Biochemical changes in the <Emphasis Type="Italic">Brassica juncea-fruticulosa</Emphasis> introgression lines after <Emphasis Type="Italic">Lipaphis erysimi</Emphasis> (Kaltenbach) infestation

Insect damage leads to changes in biochemical profile of plants. Response of three Brassica juncea-fruticulosa introgression lines (already reported resistant to Lipaphis erysimi) in terms of changes in biochemical constituents after aphid infestation was studied along with B. fruticulosa (resistant parent), B. juncea var. PBR ?210 (susceptible parent) and B. rapa ecotype brown sarson BSH-1 (susceptible check). These six genotypes were grown under aphid infested and uninfested conditions and were sampled at peak aphid infestation to analyze the biochemical changes caused by aphid feeding from top 10 cm central twig of plant. A significant reduction in glucosinolates content in aphid infested plants of three introgression lines (I₈, I₇₉ and I₈₂) was observed while opposite was observed in B. fruticulosa, PBR-210 and BSH-1. Exactly opposite trend was observed for total phenols where aphid infestation resulted in significant increase in phenols content in the three introgression lines while a decrease was observed in B. fruticulosa, PBR-210 and BSH-1. A general trend of decline in flavonols, total sugars and free amino acids content was observed after aphid infestation in all the genotypes. Glucosinolates and total phenols served as biochemical bases of resistance in the three introgression lines since there was downregulation of glucosinolates and upregulation of total phenols as against opposite trend observed in BSH-1 and PBR-210.     

Overexpression of <Emphasis Type="Italic">SlARG2</Emphasis> or <Emphasis Type="Italic">SlTD2</Emphasis> in Arabidopsis enhances resistance against <Emphasis Type="Italic">Plutella xylostella</Emphasis> L.

Tomato (Solanum lycopersicum L.) ARGINASE2 (ARG2) and THREONINE DEAMINASE2 (TD2) are involved in plant defense. These enzymes act in the midgut of herbivores fed on tomato plants to degrade the essential amino acids Arg and Thr, respectively. Although it has been demonstrated that overexpression of the SlARG2 gene in tomato enhanced its resistance against M. sexta larvae, knock-down the expression of SlTD2 reduced the resistance of tomato to lepidopteran herbivores; it remains unclear whether overexpression of SlTD2 could enhance the resistance of the host plants to herbivores, or whether combined overexpression of SlARG2 and SlTD2 could lead to synergistically enhanced resistance to insects. Here, we generated transgenic Arabidopsis plants overexpressing SlARG2 (SlARG2 OE) and SlTD2 (SlTD2 OE) individually as well as in combination (SlARG2-SlTD2 OE). Overexpression of these genes did not affect Arabidopsis development, seed yield, or Arg and Thr content. Insect-feeding bioassay was performed by feeding diamondback moth (Plutella xylostella L.) larvae on detached leaves of wild-type, SlARG2 OE, SlTD2 OE, and SlARG2-SlTD2 OE plants. Larvae fed on SlARG2 OE leaves showed approximately 31% to 35% reduction in weight and 6% to 10% reduction in survival rate compared to those fed on wild-type leaves. Although larvae fed on SlTD2 OE leaves showed no reduction in survival rate, they gained less weight. Whereas larvae fed on SlARG2-SlTD2 OE leaves showed neither reduction in weight nor reduction in survival rate. We further investigated the arginase enzymatic activity of the SlARG2 OE and SlARG2-SlTD2 OE transgenic plants. The SlARG2 OE line most resistant to diamondback moth larvae displayed the highest arginase activity. Our data indicate that overexpression of SlARG2 or SlTD2 in Arabidopsis can enhance its resistance against diamondback moth, whereas combined overexpression of SlARG2 and SlTD2 did not generate synergistically increased resistance to diamondback moth.     

