Two Xylella fastidiosa subsp. multiplex Strains Isolated from Almond in Spain Differ in Plasmid Content and Virulence Traits.

The plant-pathogenic bacterium Xylella fastidiosa is a major threat to agriculture and the environment worldwide. Recent devastating outbreaks in Europe highlight the potential of this pathogen to cause emergent diseases. X. fastidiosa subsp. multiplex ESVL and IVIA5901 strains that belong to sequence type 6 were isolated from almond orchards within the outbreak area in Alicante province (Spain). Both strains share more than 99% of the chromosomal sequences (average nucleotide identity), but the ESVL strain harbors two plasmids (pXF64-Hb_ESVL and pUCLA-ESVL). Here, virulence phenotypes and genome content were compared between both strains, using three strains from the United States as a reference for the phenotypic analyses. Experiments in microfluidic chambers, used as a simulation of xylem vessels, showed that twitching motility was absent in the IVIA5901 strain, whereas the ESVL strain had reduced twitching motility. In general, both Spanish strains had less biofilm formation, less cell aggregation, and lower virulence in tobacco compared with U.S. reference strains. Genome analysis of the two plasmids from ESVL revealed 51 unique coding sequences that were absent in the chromosome of IVIA5901. Comparison of the chromosomes of both strains showed some unique coding sequences and single-nucleotide polymorphisms in each strain, with potential deleterious mutations. Genomic differences found in genes previously associated with adhesion and motility might explain the differences in the phenotypic traits studied. Although additional studies are necessary to infer the potential role of X. fastidiosa plasmids, our results indicate that the presence of plasmids should be considered in the study of the mechanisms of pathogenicity and adaptation in X. fastidiosa to new environments. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Divergent abiotic spectral pathways unravel pathogen stress signals across species.

Plant pathogens pose increasing threats to global food security, causing yield losses that exceed 30% in food-deficit regions. Xylella fastidiosa (Xf) represents the major transboundary plant pest and one of the world's most damaging pathogens in terms of socioeconomic impact. Spectral screening methods are critical to detect non-visual symptoms of early infection and prevent spread. However, the subtle pathogen-induced physiological alterations that are spectrally detectable are entangled with the dynamics of abiotic stresses. Here, using airborne spectroscopy and thermal scanning of areas covering more than one million trees of different species, infections and water stress levels, we reveal the existence of divergent pathogen- and host-specific spectral pathways that can disentangle biotic-induced symptoms. We demonstrate that uncoupling this biotic-abiotic spectral dynamics diminishes the uncertainty in the Xf detection to below 6% across different hosts. Assessing these deviating pathways against another harmful vascular pathogen that produces analogous symptoms, Verticillium dahliae, the divergent routes remained pathogen- and host-specific, revealing detection accuracies exceeding 92% across pathosystems. These urgently needed hyperspectral methods advance early detection of devastating pathogens to reduce the billions in crop losses worldwide.

Detection of Xylella fastidiosa in almond orchards by synergic use of an epidemic spread model and remotely sensed plant traits.

The early detection of Xylella fastidiosa (Xf) infections is critical to the management of this dangerous plan pathogen across the world. Recent studies with remote sensing (RS) sensors at different scales have shown that Xf-infected olive trees have distinct spectral features in the visible and infrared regions (VNIR). However, further work is needed to integrate remote sensing in the management of plant disease epidemics. Here, we research how the spectral changes picked up by different sets of RS plant traits (i.e., pigments, structural or leaf protein content), can help capture the spatial dynamics of Xf spread. We coupled a spatial spread model with the probability of Xf-infection predicted by a RS-driven support vector machine (RS-SVM) model. Furthermore, we analyzed which RS plant traits contribute most to the output of the prediction models. For that, in almond orchards affected by Xf (n = 1426 trees), we conducted a field campaign simultaneously with an airborne campaign to collect high-resolution thermal images and hyperspectral images in the visible-near-infrared (VNIR, 400-850 nm) and short-wave infrared regions (SWIR, 950-1700 nm). The best performing RS-SVM model (OA = 75%; kappa = 0.50) included as predictors leaf protein content, nitrogen indices (NIs), fluorescence and a thermal indicator (Tc), alongside pigments and structural parameters. Leaf protein content together with NIs contributed 28% to the explanatory power of the model, followed by chlorophyll (22%), structural parameters (LAI and LIDFa), and chlorophyll indicators of photosynthetic efficiency. Coupling the RS model with an epidemic spread model increased the accuracy (OA = 80%; kappa = 0.48). In the almond trees where the presence of Xf was assayed by qPCR (n = 318 trees), the combined RS-spread model yielded an OA of 71% and kappa = 0.33, which is higher than the RS-only model and visual inspections (both OA = 64-65% and kappa = 0.26-31). Our work demonstrates how combining spatial epidemiological models and remote sensing can lead to highly accurate predictions of plant disease spatial distribution.

