Development of new bioformulations using Trichoderma and Talaromyces fungal antagonists for biological control of sugar beet damping-off disease

The aims of this study were to develop new bioformulations using Trichoderma harzianum, Trichoderma asperellum, and Talaromyces flavus and some organic and inorganic carriers and evaluate their effects against Rhizoctonia solani, the fungal causal agent of sugar beet seedling damping-off disease. Selected fungal isolates were first re-cultured and maintained on potato dextrose agar (PDA) culture medium. Antagonistic effects of eight isolates of the above-mentioned antagonistic fungi were then evaluated against R. solani, through volatile metabolites and non-volatile metabolites production mechanisms under laboratory conditions. In volatile and non-volatile metabolite experiments, five and seven isolates caused significant reduction in R. solani growth respectively. Based on the results of laboratory experiments, the most effective antagonistic isolates (one isolate from each species) were selected for development of nine bioformulations using peat, rice bran and talc as carriers. The effectiveness of developed bioformulations was then evaluated in controlling sugar beet damping-off disease in a greenhouse experiment where sugar beet seeds were coated with bioformulations and were sown in pasteurized field soil pre-inoculated with R. solani. Results of the greenhouse experiment 60 days after sowing showed that all bioformulations increased the number of healthy seedlings significantly (compared to the untreated control) with different rates. According to the results, the most effective bioformulation was Talc-T. harzianum followed by Peat-T. flavus, Talc-T. flavus and Rice bran-T. harzianum. In general, in both laboratory and greenhouse experiments, T. flavus was the most effective fungal antagonist followed by T. harzianum and T. asperellum. Based on the results of this study it is concluded that Trichoderma and Talaromyces employing different mechanisms might be potential biocontrol agents for controlling R. solani-induced sugar beet damping-off disease.     

Maize genotype susceptibility to Rhizoctonia solani and its effect on sugar beet crop rotations

Root and crown rot is the major soil-borne fungal disease in sugar beet. In Europe, the disease is mainly caused by the anastomosis group (AG) 2-2IIIB of the basidiomycete Rhizoctonia solani (Kühn). No chemical fungicide to control the disease has been registered in Europe. Therefore, agronomic measures must be optimized to keep the disease severity below an economic damage threshold and to minimize white sugar yield losses. R. solani AG 2-2IIIB infects many other crops besides sugar beet, including maize, where it causes root rot. Sugar beet and maize are frequently grown in the same crop rotation. The proportion of cultivated maize in several European sugar beet growing areas is expected to rise due to a projected increase in demand for renewable resources over the next few years. Although the susceptibility to and tolerance of the disease varies among cultivars in both crops, little is known about the effects of cultivar susceptibility in the pre-crop on a subsequent susceptible crop. The cultivation of R. solani-resistant maize genotypes in rotation with resistant sugar beet might therefore be a useful tool in an integrated control strategy against R. solani, eliminating the need to restrict the desired crop rotation for phytosanitary reasons. A crop rotation experiment with artificially inoculated R. solani was conducted in the field to investigate the pre-crop effects of maize cultivars which differed in their susceptibility to R. solani on a susceptible sugar beet cultivar. We hypothesized that the maize genotype would influence the inoculum potential and performance of a susceptible sugar beet genotype grown after a maize pre-crop, and that this would correlate with the susceptibility of the maize genotype. The results demonstrate that the susceptibility of maize genotypes is consistent over a period of years and that cultivated maize genotypes influenced the inoculum potential measured as disease severity in sugar beet. However, disease severity in sugar beet did not correlate with the disease susceptibility of the genotype of the maize pre-crop. Possible reasons for this missing relationship might be differences in the quality of maize residues for the saprophytic survival of the pathogen or a genotype-specific alteration of the antagonistic microbial community. However, our findings showed that in the presence of maize- and sugar beet-pathogenic R. solani, the most favourable maize cultivar for a crop rotation cannot be determined solely on the basis of its resistance level against Rhizoctonia root rot.     

