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

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

Le malattie batteriche emergenti o riemergenti delle colture agrarie in Italia e nel bacino del Mediterraneo

Vengono descritte alcune malattie batteriche emergenti, o riemergenti, che potrebbero diventare un serio fattore limitante le produzioni ortofrutticole italiane e mediterranee

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

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

A TRANSCRIPTOMIC APPROACH TO UNDERSTAND INDUCED RESISTANCE TO BACTERIAL CANKER OF STONE FRUITS ELICITED THROUGH TREATMENTS WITH COPPER GLUCOHUMATES.

Bacterial canker of stone fruits, caused by Xanthomonas arboricola pv. pruni, is a recurrent disease in Italian peach and plum orchards. Several field and glasshouse trials on peach and plum were done, with the aim to effectively control the disease by using some novel molecules, such as glucohumates. The most remarkable results were obtained with copper glucohumates (with a reduction of the disease by ca. 80%). Results are very promising and suggest the possibility to implement effective control strategies, where copper compounds and novel molecules are both used in commercial orchards. In order to study and understand the effect of the biomolecules used, untreated and glucohumate-treated peach plants were subject to further molecular analyses in order identify possible genes/sequences involved in the induction of disease resistance. A transcriptomic approach was developed for detecting the transcripts present in plant tissues, after elicitation of an induced protection state. Total RNA was extracted, retro-transcribed and c-DNA-AFLP was done to identify different sequence fingerprints in the protected plant tissue. Discrimination of newly expressed sequences was performed with DHPLC, and comparison of transcripts was done on the complete peach genome database in order to identify the genes or sequences involved in the elicitation of induced resistance. Preliminary data showed, in treated plants, the presence of nine putative genes like a putative senescence protein, already described as being involved in the induced resistance to fire blight in other Rosaceae. Thus, copper glucohumates might be considered possible candidates for the elicitation of resistance to bacterial diseases

Seed transmission of Acidovorax citrulli: implementation of detection in watermelon seeds and development of disinfection methods

Acidovorax citrulli is a seed-borne pathogen and the causal agent of bacterial fruit blotch of cucurbits. It is listed as an A1 quarantine pathogen by EPPO. Seed certification is based on the availability of a sensitive and specific pathogen detection in seed lots: this is a must for an effective disease management strategy. Therefore, an effective DNA extraction and purification procedure is a critical issue to ensure a robust PCR analysis. Pathogen detection in seed lots has been implemented by testing different known contamination levels by Acidovorax citrulli. Initially, two different sample preparation methods have been tested: a) Overnight soaking; b) Hammering of dry seeds, followed by three different manual DNA extraction. Each DNA sub-sample obtained has been analysed with two different primers sets, SEQID3/SEQID4 and WFB1/WFB2, to evaluate the capability to detect the pathogen. Results showed that a DNA extraction and purification procedure, based on soaking the seeds, followed by the use of the DNeasy Plant Mini kit (Qiagen) on the washing fluids gave the highest amount of DNA, sufficient to increase the detection threshold of the pathogen. This will allow the improvement of current detection procedures. Furthermore, naturally contaminated watermelon seeds were treated through different methods, in order to achieve a possible sanitation or eradication of Acidovorax citrulli: a bacterial antagonist, a microbial consortium, a plant polyphenol. Our results showed that treated seeds were only partially disinfected, and the pathogen was not eradicated after any of the methods used

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

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

Tomato and Pepper Seeds as Pathways for the Dissemination of Phytopathogenic Bacteria: A Constant Challenge for the Seed Industry and the Sustainability of Crop Production

The seed industry plays a crucial role in global food production but it faces a persistent challenge in ensuring the health and quality of seeds, particularly those of tomato and pepper seeds, which represent key seed commodities on the global market. Seeds can serve as potential pathways for the introduction and dissemination of seed-borne bacteria, which may have devastating effects on crop yield, farmers’ remunerability, and food security. Therefore, fungicides and other antimicrobial compounds are extensively used to disinfect the seeds, thus increasing the input of chemicals in the agri-environment. In this review, we address aspects that connect disease epidemiology with seed infection and health, including seed contamination, endophytic colonization, and seedborne infections. We focused on the main bacterial diseases affecting tomato and pepper seeds by discussing their official seed testing methods as requirements supporting a smooth seed trade. Moreover, we present a survey on the past and recent innovations for seed treatments, focusing on sustainable disinfection methods. Therefore, this review will be a short but indispensable guide for seed technologists and pathologists involved in the production of high-quality seeds, providing indications and suggestions to contrast seed-borne pathogen dissemination and avoid international controversies and complaints by phytosanitary authorities, extension services, and farmers

In vitro characterization of plant growth promoting and biocontrol activity of beneficial microorganisms

Plant roots are associated with numerous and diverse types of beneficial and pathogenic microorganisms. Among them, plant growth\u2013promoting (rhizo)bacteria (PGPB or PGPR) are isolated from plants crops worldwide, and many of them are used as agricultural inoculants. Agricultural biofertilization and biocontrol of pathogens are eco-friendly alternatives to chemical usage and have less energy, environmental, and economic costs. PGPB isolation and evaluation are essentials steps for determining bacteria that could improve plant development and productivity. In the present study three Streptomyces sp. strains SB14, SA51 & SL81, two Pseudomonas sp. strains PT65 & PN53, an Agrobacterium sp. strain AR39 and an internal control (IC) Pseudomonas synxantha were evaluated in vitro for different plant growth promoting and biocontrol activities. The results were aimed to identify possible antagonists able to inhibit different plant bacterial (Xanthomonas vesicatoria, Clavibacter michiganensis subsp. michiganensis, Clavibacter michiganensis subsp. sepedonicus, Acidovorax citrulli and Ralstonia solanacearum) and fungal (Rhizoctonia solani, Sclerotium sp., Fusarium oxysporum, Alternaria solani and Monilia laxa) pathogens. All the strains were screened for biocontrol activity on three different media\u2019s and AIA (average inhibition area) was calculated. Among the isolates, each strain showed different ability to inhibit the pathogens: Streptomyces sp. strain SA51 was found to be most active. The most prospective strains SA51, AR39 and DLS65 were further evaluated in the field, as possible biocontrol agents for the tomato spot disease (X. vesicatoria), singularly and as a consortium. Results will improve our understanding on the use of such microbial biocontrol agents and will implement innovative biocontrol strategies to bacterial diseases

Growth Promotion and Biocontrol Activity of Endophytic Streptomyces spp.

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

Xylella fastidiosa: il patogeno, le malattie e l’attuale situazione fitosanitaria

Xylella fastidiosa è un batterio fitopatogeno Gram negativo che appartiene alla famiglia delle Xanthomonadaceae. È l’agente causale di numerose malattie d’importanti colture arboree, erbacee, forestali e ornamentali e, a differenza di quanto molta parte dell’opinione pubblica crede, non è un problema di recente comparsa, ma ha una lunga storia che affonda le sue radici ancora alla fine del diciannovesimo secolo. L’etimologia e la semantica del nome assegnato al patogeno da parte dei ricercatori che per primi lo isolarono in coltura pura svelano due aspetti fondamentali della sua biologia: Xylella rivela la nicchia biologica del patogeno all’interno delle sue piante ospiti, cioè lo xilema; fastidiosa indica la grande difficoltà che il batteriologo incontra durante il processo d’isolamento e purificazione della coltura pura