Towards identifying pathogenic determinants of the chickpea pathogen <Emphasis Type="Italic">Ascochyta rabiei</Emphasis>

Ascochyta blight is a serious disease of cool-season grain legumes (chickpea, faba bean, lentil and pea) caused by fungal species of the anamorphic genus Ascochyta and related genera. Despite extensive studies on the biology, ecology, epidemiology and management of the disease, little is known about the pathogenic determinants of these pathogens. This research aims at using Ascochyta rabiei as a model for the genus in investigating genetic factors of pathogenicity, with the ultimate goal of elucidating pathogenic mechanisms. Three advances were made: (1) insertional mutants with altered pathogenicity were identified through in vivo screening, and genomic regions adjacent to the insertion sites in selected mutants were determined; (2) a phage library of A. rabiei genomic DNA was constructed, and the library was estimated to provide complete coverage of the A. rabiei genome. This library was used successfully to recover clones with DNA adjacent to insertional mutation sites and to isolate specific genes; (3) DNA probes specific for an acyl-CoA ligase (cps1) and a polyketide synthase gene (pks1) were developed and library clones containing the corresponding genomic regions were identified from the phage library. These advances provide the foundation and necessary tools for experimentation of ectopic complementation assays and targeted mutagenesis to elucidate the genetic mechanisms of pathogenicity of A. rabiei.     

AlgU contributes to the virulence of <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">tomato</Emphasis> DC3000 by regulating production of the phytotoxin coronatine

Pseudomonas syringae pv. tomato DC3000 (Pst DC3000), which causes bacterial speck disease of tomato, has been used as a model pathogen to investigate the molecular basis of plant–pathogen interactions. The function of many potential virulence factors encoded in the Pst DC3000 genome and their modes of action are not fully understood. P. syringae is known to produce the exopolysaccharide alginate. Although AlgU, a sigma factor, is known to regulate the expression of genes such as algD related to alginate biosynthesis, the molecular mechanisms of AlgU in the virulence of Pst DC3000 is still unclear. To investigate the function of AlgU and alginate in plant–bacterial pathogen interactions, we generated ΔalgU and ΔalgD mutants. After inoculation with ΔalgU but not ΔalgD, host plants of Pst DC3000 including tomato and Arabidopsis had milder disease symptoms and reduced bacterial populations. Expression profiles of Pst DC3000 genes revealed that AlgU can regulate not only the expression of genes encoding alginate biosynthesis, but also the expression of genes related to type III effectors and the phytotoxin coronatine (COR). We also demonstrated that the ΔalgU mutant showed full virulence in the Arabidopsis fls2 efr1 double mutant, which is compromised in the recognition of PAMPs. Further, the application of COR was able to restore the phenotype of the ΔalgU mutant in the stomatal response. These results suggest that AlgU has an important role in the virulence of Pst DC3000 by regulating COR production.     

The chemotaxis regulator <Emphasis Type="Italic">pilG</Emphasis> of <Emphasis Type="Italic">Xylella fastidiosa</Emphasis> is required for virulence in <Emphasis Type="Italic">Vitis vinifera</Emphasis> grapevines

Type IV pili of X. fastidiosa are regulated by pilG, a response regulator protein putatively involved in chemotaxis-like operon sensing stimuli through signal transduction pathways. To elucidate the roles of pilG in pathogenicity of X. fastidiosa, the pilG-deletion mutant XfΔpilG and complemented strain XfΔpilG-C were generated. While all strains had similar growth curves in vitro, XfΔpliG showed significant reduction in cell-matrix adherence and biofilm production compared with wild-type X. fastidiosa and XfΔpilG-C. The genes pilE, pilU, pilT, and pilS were down-regulated in XfΔpliG when compared with its complemented strain and wild-type X. fastidiosa. Finally, no Pierce’s disease symptoms were observed in grapevines inoculated with XfΔpilG, whereas grapevines inoculated with the wild-type X. fastidiosa and complemented strain of XfΔpilG-C developed typical Pierce’s Disease (PD) symptoms. The results indicate that pilG has a role in X. fastidiosa virulence in grapevines.     

