Identification and characterization of <Emphasis Type="Italic">Alternaria alternata</Emphasis> (Fr.) Keissler causing <Emphasis Type="Italic">Ceratonia</Emphasis> Blight disease of carob (<Emphasis Type="Italic">Ceratonia siliqua</Emphasis> L.) in Turkey

A new dagger nematode, Xiphinema tica n. sp., is described and illustrated from several populations extracted from soil associated with several crops and wild plants in Costa Rica. The new dagger nematode is characterised by a moderate body size (3276–4240 μm), a rounded lip region, ca 13.5 μm wide, separated from body contour by a shallow depression, amphidial fovea large, stirrup-shaped, a moderately long odontostyle ca 135 μm long, stylet guiding ring located at ca 122 μm from anterior end, vulva almost equatorial (50–54%), well-developed Z-organ, with heavy muscularised wall containing in the most of specimens observed two moderately refractive inclusions variable in shape (from round to star-shaped), with uterine spines and crystalloid bodies; female tail short, dorsally convex-conoid, with rounded end and a small peg, with a c’ ratio ca 0.8, bearing two or three pairs of caudal pores and male absent. The unique and novel uterine differentiation based on the coexistence of a well-developed Z-organ mixed with uterine spines and crystalloid bodies in Xiphinema prompted us to update and include this combination of characters in the polytomous key of Loof and Luc (1990). Integrative diagnosis was completed with molecular data obtained, using D2-D3 expansion segments of 28S rDNA, ITS1-rDNA, partial 18S–rDNA and the partial mitochondrial gene cytochrome c oxidase subunit 1 (coxI). The phylogenetic relationships of this species with other Xiphinema spp. indicated that X. tica n. sp. was monophyletic to the other species from the morphospecies Group 4, Xiphinema oleae.     

Integrative diagnosis and molecular phylogeny of dagger and needle nematodes of olives and grapevines in the island of Crete,Greece, with description of Xiphinema cretense n. sp. (Nematoda,Longidoridae)

The occurrence and geographic distribution of longidorid nematode species inhabiting the rhizosphere of cultivated and wild olive and grapevine in Crete Island were investigated. Morphological and morphometrical studies identified five Longidorus and six Xiphinema species, with frequencies of prevalence (for wild and cultivated olives and grapevines, respectively) as follows: Longidorus closelongatus (2.0–13.3 %), L. cretensis (1.0–6.7 %), L. moesicus (13.3 % only in grapevines), L. orientalis (3.3 % only in grapevines), L. pseudoelongatus (7.0 % only in olives), Xiphinema cretense n. sp. (3.0 % only in olives), X. index (3.0–23.3 %), X. israeliae (6.3 % only in olives), X. italiae (3.3–10.0 %), X. pachtaicum (26.7–42 %) and X. simile (3.3 % only in grapevines). Xiphinema cretense n. sp. is characterized by a body size 3,872–6,135 μm long, lip region anteriorly rounded, separated from the rest of the body by a depression, odontostyle and odontophore 140.6 and 80.3 μm long respectively, vulva position at 46.0–50.5 %, female tail 31.0–38.0 μm long, nearly hemispherical with curvature essentially dorsal and with a tip completely rounded or presenting a very short bulge, c ratio (119.1–186.9), c’ ratio (0.7–0.8). Molecular characterisation using D2-D3 expansion regions of 28S rRNA, 18S rRNA and ITS1-rRNA was carried out and maximum likelihood and Bayesian inference analysis were used to reconstruct phylogenetic relationships among these species and with other longidorids.     