Previsual symptoms of Xylella fastidiosa infection revealed in spectral plant-trait alterations.

Plant pathogens cause significant losses to agricultural yields and increasingly threaten food security1, ecosystem integrity and societies in general2-5. Xylella fastidiosa is one of the most dangerous plant bacteria worldwide, causing several diseases with profound impacts on agriculture and the environment6. Primarily occurring in the Americas, its recent discovery in Asia and Europe demonstrates that X. fastidiosa's geographic range has broadened considerably, positioning it as a reemerging global threat that has caused socioeconomic and cultural damage7,8. X. fastidiosa can infect more than 350 plant species worldwide9, and early detection is critical for its eradication8. In this article, we show that changes in plant functional traits retrieved from airborne imaging spectroscopy and thermography can reveal X. fastidiosa infection in olive trees before symptoms are visible. We obtained accuracies of disease detection, confirmed by quantitative polymerase chain reaction, exceeding 80% when high-resolution fluorescence quantified by three-dimensional simulations and thermal stress indicators were coupled with photosynthetic traits sensitive to rapid pigment dynamics and degradation. Moreover, we found that the visually asymptomatic trees originally scored as affected by spectral plant-trait alterations, developed X. fastidiosa symptoms at almost double the rate of the asymptomatic trees classified as not affected by remote sensing. We demonstrate that spectral plant-trait alterations caused by X. fastidiosa infection are detectable previsually at the landscape scale, a critical requirement to help eradicate some of the most devastating plant diseases worldwide.

Soil factors involved in the diversity and structure of soil bacterial communities in commercial organic olive orchards in Southern Spain.

Nowadays, there is a tendency in olive production systems to reduce tillage or keep a vegetative cover to reduce soil erosion and degradation. However, there is scarce information on the effects of different soil management systems (SMS) in soil bacterial community composition of olive groves. In this study, we have evaluated the effects of soil type and different SMS implemented to control weeds in the structure and diversity of bacterial communities of 58 soils in the two geographic areas that best represent the organic olive production systems in Spain. Bacterial community composition assessed by frequency and intensity of occurrence of terminal restriction profiles (TRFs) derived from terminal restriction fragment length polymorphism (T-RFLP) analysis of amplified 16S ribosomal deoxyribonucleic acid were strongly correlated with soil type/field site (Eutric/Calcaric) that differed mainly in soil particle size distribution and soil pH, followed by a strong effect of SMS, in that order. Canonical discriminant (CD) analysis of TRFs properly classified all of the olive orchard soils as belonging to their respective soil type or SMS. Furthermore, only a small set of TRFs were enough to clearly and significantly differentiate soil samples according to soil type or SMS. Those specific TRFs could be used as bioindicators to assess the effect of changes in SMS aimed to enhance soil quality in olive production systems.

A comparison of real-time PCR protocols for the quantitative monitoring of asymptomatic olive infections by Verticillium dahliae pathotypes.

