Critical factors driving aphanomyces damping‐off and root disease in clover revealed and explained using linear and generalized linear models and boosted regression trees

Subterranean clover (Trifolium subterraneum) is an important forage legume in Mediterranean regions worldwide. Aphanomyces damping‐off and root disease (Aphanomyces trifolii) poses significant threat to its persistence and productivity. Studies were conducted to define how environmental explanatory variables (temperature, soil type, moisture, nutrition) and variety influence disease severity and consequent forage productivity and persistence. Relationships were modelled using linear and generalized linear models and boosted regression trees. Linear modelling highlighted complex relationships between environmental variables and each dependent variable (emergence, tap and lateral root disease, dry shoot and root weight). All environmental variables produced significant interaction and/or main effects within each dependent variable. Boosted regression trees supported the complex nature of relationships in linear models, with temperature and either soil or variety most, and nutrition least, influential. Heat maps showed more disease for low temperatures. Least tap root disease was under high temperatures, while least lateral root disease was under medium or high temperatures, low moisture, and in sand‐based soil. These are the first studies using modelling approaches to reveal the complexities of how fluctuating soil temperature, moisture and nutrition conditions, and soil type and variety, determine aphanomyces damping‐off and root disease severity and resultant adverse impacts on forage legume productivity and persistence. Outcomes are widely applicable across soilborne oomycete pathogens of forage legumes. Studies highlighted how warming temperatures and drying climate associated with climate change should reduce future impact and importance of this and other soilborne oomycete diseases of forage legumes favoured by cold temperatures and wet and waterlogged conditions.     

Severity of phytophthora root rot and pre‐emergence damping‐off in subterranean clover influenced by moisture,temperature, nutrition,soil type,cultivar and their interactions

Studies were carried out in controlled environment rooms reflecting field situations. In the presence of the devastating soilborne pathogen Phytophthora clandestina, subterranean clover (Trifolium subterraneum) seedling emergence was significantly affected by moisture, soil type, temperature and cultivar. The level of rotting of tap and lateral roots was significantly affected by nutrition, soil type, temperature and cultivar. There were significant interactions involving temperature, moisture, soil type and cultivar; cultivar resistance, high moisture, high or medium temperature, high nutrition and sand soil all contributed towards less pre‐emergence damping‐off and tap and lateral root disease and to greater clover productivity. Host resistance of subterranean clover cultivars was critical for reducing disease severity and increasing productivity, even when favourable environmental conditions for severe disease occurred. In the presence of P. clandestina, the most resistant cultivar, Seaton Park, performed best under a high temperature, high nutrition and high moisture combination, but showed lower productivity under conditions of low nutrition or lower temperature, even when moisture level was high. In contrast, less resistant cultivars Riverina and Meteora had less disease and greater productivity under low moisture conditions. Findings reflect field observations that pre‐emergence damping‐off and root disease from P. clandestina in subterranean clover is particularly severe under colder conditions and in nutritionally impoverished sandy soils, and demonstrate how variations in soil type, nutrition, moisture, temperature and cultivar have profound effects on the expression and severity of phytophthora pre‐emergence damping‐off and root disease and the productivity of subterranean clover forages.     

Inactivation of seed-treatment fungicides by Rhizobium

2-Methoxyethylmercury chloride (MEMC) applied to cowpea seeds with Rhizobium provided little or no control of seedling rot caused by Rhizoctonia solani; similar treatment, but without Rhizobium, gave >40% control of disease. Treating seeds with quintozene and then with Rhizobium rendered the fungicide completely ineffective against R. solani infections; disease control was >70% when seeds were treated with quintozene only, and >60% when seeds were coated first with Rhizobium and then with quintozene; treatment with quintozene+Rhizobium gave about 25% control. Simultaneous treatment with Rhizobium considerably reduced the efficacy of captafol, chloroneb, carbendazim and thiophanate-methyl. Disease control by carboxin, benomyl and thiabendazole remained unaltered in the presence of Rhizobium, but only thiabendazole gave good control. Seeds treated with MEMC and Rhizobium produced seedlings without root nodules; nodulation was also decreased by captafol and by carboxin but six other fungicides had no effect. The implications of Rhizobium-fungicides interactions are discussed in the context of fungicidal control of root diseases.     