Field study of the relative susceptibility of eleven potato (Solanum tuberosum L.) varieties and the efficacy of two fungicides against Rhizoctonia solani attack

Rhizoctonia black scurf is an economically important disease of potatoes in Tunisia and around the world. It reduces the quality and yield of potatoes and has become an important impediment for export of potatoes, especially to Europe. The information on comparative susceptibility of potato cultivars will help the growers make informed decisions regarding the management of this disease. The eleven potato cultivars used in this study showed a range of susceptibility reactions to Rhizoctonia solani but none of these was completely resistant to the disease. The Spunta variety showed the least percentage of infection of progeny tubers by R. solani sclerotia at harvest, while the varieties Nicola, Santana, Labadia, Liseta and Tango showed a high percentage of infection of progeny tubers. Seed treatment using fungicides, presently registered in Tunisia, are not effective in controlling the disease to growers’ satisfaction. Field experiments conducted to evaluate the efficacy of two newly introduced fungicides pencycuron (Monceren^®) and azoxystrobin (Amistar^®) showed that they were effective for the control of the disease. Seed potatoes treatment and in-furrow application of azoxystrobin consistently provided superior protection in two years of experimentation.     

Assessing fungicide resistance in populations of Alternaria in Idaho potato fields

Early blight, caused by the fungus Alternaria solani and brown leaf spot, caused by Alternaria alternata, are important diseases of potato crops in Idaho. In recent years growers have reported a reduction in efficacy of fungicides traditionally used in the past decade to control early blight. In 2009, a collection of A. solani 39 isolates were screened for resistance to azoxystrobin, pyraclostrobin, boscalid and famoxadone. Fungicide sensitivity testing was done using spiral plate dilution gradients. Results showed that of 39 isolates screened, all were resistant to azoxystrobin and three were resistant to boscalid. None were resistant to pyraclostrobin or famoxadone. In summer 2010, more isolates were collected (9 A. alternata and 26 A. solani) and the survey was expanded to include more fungicides with four different modes of action that targeted succinate dehydrogenase (SDH), methionine biosynthesis, mitochondrial respiration and multi-site contact activity. New isolates of A. solani and A. alternata were also collected from two additional sites. The results showed that 57% of the isolates were resistant to boscalid as well as an average of 63% of the isolates being resistant to the strobilurin fungicides. Seven and 15% of isolates were resistant to penthiopyrad (an SDH inhibitor), and pyrimethanil (a methionine biosynthesis inhibitor), respectively. However, none of the isolates were resistant to fluopyram (an SDH inhibitor) or a mixture of fluopyram and pyrimethanil. Although there appears to be cross resistance developing in Alternaria spp. to some of the new SDH inhibitors like penthiopyrad, others such as fluopyram are still showing limited to no resistance development in Alternaria spp. in Idaho.     

Identification of sources of resistance to damping-off and early root/hypocotyl damage from Rhizoctonia solani in common bean (Phaseolus vulgaris L.)

Rhizoctonia solani causes economically important root and hypocotyl diseases in common bean throughout the world. Root health is a vital factor in plant development and root diseases would negatively influence water and nutrient uptake as well as cause direct stand reduction and root rot damage to the crop. An efficient common bean screening method to evaluate damping-off and early root/hypocotyl damage from R. solani was developed and used to identify dry bean lines with levels of resistance to this disease. Two sets of 163 and 111 lines previously evaluated for drought tolerance in Nebraska and Puerto Rico were evaluated for damping-off resistance and early root/hypocotyl damage under greenhouse conditions. Disease severity on plants was identified based on above-ground symptoms, seedling survival and root lesions using a rating scale of 1 (resistant) to 9 (susceptible). In the first set of lines representing commonly grown dry bean cultivars, germplasm and sources of damping-off resistance, the Rhizoctonia mean rating ranged from 1.7 to 3.9; Phaseolus vulgaris lines PI 310668 and PI 533249 had the highest damping-off resistance. In the second set of the best lines from a drought tolerance shuttle breeding program the Rhizoctonia mean rating was between 2.6 and 5.7. The availability of drought tolerant dry bean lines allowed the testing of the hypothesis that there was a correlation between selecting for drought tolerance and R. solani damping-off resistance. No correlation between mean disease rating and drought tolerance was found, but adapted dry bean lines such as NE14-08-176 released as SB-DT1, and NE14-08-225 were identified with moderate damping-off resistance and drought tolerance. Lines with both traits and other attributes will facilitate development of resistant bean cultivars to manage damping-off caused by R. solani.     