Early, specific, and accurate in planta detection and quantification of Verticillium dahliae are essential to prevent the spread of Verticillium wilt in olive using certified pathogen-free planting material and development of resistance. We comparatively assessed the accuracy, specificity, and efficiency of eight real-time quantitative polymerase chain reaction protocols published since 2002 for the specific detection and quantification of V. dahliae in various host plant species and in soil, using a background of DNAs extracted from olive roots, stems, and leaves. Results showed that some of those protocols were not specific for V. dahliae or were inhibited when using backgrounds other than water. Ranking of protocols according to a weighted score system placed protocols TAQ (based on intergenic spacer ribosomal DNA target gene) and SYBR-4 (based on the ß-tubulin 2 target gene) first in sensitivity and efficiency for the quantification of V. dahliae DNA in small amounts and different types of olive tissues (root and stem) tested. Use of TAQ and SYBR-4 protocols allowed accurate quantification of V. dahliae DNA regardless of the background DNA, with a detection limit being fixed at a cycle threshold of 36 (≈18 fg for SYBR-4 and 15 fg for TAQ) of V. dahliae. The amount of DNA from defoliating (D) and nondefoliating (ND) V. dahliae pathotypes was monitored in Verticillium wilt-resistant 'Frantoio' olive using the TAQ and SYBR-4 protocols. In the infection bioassay, higher amounts of D V. dahliae DNA were measured in olive stems, whereas the average amount of fungal DNA in roots was higher for ND-infected plants than D-infected ones. Overall, V. dahliae DNA amounts in all olive tissues tested tended to slightly decrease or remain stable by the end of the experiment (35 days after inoculation). The SYBR-4 and TAQ protocols further enabled detection of V. dahliae in tissues of symptomless plants, suggesting that both techniques can be useful for implementing certification schemes of pathogen-free planting material as well as helpful tools in breeding resistance to V. dahliae in olive.

Mycelial compatibility groups and pathogenic diversity in Sclerotium rolfsii populations from sugar beet crops in Mediterranean-type climate regions.

The population structure of Sclerotium rolfsii from autumn-sown sugar beet crops in Mediterranean-type climate regions of Chile, Italy, Portugal and Spain was determined by analyses of mycelial compatibility groups (MCGs) and pathogenicity to 11 economically important plant species. Twelve MCGs (i-xii) were identified among 459 S. rolfsii isolates. MCG iii was the most prevalent group in all countries except Italy. MCG i, the most abundant group (64·7% of isolates) was identified in Portugal and Spain. The remaining MCGs were restricted to various regions within one country (ii, vi, ix) or different countries (v), or to specific localities (iv, vii, viii, x, xi, xii). MCGs iv, vii and x each comprised one isolate. Fields extensively sampled in southern Spain were infected with one to three MCGs. Plant species differed in susceptibility to MCG tester isolates with a MCG by species interaction. Cluster analyses allowed selection into five MCG groupings and grouped plant species into species-groups 1 (broccoli, chickpea, sunflower, tomato) and 2 (cotton, pepper, sugar beet, watermelon). MCG groupings 1 (i, ix), 2 (ii, iii, vi, viii) and 5 (x, xii) were moderately virulent to species-group 1 and mildly virulent to species-group 2. MCG groupings 3 (iv, v, xi) and 4 (vii) were mildly virulent to both species-groups. Across MCG groups, species were rated highly susceptible (chickpea, sunflower), susceptible (cotton, pepper, tomato, watermelon), moderately resistant (broccoli, melon, sugar beet) and resistant (corn, wheat). Establishing the MCG population structure and virulence variability among S. rolfsii isolates should help in the management of sclerotium root rot diseases.

First Report of Southern Blight of Pepper Caused by Sclerotium rolfsii in Southern Spain.