Host resistances to Aphanomyces trifolii root rot of subterranean clover: first opportunity to successfully manage this severe pasture disease

Subterranean clover (Trifolium subterraneum) is an important pasture legume in Australia (29 million ha) and elsewhere. However, severe pasture decline occurs in association with several root pathogens, including Aphanomyces trifolii, that has been misidentified for decades as A. euteiches until recently confirmed as A. trifolii. A series of controlled environment experiments was undertaken to identify host resistance to A. trifolii in subterranean clover and to compare virulence and phylogeny of isolates. In experiment 1, Dalkeith, Bacchus Marsh, Riverina and Yarloop were the most resistant of 38 cultivars with a percentage disease index (PDI) ≤10 for both tap and lateral roots. Experiment 2 confirmed resistance of Yarloop, but a change in some relative varietal resistances suggested physiological specialization among A. trifolii isolates. Experiment 3 confirmed extensive variation in virulence and physiological specialization across 23 isolates of A. trifolii, with three distinct clades, two of which were distinct from isolates collected previously. Experiment 4 identified host resistance(s) effective against a mixture of 20 A. trifolii isolates, but the most resistant cultivars (Antas, Uniwager, Leura) still showed significant disease. This is the first study to show physiological specialization in A. trifolii and to identify host resistance. This study defines A. trifolii as a significant but largely unknown contributor to severe root disease of subterranean clover in southern Australia. Finally, development and calibration of a new soil commercial DNA test not only enables field quantification of the disease, but development of appropriate breeding, selection and farm management strategies to reduce its impact.     

Medicago truncatula as a model host for studying legume infecting Rhizoctonia solani and identification of a locus affecting resistance to root canker

The broad‐host‐range necrotizing fungal pathogen Rhizoctonia solani is responsible for economically significant diseases to crops as diverse as wheat, maize, barley, canola, sugar beet, potato, soyabean, bean, lupin and alfalfa. Germplasm screens in many of the crop hosts have not identified strong genetic resistance which, together with the lack of effective control, mean the pathogen remains a substantial problem for agriculture in many parts of the world. Following the establishment of a robust inoculation assay, a germplasm collection of the model legume Medicago truncatula was screened with various legume‐infecting isolates of R. solani. While some significant differences in susceptibility/resistance were detected between some lines, in the majority of cases M. truncatula was susceptible to R. solani. Comparison of a legume‐ and cereal‐infecting AG8 isolate with a legume‐specific AG11 isolate revealed no difference in pathogenicity between the two isolates when infecting M. truncatula. The most significant differences in susceptibility occurred with an AG6 isolate, which caused root canker. This included significant differences between the moderate resistance of the M. truncatula reference genotype A17 and the high susceptibility of line A20. The analysis of a recombinant inbred line population derived from A17 and A20 revealed a single locus contributing to the resistance in A17. Interestingly, the locus only affected the development of post‐emergent (late) symptoms, such as necrosis of cotyledons at 11 days after inoculation and root‐ and above‐ground‐weights, but not pre‐emergent seedling damping off. These findings pave the way for further studies to dissect the genetic and molecular mechanisms of resistance.     

Suppression of Rhizoctonia root-rot of tomato by Glomus mossae BEG12 and Pseudomonas fluorescens A6RI is associated with their effect on the pathogen growth and on the root morphogenesis

