Inheritance of resistance to <Emphasis Type="Italic">Meloidogyne enterolobii</Emphasis> and individual selection in segregating populations of <Emphasis Type="Italic">Psidium</Emphasis> spp

Interaction between the phytonematode Meloidogyne enterolobii and the fungus Fusarium solani has caused direct and indirect losses in the entire guava production chain and consequent extermination of guava plantations throughout Brazil. The combined action of these two pathogens is known as “guava decline”. In order to obtain and assess Psidium spp. interspecific hybrids for resistance to the nematode M. enterolobii, interspecific crosses of P. guineense (susceptible araçá) x P. cattleyanum (resistant araçá); P.guineense (susceptible araçá) x P. guajava (susceptible guava) and P. cattleyanum (resistant araçá) x P. guajava (susceptible guava) were conducted. These crosses resulted in hybrid immune, susceptible and resistant to Meloidogyne enterolobii. The chi-square test rejected the hypothesis of monogenic inheritance with incomplete dominance, which corroborates that this trait has polygenic action. Predictions of genetic values ??and parameters were obtained by the REML / BLUP procedure, at individual level. Finally, the 30 selected individuals (immune and resistant) were obtained, which will be backcrossed with guava for the recovery of the agronomic traits desired and subsequent release of a new cultivar.     

Resistance to root-knot nematodes on passion fruit genotypes in Brazil

With the expansion of passion fruit cultivation in Brazil, phytosanitary problems have increased, among them, the occurrence of root-knot nematodes. This research aimed to study the response of passion fruit genotypes to Meloidogyne incognita, M. javanica and M. enterolobii in addition to evaluating the life cycle of M. enterolobii in the passion fruit genotype ‘FB 200’. The genotype response was carried out in a greenhouse. Each pot’s soil was inoculated with 5000 eggs. Gall index, egg mass index and nematode reproduction factors were evaluated at 120 days after inoculation. All genotypes studied were resistant to M. incognita, M. javanica and M. enterolobii, except ‘Roxinho do Kênia’, which was susceptible to the three nematode species. The life cycle of M. enterolobii in “FB 200” passion fruit was studied in a growth chamber at 26 °C with a photoperiod of 12 h. Seven days after transplantation, each plant was inoculated with approximately 400 second-stage juveniles. Evaluations were done at 7, 14, 21, 28, 35, 42 and 49 days post inoculation. The nematode did not complete its life cycle.     

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.     

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.     

Host-suitability of black medick (<Emphasis Type="Italic">Medicago lupulina</Emphasis> L.) and additional molecular markers for identification of the pea cyst nematode <Emphasis Type="Italic">Heterodera goettingiana</Emphasis>

A survey was conducted in 16 fields cultivated with broad bean (Vicia faba L.) and garden pea (Pisum sativum L.) in nine localities of Apulia, southern Italy, to determine whether annual weeds were susceptible to the pea cyst nematode (PEACN), Heterodera goettingiana, and could therefore serve as alternate host for the nematode. The results of this study showed that black medick (Medicago lupulina L.) is a good host for the nematode increasing its population levels in the soil in the absence of the primary hosts. The identity of the PEACN was confirmed by integrative taxonomic approaches (classical, and molecular), resulting identical in all cases (broad bean and garden pea, as well as the spontaneous black medick infections). The phylogenetic analyses using ITS and coxI gene regions strongly support the identification of the populations of H. goettingiana from Italy. Also, ITS and coxI gene sequences were obtained from the same cyst, confirming the species identity in comparison to other nematodes and populations in the Goettingiana group, demonstrating that ITS and coxI gene regions of the PEACN are suitable molecular markers for accurate and unequivocal identification of the PEACN. Reproduction and histopathological analyses demonstrated a good host-suitability of black medick to the PEACN. This record enlarges the relatively narrow host-range of the pea cyst nematode and indicates the need to control M. lupulina to avoid the increase of the nematode population in the absence of the main host crop.     