Nonhost resistance of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> against <Emphasis Type="Italic">Colletotrichum</Emphasis> species

Arabidopsis thaliana exhibits a durable resistance called nonhost resistance against nonadapted fungal pathogens. A. thaliana activates preinvasive resistance and terminates entry attempts by nonadapted fungi belonging to the genus Colletotrichum, which cause anthracnose disease in many plants. In the interaction between A. thaliana and nonadapted C. tropicale, the preinvasive resistance involves the PENETRATION 2-related antifungal secondary metabolite pathway and the ENHANCED DISEASE RESISTANCE 1-dependent antifungal peptide pathway. The development of invasive hyphae by C. tropicale owing to the reduction of preinvasive resistance then triggers the blockage of further hyphal expansion via the activation of the second layer of resistance, i.e., postinvasive resistance, which guarantees the robustness of the nonhost resistance of A. thaliana against Colletotrichum pathogens. Both the tryptophan-derived metabolic pathway and glutathione synthesis play critical roles in the postinvasive resistance against C. tropicale, although the molecular mechanism of postinvasive resistance remains to be elucidated. In this review, we describe the current understanding of the molecular background of the Arabidopsis nonhost resistance against Colletotrichum fungi and discuss perspectives for future research on this durable resistance.     

Host-delivered RNAi-mediated root-knot nematode resistance in <Emphasis Type="Italic">Arabidopsis</Emphasis> by targeting <Emphasis Type="Italic">splicing factor</Emphasis> and <Emphasis Type="Italic">integrase</Emphasis> genes

Root-knot nematodes (RKNs) are one of the most important biotic factors limiting crop productivity in many crop plants. The major RKN control strategies include development of resistant cultivars, application of nematicides and crop rotation, but each has its own limitations. In recent years, RNA interference (RNAi) has become a powerful approach for developing nematode resistance. The two housekeeping genes, splicing factor and integrase, of Meloidogyne incognita were targeted for engineering nematode resistance using a host-delivered RNAi (HD-RNAi) approach. Splicing factor and integrase genes are essential for nematode development as they are involved in RNA metabolism. Stable homozygous transgenic Arabidopsis lines expressing dsRNA for both genes were generated. In RNAi lines of splicing factor gene, the number of galls, females and egg masses was reduced by 71.4, 74.5 and 86.6%, respectively, as compared with the empty vector controls. Similarly, in RNAi lines of the integrase gene, the number of galls, females and egg masses was reduced up to 59.5, 66.8 and 63.4%, respectively, compared with the empty vector controls. Expression analysis revealed a reduction in mRNA abundance of both targeted genes in female nematodes feeding on transgenic plants expressing dsRNA constructs. The silencing of housekeeping genes in the nematodes through HD-RNAi significantly reduced root-knot nematode infectivity and suggests that they will be useful in developing RKN resistance in crop plants.     

Root-specific expression of defensin in transgenic tobacco results in enhanced resistance against <Emphasis Type="Italic">Phytophthora parasitica</Emphasis> var. <Emphasis Type="Italic">nicotianae</Emphasis>

Phytophthora species are soil-borne pathogens that damage plants in both agro- and natural ecosystems. To suppress the devastating pathogen, we generated a root-specific expression system using a specific promoter (pPRP3) conferring elevated expression of the target gene in roots that are very susceptible to soil-borne pathogens. To verify root-specific expression, we compared β-glucuronidase (GUS) expression driven by a constitutive or root-specific promoters in shoots and roots. In histochemical and fluorometric assays, GUS activity was detected in whole tobacco plants when GUS expression was driven by p35S, but was detected only in the roots by pPRP3. We then expressed a pepper defensin (J1–1) gene in tobacco to elucidate its effect on plant resistance. The accumulation of J1–1 was also tissue-specific in transgenic tobacco plants. Finally, transgenic plants carrying GUS or J1–1 genes in combination with p35S or pPRP3 were inoculated with Phytophthora parasitica var. nicotianae and Pythium aphanidermatum. Disease symptoms were significantly suppressed in transgenic plants that accumulated J1–1, regardless of the promoter used. Furthermore, the expression of PR genes was induced in J1–1 transgenic plants, exhibiting much higher levels in p35S-driven J1–1 plants than in pPRP3::J1–1 plants. These results demonstrated that J1–1 transgenic plants were primed for enhanced expression of PR genes, which provided synergistic effects with the defensin for disease resistance.     