Rhizoctonia solani root-rot is a major soilborne disease causing growth and yield depression. The ability of Glomus mosseae BEG12 and Pseudomonas fluorescens A6RI to suppress this soilborne disease in tomato was assessed by comparing the shoot and root growth of plants infested with R. solani 1556 when protected or not by these beneficial strains. The epiphytic and parasitic growth of the pathogenic R. solani 1556 was compared in the presence and absence of the biocontrol agents by microscopical observations allowing the quantification of roots with hyphae appressed to epidermal cells (epiphytic growth) and of roots with intraradical infection (parasitic growth). The root architecture of the tomato plants under the different experimental conditions was further characterized by measuring total root length, mean root diameter, number of root tips and by calculating degree of root branching. G. mosseae BEG12 and P. fluorescens A6RI fully overcame the growth depression caused by R. solani 1556. This disease suppression was associated with a significant decrease of the epiphytic and parasitic growth of the pathogen together with an increase of root length and of the number of root tips of inoculated tomato plants. The combined effects of G. mosseae BEG12 and P. fluorescens A6RI on pathogen growth and on root morphogenesis are suggested to be involved in the efficient disease suppression.     

Plant defense activation and management of tomato root rot by a chitin-fortified <Emphasis Type="Italic">Trichoderma/Hypocrea</Emphasis> formulation

Tomato root rot caused by Rhizoctonia solani is a major soilborne disease resulting in significant yield loss. The culture filtrates of six isolates of Trichoderma/Hypocrea species were evaluated for in vitro production of hydrolytic enzymes. Results demonstrated that all the six isolates were able to produce chitinase, β-1, 3 glucanase and protease in the range of 76–235 μmol GlcNAc min^-1 mg^-1 protein, 31.90–37.72 nmol glucose min^-1 mg^-1 proteins and 63.05–86.22 μmol min^-1 mg^-1 proteins, respectively. Trichoderma/Hypocrea-based formulation(s) were prepared with chitin (1% v:v) and CMC (0.5% w:v) for root rot management in a greenhouse. Root dip application with bioformulation(s) resulted in a significant reduction of the root rot index. In addition, bioformulations increased plant growth attributing traits significantly relative to untreated control. Accumulation of total phenols, peroxidase, polyphenoloxidase and phenylalanine ammonia lyase increased in chitin-supplemented Trichoderma/Hypocrea formulation-treated plants challenged with R. solani. The results suggest that chitin-fortified bioformulation(s) could be an effective system to control root rot of tomato in an eco-compatible manner.     

Brachypodium distachyon is a pathosystem model for the study of the wheat disease rhizoctonia root rot

Brachypodium distachyon (Bd) is increasingly being used as a model for cereal diseases and to study cereal root architecture. Rhizoctonia solani AG 8 is a necrotrophic root pathogen that infects wheat soon after germination resulting in reduced plant growth and yield loss. Genetic resistance to R. solani AG 8 is not available in commercial wheat cultivars, although some quantitative levels of resistance have previously been found in mutant lines and grass relatives. Resistance mechanisms in cereals remain unknown. The ability to use Bd as a model to study the wheat–R. solani AG 8 pathosystem was investigated. The results presented show that Bd is susceptible to R. solani AG 8 and that the pathogen infects both species to a similar degree, producing comparable disease symptoms. Root length reduction was the primary indicator of disease, with shoots also affected. The second objective was to develop a repeatable phenotyping method to screen Bd populations for resistance to R. solani AG 8. Results of a preliminary experiment provide evidence for variation in resistance between Bd inbred lines. This is the first report showing the potential of Bd as a model plant for discovery of quantitative genetic variation in resistance to a necrotrophic cereal root pathogen.     

Virulence of ‘Dickeya solani’ and Dickeya dianthicola biovar‐1 and ‐7 strains on potato (Solanum tuberosum)