Downy mildew resistance evaluation in 28 grapevine hybrids promising for breeding programs in Trentino region (Italy)

Downy mildew is a major grapevine disease caused by the biotrophic oomycete, Plasmopara viticola. Numerous disease resistance studies of diverse Vitis germplasm have been previously carried out to identify downy mildew resistance sources; however, ratings were mainly reported using leaf disc in vitro testing and foliage field assessment, or upon leaf and cluster field evaluations. In the current study, 28 grapevine hybrid cultivars were screened using leaf disc bioassay, for disease resistance characterization of both existing and wild-collected materials. 16 hybrids were identified as highly resistant or resistant, and will serve as relevant resistance donors in future pre-breeding and breeding programs. All grapevine hybrids were evaluated for foliar and cluster downy mildew resistance in an untreated field trial over three successive years. This study showed that the leaf disc bioassay provided some information on the resistance level of the genotypes under scrutiny, but it was a weak predictor of their resistance level under field conditions on leaves and even more on bunches. These findings are relevant to future applications in both traditional and marker-assisted breeding programs which promote sustainable viticulture.     

Synergy between grapevine vitiviruses and grapevine leafroll viruses

An interactive relationship between vitiviruses and grapevine leafroll viruses was characterized in grapevine. Grapevine viruses A and B (GVA and GVB) were found more frequently in the presence of co-infecting Grapevine leafroll associated viruses (GLRaV-1, ?2 or ?3) than in their absence. The titers of the vitiviruses in co-infection with leafroll viruses were found to be higher than were their titers in the absence of leafroll virus infection. The occurrence of vitivirus-associated stem-pitting symptoms was correlated with leafroll virus co-infection. Specific pairing associations on the species level were found between different viti- and leafroll virus species: GVB was associated preferentially with GLRaV-2; GVA was associated preferentially with GLRaV-1 and GLRaV-3. In contrast to the increase in vitivirus titer seen with leafroll virus co-infection, the incidence and titer of grapevine leafroll virus appeared to be unaltered by vitivirus co-infection. The potential for a synergistic enhancement of grapevine disease in co-infected vines is discussed.     

Effects of light quality on conidiophore formation of the melon powdery mildew pathogen <Emphasis Type="Italic">Podosphaera xanthii</Emphasis>

The lengths of conidiophores in fungal colonies of the melon powdery mildew pathogen Podosphaera xanthii Pollacci KMP-6 N cultured under greenhouse (natural) conditions differed markedly from those cultured in a growth chamber. We hypothesized that light wavelength was responsible for the differences in conidiophore length. In this study, we examined the effects of light-emitting diode (LED) irradiation (purple, blue, green, orange, and red light) and white light on colony development and conidiophore formation in KMP-6 N using a stereomicroscope and a high-fidelity digital microscope. Colonies on leaves were flat under greenhouse conditions and under red LED light irradiation but were stacked under growth chamber conditions and under purple, blue, green, and orange LED light irradiation. In addition, KMP-6 N formed catenated conidia comprising six conidia per conidiophore under greenhouse conditions and red light but more than seven conidia per conidiophore under growth chamber conditions and purple, blue, green, and orange light. Furthermore, almost none of the conidia on top of the conidiophores grown under blue light were fully constricted. Therefore, these fungi could not scatter their conidia and spread infection. This is the first report of the effects of LED lights on conidiophore formation in the melon powdery mildew fungus P. xanthii. The results provide insight into the mechanisms underlying the responses of conidiophores to light of specific wavelengths and conidial scatter from conidiophores of melon powdery mildew fungi.     

Effect of <Emphasis Type="Italic">Metarhizium guizhouense</Emphasis> infection on mating competition and mate choice of <Emphasis Type="Italic">Bactrocera latifrons</Emphasis> (Diptera: Tephritidae)

Metarhizium guizhouense PSUM02 treated males of Bactrocera latifrons were investigated for the mating competition among males and mating choice by female flies to develop an auto-dissemination for the control of B. latifrons. In the present study, on day 1–4 of experiment, M. guizhouense–treated male flies were equally competitive with the normal male flies as we did not observe any differences in mating by treated and normal male flies of B. latifrons. Further, mating competitiveness were found low in treated adult male B. latifrons than normal male B. latifrons from 5th days of treatment until death. Kaplan-Meier survival analysis of treated male flies gave average survival times (AST) of 4.3?±?0.1 days, while the healthy female and male flies in the same cage showed AST of 9.3?±?0.3 and 8.3?±?0.4 days, respectively. The AST of untreated flies in control experiment ranged from 14.2–14.5 days. In mating preference experiment, M. guizhouense–treated male flies were chosen by virgin female than gravid female flies for mating. The treated male flies caused mortality in both virgin and gravid female flies in the same cage with AST of 4.4?±?0.1, 5.6?±?0.1 and 7.4?±?0.2 days, respectively, while untreated flies showed AST ranged from 13.9–14.3 days in control. The treated male flies could transmit the fungus infection to both untreated female and male flies as well as in virgin and gravid female flies by mating and contact. Our experiments showed the potentiality of M. guizhouense PSUM02 in management of B. latifrons by auto-dissemination with treated male flies, which transmit the fungus to a healthy population to reduce insect pest infestations.     

