Haplotypes of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Liberibacter solanacearum’ identified in Umbeliferous crops in Spain

‘Candidatus Liberibacter solanacearum’ is a phloem-limited Gram-negative bacterium that causes serious damage to different crops of the botanical families Solanaceae and Apiaceae. Five haplotypes have been described: LsoA and LsoB are present in solanaceous crops in America and vectored by the tomato/potato psyllid Bactericera cockerelli; LsoC affects carrots from Northern and Central Europe, and is transmitted by the carrot psyllid Trioza apicalis; haplotypes LsoD and LsoE are present in Southern Europe and Morocco in carrot and celery, and are associated with the psyllid Bactericera trigonica. Thirty-four ‘Ca. L. solanacearum’ isolates were collected in six different regions of Spain from distinct Apiaceae hosts (carrot, celery, parsley and parsnip) in eight consecutive years and were analysed. Their haplotypes were determined by a sequence analysis of 16S ribosomal RNA, the 16S–26S ribosomal RNA intergenic spacer, and the 23S ribosomal RNA and rplJ and rplL genes. Both haplotypes LsoD and LsoE were found across Spain, and no host specificity appeared between these two haplotypes. This is the first report of ‘Ca. L. solanacearum’ associated with parsley and parsnip.     

Molecular diversity of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma mali’ and ‘<Emphasis Type="Italic">Ca</Emphasis>. P. prunorum’ in orchards in Slovenia

The plant parasitic nematode Longidorus poessneckensis found in Austria, the Czech Republic, Germany, Poland and the Slovak Republic was molecularly characterized. Mitochondrial genes encoding cytochrome c oxidase subunit I (COI) and nicotinamide dehydrogenase subunit 4 (nad4), the D2 and D3 expansion segments of 28S rRNA and Internal transcribed spacer 1 (ITS1) rRNA were sequenced for 16 L. poessneckensis populations. Six haplotypes of COI and five haplotypes of nad4 were determined. Nucleotide intraspecific variation was up to 17.1% for the partial sequenced COI gene and up to 17.7% for the partial sequenced nad4 gene, the latter being the highest up to date known intraspecific variation in this genus. The analyses of multiple amino acid sequence alignments of mitochondrial genes revealed low variability (0–2.4%) for COI gene and high divergence (0–7.6%) for nad4 gene. Intraspecific sequence diversity for the D2-D3 of 28S rRNA gene was up to 1.2% and for ITS1 rRNA gene was up to 1.6%. It has been hypothesized, that during the Last Glacial Maximum, L. poessneckensis populations probably persisted in refuge areas in the Carpathian Mountains and subsequently expanding from these areas and colonizing other European regions.     

Recombination-like sequences in the upstream region of the phage-type DNA polymerase in ‘<Emphasis Type="Italic">Candidatus</Emphasis> Liberibacter asiaticus’

We determined and compared nucleotide sequences in the upstream region of the bacteriophage-type DNA polymerase in 15 Asian isolates of the citrus-greening bacterium ‘Candidatus Liberibacter asiaticus’ using the genomic information of American isolate Psy62. Some isolates had nucleotide changes and a large 156-bp deletion, which were concentrated in the putative bacteriophage genes ORF3 and ORF4 and corresponding intergenic region. Eight partial recombination-like patterns were observed in this variable region. Most were confirmed to have occurred between the original types 1 and 2, but a fragment, only several dozen nucleotides long and possibly derived from a third type, was also identified.     

Tachinid parasitoids (Diptera: Tachinidae) of<Emphasis Type="Italic">Spodoptera exigua</Emphasis> in cotton fields in Diyarbakır,Turkey

Tachinid parasitoidsExorista xanthaspis (Wiedemann),Nemorilla maculosa (Meigen),Palesisa maculosa (Villeneuve) andDrino imberbis (Wiedemann) were obtained fromSpodoptera exigua (Hübner) (Noctuidae) larvae collected from cotton fields in Turkey.S. exigua is a new host record forN. maculosa, andP. maculosa is recorded from Turkey for the first time. http://www.phytoparasitica.org posting July 14, 2004.     

Modelling pathogen competition and displacement– <Emphasis Type="Italic">Phytophthora infestans</Emphasis> in Scandinavia

Populations of Phytophthora infestans have displaced less fit populations in a number of regions, but not in Scandinavia. In order to study this process, a simple epidemic model was developed, based on the Lotka-Volterra model for competition. The model contains two epidemics, each with logistic growth of two separate populations that interact with each other in that they compete for the same resource base (the plant). In Scandinavia, epidemics from suspected oospore infections are thought to start earlier than those originating from tubers, so routines were added to allow the epidemics to start at different time points. A numerical solution to the model was developed using the deSolve package of the open-source statistical program ‘R’. The resulting model allows examination of the relative importance of different values for the apparent infection rates and starting parameters for the two sub-epidemics. If epidemics from oospore infections start earlier than those from tuber infections, the delay for epidemic initiation via tuber infection would require extremely high values of r in order for this population to dominate at the end of the season. This could be one reason for the lack of persistent clones in the Scandinavian Phytophthora infestans population.     

