistant to bacterial disease were analyzed, including GmBAK1 and GmFLS2. FLS2 homolog gene were Gm05g25830, Gm08g08810; BAK1 homolog gene were Gm15g05730, Gm08g07930, Gm10g36280, Gm05g24790, Gm20g31320, Gm05g24770, Gm08g19270, and Gm02g08360. By the qRT-PCR analysis we detected the gene expression pattern response to pathogen, we found the expression of Gm05g24770, Gm05g24790, and Glyma05g25830 showed significant different between Suinong14 and Hefeng35. We illustrated the GmBAK1 and GmFLS2 transcription level against bacterial speck disease pathogen infection and revealed preliminary the major gene, providing a reference direction for further research.

The Ptr1 Locus of Solanum lycopersicoides Confers Resistance to Race 1 Strains of Pseudomonas syringae pv. tomato and to Ralstonia pseudosolanacearum by Recognizing the Type III Effectors AvrRpt2 and RipBN

Race 1 strains of Pseudomonas syringae pv. tomato, which cause bacterial speck disease of tomato, are becoming increasingly common and no simply inherited genetic resistance to such strains is known. We discovered that a locus in Solanum lycopersicoides, termed Pseudomonas tomato race 1 (Ptr1), confers resistance to race 1 P. syringae pv. tomato strains by detecting the activity of type III effector AvrRpt2. In Arabidopsis, AvrRpt2 degrades the RIN4 protein, thereby activating RPS2-mediated immunity. Using site-directed mutagenesis of AvrRpt2, we found that, like RPS2, activation of Ptr1 requires AvrRpt2 proteolytic activity. Ptr1 also detected the activity of AvrRpt2 homologs from diverse bacteria, including one in Ralstonia pseudosolanacearum. The genome sequence of S. lycopersicoides revealed no RPS2 homolog in the Ptr1 region. Ptr1 could play an important role in controlling bacterial speck disease and its future cloning may shed light on an example of convergent evolution for recognition of a widespread type III effector.

The Ptr1 locus of Solanum lycopersicoides confers resistance to race 1 strains of Pseudomonas syringae pv. tomato and to Ralstonia pseudosolanacearum by recognizing the type III effectors AvrRpt2/RipBN

Race 1 strains of Pseudomonas syringae pv. tomato, which causes bacterial speck disease of tomato, are becoming increasingly common and no simply-inherited genetic resistance to such strains is known. We discovered that a locus in Solanum lycopersicoides, termed Pseudomonas tomato race 1 (Ptr1), confers resistance to race 1 Pst strains by recognizing the type III effector AvrRpt2. In Arabidopsis, AvrRpt2 degrades the RIN4 protein thereby activating RPS2-mediated immunity. Ptr1 also recognized homologs of AvrRpt2 from diverse bacteria including one in Ralstonia pseudosolanacearum and this correlated with the ability of AvrRpt2 to degrade RIN4. Using site-directed mutagenesis of AvrRpt2 we found that Ptr1 and RPS2 recognize identical features of AvrRpt2. However, the genome sequence of S. lycopersicoides revealed no RPS2 homolog in the Ptr1 region. Ptr1 could play an important role in controlling bacterial speck disease and its future cloning may shed light on an example of convergent evolution for recognition of a widespread type III effector.

Pseudomonas syringae pv. tomato Strains from New York Exhibit Virulence Attributes Intermediate Between Typical Race 0 and Race 1 Strains

Bacterial speck disease, caused by Pseudomonas syringae pv. tomato, is a persistent problem for fresh-market tomato growers in New York. Race 0 strains of this pathogen express either or both of the type III effectors AvrPto or AvrPtoB, which are recognized by tomato varieties expressing the Pto resistance gene. Pto encodes a protein kinase that activates the host immune system, thereby inhibiting bacterial multiplication and preventing disease development. Race 1 P. syringae pv. tomato strains do not express these effectors and are virulent on tomato whether or not the variety expresses Pto. Very few fresh-market tomato varieties have the Pto gene. We collected six P. syringae pv. tomato strains from naturally infected tomato plants across New York in 2015 and characterized them for their virulence and for the presence of specific effectors. In experiments conducted in the greenhouse, all strains reached population sizes in Pto-expressing tomato leaves that were intermediate between typical race 0 and race 1 strains. This phenotype has not been observed previously and suggests that the strains are recognized by Pto but such recognition is compromised by another P. syringae pv. tomato factor. The strains were found to encode avrPto, which is transcribed and translated. They also express avrPtoB although, as reported for other P. syringae pv. tomato strains, protein expression for this effector was not detectable. Deletion of avrPto from a representative New York strain allowed it to reach high populations in Pto-expressing tomato varieties, without compromising its virulence on susceptible tomato plants. Collectively, our data suggest that introgression of the Pto gene into fresh-market tomato varieties could enhance protection against extant P. syringae pv. tomato strains.