Sensitivity of Cercospora beticola isolates from Serbia to carbendazim and flutriafol

Cercospora beticola, causal agent of Cercospora leaf spot (CLS) of sugar beet, is primarily controlled by fungicides. Benzimidazole and demethylation inhibiting fungicides, including carbendazim and flutriafol, have been widely used in Serbia. Since these fungicide groups have a site-specific mode of action, there is a high risk for developing resistance in target organisms, which is the most important limiting factor in Cercospora leaf spot chemical control. A rapid identification of flutriafol and carbendazim resistance can help researchers in examining the potential of different fungicide resistance management practices, as well as in selection of fungicides for use in the areas where resistance has occurred. One hundred single-conidia isolates were collected from 70 representative locations of the sugar beet production region in Serbia. Evaluation of the isolates' sensitivity was based on the reduction of mycelial growth on medium amended with 1.25 μg mL⁻¹ flutriafol and 5 μg mL⁻¹ carbendazim. Resistance to flutriafol and carbendazim was detected in 16% and 96% of the tested isolates, respectively. All isolates resistant to flutriafol were resistant to carbendazim as well, which is the first report of a double resistance to fungicides in C. beticola. Detection of the isolates resistant to flutriafol and carbendazim using Cleaved Amplified Polymorphic Sequence (CAPS) markers confirmed the results of the in vitro tests. The efficacy of carbendazim, flutriafol, azoxystrobin, and tetraconazole at commercially recommended doses was evaluated in field trials where sugar beet plants in plots were inoculated with a mixture of isolates either sensitive and/or resistant to flutriafol and carbendazim. Carbendazim and flutriafol efficacy was very low in plots inoculated with isolates resistant to these fungicides. Presented results will contribute to development of a pathogen population sensitivity monitoring strategy that could be used for an effective CLS management in the region.     

Comprehensive Characteristics of MicroRNA Expression Profile Conferring to Rhizoctonia solani in Rice

MicroRNAs(miRNAs) are about 22 nucleotides regulatory non-coding RNAs that play versatile roles in reprogramming plant responses to biotic and abiotic stresses. However, it remains unknown whether miRNAs confer the resistance to necrotrophic fungus Rhizoctonia solani in rice. To investigate whether miRNAs regulate the resistance to R. solani, we constructed 12 small RNA libraries from susceptible and resistant rice cultivars treated with water/pathogen at 5 h post inoculation(hpi), 10 hpi and 20 hpi, respectively. By taking the advantage of next-generation sequencing, we totally collected 400–450 known mi RNAs and 450–620 novel miRNAs from the libraries. Expression analysis of mi RNAs demonstrated different patterns for known and novel miRNAs upon R. solani challenge. Thirty-four mi RNA families were identified to be expressed specifically in rice, and most of them were involved in plant disease resistance. A particular Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis result revealed that a great majority of target genes of regulated miRNAs belonged to the pathway of plant-pathogen interaction. Moreover, miR444 b.2, miR531 a, mir1861 i, novel_miR1956 and novel_miR135 conferred response to R. solani infection confirmed by Northern blot. Our global understanding of miRNA profiling revealed that the regulation of mi RNAs may be implicated in the control of rice immunity to R. solani. Analysis of the expression of miRNAs will offer the community with a direction to generate appropriate strategies for controlling rice sheath blight disease.     