Detection of <Emphasis Type="Italic">Phoma valerianellae</Emphasis> in lamb’s lettuce seeds

After the reappearance in Italy of a foliar disease of lamb’s lettuce (Valerianella olitoria L. Poll. syn. V. locusta L. Betcke) incited by Phoma valerianellae, we set out to measure the level of seed infection by this fungus, using a plating test, and to develop a molecular method for quick and reliable detection of the pathogen in seeds. All six samples of lamb’s lettuce seed tested were contaminated by P. valerianellae at levels of 0.6% to 15%. Surface disinfection of seeds did not eliminate the contamination and only reduced it to between 0.1% and 10%. The need for a sensitive, reliable and rapid diagnostic method for early identification of the fungus exists. We have developed a PCR-based method to identify the fungus in seeds. Variation within the internal transcribed spacer (ITS1, 5.8S sequences and ITS2) region of the rDNA (ITS) was used to characterize the P. valerianellae strains and to design specific primers within the ITS region.     

Distribution of the double-spined spruce bark beetle <Emphasis Type="BoldItalic">Ips duplicatus</Emphasis> in the Czech Republic: spreading in 1997–2009

Ips duplicatus (Coleoptera: Curculionidae) was reported in the Euro-Siberian taiga from Sweden to Sakhalin and in the Alps at the beginning of the 20th Century. It has been recorded in spruce stands in the north-eastern part of the Czech Republic since the beginning of the 1970s; monitoring of I. duplicatus was conducted in the Czech Republic in 9 years between 1997 and 2009. Beetles of the overwintering generation were captured in Theysohn pheromone traps baited with a lure of ipsdienol and E-myrcenol, and after 2005 in ID Ecolure lures. Surprisingly, I. duplicatus was found throughout the Czech Republic already in 1997. In 2002, there was a nationwide decrease in their numbers. Particularly since 2005, there has been a spread from the main center of occurrence in the eastern areas to the south and west into the central part of Bohemia where, until that time, its population densities had been lower and had not caused economically significant damage. Moreover, it was observed that in the north-eastern parts of the Czech Republic up to 80% of spruce trees from sanitary felling had been infested by this pest in recent years. The recorded amount of spruce trees infested by I. duplicatus is correlated significantly with the amount of spruce trees recorded as damaged by Ips typographus, Ips amitinus and Pityogenes chalcographus infestation as well as with the numbers of individuals of I. duplicatus captured during the aforementioned monitoring.     

The first detection of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma trifolii’ in <Emphasis Type="Italic">Rhododendron</Emphasis> hybridum

Four Rhododendron hybridum plants (from cvs Moravanka and Don Juan), all exhibited symptoms of shortened axillary shoots, reduced leaves with vein clearing and yellowing, undeveloped flowers, and general stunting in a rhododendron nursery garden in southern Bohemia in 2007. Electron microscopy examination of ultra-thin sections revealed the presence of numerous polymorphic phytoplasma-like bodies in the phloem tissue of leaf midribs and petioles. The phytoplasma etiology of this disease was further confirmed by polymerase chain reaction (PCR) using universal phytoplasma primers. Restriction fragment length polymorphism (RFLP) analysis of amplification products obtained with a R16F2/R16R2 primer pair from all symptomatic plants indicated the presence of phytoplasma from the 16SrVI-A subgroup. A detailed comparison of the amplified sequences and phylogenetic analysis confirmed that the phytoplasma belonged to the subgroup 16SrVI-A (clover proliferation phytoplasma group). This is the first report of the natural occurrence of ‘Candidatus Phytoplasma trifolii’ in plants of Rhododendron hybridum.     

Does <Emphasis Type="Italic">Puccinia hemerocallidis</Emphasis> regularly host-alternate between <Emphasis Type="Italic">Hemerocallis</Emphasis> and <Emphasis Type="Italic">Patrinia</Emphasis> plants in Japan?

Daylily rust fungus, Puccinia hemerocallidis, was proven to host-alternate between a wild daylily, Hemerocallis fulva var. longituba, and a patrinia, Patrinia villosa. No proof was obtained for the early belief that the fungus is pathogenic to plantainlilies, Hosta species, in addition to daylilies, Hemerocallis species. The fungus seems to alternate regularly between daylilies and patrinias in Japan because most daylily species are deciduous, and a vegetatively reproducing stage of the pathogen does not seem capable of successfully overwintering free of the living host tissue.     