Alien species of aphids (Hemiptera: Aphididae) found in Serbia,new to the Balkan Peninsula

New alien species of aphids (Illinoia liriodendri (Monell), Wahlgreniella nervata (Gillete), Takecallis arundicolens (Clarke) and Tinocallis takachihoensis Higuchi) have been found for the first time in Serbia. These records are new for the Balkan Peninsula. Data of Ericaphis scammelli (Mason), species recently mentioned in Serbia, are also presented in the text. The species have been found during the last five years at four sites (Belgrade, Zemun, Mladenovac and Bojnik) on five cultivated plants. Ericaphis scammelli is a pest of blueberry and a vector of Blueberry scorch virus. The other four develop colonies on decorative plants: I. liriodendri has been found on Liriodendron tulipifera, W. nervata on Rosa sp., T. arundicolens on Pseudosasa japonica and T. takachihoensis on Ulmus x hollandica. The origin of three of the species (E. scammelli, I. liriodendri and W. nervata) is North America and two (T. arundicolens and T. takachihoensis) have come from East Asia. Their morphology, biology and the damage they inflict are discussed here, as well as their potential economic and environmental threat. Only parthenogenetic viviparous females of all 5 species have been collected. Their biology has not been studied enough in Europe and needs further examination. Original drawings of all species are presented.     

Susceptibility of eight Algerian olive cultivars to <Emphasis Type="Italic">Bactrocera oleae</Emphasis> infestation – a pomological and nutritional quality perspective

The olive fruit fly, Bactrocera oleae (Rossi), is the major olive pest in the Mediterranean Basin, being responsible for high losses in olive production. The objective of this work was to study the influence of B. oleae infestation on the pomological parameters and nutritive value (lipids, proteins, carbohydrates and ash) of eight olive cultivars (Abani, Aellah, Blanquette de Guelma, Chemlal, Ferkani, Limli, Rougette de Mitidja and Souidi) from Algeria. The results displayed different degrees of susceptibility of the studied cultivars to this pest. Fruit size (weight) and the Fruit weight / stone weight ratio are significantly correlated with the attack. The statistical treatment of the results (PCA and HAC) highlighted different behaviors from the cultivars, concerning the attack. There are some cultivars more susceptible, being the nutritional value also affected. The lipid and protein contents were positively correlated with the attack and the carbohydrates negatively, suggesting a careful choice by the B. oleae. Ovipositional preference of olive fruit fly females and the success of larval development in different olive cultivars are crucial to establishing new approaches in Integrated Pest Management (IPM) against this pest.     

Feeding potential of <Emphasis Type="Italic">Cryptolaemus montrouzieri</Emphasis> against the mealybug <Emphasis Type="Italic">Phenacoccus solenopsis</Emphasis>

Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) is an exotic species native to the USA, damaging cotton and other plant families. The feeding potential of different development stages of Cryptolaemus montrouzieri Mulsant, a biological control agent against mealybugs, was investigated on different development stages of P. solenopsis. Fourth instar grubs and adults of C. montrouzieri were the most voracious feeders on different instars of mealybug. The number of 1^st instar nymphs of mealybug consumed by 1^st, 2^nd, 3^rd and 4^th instar larvae and adult beetles of C. montrouzieri was 15.56, 41.01, 125.38, 162.69 and 1613.81, respectively. The respective numbers of 2^nd and 3^rd instar nymphs of mealybug consumed were 11.15 and 1.80, 26.35 and 6.36, 73.66 and 13.32, 76.04 and 21.16, 787.95 and 114.66. The corresponding figures for adult female mealybugs were 0.94, 3.23, 8.47, 12.71 and 73.40, respectively. The results indicate that C. montrouzieri has the potential to be exploited as a biocontrol agent in North India; inoculative releases of 4^th instar larvae and adults may provide instant control of P. solenopsis. Field experiments should be conducted to determine the efficiency of the ladybird on this mealybug.