In May 2009, a stem rot of pepper (Capsicum annuum L.) occurred in a 20-ha field in Hacienda de Tarazona, Seville, in southern Spain. Affected plants appeared singly or were grouped in circular patches as much as 8 to 10 m in diameter. Early symptoms consisted of water-soaked lesions on crown and lower stem tissue in contact with the soil. Plant foliage became pale green and wilted, followed by a complete collapse of the plant. A dense white mycelial mat formed on the lower stem and crown with 1- to 2-mm-diameter, spherical, tan-to-dark brown sclerotia. Lower stem pieces of 12 plants with early disease symptoms were surface sterilized in 0.5% NaOCl, dried, transferred to acidified potato dextrose agar, and incubated at 25 ± 1°C in the dark. Fast-growing fungal colonies with white mycelium and abundant sclerotia developed after 6 to 10 days of incubation. On the basis of morphological characters, the fungus was identified as Sclerotium rolfsii Sacc. (2). To confirm the identity of the pathogen, the ribosomal DNA internal transcribed spacer was amplified and sequenced for two isolates (one of the two exact sequences was deposited as GenBank Accession No. GU080230). The sequence was 99% similar to sequences of Athelia rolfsii (S. rolfsii) in GenBank. Pathogenicity of two isolates was determined by placing two oat seeds colonized by each isolate 0.5 to 1 cm from the stem of 2-week-old pepper plants cv. Cristal (one plant per pot, eight replicates). Plants were incubated in a growth chamber maintained at 28 ± 1°C with a 14-h photoperiod of 360 µE·m-2·s-1 and 60 to 90% relative humidity for 10 days. By the sixth day, discoloration and blight of the foliage and stem was observed. Sclerotia formed around the crown and 88% of the plants died 7 days after inoculation. S. rolfsii was recovered from all affected pepper plants. Noninoculated control plants did not develop symptoms. In southern Spain, S. rolfsii is widely distributed in areas of sugar beet production (1). Because of the wide host range of the pathogen, southern blight could become an important disease of vegetable production in southern Spain. References: (1) R. Jordán-Ramírez et al. IOBC/WPRS Bull. 42:101. 2009. (2) J. E. M. Mordue. CMI Descriptions of Pathogenic Fungi and Bacteria. No. 410, 1974.

Changes in the redox status of chickpea roots in response to infection by Fusarium oxysporum f. sp. ciceris: apoplastic antioxidant enzyme activities and expression of oxidative stress-related genes.

Activity levels of oxidative stress-related enzymes in the root apoplast during the interaction of WR315 (resistant) and JG62 (susceptible) chickpeas (Cicer arietinum L.) with the highly virulent race 5 of Fusarium oxysporum f. sp. ciceris were compared. Because this fungus develops asymptomatic infections in the chickpea root cortex in both susceptible and resistant plants, but only intrudes into the root xylem in the susceptible variety, the interactions were compared at three specific stages during disease development in JG62: (i) before symptom development (10 days after inoculation); (ii) at the time of appearance of the first disease symptoms (15-17 days after inoculation) and (iii) when all plants had developed disease symptoms (20-22 days after inoculation). Diamine oxidase (DAO), ascorbate peroxidase (APX), glutathione reductase (GR), guaiacol-dependent peroxidase and superoxide dismutase (SOD), but not catalase (CAT), were found in the apoplast of chickpea roots. In terms of APX activity, infection by the pathogen caused a different response in the incompatible compared to the compatible plant. In the case of GR, SOD and DAO activities, the pathogen caused the same response, but it developed earlier (i.e. GR and SOD) or to higher levels (i.e. DAO) in the incompatible interaction. Expression of apx, cat, sod, lipoxygenase (lox) and actin genes was also analysed in infected roots. Infection by F. oxysporum f. sp. ciceris race 5 only caused a significant change in the root expression of lox and actin genes. This up-regulation was earlier (lox) or higher (actin) in the incompatible than in the compatible interaction. Thus, changes in oxidative metabolism differ in compatible and incompatible interactions in Fusarium wilt of chickpea.

Spatiotemporal analysis of spread of infections by Verticillium dahliae pathotypes within a high tree density olive orchard in southern Spain.

The development of Verticillium wilt epidemics in olive cv. Arbequina was studied from November 1999 to May 2003 in a drip-irrigated, nontillage orchard established in a soil without a history of the disease at Córdoba, southern Spain. Disease incidence measured at 1-month-intervals increased from 0.2 to 7.8% during this period. Verticillium dahliae infecting the trees was characterized as defoliating (D) or nondefoliating (ND) pathotypes by a specific, multiplex-polymerase chain reaction (PCR) assay. Of the symptomatic trees, 87.2 and 12.8% were infected by the D or ND pathotypes, respectively. Dynamics of disease incidence were described by a generalized logistic model with a multiple sigmoid pattern. In the fitted model, the infection rate was highest in the winter to spring period and decreased to minimum values in the summer to fall period. Binary data of disease incidence was analyzed for point pattern and spatial correlation, either directly or after parsing them in contiguous quadrats. Overall, ordinary runs analysis indicated a departure from randomness of disease within rows. The binomial index of dispersion, interclass correlation, and Taylor's power law for various quadrat sizes suggested aggregation of diseased trees within the quadrat sizes tested. Spatial analysis by distance indices showed a nonrandom arrangement of quadrats containing infected trees. Spatial pattern was characterized by the occurrence of several clusters of infected trees. Increasing clustering over time was generally suggested by stronger values of clustering index over time and by the increase in the size of patch clusters. Significant spatial association was found in the clustering of diseased trees over time across cropping seasons; however, clustering was significant only for infections by D V. dahliae, indicating that infections by the D pathotype were aggregated around initial infections. The number and size of clusters of D V. dahliae-infected trees increased over time. Microsatellite-primed PCR assays of a representative number of V. dahliae isolates from diseased trees indicated that the majority of infecting D isolates shared the fingerprinting profile with D V. dahliae isolated from soil of a naturally infested cotton field in close proximity to the orchard, suggesting that short distance dispersal of the pathogen from this soil to the olive orchard may have occurred.