Relative potency of culture supernatants of <Emphasis Type="Italic">Xenorhabdus</Emphasis> and <Emphasis Type="Italic">Photorhabdus</Emphasis> spp. on growth of some fungal phytopathogens

In previous research, concentrated metabolites produced by bacteria of the genera Xenorhabdus and Photorhabdus (which are symbionts of entomopathogenic nematodes) were reported to be highly suppressive to fungal and oomycete plant pathogens. Conceivably, application of non-concentrated bacterial filtrates would be more economically feasible compared to using concentrated metabolites. We evaluated the potency of 10 % v/v cell-free supernatants of the bacteria X. bovienii, X. nematophila, X. cabanillasii, X. szentirmaii, P. temperata, P. luminescens (VS) and P. luminescens (K22) against Fusicladium carpophilum (peach scab), F. effusum (pecan scab), Monilinia fructicola (brown rot), Glomerella cingulata (anthracnose) and Armillaria tabescens (root rot). A bioactive compound derived from Photorhabdus bacteria, trans-cinnamic acid (TCA), was also compared with the bacterial filtrates. Fungal colony size based on manual measurements was compared for accuracy to measurements taken by image analysis. Supernatants of Xenorhabdus spp. exhibited stronger suppressive effects on spore germination and vegetative growth when compared with Photorhabdus spp. Overall, TCA was the most effective treatment; vegetative growth was completely inhibited by TCA (1.27 mg/ml). TCA treatments also suppressed spore germination of F. carpophylium and F. effussum by approximately 90 %. The efficacy of supernatants varied among Xenorhabdus species depending on the species tested, but X. szentirmaii filtrates tended to cause greater inhibition relative to the other bacteria supernatants. Manual measurement of colony diameter required at least two replicate estimates of the colony to avoid a type II error. Area measurements were slightly overestimated based on ruler measurements, but did not affect the outcome of the analysis. Supernatants of Xenorhabdus spp., Photorhabdus spp., or TCA, did not cause any phytotoxic effects when applied to various plant species in the greenhouse. Our results indicate the potential of using TCA or Xenorhabdus cell free supernatants as bio-fungicides. Such a product, based on bacterial culture supernatants, would be economically viable, marketable and easily applicable by the end-users in many situations.     

Natural occurrence of <Emphasis Type="Italic">Fusarium</Emphasis> species and fumonisin on maize grains in Ethiopia

Fusarium species causing maize kernel rot are major threats to maize production, due to reduction in yield as well as contamination of kernels by mycotoxins that poses a health risk to humans and animals. Two-hundred maize kernel samples, collected from 20 major maize growing areas in Ethiopia were analyzed for the identity, species composition and prevalence of Fusarium species and fumonisin contamination. On average, 38 % (range: 16 to 68 %) of maize kernels were found to be contaminated by different fungal species. Total of eleven Fusarium spp. were identified based on morphological characteristics and by sequencing the partial region of translation elongation factor 1-alpha (EF-1α) gene. Fusarium verticillioides was the dominant species associated with maize kernels (42 %), followed by F. graminearum species complex (22.5 %) and F. pseudoanthophilium (13.4 %). The species composition and prevalence of Fusarium species differed among the areas investigated. Fusarium species composition was as many as eight and as few as four in some growing area. The majority of the maize samples (77 %) were found positive for fumonisin, with concentrations ranging from 25 μg kg^?1 to 4500 μg kg^?1 (mean: 348 μg kg^?1 and median: 258 μg kg^?1). Slight variation in fumonisin concentration was also observed among areas. Overall results indicate widespread occurrence of several Fusarium species and contamination by fumonisin mycotoxins. These findings are useful for intervention measures to reduce the impact of the main fungal species and their associated mycotoxins, by creating awareness and implementation of good agricultural practices.     