Presence of less-preferred hosts of the aphid parasitoids <Emphasis Type="Italic">Aphidius ervi</Emphasis> and <Emphasis Type="Italic">Praon volucre</Emphasis> reduces parasitism efficiency

Parasitoids are characterized by a defined range of hosts, either more specialist or generalist. Under natural conditions, females may encounter different host species on the same plant or in the same location. In this case, their preference for one host could influence their choice. However, the presence of less suitable hosts may also affect their choice and, in some cases, may reduce their interest in a patch where both preferred and less preferred hosts are available. The aim of the present study was to test the consequences of the simultaneous presence of three cereal aphids (Sitobion avenae Fabricius, Metopolophium dirhodum Walker, and Rhopalosiphum padi Linnaeus) on the parasitism by two of their parasitoids, Aphidius ervi Haliday and Praon volucre Haliday. Firstly, in the no-choice experiment, A. ervi parasitized on S. avenae at a significantly higher rate as compared to M. dirhodum, whereas no parasitism on R. padi was observed. P. volucre parasitized the three species of cereal aphids with a significant preference for S. avenae. Interestingly, when two or three host species were offered simultaneously in the same quantity to pairs of parasitoids, the level of parasitism was less than that observed for one host species alone. This observation exhibits a distractive effect on non-host species, from the defense mechanism of a non-suitable host or from the perception of bad quality patches. These results raise the question of the practical application of inundative release of parasitoids for biocontrol when several hosts are available simultaneously.     

Conidial density of <Emphasis Type="Italic">Monilinia</Emphasis> spp. on peach fruit surfaces in relation to the incidences of latent infections and brown rot

To evaluate the effect of conidial density of Monilinia spp. on the fruit surface on the incidence of latent infection and brown rot in peaches, eleven field surveys were performed in commercial orchards located in Cataluña, Spain over four growing seasons from 2002 to 2005, and nine surveys were conducted to determine the sources of overwintered Monilinia spp. inoculum. There was a significant positive relationship (r?=?0.69) between the numbers of conidia of Monilinia spp. on the fruit surface and the incidence of latent infections, but not with brown rot at harvest. Although mummified fruit, twigs and pits have been identified as being able to carry the pathogen from year to year in peaches grown in Spanish orchards, no relationships between any of these sources and the numbers of conidia on the fruit surface, or incidence of latent infection or brown rot were found. The effect of temperature (T), solar radiation (SR), rainfall (R) and wind speed (WS) on the area under the number of conidia of Monilinia spp. curve (AUncC) on peach surfaces was analysed. Regression analysis revealed that T, SR, R, and WS could account for 99% of the total variation in the area of the AUncC on peach surfaces. Thus, in order to reduce the incidence of latent infection and brown rot it is essential not only to remove the sources of primary inoculum but also to reduce the number of Monilinia spp. conidia on the fruit surface. Furthermore, the sources of airborne conidia of Monilinia spp. should be taken into consideration in disease management programmes in Spain.     

<Emphasis Type="Italic">Colletotrichum acutatum</Emphasis> occurs asymptomatically on apple leaves

Fungi within the Colletotrichum acutatum species complex occur asymptomatically on plant parts of many different plant species. Leaves from apple orchards in southern Norway were sampled, frozen for five hours and incubated for six days to reveal presence of asymptomatic infections of C. acutatum. Number of leaves (incidence) and leaf area covered (severity) with conidial masses of C. acutatum were assessed biweekly on cv. Aroma from late May to late September during three growing seasons. The first finding of conidial masses occurred in the second half of July, and there was a higher incidence occurring in August and September. Sampling of leaves from fruit spurs and vegetative shoots of cvs. Aroma and Elstar showed that conidial masses of C. acutatum developed on leaves on both shoot types, and there was no difference in incidence between these two types. The fungus was detected on leaves from six of eight commercial orchards of cv. Aroma over three years, with a mean incidence of 5.5 %. After storage, bitter rot was found on apple fruit from all eight orchards. There was no correlation between incidence of conidial masses of C. acutatum on leaves and on fruit. In all orchards and seasons investigated, incidence and severity on leaves varied from 0 to 67 % and 0 to 85 %, respectively. The discovery of apple leaves containing conidial masses of C. acutatum clearly indicate for leaves as a potential source of inoculum for fruit infections.     

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.     

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.     

Tight regulation of allene oxide synthase (AOS) and allene oxide cyclase-3 (AOC3) promote <Emphasis Type="Italic">Arabidopsis</Emphasis> susceptibility to the root-knot nematode <Emphasis Type="Italic">Meloidogyne javanica</Emphasis>