Population features of <Emphasis Type="Italic">Xanthomonas arboricola</Emphasis> pv. <Emphasis Type="Italic">pruni</Emphasis> from <Emphasis Type="Italic">Prunus spp.</Emphasis> orchards in northern Italy

Bacterial leaf/fruit spot and canker of stone fruits, caused by Xanthomonas arboricola pv. pruni, is a recurrent disease in Italy. A set of 23 strains has been isolated in peach and plum orchards in an intensively stone fruit cultivated area located in north-eastern Italy. They were all identified as X. arboricola pv. pruni by means of phytopathological and serological features: hypersensitive reaction on bean pods, pathogenicity test on immature peach or plum fruitlets, identification by immunofluorescence assay and conventional PCR. Phylogenetic analysis based on sequencing of the gyrB housekeeping gene of the isolates showed that they formed a unique clade, well characterised and separated from other xanthomonads. An insight into the genetic population features was attempted by rep-PCR analysis, using the ERIC, REP and BOX primers. The combined rep-PCR fingerprints showed a slight intra-pathovar variation within our isolates, which grouped in five close clusters. Copper resistance has been assessed in vitro for our whole X. arboricola pv. pruni collection, highlighting that two isolates show a level of resistance in vitro up to 200 ppm of copper. Nonetheless, the copLAB gene cluster, present in many other species of Xanthomonads, was not detected in any isolate, confirming the presence of a still unknown mechanism of copper detoxification in our Xanthomonads arboricola pv. pruni tolerant/resistant strains.     

The effects of <Emphasis Type="Italic">Gibberella zeae</Emphasis>, <Emphasis Type="Italic">Barley Yellow Dwarf Virus</Emphasis>, and co-infection on <Emphasis Type="Italic">Rhopalosiphum padi</Emphasis> olfactory preference and performance

Insect-borne viruses promote several changes in plant phenotype, which can modify plant-vector interactions in favor of virus survival and dissemination. Although co-infections commonly occur in the field, little is known about their effects on interactions with the vector. The ecological interactions between Barley Yellow Dwarf Virus (BYDV) and its aphid vector, Rhopalosiphum padi, have been investigated extensively, but the vector’s behavior in more complex scenarios has yet to be examined. We assessed olfactory response and performance of R. padi to wheat singly and doubly infected by the pathogenic fungus Giberella zeae and BYDV. Non-viruliferous aphids preferred odors of BYDV-infected wheat over healthy wheat, as previously reported in the literature, and they were still preferentially attracted to BYDV-infected plant during co-infection. However, around 35% more non-viruliferous aphids chose healthy wheat over G. zeae-infected wheat. Viruliferous aphids did not show any preference to the treatments. BYDV-infected wheat was a superior host than healthy wheat for the aphids whose population increased in 25%. We observed a synergistic effect of the co-infected wheat, which was the best host for aphids, and promoted an elevation of 42% on population growth. Our results indicate that co-infection might be beneficial for virus spread as does not interfere with aphid olfactory preference and provides greater colony growth than in singly infected plants.     

Genetic structure of <Emphasis Type="Italic">Pyrenophora teres</Emphasis> f. <Emphasis Type="Italic">teres</Emphasis> and <Emphasis Type="Italic">P. teres</Emphasis> f. <Emphasis Type="Italic">maculata</Emphasis> populations from western Canada

Infection by Pyrenophora teres f. teres (Ptt) or P. teres f. maculata (Ptm), the causal agents of the net and spot forms of net blotch of barley, respectively, can result in significant yield losses. The genetic structure of a collection of 128 Ptt and 92 Ptm isolates from the western Canadian provinces of Alberta (55 Ptt, 27 Ptm), Saskatchewan (58 Ptt, 46 Ptm) and Manitoba (15 Ptt, 19 Ptm) were analyzed by simple sequence repeat (SSR) marker analysis. Thirteen SSR loci were examined and found to be polymorphic within both Ptt and Ptm populations. In total, 110 distinct alleles were identified, with 19 of these shared between Ptt and Ptm, 75 specific to Ptt, and 16 specific to Ptm. Genotypic diversity was relatively high, with a clonal fraction of approximately 10 % within Ptt and Ptm populations. Significant genetic differentiation (PhiPT = 0.230, P = 0.001) was found among all populations; 77 % of genetic variation occurred within populations and 23 % between populations. Lower, but still significant genetic differentiation (PhiPT = 0.038, P = 0.001) was detected in Ptt, with 96 % of genetic variation occurring within populations. No significant genetic differentiation (PhiPT = 0.010, P = 0.177) was observed among Ptm populations. Isolates clustered in two distinct groups conforming to Ptt or Ptm, with no intermediate cluster. The high number of haplotypes observed, combined with an equal mating type ratio for both forms of the fungus, suggests that P. teres goes through regular cycles of sexual recombination in western Canada.     