Pathogenicity and Fungicide Sensitivity of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> and <Emphasis Type="Italic">R. cerealis</Emphasis> Isolates

The Rhizoctonia solani species consists of multinucleate isolates that belong to anastomosis groups AG1–AG3 and differ in virulence and host affinity. R. cerealis is a binucleate species of anastomosis group AG-D which causes sharp eyespot, a common plant disease in Poland. Rhizoctonia spp. is a ubiquitous soil pathogen that poses a significant threat for global crop production due to the absence of effective crop protection products. The aim of this study was to determine the virulence of R. solani and R. cerealis isolates towards Beta vulgaris, Zea mays, Triticum spelta and T. aestivum seedlings, to confirm the presence of endopolygalacturonase genes pg1 and pg5 in the genomes of the tested isolates and to evaluate the tested isolates’ sensitivity to triazole, strobilurin, imidazole and carboxamide fungicides. All tested isolates infected B. vulgaris seedlings. but none of them were virulent against Z. mays plants. R. solani isolates AG4 PL and AG2-2IIIB PL were characterized by the highest virulence (average infestation score of 2.37 and 2.53 points on a scale of 0–3 points) against sugar beet seedlings. The prevalence of infections caused by most of the analysed isolates (in particular R. solani AG4 J—11.8, and R. cerealis RC2—0.78) was higher in spelt than in bread wheat. The virulence of the analysed isolates was not correlated with the presence of pg1 and pg5 genes. The efficacy of the tested fungicides in controlling Rhizoctonia spp. infections was estimated at 100% (propiconazole + cyproconazole), 98.8% (penthiopyrad), 95.4% (tebuconazole) and 78.3% (azoxystrobin).     

Inhibitory effect of <Emphasis Type="Italic">Lycium europaeum</Emphasis> extracts on phytopathogenic soil-borne fungi and the reduction of late wilt in maize

The ability to control soil-borne pathogens in agriculture is highly conditioned by the restricted use of synthetic pesticides. Allelopathy, the antimicrobial activity of plant extracts, is a promising option against crop pathogens. Extracts from Lycium spp. such as L. barbarum, L. chinense and L. intricatum possess biological and therapeutic properties. Individual methanolic extracts from leaves and stems of the Mediterranean medicinal species L. europaeum collected in two locations of Tunisia were each evaluated in vitro against Verticillium dahliae (Vd), Sclerotinia sclerotiorum (Ss) and Harpophora maydis (Hm). The mycelial growth of the three fungi was significantly reduced by all the extracts at doses of 10 and 30 μl mL^?1 (equivalent to 1 and 3 mg plant tissue mL^?1). The sporulation of Hm was almost completely inhibited in all the amendments, but that of Vd was stimulated by one of the leaf extracts when 1 and 3 mg dried plant tissue mL^?1 were used. Sclerotia of Ss were formed in a smaller number, their total weight increasing at extract doses equivalent to 1 mg plant tissue mL^?1 and higher. In greenhouse, the pathogenicity of Hm was confirmed as early as 6 weeks after inoculation, since it caused significant decreases of weights in both roots and aboveground parts of maize. The detrimental effect of Hm on maize root weight in greenhouse was significantly counteracted by one of the leaf extracts added by watering. In total, 11 phenolic compounds were separated in the four extracts. The hydroxycinnamic acid family, including chlorogenic acid as a major compound, represented more than 50% of the total content in all the samples. Rutin was the most abundant flavonoid. The results of this work show the detrimental effect of L. europaeum extracts against the soil-borne pathogens Hm, Ss and Vd, and highlight their potential in crop protection if adequately developed into final products and used in combination with other tools.     

Antifungal activity of <Emphasis Type="Italic">Boerhavia diffusa</Emphasis> L. extract against <Emphasis Type="Italic">Phytophthora</Emphasis> spp. in tomato and pepper