Infection by Meloidogyne artiellia does not break down resistance to races 0, 1a, and 2 of Fusarium oxysporum f. sp. ciceris in chickpea genotypes.

Fusarium oxysporum f. sp. ciceris, and the root-knot nematode Meloidogyne artiellia, coinfect chickpea crops in several countries of the Mediterranean Basin. The influence of root infection by M. artiellia on the reactions of chickpea genotypes with different reaction to infection with F. oxysporum f. sp. ciceris races 0, 1A, and 2 was investigated under controlled environmental conditions. Results demonstrated that co-infection of chickpea genotypes resistant to specific fungal races by M. artiellia did not influence the Fusarium wilt reaction of the plant, irrespective of the F. oxysporum f. sp. ciceris race assayed. However, in some of the assayed combinations, coinfection by both pathogens significantly affected the level of colonization by the fungus or reproduction of the nematode in the root system. Thus, coinfection of chickpea plants with Foc-0 and M. artiellia significantly decreased the level of colonization of the root system by F. oxysporum f. sp. ciceris in genotypes 'CA 336.14.3.0' and 'PV 61', but not in 'ICC 14216 K' and 'UC 27'. Similarly, the nematode reproduction index was also significantly reduced by coinfection with Foc-0 in the four chickpea genotypes tested and inoculated with this race. Conversely, coinfection of chickpea plants with Foc-1A and M. artiellia significantly increased colonization of the root system by the fungus in all genotypes inoculated with this race, except for line BG 212. Altogether, we confirmed the complete resistance phenotype of 'UC 27' and 'ICC 14216 K' to Foc-0, and of 'ICC 14216 K' to Foc-1A and Foc-2, and demonstrated that this resistance was not modified by coinfection of the resistant plant with M. artiellia.

First Report of Meloidogyne arenaria Parasitizing Lettuce in Southern Spain.

During the 2005-2006 autumn to winter lettuce-growing (Lactuca sativa cv. Iceberg) season, severely stunted and yellowing lettuce plants with disease incidence ranging from 80 to100% were observed in four commercial, fall-sown fields at Almodóvar del Río (Córdoba Province) in southern Spain. Early symptoms consisted of severely reduced growth of the plants that continued with extensive leaf yellowing and the absence of tight-head formation. Attacks by the disease were estimated to cause near complete loss of the crop yields since the lettuce head produced in affected fields were unmarketable. Observations of affected lettuce plants revealed high parasitism of the root system by a root-knot nematode (Meloidogyne sp.) in the main and feeder roots as well as heavy soil infestations by the nematode. The nematode was identified by the female perineal pattern, esterases phenotype, and a sequence-characterized amplified region polymerase chain reaction (SCAR-PCR) technique (1,2,4). Measurements and morphological observations of 20 second-stage juveniles (J2s) (body length = 463 ± 28 µm, dorsal gland orifice from stylet base = 2.8 ± 0.6 µm, stylet length = 10.4 ± 0.5 µm, tail length = 54.4 ± 0.6 µm; hyaline tail terminus = 9.4 ± 0.6 µm) and 10 adult females (stylet length = 14.5 ± 0.7 µm, dorsal gland orifice from stylet base = 4.7 ± 0.5 µm, and perineal pattern with low and rounded dorsal arch with coarse striae) conformed to the description of Meloidogyne arenaria (3). On the basis of the characteristics of the perineal pattern, the 2-band esterase phenotype, and the 420-bp SCAR fragment, the causal agent was identified as the peanut root-knot nematode M. arenaria. Nematodes were extracted from soil and root samples by standard procedures and their populations quantified. M. arenaria was detected in nearly all soil and root samples assessed, with nematode population densities ranging from 206 to 1,072 eggs and J2s per 5 g of fresh roots. Different Meloidogyne spp. have been reported parasitizing lettuce roots, especially M. hapla in northern areas (2); however, to our knowledge this is the first time that M. arenaria is reported parasitizing lettuce roots in Spain and elsewhere. References: (1) P. R. Esbenshade and A. C. Triantaphyllou. J. Nematol. 22:10, 1990. (2) N. A. Mitkowski et al. Plant Dis. 86:840, 2002. (3) K. J. Orton Williams. Meloidogyne arenaria. CIH Descriptions of Plant-Parasitic Nematodes. Set 5, No. 62. Commonwealth Institute of Helminthology, St. Albans, 1975. (4) C. Zijlstra et al. Nematology 2:847, 2000.

