The origins of crop diseases are linked to domestication of plants. Most crops were domesticated centuries – even millennia – ago, thus limiting opportunity to understand the concomitant emergence of disease. Kiwifruit (Actinidia spp.) is an exception: domestication began in the 1930s with outbreaks of canker disease caused by P. syringae pv. actinidiae (Psa) first recorded in the 1980s. Based on SNP analyses of two circularized and 34 draft genomes, we show that Psa is comprised of distinct clades exhibiting negligible within-clade diversity, consistent with disease arising by independent samplings from a source population. Three clades correspond to their geographical source of isolation; a fourth, encompassing the Psa-V lineage responsible for the 2008 outbreak, is now globally distributed. Psa has an overall clonal population structure, however, genomes carry a marked signature of within-pathovar recombination. SNP analysis of Psa-V reveals hundreds of polymorphisms; however, most reside within PPHGI-1-like conjugative elements whose evolution is unlinked to the core genome. Removal of SNPs due to recombination yields an uninformative (star-like) phylogeny consistent with diversification of Psa-V from a single clone within the last ten years. Growth assays provide evidence of cultivar specificity, with rapid systemic movement of Psa-V in Actinidia chinensis. Genomic comparisons show a dynamic genome with evidence of positive selection on type III effectors and other candidate virulence genes. Each clade has highly varied complements of accessory genes encoding effectors and toxins with evidence of gain and loss via multiple genetic routes. Genes with orthologs in vascular pathogens were found exclusively within Psa-V. Our analyses capture a pathogen in the early stages of emergence from a predicted source population associated with wild Actinidia species. In addition to candidate genes as targets for resistance breeding programs, our findings highlight the importance of the source population as a reservoir of new disease.

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Genomic Analysis of the Kiwifruit Pathogen Pseudomonas syringae pv. actinidiae Provides Insight into the Origins of an Emergent Plant Disease

The grapevine (Vitis spp.) undoubtedly represents one of the woody crops most widely grown in temperate climates, and a highly valuable agricultural commodity. As most of the vegetatively propagated crops, grapevines are exposed to the attacks of a variety of pests and pathogens among which infectious intracellular agents (viruses, viroids, phloem‑ and xylem‑limited prokaryotes) play a major role, causing heavy losses, shortening the productive life of vineyars, and endangering the survival itself of affected vines. The importance of the grapevine industry and the magnitude of the problems caused by these pathogens has generated wide interest which, in turn, has fostered intensive research which has been especially active at the international scale from the late 1950’s onwards.

Directory of virus and virus-like diseases of the grapevine and their agents

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.

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

Leaves from gold kiwifruit plants, Actinidia chinensis, with dark brown angular spots and flowers that were brown and wilted, first yielded non-fluorescent bacterial colonies following isolation. These bacterial colonies were identified by diagnostic polymerase chain reaction (PCR) as Pseudomonas syringae pv. actinidiae. These samples were obtained from the Te Puke region of New Zealand. All isolates were Gram negative and were levan positive, oxidase negative, potato soft rot negative, arginine dehydrolase negative and tobacco hypersensitivity positive (LOPAT 1a). Sequences of the gyrB and the rpoD genes of these isolates were 100% homologous to sequences of P.s. pv. actinidiae deposited in GenBank including the type strain. Koch’s postulates were proven by pathogenicity tests on kiwifruit seedlings.

https://link.springer.com/content/pdf/10.1007%2Fs13314-011-0023-9.pdf

First report of Pseudomonas syringae pv. actinidiae causing kiwifruit bacterial canker in New Zealand.

Animal communication is a dynamic field that promotes cross-disciplinary study of the complex mechanisms of sending and receiving signals, the neurobiology of signal detection and processing, and the behaviors of animals creating and responding to encoded messages. Alongside visual signals, songs, or pheromones exists another major communication channel that has been rather neglected until recent decades: substrate-borne vibration. Vibrations carried in the substrate are considered to provide a very old and apparently ubiquitous communication channel that is used alone or in combination with other information channels in multimodal signaling. The substrate could be ‘the ground’, or a plant leaf or stem, or the surface of water, or a spider’s web, or a honeybee’s honeycomb. Animals moving on these substrates typically create incidental vibrations that can alert others to their presence. They also may use behaviors to create vibrational waves that are employed in the contexts of mate location and identification, courtship and mating, maternal care and sibling interactions, predation, predator avoidance, foraging, and general recruitment of family members to work. In fact, animals use substrate-borne vibrations to signal in the same contexts that they use vision, hearing, touch, taste, or smell. Study of vibrational communication across animal taxa provides more than just a more complete story. Communication through substrate-borne vibration has its own constraints and opportunities not found in other signaling modalities. Here, I review the state of our understanding of information acquisition via substrate-borne vibrations with special attention to the most recent literature.