A new longidorid nematode, Longidorus persicus n. sp., is described and illustrated from a population extracted from soil associated with wild rose (Rosa sp.) naturally growing in the mountains close to the village of Cheshmeh-e-Nezamei near the city of Gilan-e-Gharb, Kermanshah province, western Iran. The new needle nematode is characterised by a large body size (6550–7763 μm), an anteriorly flattened lip region, ca 13 μm wide, distinctly set off from body contour by a depression, amphidial fovea large, pocket shaped, slightly asymmetrically bilobed, a moderately long and flexible odontostyle ca 86 μm long, stylet guiding ring located at ca 25 μm from anterior end, posterior arrangement of the pharyngeal gland nuclei, vulva almost equatorial (49–54 %), tail short, about 3/4 of its width, dorsally convex-conoid, with rounded terminus, with a c’ ratio ca 1.3, bearing three pairs of caudal pores and male rare (ratio 1:10 females) with spicules ca 46 μm long. Integrative diagnosis was completed with molecular data obtained using D2-D3 expansion segments of 28S rDNA, ITS1-rDNA, and partial 18S-rDNA. The phylogenetic relationships of this species with other Longidorus spp. using D2-D3 expansion segments and partial 18S-rDNA indicated that L. persicus n. sp. clustered together with L. perangustus and L. euonymus: both of them sharing an anteriorly flattened lip region and distinctly set off from body contour by a depression.     

Feeding preference of Thaumastocoris peregrinus on several Eucalyptus species and the relationship with the profile of terpenes in their essential oils

The feeding preference by Thaumastocoris peregrinus, Carpintero and Dellapé (Heteroptera: Thaumastocoridae) on different Eucalyptus species was evaluated through choice tests in a paired comparisons design. Twenty individuals per combination were tested over a 48 h period, and each one was offered leaves of two species at a time, in all possible combinations. Preference was determined by counting the number of insect excrements on each plant as a proxy for effective feeding, and then ranked through a paired comparison matrix. A linearity index was calculated to test for hierarchy preferences and, after that, a cardinal index (David’s score) was calculated to assign a preference value for each species. The essential oils were extracted from leaves of each Eucalyptus species, and their chemical composition determined through GC-MS. The results show a significant and strictly linear feeding preference ranking between the species tested. The order of preference between species was E. viminalis > E. grandis > E. tereticornis > E. dunnii > E. maidenii > E. globulus. The qualitative and quantitative chemical compositions of the essential oils of these Eucalyptus species differed. A significant correlation was found between the feeding preferences and the concentrations of 1,8-cineole as a repellent and p-cymene as a feeding stimulant. These results underline the importance of selecting the right Eucalyptus species in areas infested by T. peregrinus.     

Environmental and inoculum effects on epidemiology of bacterial spot disease of stone fruits and development of a disease forecasting system

Bacterial spot disease of stone fruits, caused by Xanthomonas arboricola pv. pruni, is of high economic importance in the major stone-fruit-producing areas worldwide. A better understanding of disease epidemiology can be valuable in developing disease management strategies. The effects of weather variables (temperature and wet/dry period) on epiphytic growth of X. arboricola pv. pruni on Prunus leaves were analyzed, and the relationship between inoculum density and temperature on disease development was determined and modeled. The information generated in this study, performed under controlled environmental conditions, will be useful to develop a forecasting system for X. arboricola pv. pruni. Optimal temperature for growth of epiphytic populations ranged from 20 to 30 °C under leaf wetness. In contrast, multiplication of epiphytic populations was not only interrupted under low relative humidity (RH) (< 40%) at 25 °C, but also resulted in cell inactivation, with only 0.001% initial cells recovered after 72 h incubation. A significant effect of inoculum density on disease severity was observed and 10⁶ CFU/ml was determined as the minimal infective dose for X. arboricola pv. pruni on Prunus. Infections occurred at temperatures from 15 to 35 °C, but incubation at 25 and 30 °C gave the shortest incubation periods (7.7 and 5.9 days respectively). A model for predicting disease symptom development was generated and successfully evaluated, based on the relationship between disease severity and the accumulated heat expressed in cumulative degree day (CDD). Incubation periods of 150, 175 and 280 CDD were required for 5, 10 and 50% of disease severity, respectively.     