Studies were conducted to explain the relative success of ‘Dickeya solani’, a genetic clade of Dickeya biovar 3 and a blackleg‐causing organism that, after recent introduction, has spread rapidly in seed potato production in Europe to the extent that it is now more frequently detected than D. dianthicola. In vitro experiments showed that both species were motile, had comparable siderophore production and pectinolytic activity, and that there was no antagonism between them when growing. Both ‘D. solani’ and biovar 1 and biovar 7 of D. dianthicola rotted tuber tissue when inoculated at a low density of 10³ CFU mL^?1. In an agar overlay assay, D. dianthicola was susceptible to 80% of saprophytic bacteria isolated from tuber extracts, whereas ‘D. solani’ was susceptible to only 31%, suggesting that ‘D. solani’ could be a stronger competitor in the potato ecosystem. In greenhouse experiments at high temperatures (28°C), roots were more rapidly colonized by ‘D. solani’ than by biovar 1 or 7 of D. dianthicola and at 30 days after inoculation higher densities of ‘D. solani’ were found in stolons and progeny tubers. In co‐inoculated plants, fluorescent protein (GFP or DsRed)‐tagged ‘D. solani’ outcompeted D. dianthicola in plants grown from vacuum‐infiltrated tubers. In 3 years of field studies in the Netherlands with D. dianthicola and ‘D. solani’, disease incidence varied greatly annually and with strain. In summary, ‘D. solani’ possesses features which allow more efficient plant colonization than D. dianthicola at high temperatures. In temperate climates, however, tuber infections with ‘D. solani’ will not necessarily result in a higher disease incidence than infections with D. dianthicola, but latent seed infection could be more prevalent.     

Suppressive activity of some glucosinolate enzyme degradation products on Pythium irregulare and Rhizoctonia solani in sterile soil

Glucosinolate degradation products (GLDPs), mainly isothiocyanates, obtained by myrosinase‐catalysed hydrolysis of glucosinolates (GLs), are an important group of natural bioactive substances. The fungitoxic activity of four GLDPs, chosen on the basis of their side‐chain structures, was assayed against Pythium irregulare and Rhizoctonia solani. The effectiveness of the degradation products (DPs) of sinigrin (alkenyl GL), epi‐progoitrin (hydroxy‐alkenyl GL), glucoiberin and glucoerucin (thiofunctionalised GLs) in inhibiting P irregulare oospore germination and R solani soil colonisation were tested in a closed system, using an artificially infected soil. The fungitoxic activity of these GLDPs varied according to their side‐chain structure. As in previous in vitro studies, the thiofunctionalised GLDPs were found the most effective, producing complete inhibition of P irregulare oospore germination (0.01 µmole g⁻¹ soil) and R solani soil colonisation (0.5 µmole g⁻¹ soil), but even sinigrin DP showed a fungitoxic activity higher than that of epi‐progoitrin DP. © 2000 Society of Chemical Industry     

Rhizoctonia solani: taxonomy,population biology and management of rhizoctonia seedling disease of soybean

Rhizoctonia solani, the most important species within the genus Rhizoctonia, is a soilborne plant pathogen with considerable diversity in cultural morphology, host range and aggressiveness. Despite its history as a destructive pathogen of economically important crops worldwide, our understanding of its taxonomic relationship with other Rhizoctonia‐like fungi, incompatibility systems, and population biology is rather limited. Among the host of diseases it has been associated with, seedling diseases inflicted on soybean are of significant importance, especially in the soybean growing regions of North America. Due to the dearth of resistant soybean genotypes, as well as the paucity of information on the mechanisms of host–pathogen interactions and other molecular aspects of pathogenicity, effective management options have mostly relied upon a combination of cultural and chemical control options. The first section of this review summarizes what is currently known about the taxonomy and systematics, population biology and molecular genetics of R. solani. The second section provides an overview of the pathology and management of rhizoctonia root and hypocotyl rot of soybean, a seedling disease of importance in North America.     