Remote sensing to detect plant stress induced by Heterodera schachtii and Rhizoctonia solani in sugar beet fields

The characteristically clustered occurrence and low level of mobility of Heterodera schachtii and Rhizoctonia solani in the soil and the induction of stress symptoms in the sugar beet canopy make them ideal targets for site-specific arrangements with precision agriculture tools. A field site infested with H. schachtii and R. solani was investigated in 2009 with near-range and aerial hyperspectral sensors during the growing season. At 31 sample points ground truth data for incidence and severity of the two organisms were collected and geo-referenced. Spectral vegetation indices computed from reflectance measurements obtained from two flight campaigns (AISA, 17th of June; HyMap, 28th of August) and the near-range spectroradiometers were significantly correlated (P < 0.01) with symptoms caused by the nematode or Rhizoctonia crown and root rot. A supervised classification with Spectral Angle Mapper of leaf symptoms induced by the organisms resulted in a classification accuracy of 72 and 64% for the AISA and HyMap data, respectively. The results demonstrated that remote sensing in combination with geographic information system technologies can be used effectively for the detection and mapping of symptoms caused by beet cyst nematode and Rhizoctonia crown and root rot.     

Effect of Brassicaceae seed meals with different glucosinolate profiles on Rhizoctonia root rot in wheat

The soil-borne pathogen Rhizoctonia solani AG 8 causes major yield losses in wheat (Triticum aestivum. L) production worldwide. Plant tissues of Brassicaceae species contain glucosinolates that are hydrolyzed in the presence of the enzyme myrosinase into products with pesticidal properties. Growth chamber studies were conducted to determine the effect of the Brassicaceae seed meals (SMs) from Brassica juncea, Brassica napus and Sinapis alba on the suppression of the R. solani AG 8 infection of winter wheat. Pasteurized sandy soils were amended with intact and denatured SMs of rape seed and mustard at a rate of 0.5% by soil weight. Regardless of the glucosinolate type and content, all intact and denatured Brassicaceae significantly reduced the infection of winter wheat seedlings by R. solani AG 8 compared to the un-amended control. However, soils amended with S. alba SMs had the lowest severity of Rhizoctonia root rot relative to other amended soils. Phytotoxicity arising from the use of Brassicaceae SMs was observed particularly in soils amended with high glucosinolate-containing SMs. These studies demonstrate that Brassicaceae SMs can be used to manage disease caused by R. solani AG-8. However, future studies will need to focus on strategies for diminishing the crop growth-reducing effects associated with Brassicaceae SM amendment to fully maximize these fungicidal benefits.     

Effect of inoculum density and quantitative PCR-based detection of Rhizoctonia solani AG-2-1 and Fusarium avenaceum on canola

Canola seedling blight, caused by Rhizoctonia solani, and Fusarium spp., can result in large yield losses to canola (Brassica napus) at high inoculum pressure. The effect of inoculum density was studied by mixing different amounts of R. solani AG-2-1 and Fusarium avenaceum into a sterilized natural soil and soil-less mix (2:1, v:v) separately, and recording seedling emergence, damping-off and seedling height within ten days after seeding; root rot severity at 12 days after seeding and seed yield at harvest on canola cultivars ‘45H29’ and ‘73-77RR’. Root rot severity increased and emergence, plant height and seed yield decreased with increased inoculum density of both R. solani and F. avenaceum. For quantification of R. solani AG-2-1, a primer and TaqMan probe set (Rs21F/Rs21R/Rs21P) was designed based on the nuclear ribosomal internal transcribed spacer (ITS) region of R. solani AG-2-1. From a conventional PCR amplification, an 88-bp product was amplified from all isolates classified as AG-2-1 with the primers Rs21F and Rs21R. No product was amplified with DNA from isolates belonging to other anastomosis groups of R. solani, other pathogens or the host plant. By using quantitative PCR, DNA amounts as low as 100 fg of R. solani AG-2-1 were detected. The quantity of DNA from soil samples with different inoculum densities estimated using qPCR was highly correlated to the number of colony-forming units (cfu) obtained from the same soil samples for both R. solani AG-2-1 and F. avenaceum.     