Insecticidal activity of plant-derived extracts against different economically important pest insects

With the aim of selecting potential botanical insecticides, seven plant extracts (Daphne mucronata (Family: Thymelaeaceae), Tagetes minuta (Asteraceae), Calotropis procera (Apocynaceae), Boenninghausenia albiflora (Rutaceae), Eucalyptus sideroxylon (Myrtaceae), Cinnamomum camphora (Lauraceae) and Isodon rugosus (Lamiaceae)) were screened for their toxic effects against four important agricultural pest insects, each representing a separate insect order; pea aphids of Acyrthosiphon pisum (Hemiptera), fruit flies of Drosophila melanogaster (Diptera), red flour beetles of Tribolium castaneum (Coleoptera), and armyworms of Spodoptera exigua (Lepidoptera). Aphids were the most susceptible insect with 100% mortality observed after 24 h for all the plant extracts tested. Further bioassays with lower concentrations of the plant extracts against aphids, revealed the extracts from I. rugosus (LC₅₀ 36 ppm and LC₉₀ 102 ppm) and D. mucronata (LC₅₀ 126 ppm and LC₉₀ 198 ppm) to be the most toxic to aphids. These most active plant extracts were further fractionated into different solvent fractions on polarity basis and their insecticidal activity evaluated. While all the fractions showed considerable mortality in aphids, the most active was the butanol fraction from I. rugosus with an LC₅₀ of 18 ppm and LC₉₀ of 48 ppm. Considering that high mortality was observed in aphids within 24 h of exposure to a very low concentration of the butanol fraction from I. rugosus, we believe this could be exploited and further developed as a potential plant-based insecticide against sucking insect pests, such as aphids.     

Control of infection of tomato fruits by <Emphasis Type="Italic">Alternaria</Emphasis> and mycotoxin production using plant extracts

Tomato fruits are susceptible to infection by Alternaria species. In addition, Alternaria species may contaminate the fruits with mycotoxins. There is thus interest in control systems to minimise pathogenicity and control toxin production. The objectives of this study were to examine the effect of plant extracts of Eucalyptus globulus and Calendula officinalis on the growth of strains of Alternaria alternata and A. arborescens, on pathogenicity of tomato fruits and mycotoxin production. The growth bioassays showed that the ethanolic and chloroformic fractions of E. globulus were the most effective in reducing growth of A. alternata (66–74 %) and A. arborescens (86–88 %), respectively at 2500 μg/g. The effects of plant extracts on mycotoxin biosynthesis were variable and strain dependent. The most effective fractions in decreasing mycotoxin accumulation were the ethanolic and chloroformic extracts of E. globulus, which reduced tenuazonic acid by 89 %, alternariol by 75–94 % and almost complete inhibition of alternariol monomethyl ether. All the tested fractions reduced percentage of infected tomato fruits when compared to the controls. The ethanolic and chloroformic fractions of E. globulus completely inhibited growth of A. alternata and A. arborescens on unwounded fruits and reduced the aggressiveness on wounded fruits of strains of both species significantly.     

Disease development and mycotoxin production by the <Emphasis Type="Italic">Fusarium graminearum</Emphasis> species complex associated with South African maize and wheat

Fungal species comprising the Fusarium graminearum species complex (FGSC) may cause disease in maize and wheat. Host preference within the FGSC has been suggested, in particular F. boothii towards maize ears. Therefore, the disease development and mycotoxin production of five FGSC species in maize and wheat grain was determined. Eighteen isolates representing F. acaciae-mearnsii, F. boothii, F. cortaderiae, F. graminearum and F. meridionale were used. Each isolate was inoculated on maize ears and wheat heads to determine host preferences. Disease severity and disease incidence was measured for maize and wheat, respectively. Fungal colonisation and mycotoxins, deoxynivalenol (DON), nivalenol and zearalenone, was also quantified. Isolates differed significantly (P < 0.05) in their ability to produce symptoms on maize ears, however, no significant differences between FGSC species were determined. Similarly, significant differences (P < 0.05) between isolates but not between FGSC species in disease incidence on wheat were determined. The isolates also differed significantly (P < 0.05) in their ability to colonise maize and wheat grain. No significant differences in fungal colonisation, among the five FGSC species, were determined in field grown maize. However, under greenhouse conditions, F. boothii was the most successful coloniser of maize grain (P < 0.05). In wheat, F. graminearum colonised the grain more successfully and produced significantly more (P < 0.05) DON than the other species. Fusarium boothii isolates were the best colonisers and mycotoxin producers in maize, and F. graminearum isolates in wheat. The selective advantage of F. boothii to cause disease on maize was supported in this study.     