Soil properties related to suppression of Rhizoctonia solani on tobacco fields from northwest Argentina

Biotic and abiotic factors from soils have been implicated in the disease suppression of Rhizoctonia solani. This study included a Eucalyptus twig baiting assay, disease index and qPCR quantification of R. solani, and physicochemical analysis of 10 tobacco soils from five different locations (V: Vaqueros, C: Cerrillos, R: Rosario de Lerma, SA: San Agustín, CH: Chicoana) in the northwest of Argentina. Levels of Rhizoctonia soil inoculum quantified by baiting assay and qPCR were positively correlated. However, there was no correlation with root rot disease index in tobacco fields. Soils from V1, SA2 and CH2 fields, which reduced root rot disease on tobacco plants, were suppressive to R. solani infection. High clay, pH, organic matter content and physical stability in tobacco soils were the main physicochemical properties that limited Rhizoctonia development. Interestingly, growth of R. solani subgroups AG4-HGI and AG4-HGIII was highly suppressed in V1 and CH2 fields, and in SA2 fields, respectively. Undisturbed soil from a local forested mountain also resulted in reduction of growth of AG4-HGIII and AG4-HGI, while AG2-1 was less affected, suggesting that high soil organic matter contributed to suppression of R. solani. Soils highly suppressive of R. solani had significantly different populations of culturable bacteria, Pseudomonas and fungi, but populations of actinobacteria and Trichoderma spp. did not differ. These different populations may be involved in the inhibition of fungal growth. The results demonstrated that physicochemical and biological properties of soil suppressive to R. solani could act as an alternative for controlling Rhizoctonia diseases on tobacco.     

Interaction ofPseudomonas fluorescens withRhizobium for their effect on the management of peanut root rot

A biocontrol agent (Pseudomonas fluorescens) and a phytostimulator (Rhizobium) have been shown to have beneficial effects on plant growth and health. The study of plants inoculated withPseudomonas andRhizobium requires special attention because of the possibility that these agents may influence each other. Our study was conducted to test the effect of these inoculants on co-inoculation in peanut to control root rot, a severe soilborne disease caused byMacrophomina phaseolina. One fluorescent pseudomonad strain, Pf 1, which effectively inhibited the mycelial growth ofM. phaseolina underin vitro conditions, was studied for its compatibility with the biofertilizer bacterial strainRhizobium TNAU 14. Dual culture and colorimetric studies indicated the existence of a positive interaction between the microbial inoculants. However, glasshouse and field studies showed seed treatment and soil application ofPseudomonas fluorescens Pf 1 to be the most effective treatment in reducing root rot incidence and improving the crop vigor index, in comparison with treatments in which both inoculants were applied. http://www.phytoparasitica.org posting Feb. 11, 2002.     

Effects of plant growth-promoting rhizobacteria on nodulation of <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L. are dependent on plant P nutrition

Several plant growth-promoting rhizobacteria (PGPR) have shown potential to enhance nodulation of legumes when coinoculated with Rhizobium. To optimize the efficiency of these Rhizobium-PGPR-host plant interactions, unravelling the underlying mechanisms and analyzing the influence of specific environmental conditions is crucial. In this work the effect of four PGPR strains on the symbiotic interaction between Rhizobium and common bean (Phaseolus vulgaris) was studied under deficient versus sufficient phosphorus supply. It was observed that the effect on nodulation of three out of four PGPR tested was strongly dependent on P nutrition. Further, the use of specific PGPR mutant strains indicated that bacterial indole-3-acetic-acid production (IAA) and 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity play an important role in the host nodulation response, particularly under low P conditions. Moreover, it was shown that the differential response to PGPR under low versus high P conditions was associated with changes in the host hormone sensitivity for nodulation induced under P deficiency. These findings contribute to the understanding of the interplay between Rhizobium, PGPR and the plant host under different environmental settings.     

Pathogenicity and reproductive fitness of <Emphasis Type="Italic">Pratylenchus coffeae</Emphasis> and <Emphasis Type="Italic">Radopholus arabocoffeae</Emphasis> on Arabica coffee seedlings (<Emphasis Type="Italic">Coffea arabica</Emphasis> cv. Catimor) in Vietnam