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How do animals use substrate-borne vibrations as an information source?

In May 2013, 20 plants in a production orchard of kiwifruit (Actinidia deliciosa) cv. Hayward in the seaside area of Primorska showed small, angular, coalescing necrotic leaf spots and cankers on green shoots. In the following 2 weeks, disease progressed to wilting and shoot dieback with exudates. Symptoms were consistent with Pseudomonas syringae pv. actinidiae. Circular, flat, granulated colonies with entire margins were isolated from leaf spots on King's medium B (KB) and on sucrose nutrient agar with boric acid, cephalexine, and cycloheximide. Strains were purified on KB and showed weak fluorescence upon a prolonged incubation (>10 days) and belonged to P. syringae LOPAT group Ia (+---+). DNA was extracted from strains and plant extracts with Chelex 100 resin and Bio-Nobile QuickPick Plant Kit (Turku, Finland), respectively. PCR products of expected sizes were generated by PCR assays (2,4) from all strains and plant extract, supporting the strains as being P. syringae pv. actinidiae. Two strains (NIB Z 1870 and 1871) were further identified by cytochrome C oxidase (negative), glucose metabolism (oxidative), aesculine (negative), and nitrate (negative). Their partial rpoD gene sequences (GenBank Accession Nos. KJ724117 and KJ724118) (3) were identical to the sequence of the P. syringae pv. actinidiae pathotype strain NCPPB 3739 (FN433222, 100% coverage) and to the sequence of P. syringae pv. theae at 96% coverage (FN433271). BOX-PCR fingerprinting and multilocus sequence analysis (MLSA) based on four housekeeping genes gapA (KJ733923 and KJ733924), gltA (KJ733925 and KJ733926), gyrB (KJ733927 and KJ733928), and rpoD identified both strains as biovar 3, a highly virulent biovar of P. syringae pv. actinidiae (5). The pathogenicity of the two strains was confirmed on four plants of A. deliciosa 'Hayward' for each strain. Six-month-old plants were sprayed on the abaxial sides of leaves with 30 ml cell suspension prepared from a 72-h-old culture of the appropriate strain (~8 × 106 CFU/ml in 0.01 M MgSO4), covered with plastic bags for 24 h, and incubated under high relative humidity (80%) with 14 h daylight and 24/21°C day/night temperature. Three positive and three negative control plants were inoculated with the Italian P. syringae pv. actinidiae virulent strain K9 (kindly provided by Dr. Gian Luca Bianchi of the Plant Health Service of Friuli Venezia Giulia region) and 0.01 M MgSO4, respectively. After 7 days, water-soaked brown spots with pale green halos were observed on all plants inoculated with bacteria. Re-isolated bacteria were identical to the original strains in their morphology, PCR products, and rpoD sequences. Negative control plants did not develop symptoms, and no growth was observed on media. This is the first laboratory confirmation of bacterial canker of kiwifruit in Slovenia. Visual inspections carried out by the plant health authorities in 2013 and laboratory analysis confirmed additional infection with P. syringae pv. actinidiae in a single, nearby orchard. The pest status of P. syringae pv. actinidiae in Slovenia is officially declared as present, subject to official control (1). References: (1) EPPO Reporting Service. Online publication: http://archives.eppo.int/EPPOReporting/2014/Rse-1402.pdf . No. 02 2014/026, 2014. (2) A. Gallelli et al. J. Plant Pathol 93:425, 2011. (3) N. Parkinson et al. Plant Pathol. 60:338, 2011. (4) J. Rees-George et al. Plant Pathol. 59:453, 2010. (5) J. L. Vanneste et al. Plant Dis. 97:708, 2013.

First Report of Pseudomonas syringae pv. actinidiae, the Causal Agent of Bacterial Canker of Kiwifruit in Slovenia.

Vibrational communication of the Nearctic flatid planthopper Metcalfa pruinosa (Say) (Hemiptera: Flatidae) was investigated to determine the role of substrate-borne vibrational signals in mating behavior. Signaling activity changes throughout the day and vibrational communication in this planthopper begins at night. All recorded male and female signals consisted of a series of single pulses. Most of these signals were classified as calling signals. They were emitted spontaneously or were evoked by playback stimulation with prerecorded conspecific signals. Male calling signals are simple, and the main difference between male and female calls was in the pulse repetition time. Pulse repetition time in the middle section of the calling signal was the least variable property of the vibrational signals. During later stages of pair formation, males emit a phrase that is a sequence of pulse trains that has a distinct time and amplitude pattern, whereas females produce continuing monotonous substrate vibrations that could last for several hours. Males participated in continued exchange of vibrational signals. Signals exchanged during male–male interaction did not differ structurally from male calling signals and males never overlapped their signals.