Fungicide resistance profile and genetic structure of <Emphasis Type="Italic">Botrytis cinerea</Emphasis> from greenhouse crops in Cyprus

Botrytis cinerea is a complex species prone to fungicide resistance and characterized by enormous genetic diversity. During 2013, 220 B. cinerea isolates causing gray mold were collected from greenhouse-grown crops in the regions of Ammochostos, Larnaca, and Limassol (Cyprus). Sensitivities of the sampled populations to seven botryticides with different modes of action were screened in vitro. The results of this in vitro screening highlighted the widespread phenomenon of fungicide resistance in greenhouses, since only 8.6 % of the isolates were sensitive to all botryticides. Resistance to thiophanate-methyl was the most prevalent, with frequencies ranging from 53.8 % to 80 %. Similarly, high resistance frequencies were observed for pyraclostrobin (27.1 to 78.9 %) and boscalid (28.2 to 66.2 %). Multiple fungicide resistant phenotypes were predominant, covering 67.3 % of the population, with frequencies of 80.0, 37.5, 53.8, 83.1, and 60.2 % in cucumber, eggplant, green bean, strawberry, and tomato, respectively. No fludioxonil-resistant isolates were observed. Botrytis cinerea and Botrytis group S genotypes comprised the gray mold population. B. cinerea was predominant within cucumber, eggplant and strawberry, whereas both genotypes were in equilibrium in green bean and tomato. However, Botrytis group S was found in all hosts. B. cinerea was the most prevalent in the majority of fungicide resistance phenotypes from strawberry, while genotype distributions within tomato were generally more balanced. B. pseudocinerea was not detected in the sampled population. Overall, frequency of the mating type allele MAT1–1 was higher to MAT1–2, underlying their unequal distribution in the population. However, cases of 1:1 distribution were apparent within particular subpopulations, suggesting that mating in the field cannot be excluded.     

Influence of exogenous cholesterol in Pythiaceae resistance to inhibition by <Emphasis Type="Italic">Trichoderma</Emphasis> antibiosis

The purpose of this study was to determine if exogenous cholesterol availability influenced Pythiaceae resistance to antibiosis. Characterisation of an isolate of Phytophthora erythroseptica and Pythium ultimum for tolerance to antibacterial compounds found that 0.05 g.l^?1 chloramphenicol inhibited mycelial growth by 96.6 % and 23.5 % respectively. However, the addition of cholesterol (0.01 g l^?1) to potato dextrose agar (PDA) containing 0.05 g l^?1 chloramphenicol was found to increase mycelial growth of P. erythroseptica, indicating a role for cholesterol in tolerance to inhibitory antibacterial compounds. To determine if this property extended to suppressive effects of a potential biocontrol agent, P. erythroseptica and P. ultimum were then tested against a cell-free filtrate of diffusible metabolites produced by a suppressive Trichoderma harzianum isolate in the presence and absence of cholesterol in PDA. In the absence of cholesterol, diffusible metabolites of the T. harzianum isolate were found to inhibit mycelial growth of P. erythroseptica and P. ultimum on PDA by 98 % and 63.6 % respectively (P?<?0.0001). However, the inhibitory effect of the metabolites was mitigated when 0.005 g l^?1 of cholesterol was present in PDA, with mycelial growth of P. ultimum and P. erythroseptica reduced by only 60.4 % and 41.8 %, respectively (P?<?0.0001), much less inhibition than was observed in the absence of cholesterol. These results demonstrated that access to exogenous cholesterol can influence the sensitivity of Pythiaceae species to antibiosis by positively influencing mycelial growth.     