Biosynthesis of the oxylipin jasmonic acid (JA) in Arabidopsis thaliana is catalyzed by a single allene oxide synthase (AOS)-encoding gene and four genes encoding four functional allene oxide cyclase (AOC) polypeptides (AOC1, AOC2, AOC3, and AOC4). To elucidate the biological activities of the JA pathway in regulating the plant defense response to plant-parasitic nematodes, transgenic lines carrying the GUS reporter gene under the control of individual AOC or AOS promoters were examined. Upon penetration by second-stage juveniles (J2 s), promoter activities of AOC1, AOC3 and AOC4 appeared in the root tip and root-elongation zone, with AOC3 demonstrating highest induction. At 5 days AOC3 activity continued to be highly pronounced in the stele and root cortex, associated with nematode invasion throughout gall initiation and maturation. AOS expression appeared 3 days postinfection and accompanied all later infection stages. Mutant lines were analyzed: disruption in AOS rendered plants more resistant to nematode infection, as reflected by the decreased number of females produced on this line; loss-of-function of AOC3 rendered plants more susceptible to nematode infection. Oxylipins derived from the 9- and 13-lipoxygenase pathways were assayed for their direct inhibitory activity toward M. javanica J2 s. Clear nematicidal activity of the bioactive 9- and 13-hydroperoxides was observed. Oxylipins produced by divinyl ether synthase, colneleic acid, colnelenic acid and ω5(Z)-etherolenic acid demonstrated strong inhibitory activity. These data, along with those of other assayed oxylipins, suggest that temporal and spatial fine tuning of the JA route allowing nematodes parasitism on plant host.     

Fungicide sensitivity,growth rate,aggressiveness and frost hardiness of <Emphasis Type="Italic">Monilinia fructicola</Emphasis> and <Emphasis Type="Italic">Monilinia laxa</Emphasis> isolates

Monilinia fructicola, the most destructive pathogen of the genus Monilinia, has recently been introduced into Serbia and many other European countries. Since then, many studies have been conducted to evaluate the characteristics of Monilinia species that have a role in the establishment and survival of the pathogen in new areas. The present study assessed the capacity of M. fructicola to repress and replace Monilinia laxa in Serbia based on: fungicide sensitivity, growth rate and aggressiveness at different temperatures, as well as frost hardiness of the isolates of both species. The results showed that the isolates of M. fructicola, compared to M. laxa, were significantly less sensitive to the following fungicides: iprodione, tebucanozole, chlorothalonil, azoxystrobin, fluopyram, and boscalid. In addition, M. laxa isolates exhibited little variation in sensitivity to all of the tested fungicides, whereas M. fructicola isolates displayed a wide range of sensitivity. The temperature of 5°C favored M. laxa growth and aggressiveness, while at 30°C M. fructicola grew faster and had higher lesion expansion rate. These results support an assumption that M. fructicola will continue to spread in Serbian orchards in coming years, particularly on stone fruits harvested during hot summer weather.     

Characterization of <Emphasis Type="Italic">Phaeoacremonium</Emphasis> isolates associated with Petri disease of table grape in Northeastern Brazil,with description of <Emphasis Type="Italic">Phaeoacremonium nordesticola</Emphasis> sp. nov.

Severe outbreaks of Alternaria leaf blotch and fruit spot were recently observed in cv. Pink Lady apples in northern Israel, especially on fruit. Such severe outbreaks have not been reported from other countries. Symptoms involved cracks and rot around the calyx and external rot of the fruit body. Up to 80 % of the fruit in some orchards were affected by the disease. Microscopic examinations, fulfillment of Koch’s postulates and molecular (genetic) analyses confirmed the causal agent as Alternaria alternata f. sp. mali. The incidence of Alternaria increased as the degree of calyx cracking increased, or if fruit were both cracked and rotted. Injecting spore suspensions into the fruit produced typical rot symptoms. Injection assays of detached fruit of eight apple cultivars showed that cvs. Pink Lady and Golden Delicious were susceptible whereas cv. Jonathan was resistant. Pink Lady and Golden Delicious produced more fruit rot as the inoculum concentration increased. Rot in all three cultivars was moderate close to the skin but more severe close to the seed locule. Aqueous extracts taken from Jonathan fruit peel inhibited germ tube elongation of A. alternata f. sp. mali in vitro. This is the first report on heavy infection of Pink Lady fruit in Israel caused by A. alternata f. sp. mali.     

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.     

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.     

Field Monitoring and Efficacy of Entomopathogenic Nematodes Against Chamomile Stem Weevil Microplontus rugulosus and Chamomile Smooth Beetle Olibrus aeneus Under Greenhouse Conditions

Chamomile Matricaria recutita (L.) is an ancient healing plant; it is used for a sore stomach, a mild laxative, anti-inflammatory and a gentle sleep aid. Chamomile plants are infested by chamomile stem-weevil Microplontus rugulosus (Col., Curculionidae) and chamomile smooth beetle Olibrus aeneus (Col., Phalacridae), and can lead to a high reduction in chamomile yield. This work aimed to survey M. rugulosus and O. aeneus by different monitoring methods as well as to manage them by using entomopathogenic nematodes.