Inheritance of resistance to Meloidogyne enterolobii in <Emphasis Type="Italic">Psidium guajava</Emphasis> x <Emphasis Type="Italic">P. guineense</Emphasis> hybrid

Inheritance of the resistance to nematodes has been studied on many different crops, however to our knowledge, no data are available for guava species. The basic genetic resistance parameters to Meloidogyne enterolobii are estimated in the current research in order to guide the development of genotypes resistant to the pathogen. The parental plants, F1 and F2 from a Psidium guajava x P. guineense cross were assessed for the presence or absence of galls and for the number of eggs and juveniles in the root system at the 120th and 240th days after inoculation with 10,000 eggs and juveniles of the nematode. At the age of nine years, the P. guineense plant remained without nematode attack symptoms, whereas the maternal plant was destroyed by the pathogen. The F1 generation showed 270 plants with reproduction factor (RF) <0.322, and there were tiny galls in only 16 plants. The segregation for the presence or absence of galls in the root system in generation F2 was 9:7, wherein the χ² values were 0.78 and 2.66, respectively, at the 120th and 240th days after inoculation, whereas the segregation for RF was 15:1, wherein the χ² values were 2.76 and 1.18, respectively, at the 120th and 240th days. These results indicate epistatic interaction between two genes: in RF < 1 only one allele sets the resistance to the pathogen. The broad sense heritability of RF, estimated to the two assessments was 0.97, and it also indicates a simple inheritance of resistance to M. enterolobii.     

Host status of selected cultivated fruit crops to <Emphasis Type="Italic">Meloidogyne enterolobii</Emphasis>

Meloidogyne enterolobii (syn. M. mayaguensis) has been reported to cause severe damage in commercial guava orchards and other plants in Central and South American countries. Considering the risk of introduction and dissemination of this pest in the European region, M. enterolobii was placed on the EPPO A2 list in 2010. The use of non-host fruit species is a recommended strategy to manage root-knot nematodes in infested guava orchards. This study screened 89 plant genotypes from 25 fruit plants of economic importance, plus two susceptible controls (guava and tomato) for its host status to M. enterolobii. Three to eight months after inoculation, nematode reproduction factor (RF) was used to characterize host suitability of fruit crops to this nematode. Ten banana genotypes, six Barbados cherries, one fig, two grape rootstocks and six melons were rated as good hosts for this nematode. Sixteen fruit plants behaved either as non-hosts or poor hosts to M. enterolobii, including assaí, atemoya, avocado, cashew nut, citrus, coconut, grape, jabuticaba, mango, mulberry, papaya, passion fruit, sapodilla, soursop, starfruit and strawberry. For the future, field experiments in areas infested by this nematode are essential to confirm the greenhouse results. These non-host fruit species can replace in the future eradicated guava trees in fields severely infested by this nematode and become an economic option for growers where M. enterolobii is considered a serious problem.     

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.     

Infection of <Emphasis Type="Italic">Sorghum bicolor</Emphasis>, selected grass species and <Emphasis Type="Italic">Zea mays</Emphasis> by G<Emphasis Type="Italic">loeocercospora sorghi</Emphasis>, causal pathogen of zonate leaf spot

Zonate leaf spot (Gloeocercospora sorghi) is a common disease in Sorghum bicolor producing areas of the U.S., but little is known about its biology, virulence and severity on S. bicolor, Zea mays, and related crop grassweeds. Greenhouse studies were conducted to determine and compare the virulence and severity of G. sorghi on 10 commercially available sorghum hybrids, four Z. mays hybrids and selected grassweed species including Sorghum bicolor (grain sorghum and shattercane biotypes) and Sorghum halepense (Johnsongrass), two of the most problematic arable weeds. Plants from the respective species were inoculated with a local G. sorghi isolate and maintained in a dew-chamber at 24 °C for 24 h and then incubated under greenhouse conditions for 4 weeks. Plants were observed for lesion expression and rated using a modified Horsfall-Barrett scale (0–10). The first symptoms of infection were visible within 24 h following inoculation on shattercane and S. bicolor hybrids. Symptoms consisted of small, non-diagnostic purple lesions on the leaves. Results showed that S. bicolor, S. halepense and shattercane were susceptible to G. sorghi. All other species tested in this study were not infected. More particularly, disease severity, increased from a rating of 3 to 10 on sorghum and from 2 to 7 on S. halepense between 2 and 23 days after inoculation, respectively. However, disease severity on shattercane increased rapidly from 3.5 to 10 between 2 and 8 days after inoculation, respectively. Among the sorghum hybrids tested, FFR-322 appeared to be the most resistant to G. sorghi while Pioneer 83G66 appeared to be the most susceptible. Z. mays hybrids were not infected by the fungus used in this study. G. sorghi could be used effectively to manage shattercane and S. halepense infestations occurring in Z. mays and S. bicolor fields consisting of specific G. sorghi-resistant hybrids.     