The pathogenicity and reproductive fitness of Pratylenchus coffeae and Radopholus arabocoffeae from Vietnam on coffee (Coffea arabica) seedlings cv. Catimor were evaluated in greenhouse experiments. The effect of initial population densities (Pi = 0, 1, 2, 4, 8, 16, 32, 64, 128, and 256 nematodes per cm³ soil) was studied for both species at different days after inoculation (dai). The data were adjusted to the Seinhorst damage model Y = m + (1-m).z^Pi-T. Tolerance limit (T) for P. coffeae was zero for the height and the diameter of the coffee plants. For the diameter, the T-value for R. arabocoffeae was 25.6 for 30 and 60 dai and 12.8 for 90 and 120 dai. After 4 months T was zero. The low tolerance limits indicate that Arabica coffee is highly intolerant to both nematode species. At the end of the experiment (180 dai), all plants were infected and most were dead when inoculated with R. arabocoffeae at initial densities of 32, 64, 128 and 256 nematodes/cm³ soil. For P. coffeae plant death was already observed at the lowest inoculation densities. Growth of coffee was reduced at all inoculation levels for both species. Pratylenchus coffeae and R. arabocoffeae caused intense darkening of the roots, leaf chlorosis and a strong reduction of root and shoot growth. It was observed that P. coffeae mainly destroyed lateral roots rather than tap roots, whereas R. arabocoffeae reduced tap root length rather than the lateral roots. At the lowest inoculum densities, the reproduction factor of P. coffeae was 2.38 and 2.01 for R. arabocoffeae, indicating that arabica coffee is a host for both species. Plant growth as expressed by shoot height and shoot and root weight measured 60 dai was negatively correlated with nematode (both species) density as expressed by the geometric mean of nematode numbers at 30 and 60 dai.     

Virulence of Pectobacterium carotovorum subsp. brasiliense on potato compared with that of other Pectobacterium and Dickeya species under climatic conditions prevailing in the Netherlands

In western Europe, Pectobacterium carotovorum subsp. brasiliense is emerging as a causal agent of blackleg disease. In field experiments in the Netherlands, the virulence of this pathogen was compared with strains of other Dickeya and Pectobacterium species. In 2013 and 2014, seed potato tubers were vacuum infiltrated with high densities of bacteria (10⁶ CFU mL^?1) and planted in clay soil. Inoculation with P. carotovorum subsp. brasiliense and P. atrosepticum resulted in high disease incidences (75–95%), inoculation with D. solani and P. wasabiae led to incidences between 5% and 25%, but no significant disease development was observed in treatments with P. carotovorum subsp. carotovorum, D. dianthicola or the water control. Co‐inoculations of seed potatoes with P. carotovorum subsp. brasiliense and D. solani gave a similar disease incidence to inoculation with only P. carotovorum subsp. brasiliense. However, co‐inoculation of P. carotovorum subsp. brasiliense with P. wasabiae resulted in a decrease in disease incidence compared to inoculation with only P. carotovorum subsp. brasiliense. In 2015, seed potatoes were inoculated with increasing densities of P. carotovorum subsp. brasiliense, D. solani or P. atrosepticum (10³–10⁶ CFU mL^?1). After vacuum infiltration, even a low inoculum density resulted in high disease incidence. However, immersion without vacuum caused disease only at high bacterial densities. Specific TaqMan assays were evaluated and developed for detection of P. carotovorum subsp. brasiliense, P. wasabiae and P. atrosepticum and confirmed the presence of these pathogens in progeny tubers of plants derived from vacuum‐infiltrated seed tubers.     

Characterization of fungi (Fusarium and Rhizoctonia) and oomycetes (Phytophthora and Pythium) associated with apple orchards in South Africa