First Report of Broomrape (Orobanche crenata) Infecting Lettuce in Southern Spain.

Broomrapes (Orobanche spp., Orobanchaceae) are chlorophyll-lacking, obligately parasitic flowering plants that infect roots of many dicotyledoneous species and cause severe damage to vegetable and field crops worldwide, but particularly in North Africa, southern and eastern Europe, and the Middle East. (1). Orobanche crenata is one of the most important broomrapes and mainly infects legume crops (2). In January 2006, we observed severe broomrape attacks in four commercial fields of fall-sown lettuce (Lactuca sativa cv. Iceberg) crops at Almodóvar del Río (Córdoba Province) in southern Spain. Infected lettuce plants showed severe stunting, foliar yellowing, and had loose-formed heads. Infection of lettuce plants by Orobanche sp. was confirmed by removing plants to verify the attachment of broomrapes to lettuce roots. There were one to four broomrapes per lettuce plant. Incidence of infected lettuce ranged from 10 to 20% in different areas of the fields. Morphological observations of broomrape plants identified the parasite as O. crenata. The main botanical features were as follows: plants 20 to 40 cm tall; corolla 20 to 28 mm, white, lips with lilac, divergent veins, lower lip large with suborbicular lobes, not ciliate; filaments hairy, obliquely inserted 2 to 4 mm above the base of corolla, with short glandular hairs in the upper third; anthers glabrous, 2 to 2.5 mm in length, and stigma yellow or pinkish at anthesis (2). O. crenata also was observed infecting faba bean (Vicia faba) plants in a field in close proximity to the affected lettuce fields. The complete 5.8S ribosomal DNA gene and internal transcribed spacers (ITS) 1 and 2 of O. crenata were sequenced using adventitious roots and stem tissues sampled from infected faba bean and lettuce plants (Genbank Accession Nos. DQ458908 and DQ458909) by standard protocols (3). A nucleotide BLAST search revealed that both sequences were identical and share 100% similarity with three reported ITS1-5.8S-ITS2 sequences from two Orobanche spp. (O. crenata and O. minor; Genbank Accession Nos. AY209267, AY209266, and AY209272). On the basis of the morphological characters described above, the parasite was O. crenata and not O. minor. O. crenata has been reported infecting many legume crops in southern Spain, including faba bean, pea, lentil, and vetch. To our knowledge, this is the first report of O. crenata infecting lettuce in Spain and elsewhere. The high incidence of O. crenata on legume crops, and the severe infections found on lettuce plants suggest that this parasitic plant may be an important constraint for fall-sown lettuce in southern Spain. References: (1) A. O. Chater and D. A. Webb. Orobanchaceae. In: Flora Europaea, T. G. Tutin et al., eds. Vol. 3. Cambridge University Press, Cambridge, 1972. (2) A. J. Pujadas-Salvà. Orobanchaceae L. In: Plantas Parásitas de la Península Ibérica y Baleares. J. A. López Sáez et al., eds. Mundi-Prensa, Madrid, 2002. (3) G. M. Schneeweiss et al. Mol. Phylogenet. Evol. 30:465, 2004.