Toxicity of soybean-registered agrochemicals to <Emphasis Type="Italic">Telenomus podisi</Emphasis> and <Emphasis Type="Italic">Trissolcus basalis</Emphasis> immature stages

Biological control of phytophagous bugs in soybean crops is efficiently performed by egg parasitoids, such as Telenomus podisi and Trissolcus basalis. Based on this, the use of agrochemicals in these crops must be managed consciously, making use of pesticides that are selective to the egg of these parasitoids, in order to ensure a balanced ecosystem. The aim of this study was to assess the selectivity of 15 registered pesticides to the immature stages (pre and post-parasitism) of T. podisi and T. basalis, following the method proposed by the “International Organization for Biological and Integrated Control” (IOBC). Pesticides were classified as class 1 – harmless (RP?<?30%); class 2 – slightly harmful (30%?≤?RP?≤?79%); class 3 – moderately harmful (80%?≤?RP?≤?99%); and class 4 – harmful (RP?>?99%). During pre-parasitism, the insecticides imidacloprid+beta-cyfluthrin, deltamethrin, lambda-cyhalothrin+thiamethoxam, acephate, and fenitrothion reduced parasitism of both parasitoids. The others: flubendiamide, diflubenzuron, Bacillus thuringiensis, lufenuron, and the herbicide isopropylamine were selective, i.e. harmless (class 1), to both parasitoids, except for pyraclostrobin+metconazole, which significantly reduced T. basalis parasitism, being considered slightly harmful (class 2). In post parasitism, all the aforementioned pesticides were harmless to T. podisi and T. basalis. Moreover, in pre-parasitism, T. basalis was found to be more sensitive to the tested pesticides when compared to T. podisi. Still, more studies must be conducted to provide a better understanding of the impact of agrochemicals on these parasitoid species in semi-field conditions.     

Competence of <Emphasis Type="Italic">Frankliniella occidentalis</Emphasis> and <Emphasis Type="Italic">Frankliniella intonsa</Emphasis> strains as vectors for <Emphasis Type="Italic">Chrysanthemum stem necrosis virus</Emphasis>

The vector competence of Frankliniella occidentalis for Chrysanthemum stem necrosis virus (CSNV) was evaluated. Three vector strains with distinct competences for Tomato spotted wilt virus (TSWV) transmission were investigated, including an artificially selected strain (TsH) that has a particularly high competence (>90 %). Newly hatched larvae of F. occidentalis were given an acquisition access period of 5 days on CSNV-infected D. stramonium leaves, and reared to maturity. Their transmission efficiencies were examined using a leaf disk assay using Petunia x hybrida leaves. Following the leaf disk assay, the virus accumulation in the vectors was examined via a double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) of their bodies. The results showed that the CSNV acquisition and transmission efficiency of the TsH strain did not differ from those of the others, indicating that the competence of F. occidentalis as a vector for CSNV is not related to that for TSWV. The CSNV transmission and acquisition efficiencies of two F. intonsa strains (Hiroshima and Fukuoka) were also evaluated. In Hiroshima strain, 35 % of adults were viruliferous, but only two transmitters (3 %) were observed. In Fukuoka strain, 6 % were viruliferous, and no transmitters were observed. These results indicate that F. intonsa cannot be a major vector for CSNV. The accumulation of CSNV in the adults of F. occidentalis and F. intonsa evaluated using DAS-ELISA showed a significant difference in ELISA values among transmitter, viruliferous non-transmitter, and non-viruliferous individuals. These results clearly demonstrated that only transmitters that accumulated a threshold quantity of virus can transmit CSNV to plants.     