Effect of simultaneous and sequential inoculations of <Emphasis Type="Italic">Meloidogyne incognita</Emphasis>, <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> and <Emphasis Type="Italic">Phomopsis vexans</Emphasis> on eggplant in sand mix and fly ash mix soil

Effects simultaneous and sequential inoculations of Meloidogyne incognita, Ralstonia solanacearum and Phomopsis vexans were studied on the growth, chlorophyll and carotenoid contents of eggplants grown in 25% fly ash and 25% sand mix soil. Plants grown in 25% fly ash mix soil had lesser plant growth than grown in 25% sand ash mix soil. Inoculation of M. incognita / R. solanacearum or P. vexans caused reduction in plant growth, chlorophyll and carotenoid contents in both types of soils but these pathogens in combination caused a greater reduction in than individual inoculation. Inoculation of M. incognita 20 days prior to R. solanacearum caused a greater reduction in plant growth than inoculation of M. incognita prior to P. vexans. Inoculation of P. vexans prior to R. solanacearum caused a lesser reduction in plant growth, chlorophyll and carotenoid contents than inoculation of P. vexans prior to M. incognita. Inoculation of R. solanacearum 20 days prior to M. incognita caused a greater reduction in plant growth, chlorophyll and carotenoid contents than inoculation of R. solanacearum prior to P. vexans. Galling and multiplication of M. incognita was higher in plants grown in 25% sand amended soil than with 25% fly ash soil. R. solanacearum and P. vexans had adverse effects on galling and nematode multiplication. Wilt and blight indices caused by R. solanacearum and P. vexans were 3 respectively. Wilt and blight indices were 4 when two pathogens were inoculated together.     

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.     

Temperature and plant age drive downy mildew disease epidemics on oilseed <Emphasis Type="Italic">Brassica napus</Emphasis> and <Emphasis Type="Italic">B. juncea</Emphasis>

Studies were undertaken on the effects of temperature (14/10 °C and 22/17 °C day/night) and plant age (15, 23, 31 and 40 day-old-plants) on the severity of downy mildew (Hyaloperonospora parasitica) on oilseed Brassica cultivars (temperature: Brassica juncea Montara, B. napus Atomic, ATR-Hyden, Hyola 432, Hyola 450 TT, Thunder TT; plant age: B. juncea Dune, B. napus Surpass 402 and Hyola 450 TT). For temperature studies, there were significant (P?<?0.001) effects of temperature, cultivar, and cultivar x temperature interaction. On cotyledons of susceptible cultivars (B. napus Hyola 450 TT and Thunder TT), plants were symptomatic at 22/17 °C by 48 h post inoculation (hpi) and with abundant sporulation evident by 72 hpi, and with all cotyledons of B. napus Thunder TT collapsed by 7 days post inoculation (dpi). However, at 14/10 °C, there were no symptoms on the same cultivars until 5 dpi, and sporulation only observed at 7 dpi. Percent disease index values (DI%) at 22/17 °C of B. juncea Montara and B. napus ATR-Hyden, Hyola 432, Atomic, Hyola 450 TT and Thunder TT were 4.5, 49.0, 51.4, 65.8, 86.3 and 96.0, respectively, with all except B. juncea Montara having significantly lower (P?<?0.001) disease at 14/10 °C with DI% values of 2.8, 30.4, 27.9, 31.1, 44.4 and 76.4, respectively. For plant age studies, there were significant (P?<?0.001) effects of plant age, cultivar, and cultivar x plant age interaction. DI% was significantly higher at 15 compared to 40 day-old-plants (dop) across all cultivars. B. juncea Dune showed greatest resistance, particularly on 40 dop, with DI% values of 25.8, 24.6, 22.9 and 7.5, for 15, 23, 31 and 40 dop, respectively. B. napus Surpass 402 showed high susceptibility on cotyledons of 15 dop but moderate resistance on leaves of other ages, with DI% values of 59.0, 31.2, 27.1 and 26.2 for 15, 23, 31 and 40 dop, respectively. B. napus Hyola 450 TT showed very high susceptibility at the cotyledon stage on 15 dop, but some resistance on 23 dop and more so on 31 and 40 dop, with DI% values of 84.0, 41.2, 35.4 and 32.9 for 15, 23, 31 and 40 dop, respectively. Together, these findings explain for the first time why development of downy mildew epidemics on susceptible cultivars occurs early in the growing season when warmer seasonal temperatures in autumn coincide with presence of seedlings; in contrast to later in the growing season on less susceptible older plants coinciding with cooler and less favourable winter temperatures. Increasing maximum and minimum temperatures associated with climate change have likely fostered the increased severity of downy mildew over the past 15 years.     