Effects of moldboard plowing,chisel plowing and rotation crops on the Rhizoctonia disease of white potato

Two tillage practices, chisel plowing (30 cm) and deep moldboard plowing (22 cm), and five rotation crops (oats, lupine, buckwheat, broccoli and peas) were studied for their effects on the soil population ofRhizoctonia solani AG-3 and on Rhizoctonia disease on potato. All rotation crops were harvested except buckwheat, which was treated as a green manure crop. Chisel plowing significantly reduced (p = 0.05) the incidence and severity of stem lesions on potato caused byR. solani AG-3. In 1990, oats after moldboard plowing significantly increased disease when compared to other crops and broccoli after chisel plowing decreased disease severity. Soil populations ofR. solani AG-3 were significantly lower with chisel plowing. No interactions between tillage and rotation crops were observed. Rhizoctonia solani Kühn is a soil inhabiting plant pathogen found worldwide that affects many plant species including white potato (Solanum tuberosum L.).R. solani attacks potato at one or more stages in development resulting in distinct disease symptoms (25) often termed the Rhizoctonia disease complex of potato. In Maine, only strains AG-3 and AG-5 ofR. solani (4, 5) have been identified as attacking potato and causing four distinct types of symptoms: 1) black scurf (sclerotia) on tubers, 2) stem cankers, 3) aerial tubers and top rosetting, and 4) killing of sprouts. Crop rotation has been reported to reduce the incidence and severity ofR. solani on potato, but no single rotation method controls completely or to a high degree of reliability (11, 23, 24, 26). Deep moldboard plowing has been shown to reduce diseases caused byR. solani andSclerotium rolfsii Sacc. in crops other than potatoes (2, 7, 15, 19, 20). However, Gudmestadet al. (6) reported that deep moldboard plowing increased the severity ofR. solani on stems and stolons of potatoes. The reduction of diseases caused byR. solani andS. rolfsii by moldboard plowing is attributed to low inoculum densities in the upper soil layer by the burial of sclerotia to depths where germination and infection were prevented (14, 15, 19, 20). However, disking to a depth of 5–7 cm did not affect disease as the inoculum remained in the root zone (14, 15). Gurkin (7) states that the rationale for deep moldboard plowing is to promote decay of organic matter, remove organic matter from the infection court and to bury the sclerotia below the infection court. Cultural control techniques are largely preventive and are designed to reduce the quantity or the activity of inoculum by means of crop rotation, tillage practices, green manure crops, etc. (22). This study was conducted to determine individual effects and possible interactions of deep moldboard plowing versus chisel plowing in various rotation crops on the presence ofR. solani AG-3 in the soil and on the incidence of Rhizoctonia disease complex of potato.     

Rhizoctonia disease of potatoes (Rhizoctonia solani): Fungicidal efficacy and cultivar susceptibility

Rhizoctonia stem canker and black scurf is an economically important disease of potatoes in Alberta and around the world. It reduces the quality and yield of potatoes and has become an important impediment for export of seed potatoes, especially to Mexico. Seed treatment using fungicides, presently registered in Canada, are not effective in controlling the disease to growers’ satisfaction. Field experiments conducted to evaluate the efficacy of various fungicides showed that captan (Captan), iprodione (Rovral), mancozeb (Tuberseal), and fludioxonil (Maxim) provide effective control of the disease. Of these fungicides, fludioxonil consistently provided superior protection. Based on the results of this study, Maxim was registered in 2000 for control of this disease in Canada. Potato cultivars/breeding selections showed a range of susceptibility reactions toR. solani but none of these was completely resistant to the disease. The information on comparative susceptibility of potato cultivars will help the growers make informed decisions regarding the management of this disease. Of six plant species (Beta vulgaris,Brassica compestris, Hordeum vulgare, Pisum sativum,Triticum aestivum, Zea mays) most commonly used in rotation with potatoes in Alberta, none was infected by potato isolates ofR. solani. The results suggested that use of these plant species as rotation crops with potatoes in Alberta will help in reducing the development of rhizoctonia disease in potatoes.     