Construction of a recombinant strain of <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis> producing both phenazine-1-carboxylic acid and cyclic lipopeptide for the biocontrol of take-all disease of wheat

Four Neofabraea species are responsible for bull’s eye rot, which is an important postharvest disease of apples and pears. The species diversity of its causal agents in Europe has not been thoroughly explored using molecular genetic methods. Eighty-one Neofabraea isolates were obtained mostly from apples with bull’s eye rot symptoms in the Czech Republic over a two year period. The isolates were identified using PCR fingerprinting and DNA sequencing of the ITS rDNA region, the mitochondrial SSU rDNA and the β-tubulin and EF1α genes. The most common species was N. alba (89 %), followed by N. perennans (5 %) and N. kienholzii (5 %). This is the third published record of N. kienholzii in Europe. The species identity of the isolate CPPF507, which was placed close to N. kienholzii, remains unclear. EF1α was shown to be a suitable marker for the identification of species of the genus Neofabraea and was comparable to the previously used β-tubulin gene. Furthermore, the aggressiveness of individual species was compared and species distribution across Europe was summarized. N. perennans and isolate CPPF507 proved to be the most aggressive, whereas the least aggressive was N. kienholzii. Two N. alba isolates isolated from symptomless apple fruits and leaves were pathogenic to apples in the infection tests.     

Rutin promoted resistance of tomato against <Emphasis Type="Italic">Xanthomonas perforans</Emphasis>

Xanthomonas perforans is the causal agent of bacterial spot, one of the most devastating diseases of tomato that results in considerable yield losses worldwide. Rutin, as a polyphenolic substance, was used to induce resistance in tomato against X. perforans. Rutin at concentration of 2 mM had ability to reduce the disease severity of bacterial spot. On the other hand, 2 mM rutin had no antibacterial activity in vitro. Expression profiling of pathogenesis-related gene 5 (PR-5), Phenylalanine ammonia-lyase (PAL) and lipoxygenase (LOX) was probed during the enhanced resistance by rutin. Pretreatment with rutin (rutin/ X. perforans) led to induction of PR-5, PAL and LOX compared to controls (water/ X. perforans). Our results suggest that rutin-induced resistance against X. perforans in tomato might be mediated through stimulation of some defense genes such as PR-5, PAL and LOX.     

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.     

Attempts to eradicate graft-transmissible infections through somatic embryogenesis in <Emphasis Type="Italic">Citrus</Emphasis> ssp. and analysis of genetic stability of regenerated plants

Ability to detect Pseudocercospora macadamiae infection in macadamia husk at least four months before symptoms become visible will aid the development of disease control measures. This study examined the distinctness of P. macadamiae within the phylogenetic lineages of the genus Pseudocercospora. In addition, we developed two quantitative PCR (qPCR) assays, as rapid diagnostic tools, for early detection and quantification of P. macadamiae in planta. Phylogenetic analysis of concatenated sequences of four gene loci (large subunits, internal transcribed spacer (ITS), translation elongation factor 1-alpha (TEF-1α) and actin of 47 P. macadamiae isolates showed that P. macadamiae is a distinct species in the genus Pseudocercospora. P. macadamiae isolates were partitioned into subunits in the cluster but the grouping of the isolates was regardless of location. Nucleotide diversity (0.02) and the coefficient of genetic differentiation (0.07) were low in the P. macadamiae population. Two qPCR primer sets, based on ITS (PMI) and TEF-1α (PME) were designed that consistently amplified P. macadamiae in fungal cultures (Ct = 16.93 ± 0.11 and Ct = 21.20 ± 0.11, respectively) and in planta (Ct = 32.36 ± 0.28 and Ct = 38.07 ± 1.20, respectively). The PMI primers also detected species in the genus Pseudocercospora, while PME was more specific and robust for quantification of P. macadamiae. Both primer sets detected P. macadamiae in asymptomatic tissue samples and strongly differentiated various stages of disease progression, which revealed approximately 10-fold increase in fungal biomass between each consecutive stage of symptom development.