Influence of Temperature and Inoculum Density of Fusarium oxysporum f. sp. ciceris on Suppression of Fusarium Wilt of Chickpea by Rhizosphere Bacteria.

The effects of temperature and inoculum density of Fusarium oxysporum f. sp. ciceris race 5 on suppression of Fusarium wilt in chickpea (Cicer arietinum) cv. PV 61 by seed and soil treatments with rhizobacteria isolated from the chickpea rhizosphere were studied in a model system. Disease development over a range of temperatures (20, 25, and 30 degrees C) and inoculum densities (25 to 1,000 chlamydospores per gram of soil) was described by the Gompertz model. The Gompertz relative rate of disease progress and final amount of disease increased exponentially and monomolecularly, respectively, with increasing inoculum densities. Disease development was greater at 25 degrees C compared with 20 and 30 degrees C. At 20 and 30 degrees C, disease development was greater at 250 to 1,000 chlamydospores per gram of soil compared with 25 to 100 chlamydospores per gram of soil. At 25 degrees C, increasing inoculum densities of the pathogen did not influence disease. Nineteen Bacillus, Paenibacillus, Pseudomonas, and Stenotrophomonas spp. out of 23 bacterial isolates tested inhibited F. oxysporum f. sp. ciceris in vitro. Pseudomonas fluorescens RGAF 19 and RG 26, which did not inhibit the pathogen, showed the greatest Fusarium wilt suppression. Disease was suppressed only at 20 or 30 degrees C and at inoculum densities below 250 chlamydospores per gram of soil. Bacterial treatments increased the time to initial symptoms, reduced the Gompertz relative rate of disease progress, and reduced the overall amount of disease developed.

Yield loss in chickpeas in relation to development of fusarium wilt epidemics.

ABSTRACT Development of 108 epidemics of Fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris were studied on cvs. P-2245 and PV-61 in field microplots artificially infested with races 0 and 5 of F. oxysporum f. sp. ciceris in 1986 to 1989. Disease progression data were fitted to the Richards model using nonlinear regression. The shape parameter was influenced primarily by date of sowing and, to a lesser extent, by chick-pea cultivars and races of F. oxysporum f. sp. ciceris. Fusarium wilt reduced chickpea yield by decreasing both seed yield and seed weight. These effects were related to sowing date, chickpea cultivar, and virulence of the prevalent F. oxysporum f. sp. ciceris race. Regression models were developed to relate chickpea yield to Fusarium wilt disease intensity with the following independent variables: time to initial symptoms (t(is)), time to inflection point (t(ip)) of the disease intensity index (DII) progress curve, final DII (DII(final)), standardized area under DII progress curve (SAUDPC), and the Richards weighted mean absolute rate of disease progression (rho). Irrespective of the chickpea cultivar x pathogen race combination, the absolute and relative seed yields decreased primarily by delayed sowing. The relative seed yield increased with the delay in t(is) and t(ip) and decreased with increasing DII(final), SAUDPC, and rho. A response surface as developed in which seed yield loss decreased in a linear relationship with the delay in t(is) and increased exponentially with the increase of rho.

Effect of Sowing Date, Host Cultivar, and Race of Fusarium oxysporum f. sp. ciceris on Development of Fusarium Wilt of Chickpea.

ABSTRACT Microplots experiments were carried out at Córdoba, southern Spain, from 1986 to 1989 to determine the effects of sowing date in the management of Fusarium wilt of chickpea as influenced by virulence of the pathogen race and by cultivar susceptibility. A total of 108 epidemics of the disease were described, analyzed, and compared to assess the degree of disease control. The epidemics were characterized by five curve elements: final disease intensity index (DII), standardized area under DII progress curve, time to epidemic onset, time to inflection point (t(ip)), and the DII value at t(ip), the last two parameters being estimates from the Richards function adjusted by nonlinear regression analysis. The structure of Fusarium wilt epidemics was examined by conducting multivariate principal components and cluster analyses. From these analyses, three factors accounting for 98 to 99% of the total variance characterized the DII progress curves and provided plausible epidemiological interpretations. The first factor included the t(ip) and the time to disease onset and can be interpreted as a positional factor over time. This factor accounted for the largest proportion of the total variance and may, therefore, be considered as the main factor for analysis of Fusarium wilt epidemics. The second factor concerns the standardized area under DII progress curves and the final DII of the epidemics. The third factor identified the uniqueness of the estimated value for the point of inflection of the DII progress curve over time. Our results indicate that for each year of experiment epidemic development was related mainly to the date of sowing. Thus, for chickpea crops in southern Spain, advancing the sowing date from early spring to early winter can slow down the development of Fusarium wilt epidemics, delay the epidemic onset, and minimize the final amount of disease. However, the net effect of this disease management practice may also be influenced, though to a lesser extent, by the susceptibility of the chickpea cultivar and the virulence and inoculum density of the Fusarium oxysporum f. sp. ciceris race.