Induced resistance in potato to <Emphasis Type="Italic">Phytphthora infestans</Emphasis>—effects of BABA in greenhouse and field tests with different potato varieties

We have investigated to what degree induced resistance with β-aminobutyric acid (BABA) can protect potato from late blight infection under Swedish field conditions and if synergistic interactions occur if BABA is applied in combination with a commonly used fungicide, Shirlan. In greenhouse experiments we also investigated the durability of BABA induced resistance, the dose-response relationships in susceptible (Bintje) and partially resistant (Ovatio, Suberb) cultivars and effects of combined applications of BABA and fungicides. We found a clear effect of BABA on P. infestans infection of greenhouse grown potato plants. The lesion sizes were reduced by on average 40–50% compared to untreated control. However, this effect lasted for only 4–5 days after BABA treatment and then the efficacy was lower. When BABA was given in combination with the fungicides it appeared to have an additive effect both in greenhouse and field experiments. Higher concentrations of BABA gave a stronger protective effect. The partially resistant cultivars Ovatio and Superb reacted to lower concentrations of BABA where no effect was found in susceptible Bintje. According to our field data, 20–25% reduction of the fungicide dose in combination with BABA gave on average the same result on late blight development as full dose Shirlan alone; while reduced dose of Shirlan alone sometimes resulted in less effective protection. Our results indicate that induced resistance could be used in practice in combinations with fungicides in order to reduce the amount of toxic compounds under north European conditions.     

Does genetic variability in weeds respond to non‐chemical selection pressure in arable fields?

At a time when herbicide use is being challenged by the selection of herbicide‐resistant weeds, reliance on other, innovative weed control strategies is becoming increasingly necessary. However, one may question the sustainability of these novel farming practices if weeds adapt rapidly to these non‐chemical selection pressures. Although farmers and agronomists impose many selective processes through farming practices, there is a paucity of literature demonstrating these selection cases in arable fields. In contrast to the relatively simple case of herbicide resistance, random trait association and variability in selection pressures in field conditions could explain why there are so few clear examples of adaptive processes to non‐chemical control in arable fields.     

Mechanisms of induced resistance in lettuce against <Emphasis Type="Italic">Bremia lactucae</Emphasis> by DL<Emphasis Type="Bold">-</Emphasis>β<Emphasis Type="Bold">-</Emphasis>amino<Emphasis Type="Bold">-</Emphasis>butyric acid (BABA)

BABA induced local and systemic resistance in lettuce (Lactuca sativa) against the Oomycete Bremia lactucae. Structure-activity analysis showed no induced resistance by related amino-butanoic acids or β-alanine. The R-enantiomer of BABA induced resistance whereas the S-enantiomer did not, suggesting binding to a specific receptor. Other compounds known to be involved in SAR signaling, including abscisic acid, methyl-jasmonate, ethylene, sodium-salicylate and Bion® (BTH) did not induce resistance. Systemic translocation of ¹⁴C-BABA and systemic protection against downy mildew were tightly correlated. BABA did not affect spore germination, appressorium formation, or penetration of B. lactucae into the host. Epifluorescence and confocal microscopy revealed that BABA induced rapid encasement with callose of the primary infection structures of the pathogen, thus preventing it from further developing intercellular hyphae and haustoria. Invaded host cells treated with BABA did not accumulate phenolics, callose or lignin, or express HR. In contrast, cells of genetically-resistant cultivars accumulated phenolics, callose and lignin and exhibited HR within one day after inoculation. The callose synthesis inhibitor DDG did not inhibit callose encasement nor compromised the resistance induced by BABA. PR-proteins accumulated too late to be responsible for the induced resistance. DAB staining indicated that BABA induced a rapid accumulation of H₂O₂ in the penetrated epidermal host cells. Whether H₂O₂ stops the pathogen directly or via another metabolic route is not known.     

Epigenetic regulation – contribution to herbicide resistance in weeds?

Continuous use of herbicides has resulted in the evolution of resistance to all major herbicide modes of action worldwide. Besides the well‐documented cases of newly acquired resistance through genetic changes, epigenetic regulation may also contribute to herbicide resistance in weeds. Epigenetics involves processes that modify the expression of specific genetic elements without changes in the DNA sequence, and play an important role in re‐programming gene expression. Epigenetic modifications can be induced spontaneously, genetically or environmentally. Stress‐induced epigenetic changes are normally reverted soon after stress exposure, although in specific cases they can also be carried over multiple generations, thereby having a selective benefit. Here, we provide an overview of the basis of epigenetic regulation in plants and discuss the possible effect of epigenetic changes on herbicide resistance. The understanding of these epigenetic changes would add a new perspective to our knowledge of environmental and management stresses and their effects on the evolution of herbicide resistance in weeds. © 2017 Society of Chemical Industry     

