Introduction to Soil Microbiology

Antioxidant and antibacterial lignin nanoparticles in polyvinyl alcohol/chitosan films for active packaging

SUMMARY Pseudomonas syringae pv. actinidiae is the causal agent of bacterial canker of green-fleshed kiwifruit (Actinidia deliciosa) and yellow-fleshed kiwifruit (A. chinensis). A recent, sudden, re-emerging wave of this disease has occurred, almost contemporaneously, in all of the main areas of kiwifruit production in the world,suggesting that it can be considered as a pandemic disease. Recent in-depth genetic studies performed on P. syringae pv. actinidiaestrains have revealed that this pathovar is composed of four genetically different populations which, to different extents, can infect crops of the genus Actinidia worldwide. Genome comparisons of these strains have revealed that this pathovar can gain and lose the phaseolotoxin gene cluster, as well as mobile genetic elements, such as plasmids and putative prophages, and that it can modify the repertoire of the effector gene arrays. In addition, the strains currently causing worldwide severe economic losses display an extensive set of genes related to the ecological fitness of the bacterium in planta, such as copper and antibiotic resistance genes, multiple siderophore genes and genes involved in the degradation of lignin derivatives and other phenolics.This pathogen can therefore easily colonize hosts throughout the year. Taxonomy: Bacteria; Proteobacteria, gamma subdivision; Order Pseudomonadales; Family Pseudomonadaceae; Genus Pseudomonas; Pseudomonas syringae species complex, genomospecies 8; Pathovar actinidiae.

/pdf/pseudomonas-syringae-pv-actinidiae-a-re-emerging-multi-1mxuh5m7p1.pdf

Pseudomonas syringae pv. actinidiae: a re‐emerging, multi‐faceted, pandemic pathogen

Use of endophytes as biocontrol agents

Xanthomonas is a well-studied genus of bacterial plant pathogens whose members cause a variety of diseases in economically important crops worldwide. Genomic and functional studies of these phytopathogens have provided significant understanding of microbial-host interactions, bacterial virulence and host adaptation mechanisms including microbial ecology and epidemiology. In addition, several strains of Xanthomonas are important as producers of the extracellular polysaccharide, xanthan, used in the food and pharmaceutical industries. This polymer has also been implicated in several phases of the bacterial disease cycle. In this review, we summarise the current knowledge on the infection strategies and regulatory networks controlling virulence and adaptation mechanisms from Xanthomonas species and discuss the novel opportunities that this body of work has provided for disease control and plant health.

Mechanistic insights into host adaptation, virulence and epidemiology of the phytopathogen Xanthomonas.

There has been many recent studies on the use of microbial antagonists to control diseases incited by soilborne and airborne plant pathogenic bacteria and fungi, in an attempt to replace existing methods of chemical control and avoid extensive use of fungicides, which often lead to resistance in plant pathogens. In agriculture, plant growth-promoting and biocontrol microorganisms have emerged as safe alternatives to chemical pesticides. Streptomyces spp. and their metabolites may have great potential as excellent agents for controlling various fungal and bacterial phytopathogens. Streptomycetes belong to the rhizosoil microbial communities and are efficient colonizers of plant tissues, from roots to the aerial parts. They are active producers of antibiotics and volatile organic compounds, both in soil and in planta, and this feature is helpful for identifying active antagonists of plant pathogens and can be used in several cropping systems as biocontrol agents. Additionally, their ability to promote plant growth has been demonstrated in a number of crops, thus inspiring the wide application of streptomycetes as biofertilizers to increase plant productivity. The present review highlights Streptomyces spp.-mediated functional traits, such as enhancement of plant growth and biocontrol of phytopathogens.

https://www.mdpi.com/1422-0067/19/4/952/pdf

Plant Growth Promoting and Biocontrol Activity of Streptomyces spp. as Endophytes.

Presence of Pseudomonas syringae pv. actinidiae (Psa), the causal agent of bacterial canker of kiwifruit, in pollen samples collected from infected and non infected orchards in Italy and in New Zealand was determined by polymerase chain reaction (PCR) and by direct bacterial isolation. Psa was isolated only from pollen samples collected in Italy, including pollen collected from two uninfected orchards, which the following year showed signs of infection. Psa was also detected in pollen collected from male and female vines in an Italian infected orchard. Pollen samples from Italy, but not from New Zealand, were collected with a vacuum device. Psa could not be isolated from any of the 25 New Zealand pollen samples analysed. This is the first report of Psa being associated with pollen. There is currently no evidence that artificial pollination leads to increased infection or that pollen has been responsible for the introduction of Psa in a previously Psa-free area.

/pdf/detection-of-pseudomonas-syringae-pv-actinidiae-in-kiwifruit-4lcxgr2vip.pdf

Detection of Pseudomonas syringae pv. actinidiae in kiwifruit pollen samples.