First report of leaf spot caused by <Emphasis Type="Italic">Alternaria alternata</Emphasis> on <Emphasis Type="Italic">Drimia maritima</Emphasis>

Drimia maritima (squill) is a historically important medicinal plant. During the spring of 2016, small, yellow leaf spots, which became brown and finally necrotic, were observed on squill plants in Kohgiluyeh and Boyer-Ahmad Provinces in Iran. A fungus was consistently isolated from infected leaves and identified as Alternaria alternata based on morphological and phylogenetic analyses. Pathogenicity tests confirmed A. alternata to be the causal agent of the newly observed leaf spot disease. This is the first report of leaf spot on D. maritima caused by A. alternata in the world.     

<Emphasis Type="Italic">Fusarium oxysporum</Emphasis> and <Emphasis Type="Italic">Fusarium avenaceum</Emphasis> associated with yield-decline of pyrethrum in Australia

Fusarium wilt, one of the destructive diseases of cucumber can be effectively controlled by using biocontrol agents such as Trichoderma harzianum. However, the mechanisms controlling T. harzianum-induced enhanced resistance remain largely unknown in cucumber plants. Here we screened the potent T. harzianum isolate TH58 that could effectively control F. oxysporum (FO). Glasshouse efficacy trials also showed that TH58 decreased disease incidence by 69.7 %. FO induced ROS over accumulation, while TH58 inoculation suppressed ROS over accumulation and improved root cell viability under F. oxysporum infection. TH58 inoculation could reverse the FO-induced cell division block and regulate the proportional distribution of nuclear DNA content through inducing 2C fraction. Moreover, the expression levels of cell cycle-related genes such as CDKA, CDKB, CycA, CycB, CycD3;1 and CycD3;2 in TH58 - pre-inoculated seedlings were up-regulated compared with those infected with FO alone. Taken together, these results suggest that T. harzianum improved plant resistance against Fusarium wilt disease via alterations in nuclear DNA content and cell cycle-related genes expression that might maintain a lower ROS accumulation and higher root cell viability in cucumber seedlings.     

Natural infection of <Emphasis Type="Italic">Nicandra physaloides</Emphasis> by <Emphasis Type="Italic">Tomato severe rugose virus</Emphasis> in Brazil

Nicandra physaloides, a common weed in South America, was found to be infected by an isolate of Tomato severe rugose virus (ToSRV), a bipartite begomovirus. The plants developed severe yellow rugose mosaic and were collected in São Paulo State, Brazil. This isolate of ToSRV was transmitted by Bemisia tabaci B biotype from infected plants of N. physaloides to healthy plants of N. physaloides and tomato in a glasshouse. This is the first report of natural infection of N. physaloides by ToSRV in Brazil.     

Chitin elicitor receptor kinase 1 (CERK1) is required for the non-host defense response of <Emphasis Type="Italic">Arabidopsis</Emphasis> to <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. Sp. <Emphasis Type="Italic">cubense</Emphasis>

Banana wilt disease is a typical vascular disease caused by the fungal pathogen Fusarium oxysporum f. sp. cubense 4 (Foc 4). Pattern recognition receptors in the plant cell membrane can recognize pathogen-associated molecular patterns (PAMPs) to activate multi-layer defense responses, including defense gene expression, stomatal closure, reactive oxygen species (ROS) burst and callose deposition, to limit pathogen growth. In the present study, we found that chitin elicitor receptor kinase 1 (CERK1) was required for the non-host resistance of Arabidopsis thaliana to Foc B2 (a strain of Foc 4). The cerk1 mutant had weaker defense responses after Foc B2 treatment, including lower expression of PAMP- and salicylic acid-responsive genes, no stomatal closure, lower ROS level and less callose deposition, than that of the wild-type plant. Consistent with this, the cerk1 mutant plants exhibited higher susceptibility to non-host pathogen Foc B2. These results suggest the crucial importance of CERK1 in Foc B2-triggered non-host resistance.     