Structure and variation in the wild-plant pathosystem: <Emphasis Type="Italic">Lactuca serriola–Bremia lactucae</Emphasis>

Over the past decade, extensive research on the wild-plant pathosystem, Lactuca serriola (prickly lettuce)–Bremia lactucae (lettuce downy mildew), has been conducted in the Czech Republic. Studies focused on pathogen occurrence and distribution, host range, variation in symptom expression and disease severity, interactions of B. lactucae with another fungal species (Golovinomyces cichoracearum) on L. serriola, variation in resistance within natural populations of L. serriola, the structure and dynamics of virulence within populations of B. lactucae, sexual reproduction of B. lactucae, and a comparison of virulence structure and changes in B. lactucae populations occurring in wild (L. serriola) and crop (L. sativa) pathosystems. The incidence of B. lactucae on naturally growing L. serriola and other Asteraceae was recorded. Lactuca serriola was the most commonly occurring host species, followed by Sonchus oleraceus. Over the duration of these studies, the incidence of B. lactucae in L. serriola populations varied between 45–87%. Disease incidence and disease prevalence were partly related to the size, density and different habitats of L. serriola populations. In addition to B. lactucae infection, infection by the lettuce powdery mildew fungus (Golovinomyces cichoracearum) was quite common, including co-infection. Variation in resistance to B. lactucae was studied by using ten isolates (NL and BL races with known virulence patterns) at a metapopulation level, i.e. 250 L. serriola samples representing 16 populations from the Czech Republic (CZ). Our comparisons revealed broad variation in host resistance among host populations and also intrapopulation variability. In the CZ populations, 45 resistance phenotypes were recorded, the most frequent were race-specific reaction patterns. Structural and temporal changes in virulence variation of B. lactucae populations on L. serriola were studied during 1998–2005. Altogether, 313 isolates of B. lactucae originating from the Czech Republic were examined for the presence of 32 virulence factors (v-factors), and 93 different virulence phenotypes (v-phenotypes) were recorded. A study of v-factor frequency showed that common v-factors in B. lactucae populations match some of the race-specific resistance genes/factors (Dm genes or R-factors) originating from L. serriola. The highest frequency was recorded by v-factors v7, v11, v15–17, and v24–30. In contrast, v-factors (e.g. v1–4, 6, and 10) matching Dm genes originating from L. sativa were very rare. This demonstrates the close adaptation of B. lactucae virulence to the host (L. serriola) genetic background. Temporal changes in virulence frequencies over the period were recorded. In many v-factors (v11, v14, v16, and v25–28), fluctuations were observed, some (v14 and v17) shifting to higher frequencies, and others (v5/8 and v23) decreasing. The occurrence of mating types was studied (1997–1999) in a set of 59 B. lactucae isolates. Both compatibility types (B1 and B2) were recorded; however the majority of the isolates (96%) were type B2. A comparative study of B. lactucae virulence variation between the wild (L. serriola) and crop (L. sativa) pathosystems showed major differences. Migration and gene flow between both pathosystems and the potential danger of wild B. lactucae populations for cultivated lettuce are discussed. This paper summarizes comprehensive and unique research on an oomycete pathogen (B. lactucae) that is shared between a crop (lettuce, L. sativa) and its close wild relative (prickly lettuce, L. serriola). The data demonstrate clear evidence about race-specific interactions, variation and changes in virulence, and coevolutionary relationships in the wild pathosystem L. serriola–B. lactucae. Conclusions contribute to the broadening and better understanding of gene-for-gene systems in natural host–pathogen populations and their relationships to crop pathosystems.     

Reduced BYDV–PAV transmission by the grain aphid in a <Emphasis Type="Italic">Triticum monococcum</Emphasis> line

The luteovirus Barley yellow dwarf virus–PAV (BYDV–PAV) and its vector, the aphid Sitobion avenae are two major sources of yield losses in cereal crops. We report in this paper the effects of a Triticum monococcum line (TM44), resistant by antibiosis to S. avenae, on the different steps of transmission of one BYDV–PAV isolate by the aphids. First, it was shown that TM44 is strongly resistant to BYDV–PAV transmission, but exclusively when S. avenae is the vector. Second, that TM44 is resistant (1) to BYDV–PAV acquisition by S. avenae and (2) to its inoculation, whatever the respective duration of these two periods. Third, that both resistances have partially additive effects. In the discussion, several lines of evidence are given to support the hypothesis that resistance of TM44 to PAV transmission is due to the same disturbances to S. avenae feeding behaviour that are involved in its antibiosis against this aphid species. Reasons for caution in releasing resistant material in the field are presented.