The role of pollen in disseminating Pseudomonas syringae pv. actinidiae ( Psa ) in kiwifruit orchards was investigated and the survival of the pathogen as an epiphyte on leaves and fruits was followed, from pollination time until the pre-harvest season. Pollen with natural inoculum was obtained from an infected orchard and pollen harvested without any contamination by Psa was experimentally inoculated at approximately the same contamination level. Two pollination techniques were used in glasshouse experiments: dusting and spraying. In parallel, field trials in commercial orchards were carried out: two plots were designed in two orchards, where bacterial canker was present at low incidence. From petal fall to 3 weeks before harvesting, leaf and fruit samples were taken and analysed for the presence of Psa , using two different PCR protocols and direct isolation. Results confirmed the dissemination of Psa through pollen, especially when using the aqueous suspensions. Both in glasshouse experiments and in the orchards Psa was found as an epiphyte for several weeks after pollination. Pathogen populations on leaves were, initially, 10 to 100 times less than on fruitlets. As the summer continued, the epiphytic contamination levels of fruits decreased constantly, being no more detectable from early August, whereas Psa was present at detectable levels on leaves until early October, approx. 20 days before harvesting time. Our results confirmed the role of pollen in disseminating Psa , the long epiphytic survival of the pathogen on kiwifruit leaves and the increasing unsuit­ability of fruits to harbour detectable, epiphytic populations of the bacterium through the summer season in commercial orchards with low disease incidence, when they are reaching the final development stage. Thus, kiwifruit surfaces do not appear to be a suitable niche for a long term survival of Psa as an epiphyte and, therefore, kiwifruits should not represent a pathway for Psa dissemination and pose a negligible risk for the introduction of the pathogen into new areas.

Dissemination of Pseudomonas syringae pv. actinidiae through pollen and its epiphytic life on leaves and fruits

Seventy-seven Xanthomonas arboricola pv. juglandis isolates, originating from a small region (Romagna, Italy) within 4 years, were phenotypically typed, in order to study their population features. Assessment of phenotypes resulted in the identification of three different groups of morphotypes, in the assessment of different virulence on walnut fruitlets, and in the evidence that all isolates were able to grow on Mannitol-glutamate-yeast agar containing 50 ppm of copper sulphate. Moreover, several isolates showed to be highly copper resistant in vitro, up to 500 ppm. Forty-one isolates, selected considering year/origin of plant material and phenotypic features, were molecularly studied by rep-PCR fingerprinting using BOXA1R primer. These strains showed a clear intra-pathovar variation by the presence of eight different haplotypes. Twenty isolates, representative of different BOX profile, were studied by means of variable number of tandem repeats (VNTR) on the locus TR5b. Such analysis highlighted five different sequence types. Eight polymorphic strains on this flanking region isolated in between 2007 and 20 and one isolated in 2010 were subject to multilocus sequence analysis (MLSA) using atpD, dnaK, efP, fyuA, glnA, gyrB, and rpoD housekeeping genes. A diversity level in Italian isolates was highlighted in the same range as in reference strains from a worldwide origin. Finally, the gene cluster copLAB presence was confirmed for all isolates. In this study, the high phenotype and genotype variability inside Xanthomonas arboricola pv. juglandis, was explained by the different origin of the propagation material. Information provided in this study on an Italian Xanthomonas arboricola pv. juglandis collection allows a better understanding of the walnut bacterial blight epidemiology.

Morphological and genotypic features of Xanthomonas arboricola pv. juglandis populations from walnut groves in Romagna region, Italy

Pollen collected in a kiwifruit orchard with symptoms of bacterial canker and naturally contaminated by Pseudomonas syringae pv. actinidiae (Psa), was used to pollinate an experimental orchard, in order to confirm its role, under commercial orchard conditions, in disseminating the pathogen and, possibly, contributing to disease spread. A pollen lot, certified free from Psa, was used with the same methods as a control. Two pollination techniques were used: dusting (dry pollen) and spraying (pollen suspension in water). The orchard was monitored during 2 years from experimental pollination, with regular sampling of flowers, fruits, leaves, and vines, to check for Psa as an epiphyte or endophyte, and for bacterial canker symptoms. Psa was recovered from flowers, fruitlets and leaves during the first season, mainly in plots where contaminated pollen had been sprayed in water suspension. From early August until harvesting time (mid-October), Psa detection was possible only on leaves. No symptoms developed during the first season after pollination. No endophytic Psa was detected in pruned vines in the following winter. During the second season, detection and isolation of Psa was erratic, but direct isolation was achieved from four plots. During the second season after pollination, typical leaf symptoms were observed on a few vines, and Psa was isolated and identified. Our results suggest that Psa could be disseminated via contaminated kiwifruit pollen as a pathway for spread of bacterial canker. However, further pollination experiments are needed to establish, beyond any doubt, whether contaminated pollen may contribute to possible disease outbreaks.

Davide Giovanardi

Papers

Plant Growth Promoting and Biocontrol Activity of Streptomyces spp. as Endophytes.

Detection of Pseudomonas syringae pv. actinidiae in kiwifruit pollen samples.

Dissemination of Pseudomonas syringae pv. actinidiae through pollen and its epiphytic life on leaves and fruits

Morphological and genotypic features of Xanthomonas arboricola pv. juglandis populations from walnut groves in Romagna region, Italy

Pollen as a possible pathway for the dissemination of Pseudomonas syringae pv. actinide and bacterial canker of kiwifruit