Hydrogen sulfide causes excision of a genomic island in <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">phaseolicola</Emphasis>

Hydrogen sulfide (H₂S) is known to be an important signalling molecule in both animals and plants, despite its toxic nature. In plants it has been seen to control stomatal apertures, so altering the ability of bacteria to invade plant tissues. Bacteria are known to generate H₂S as well as being exposed to plant-generated H₂S. During their interaction with plants pathogenic bacteria are known to undergo alterations to their genomic complement. For example Pseudomonas syringae pv. phaseolicola (Pph) strain 1302A undergoes loss of a section of DNA known as a genomic island (PPHGI-1) when exposed to the plants resistance response. Loss of PPHGI-1 from Pph 1302A enables the pathogen to overcome the plants resistance response and cause disease. Here, with the use of H₂S donor molecules, changes induced in Pph 1302A genome, as demonstrated by excision of PPHGI-1, were investigated. Pph 1302A cells were found to be resistant to low concentrations of H₂S. However, at sub-lethal H₂S concentrations an increase in the expression of the PPHGI-1 encoded integrase gene (xerC), which is responsible for island excision, and a subsequent increase in the presence of the circular form of PPHGI-1 were detected. This suggests that H₂S is able to initiate excision of PPHGI-1 from the Pph genome. Therefore, H₂S that may emanate from the plant has an effect on the genome structure of invading bacteria and their ability to cause disease in plants. Modulation of such plant signals may be a way to increase plant defence responses for crops in the future.     

Antagonistic activity of <Emphasis Type="Italic">Trichoderma</Emphasis> spp. against <Emphasis Type="Italic">Scytalidium lignicola</Emphasis> CMM 1098 and antioxidant enzymatic activity in cassava

Trichoderma spp. are used as antagonists against different pathogens. Despite many possibilities of using Trichoderma as an antagonist, there are gaps in the knowledge of the interaction between Trichoderma, cassava and Scytalidium lignicola. This fungus causes cassava black root rot and is an inhabitant of the soil, so it is difficult to control. Antagonists may contribute to the possible induction of resistance of plants because, when exposed to such pathosystems, plants respond by producing antioxidative enzymes. The test for potential inhibition of growth of S. lignicola CMM 1098 in vitro was performed in potato-dextrose-agar with two Trichoderma strains T. harzianum URM3086 and T. aureoviride URM 5158. We evaluated the effect of the two selected Trichoderma to reduce the severity of cassava black root rot and shoots. Subsequently, the production of enzymes (ascorbate peroxidase, catalase, peroxidase and polyphenol oxidase) was evaluated in cassava plants. All two Trichoderma strains show an inhibition of the growth of S. lignicola CMM 1098. The most efficient was T. harzianum URM 3086, with 80.78% of mycelial growth inhibition. T. aureoviride URM 5158 was considered the best chitinase producer. All treatments were effective in reducing severity, especially treatments using Trichoderma. Cassava plants treated with T. aureoviride URM 5158 had the highest enzyme activity, especially peroxidase and ascorbate peroxidase. Trichoderma harzianum URM3086 and Trichoderma aureoviride URM 5158 were effective in reducing the severity of cassava black root rot caused by S. lignicola CMM 1098.     

Rice <Emphasis Type="Italic">OsPBL1</Emphasis> (ORYZA SATIVA ARABIDOPSIS PBS1-LIKE 1) enhanced defense of <Emphasis Type="Italic">Arabidopsis</Emphasis> against <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> DC3000

The receptor-like cytoplasmic kinases (RLCK family VII) are required for plant defense against various pathogens. Previously, OsPBL1 (ORYZA SATIVA ARABIDOPSIS PBS1-LIKE 1) was isolated from rice as a potential RSV (rice stripe virus) resistant factor, but its physiological roles in plant defense are yet to be investigated. In this study, we demonstrated that OsPBL1increased defense against P. syringae in transgenic Arabidopsis. To ascertain the role of OsPBL1 gene in plant defense, OsPBL1 tagged with HA (i.e. Hemagglutinin) was overexpressed in Arabidopsis and examined for the resistance against Pseudomonas syringae pv. tomato DC3000 (i.e. Pst DC3000). At 3 dpi of Pst DC3000, transgenic Arabidopsis lines exhibited the reduced chlorotic lesion and propagation of P. syringae, compared to wild type. Elevated pathogen resistance of transgenic lines was correlated with increased H₂O₂ accumulation and callose deposition on the infected leaves. It was also revealed that expression levels of salicylic acid dependent genes such as PR1, PR2, and PR5, were induced higher in transgenic lines than wild type. Taken together, our data suggested that OsPBL1 exerted the role in defense against pathogen attacks in plant via mainly facilitating salicylic acid dependent pathway.     

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.