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Vibrational Communication of Metcalfa pruinosa (Hemiptera: Fulgoroidea: Flatidae)

Unusual virus-like symptoms were first observed in 2001 on grapevine cvs. Pinot gris and Sauvignonasse in vineyards from the western part of Slovenia. Symptomatic plants showed shortened internodes, poor leaf development, mottling, and deformations of leaves that resulted in poor growth of symptomatic plants. In 2003 and 2004, several samples were tested for Arabis mosaic virus, Cherry leafroll virus, Grapevine fanleaf virus, Raspberry bushy dwarf virus, Strawberry latent ringspot virus, Tomato black ring virus, Tomato ringspot virus, and Tobacco ringspot virus by DAS-ELISA, but none of them could be confirmed as the cause of the observed symptoms. During intensive visual inspections between 2002 and 2006, the symptoms were observed on most grapevine cultivars grown in the Primorska region but predominantly on the two previously mentioned cultivars. In Trentino, northern Italy, similar virus-like symptoms, i.e., chlorotic mottling, puckering and deformation of the leaves, reduced yield, and low quality of the berries were observed in grapevine plants cv. Pinot gris in 2003 and in cvs. Traminer and Pinot noir in 2009 (2). No common grapevine viruses could be associated with the disease. In 2012, a new trichovirus named Grapevine Pinot gris virus (GPGV) was found in Pinot gris plants using deep sequencing. The virus was also detected in symptomless plants (2). GPGV was later reported also from Korea causing inner necrosis of berries and poor fruit set in grapevine cv. Tamnara (1). In 2012, 42 leaf samples from mostly symptomatic grapevine plants of cvs. Pinot gris, Pinot noir, and Muscat blanc were collected at three locations in the Primorska region. Total RNA was extracted from leaves using the MagMAX Express magnetic particle processor with MagMAX-96 Total RNA Isolation Kit and Plant RNA Isolation Aid in Lysis Binding Solution Concentrate (all by Life Technologies, Grand Island, NY). DNA fragments of 1,049 bp corresponding to the movement protein gene were successfully amplified by RT-PCR from 40 samples using primers GPgV5619 and GPgV6668 (2). Amplification products from three plants were cloned into the pGEM-T Easy vector (Promega, Madison, WI) and sequenced. The sequences were deposited in the EBI database under the accession numbers HG738850 to 52. All the nucleotide sequences shared 97.4 to 97.6% identity with GPGV from Italy (sequence FR877530) and 97.1 to 98.2% amino acid identity within the translated region. To our knowledge, this is the first report of GPGV in Slovenia. The disease seems to be spreading extensively in the Primorska region, causing considerable economic losses, and in 2013 it was also observed in other regions of Slovenia. Since the virus could be found in symptomless plants in Italy and in Slovenia, its role in the development of the disease should be further investigated. References: (1) I. S. Cho et al. New Dis. Rep. 27:10, 2013. (2) A. Giampetruzzi et al. Virus Res. 163:262, 2012.

First Report of Grapevine Pinot gris virus Infecting Grapevine in Slovenia

First report of ‘Candidatus Phytoplasma mali’ in Prunus avium, P. armeniaca and P. domestica

In 2002, severe vein yellowing and partial or complete yellowing of leaves was observed on some shoots of red raspberry (Rubus idaeus) cvs. Golden Bliss and Autumn Bliss. Sap of infected plants of cv. Golden Bliss was inoculated onto Chenopodium quinoa and Nicotiana benthamiana. Faint chlorotic spots were observed on inoculated leaves of C. quinoa approximately 14 days after inoculation but no systemic symptoms appeared. No symptoms were observed on N. benthamiana. Raspberry bushy dwarf virus (RBDV) was detected in the original raspberry plant using double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) with polyclonal antiserum (Loewe Biochemica, Sauerlach, Germany). Systemic infections of inoculated C. quinoa and N. benthaminana were confirmed using DAS-ELISA. In 2001 and 2002, unusual virus symptoms were observed on grapevine grafts (Vitis vinifera) of cv. Laski Rizling. Symptoms appeared as curved line patterns and yellowing of the leaves. No nepoviruses were found in symptomatic plant...

I. Žežlina

Papers

First Report of Pseudomonas syringae pv. actinidiae, the Causal Agent of Bacterial Canker of Kiwifruit in Slovenia.

Vibrational Communication of Metcalfa pruinosa (Hemiptera: Fulgoroidea: Flatidae)

First Report of Grapevine Pinot gris virus Infecting Grapevine in Slovenia

First report of ‘Candidatus Phytoplasma mali’ in Prunus avium, P. armeniaca and P. domestica

First Report of Raspberry bushy dwarf virus on Red Raspberry and Grapevine in Slovenia