Phenology of Didymella rabiei Development on Chickpea Debris Under Field Conditions in Spain.

ABSTRACT The development of Didymella rabiei on debris of naturally infected chickpea was investigated in four chickpea-growing areas with different climatic conditions in Spain during 1987 to 1992. D. rabiei extensively colonized chickpea debris and formed pseudothecia and pycnidia. Differentiation of pseudothecial initials occurred regularly across experimental locations by November, 1 month after placement of debris on the soil. Ascospore maturation occurred mainly from late January to late March, depending on location and year. Maximum ascospore discharge from sampled debris pieces placed under suitable environmental conditions occurred 2 to 4 weeks after ascospore maturation, after which ascospore release decreased sharply. Pseudothecia were exhausted, due to ascospore discharge, by the beginning of summer. New asci did not develop in empty pseudothecia and no pseudothecia formed in tissues after the first season. Ascospore maturation and liberation in cooler locations were more uniform and occurred later compared to maturation in warmer locations. Also, production of asci and ascospores per pseudothecium was much higher in cooler than in warmer locations. A similar relationship was found for density of pseudothecia and pycnidia and conidia production per pycnidium. The percentage of mature pseudothecia increased according to the logistic model, with the cumulative number of Celsius degree days calculated by computing the mean of the maximum and minimum daily air temperatures on rainy days from the date of debris placement on the soil. There were significant differences among model parameter estimates between cooler and warmer locations, but minor differences were found among parameters for locations with similar environmental conditions. There was an inverse linear relationship between the average temperature during the period of pseudothecia maturation and the number of asci produced per pseudothecium.

Interactions of Pratylenchus thornei and Fusarium oxysporum f. sp. ciceris on Chickpea.

ABSTRACT Fusarium oxysporum f. sp. ciceris and the root-lesion nematode Pratylenchus thornei coinfect chickpeas in southern Spain. The influence of root infection by P. thornei on the reaction of Fusarium wilt-susceptible (CPS 1 and PV 61) and wilt-resistant (UC 27) chickpea cultivars to F. oxysporum f. sp. ciceris race 5 was investigated under controlled and field conditions. Severity of Fusarium wilt was not modified by coinfection of chickpeas by P. thornei and F. oxysporum f. sp. ciceris, in simultaneous or sequential inoculations with the pathogens. Root infection with five nematodes per cm(3) of soil and 5,000 chlamydospores per g of soil of the fungus resulted in significantly higher numbers of propagules of F. oxysporum f. sp. ciceris with the wilt-susceptible cultivar CPS 1, but not with the wilt-resistant one. However, infection with 10 nematodes per cm(3) of soil significantly increased root infection by F. oxysporum f. sp. ciceris in both cultivars, irrespective of fungal inoculum densities (250 to 2,000 chlamydospores per g of soil). Plant growth was significantly reduced by P. thornei infection on wilt-susceptible and wilt-resistant chickpeas in controlled and field conditions, except when shorter periods of incubation (45 days after inoculation) were used under controlled conditions. Severity of root necrosis was greater in wilt-susceptible and wilt-resistant cultivars when nematodes were present in the root, irrespective of length of incubation time (45 to 90 days), densities of nematodes (5 and 10 nematodes per cm(3) of soil), fungal inocula, and experimental conditions. Nematode reproduction on the wilt-susceptible cultivars, but not on the wilt-resistant one, was significantly increased by F. oxysporum f. sp. ciceris infections under controlled and field conditions.