Interspecific variation of constitutive chemical compounds in <Emphasis Type="Italic">Pinus</Emphasis> spp. xylem and susceptibility to pinewood nematode (<Emphasis Type="Italic">Bursaphelenchus xylophilus</Emphasis>)

Pinewood nematode (PWN), Bursaphelenchus xylophilus, the causal agent of pine wilt disease (PWD), was detected in Spain in 2008. This gives rise to serious concern, as the disease has caused severe environmental and economic losses in Portugal and in Asian countries. We studied interspecific variation in susceptibility to pine wilt disease and differences in constitutive chemical compounds in the xylem tissue of the seven pine species -P. canariensis, P. halepensis, P. pinaster, P. pinea, P. sylvestris, P. radiata and P. taeda. Two-year-old trees were inoculated with B. xylophilus. Water potential and nematode densities were measured for each species on specific dates; whereas, wilting symptoms were recorded weekly until the end of the assay. Chemical compounds in the xylem were determined prior to inoculation. Three different resistance groups can be established in terms of the pine species susceptibility to PWN: non- to slightly-susceptible (P. canariensis, P. halepensis, P. taeda and P. pinea), susceptible (P. pinaster and P. radiata), and highly-susceptible (P. sylvestris). Nematodes migrated downward to the roots in all seven species. Constitutive xylem nitrogen, total polyphenols, and marginally phosphorus were negatively correlated with mortality caused by PWN. The most susceptible species, Pinus sylvestris, presented high levels of constitutive lipid-soluble substances and low levels of manganese, pointing to a possible relation between these components and PWN susceptibility. The results suggest P. sylvestris, P. pinaster and P. radiata forests could be severely damaged by PWN in Spain and highlight how constitutive chemical compounds such as nitrogen might play a role in resistance mechanisms against PWN.     

Host status of cultivated plants to <Emphasis Type="Italic">Meloidogyne hispanica</Emphasis>

The reproduction of a Meloidogyne hispanica isolate from Portugal was evaluated in 63 plant species/cultivars, in pot assays at 25?±?2.0°C, on the basis of root gall index (GI) and reproduction factor (Rf?=?final/initial egg density) at 60 days after inoculation. Cultivars of aubergine, bean, beetroot, broccoli, carnation, corn, cucumber, French garlic, lettuce, melon, onion, parsley, pea, potato, spinach, and tobacco and two of cabbage were susceptible (3?≤?GI?≤?5; 1.15?≤?Rf?≤?262.86). Cabbage cv. Bacalan, cauliflower cv. Temporão and pepper cv. Zafiro R2 were hypersusceptible or poor hosts (Rf??2) and pepper cvs. Aurelio and Solero were resistant (0.0?≤?GI?≤?0.4; 0.00?≤?Rf?≤?0.03). The response of the pepper cultivars and the Mi-1 resistant tomato cv. Rossol was also conducted in pots using two inoculum levels and four temperatures, three growth chamber (25?±?2.7°C, 29.3?±?1.8°C and 33.6?±?1.2°C) and one outdoors (24.4?±?8.2°C). At 24.4?±?8.2°C and 25?±?2.7°C, the reproduction on the resistant tomato was significantly lower compared to the susceptible cv. Easypeel. At all temperatures, resistance was evident for the pepper cultivars, despite the fact they were not found to contain any of the Me1, Me3, Me7 and N genes. The eggs obtained on cv. Aurelio at 33.6?±?1.2°C were used to get a selected resistance breaking isolate of M. hispanica that was able to reproduce on the three pepper cultivars. Our results suggest that the initial M. hispanica isolate is a mixture of virulent and avirulent individuals. The pepper cultivars tested, have potential to reduce M. hispanica populations in agro-ecosystems under certain conditions, but they should be used as a part of an integrated management strategy in order to prevent the development of virulent populations.     