Several species of fungi and oomycetes including Fusarium, Rhizoctonia, Phytophthora and Pythium have been reported as root pathogens of apple where they contribute to a phenomenon known as apple replant disease. In South Africa, little is known about specific species in these genera and their pathogenicity toward apple. Therefore, these aspects were investigated along with the development and optimization of qPCR tests for detection and quantification of the most virulent oomycete species. In eight investigated orchards, the oomycete Phythophthora cactorum was widely distributed, while nine Pythium species were differentially distributed among the orchards. Pythium irregulare was the most widely distributed and the most virulent species along with P. sylvaticum, P. vexans and Ph. cactorum. Seven binucleate Rhizoctonia anastomosis groups (AGs) were also differentially distributed among the orchards, with the majority appearing to be non-pathogenic while certain AG-I and AG-F isolates exhibited low virulence on apple. In the genus Fusarium, F. oxysporum was widely distributed, but isolates were non-pathogenic. Fusarium solani and F. avenaceum were less frequently encountered, with only some isolates having low virulence. qPCR data obtained from seedling roots inoculated with the most virulent Pythium species (P. irregulare, P. sylvaticum and P. vexans) and the genus Phytophthora were not always reproducible between trials, or isolates of the same species. In general, seedling growth inhibition was associated with the presence of a low amount of pathogen DNA (±40 fg μl⁻¹ to 2 pg μl⁻¹) in roots. Pythium irregulare, although having the lowest DNA concentrations in roots, was the only species for which a significant negative correlation was found between seedling weight and pathogen DNA concentration.     

Genetic structure of populations of Rhizoctonia solani AG‐4 from five provinces in Iran

Rhizoctonia solani anastomosis group 4 (AG‐4) is a serious pathogen causing damping off and root rot in many important crop plants. A total of 190 isolates of R. solani AG‐4 HG‐I were collected from host fields in five provinces of Iran. The genetic structure of this pathogen was evaluated using seven microsatellite loci, focusing particularly on geographic differentiation. Most of the multilocus genotypes (MLGTs) were unique, with few MLGTs shared among populations. High to moderate levels of gene flow among populations was indicated by low to moderate differentiation between pairs of populations based on the fixation index (F_ST). Gametic equilibrium of most pairs of microsatellite loci and moderate genotypic diversity were found for two out of five populations, indicating that these populations were sexually recombining in structure. High genotypic diversity, moderate clonal fractions and site‐specific genotypes were consistent with mixed reproductive systems for the remaining populations. The findings of departures from Hardy–Weinberg (HW) equilibrium, gametic disequilibrium and a significant excess of homozygotes in half or more than half of the loci were probably caused by the presence of null alleles and the Wahlund effect. This is the first study to consider the population genetics of the root and crown rot pathogen R. solani AG‐4.     

<Emphasis Type="Italic">Trichoderma harzianum</Emphasis>- mediated reprogramming of oxidative stress response in root apoplast of sunflower enhances defence against <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

Trichoderma harzianum is an effective biocontrol agent against the devastating plant pathogen Rhizoctonia solani. Despite its wide application in agriculture, the mechanisms of biocontrol are not yet fully understood. Mycoparasitism and antibiosis are suggested, but may not be sole cause of disease reduction. In the present study, we investigated the role of oxidant-antioxidant metabolites in the root apoplast of sunflower challenged by R. solani in the presence/absence of T. harzianum NBRI-1055. Analysis of oxidative stress response revealed a reduction in hydroxyl radical concentration (^•OH; 3.6 times) at 9 days after pathogen inoculation (dapi), superoxide anion radical concentration (O₂^•−; 4.1 times) at 8 dapi and hydrogen peroxide concentration (H₂O₂; 2.7 times) levels at 7 dapi in plants treated with spent maize-cob formulation of T. harzianum NBRI-1055 (MCFT), as compared to pathogen-inoculated plants. The protection afforded by the biocontrol agent was associated with the accumulation of the ROS gene network: the catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and ascorbate peroxidase (APx), maximum activity of CAT (11.0 times) was observed at 8 dapi, SOD (7.0 times) at 7 dapi, GPx (5.4 times) and APx (8.1 times) at 7 dapi in MCFT-treated plants challenged with the pathogen. This was further supported by the inhibition of lipid and protein oxidation in Trichoderma-inoculated plants. MCFT stimulated the accumulation of secondary metabolites of phenolic nature that increased up to five-fold and also exhibited strong antioxidant activity at 8 dapi, eventually leading to the systemic accumulation of phytoalexins. These results suggest that T. harzianum–mediated biocontrol may be related to alleviating R. solani-induced oxidative stress.