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.     

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.     

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.     

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 Eurotiomycete <Emphasis Type="Italic">Apinisia graminicola</Emphasis> as the causal agent of a leaf spot disease on the energy crop <Emphasis Type="Italic">Miscanthus</Emphasis> x <Emphasis Type="Italic">giganteus</Emphasis> in Northern Germany

Miscanthus x giganteus is a fast growing, perennial energy crop for temperate climates. Because of its high annual biomass production rates and its characteristics as a low-input crop, an expansion of field cultivation can be anticipated to cover increasing demands for sustainable biomass production. However, knowledge about pathogens that could have an impact on biomass production is still limited for M. giganteus. Here, we report about the isolation of the filamentous fungus Apinisia graminicola from necrotic leaf lesions of M. giganteus grown on a field trial plot in Northern Germany. Inoculation assays with the isolated A. graminicola strain confirmed its capacity to cause a leaf spot disease on M. giganteus. Additional inoculation assays revealed that A. graminicola also caused necrotic lesions on leaves of the model grass Brachypodium distachyon. Generally, symptoms of A. graminicola-caused leaf spot disease were stronger on B. distachyon compared to M. giganteus. Incubation temperatures above 22 °C during A. graminicola infection resulted in stronger disease symptoms on both, M. giganteus and B. distachyon leaves. Microscopic analysis of cross sectioned, infected leaf tissue revealed an epiphytic mycelium formation on the surface and an endophytic colonization of the mesophyll leave tissue, especially in M. giganteus. Our results revealed that the isolated A. graminicola strain is a causal agent of a leaf spot disease on grass leaves. Its potential on endophytic growth in M. giganteus might open new possibilities in studying this type of plant-fungal interaction on a cellular and molecular level in an energy crop.     

Locomotory behaviour of susceptible and resistant <Emphasis Type="Italic">Plutella xylostella (Linnaeus)</Emphasis> (Lepidoptera: Plutellidae)

Locomotory behaviour of insecticide-resistant and susceptible populations of diamondback moth, Plutella xylostella (Linnaeus), a challenging insect-pest of cruciferous vegetable crops round the globe, was studied with the help of Ethovision. Larvae from the susceptible population of P. xylostella travelled more distance on insecticide-treated or untreated surface and consequently their speed was also more as compared to those taken from the insecticide-resistant population.Whereas, the turn angle of larvae from insecticide-resistant population was significantly higher as compared to those from susceptible population. The resistant larvae travelled significantly less distance (260.68 cm/5 min) on treated-surface (with LC₅₀ value of endosulfan, quinalphos, fenvalerate and spinosad) with more value of turn angle (231.16°) than the susceptible ones which moved faster (517.23 cm/5 min) with less value of turn angle (100.63°). Influence of varying temperatures on locomotory behaviour of larvae from the susceptible and resistant population of P. xylostella was also studied and observed that susceptible P. xylostella larvae travelled significantly greater distance as compared to the larvae from insecticide-resistant population, at different temperatures. Locomotory behaviour of larvae of P. xylostella also changes with food conditions. Resistant larvae starved for 24 h travelled significantly lesser distance (147.29 cm) as compared to unstarved ones (332.50 cm). Similar trend in behaviour was also recorded with respect to speed of larvae of P. xylostella. Larvae turn more frequently when kept without food (starved) than those fed normally; the turn angle was significantly higher (290.07°) for resistant larvae and for susceptible larvae it was lesser i.e. 151.55°, when kept at starved conditions. Hence, certain pronounced behavioural differences were registered in locomotion of insecticide-resistant P. xylostella as compared to the susceptible one and this knowledge would help to find effective management solutions to P. xylostella.     

Characterization of <Emphasis Type="Italic">Myoviridae</Emphasis> and <Emphasis Type="Italic">Podoviridae</Emphasis> family bacteriophages of <Emphasis Type="Italic">Erwinia amylovora</Emphasis> from Hungary - potential of application in biological control of fire blight

The virulence spectrum of 300 isolates of Xanthomonas oryzae pv. oryzae (Xoo), representing 17 districts of Punjab, Pakistan was elucidated through inoculation on a set of six rice IRRI-differentials. The virulence level was assessed by using principal component and cluster analysis. Among six principal components (PCs), PC-1 exhibited 59.3 % of the total variance. The highly virulent isolates clusters on the positive side of the ordination away from the point of intersection of PC1 and PC2 and classifies the Xoo isolates from slow disease to the highest disease causing entities. The 300 isolates were categorized into 29 pathotypes (Pt1-29) wherein the highly virulent pathotype (Pt-1), comprises of 39 Xoo isolates were widespread in 12 districts. The majority of Xoo isolates were moderately to least virulent (21.7–43 %) and average disease progress curves confirmed the field reactions of these pathotype clusters for an efficient recognition of Xoo isolates. Interaction of the pathogen with differentials harboring different resistant genes was well investigated in the current study for future management approaches for which the surveillance of the new Xoo pathotypes may expedite the disease resistant rice breeding programme in the country.     

Conidial sporulation of <Emphasis Type="Italic">Stemphylium solani</Emphasis> under laboratory conditions and infectivity of the inoculum produced <Emphasis Type="Italic">in vitro</Emphasis>

Potato virus Y (PVY) is the type-species of the genus Potyvirus, family Potyviridae, being reported as a major tomato (Solanum lycopersicum L.) pathogen in several regions of the world. Pepper yellow mosaic virus (PepYMV) was originally described as a resistance-breaking Potato virus Y (PVY) isolate on Capsicum annuum L. cultivars, and afterwards it was also reported infecting tomatoes in Brazil. In the present work, a search for sources of resistance to both PepYMV and PVY was conducted in a collection of 119 accessions belonging to seven Solanum (section Lycopersicon) species. This germplasm was initially evaluated to PepYMV reaction by mechanical inoculation followed by symptom observations and ELISA. Potential PepYMV resistance sources were identified for the first time in S. habrochaites, S. peruvianum, S. corneliomuelleri, S. chilense, S. pimpinellifolium, and one accession derived from an interspecific cross (S. lycopersicum x S. peruvianum). A sub-group of 24 accessions with negative serology for PepYMV was also challenged with a PVY isolate, followed by serological and molecular detection with universal primers. Solanum habrochaites ‘L.03683’ and ‘L.03684’ were the only accessions found with stable resistance to both viruses. These results confirm S. habrochaites as the most important source of multiple resistance factor(s) to distinct Potyvirus species.     

Vertical distribution of resting spores of <Emphasis Type="Italic">Plasmodiophora brassicae</Emphasis> in soil

Pratylenchus zeae parasitizes various crops and damages the host roots, resulting in decreased yield and quality of the host plants. Alignments of mitochondrial DNA (mtDNA) Cytochrome Oxidase I (COΙ) sequences revealed the genetic variation among Pratylenchus species. The results indicated 0.2–2.4% intraspecific variations for mtDNA COI sequences among eight P. zeae populations, and 25.4–35.1% interspecific variations between P. zeae and other Pratylenchus species. Based on the mtDNA COΙ region, a loop-mediated isothermal amplification (LAMP) assay was developed for the rapid and specific detection of P. zeae. The optimal conditions for the LAMP assay were 64 °C for 40 min. The LAMP products were confirmed using conventional polymerase chain reaction (PCR), analysis with the restriction enzyme Bam HI and visual inspection by adding SYBR Green I to the products. The LAMP assay could detect P. zeae populations from different hosts and different geographical origins specifically. The LAMP assay was also sensitive, detecting 0.1 individual P. zeae, which was 10 times more sensitive than conventional PCR. This is the first report of the detection of Pratylenchus spp. using LAMP. In addition, the results also suggested that use of the COI gene might allow for good resolution at the Pratylenchus species level.     

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.