Increased severity of Rhizoctonia root rot in sugar beet treated with systemic insecticides

The systemic insecticides aldicarb 15G (15% granules) at 2·6 kg active ingredient (a.i.)/ha and phorate 1OG (10% granules) at 1·7 kg a.i./ha, applied as side dressings about 1 month after planting in 1979 and 1980, significantly increased the severity of root rot, caused by Rhizoctonia solani Kühn, in sugar beet (Beta vulgaris L.). Carbofuran 10G (10% granules) at 2·2 kg a.i./ha also increased root rot, but not significantly. Numbers of harvestable roots were reduced by all treatments but significantly by phorate only. Both aldicarb and phorate were slightly fungistatic to R. solani when the pathogen was grown on potato-dextrose agar incorporating 0·5, 5·0, and 25 μg a.i./ml. Trichoderma sp., a potential antagonist of Rhizoctonia, was slightly inhibited by aldicarb and phorate initially, but soon overcame the effect. Thus, the increase in disease severity in the field may be attributable to some metabolic or physiological effect of the chemicals either on the host or on the infection process of the pathogen. Indiscriminate use of these insecticides should therefore be avoided in areas where root rot is prevalent but where insects are not a problem.     

Suppression of rice sheath blight disease using a heat stable culture filtrate from Streptomyces philanthi RM-1-138

Streptomyces philanthi RM-1-138, isolated from the rhizosphere soil of chili pepper in southern Thailand, was investigated for its antagonistic activity against phytopathogenic fungi. In dual culture on glucose yeast-malt extract (GYM) agar plates, this strain suppressed the mycelial growth of all seven plant pathogenic fungi tested (Rhizoctonia solani PTRRC-9, Pyricularia grisea PTRRC-18, Colletotrichum gloeosporioides NBCRSR-3, Colletotrichum capcisi NBCRSR-15, Ganoderma boninense NBCRSR-26, Fusarium fujikuroi PTRRC-16 and Bipolaris oryzae PTRRC-36) with an 82.2–89.2% inhibition and that was most pronounced on R. solani PTRRC-9. Heat treatment of the culture filtrate from growing R. solani PTRRC-9 at 40 °C, 60 °C, 80 °C, and 100 °C for 30 min and 121 °C for 15 min had no negative effect on the inhibitory activity against R. solani PTRRC-9 tested on both solid and liquid culture. The effective dose (>80% inhibition) of culture filtrate in liquid culture was at 5.0% (v/v) while it was at 10% (v/v) on the solid medium. This effectiveness was similar to those of the four chemical fungicides tested. The effect of S. philanthi RM-1-138 against R. solani PTRRC-9 was investigated using SEM and TEM. The compounds produced by S. philanthi RM-1-138 induced alterations to the cell-wall structure of R. solani PTRRC-9, that resulted in the loss of cytoplasm materials by partial lysis. The greenhouse experiment revealed that using either the culture filtrate or the autoclaved culture filtrate from S. philanthi RM-1-138 effectively suppressed rice sheath blight disease by up to 65.6 and 60.8%, respectively.     

Combining residues of herb crops with soil heating for control of soilborne pathogens in a controlled laboratory system

Plant residues from herb crops were examined as organic amendments for the control of soilborne pathogens with and without soil heating for a short exposure of 2 weeks. Residues of herbaceous plants of oregano, sage, rosemary, tarragon, bay, wild rocket, spearmint and thyme were incubated for 14 days in a controlled laboratory system which emulates soil solarization. Green cabbage residues were included for comparison. The tested pathogens included Fusarium oxysporum f. sp. radicis-lycopersici, Macrophomina phaseolina, Rhizoctonia solani and Meloidogyne javanica. Heating soil amended with herb residues, particularly tarragon, spearmint and wild rocket, resulted in high mortality of the tested pathogens. Volatile compounds which were generated in heated soils amended with wild rocket or tarragon showed significant toxicity to M. phaseolina and R. solani. Only oregano, thyme, bay, wild rocket and tarragon amendments were effective in pathogen control without heating. Incorporation of leaf and stem residues of thyme, sage, tarragon, rosemary or wild rocket in soil and exposure to solarization under field conditions resulted in 95-100% mortality of the tested pathogen. This study demonstrates that the residues from herb crop production can serve as organic amendments for the control of soilborne pathogens, especially when combined with heating or solarization.     

Evaluation of potato cultivars,clones and a true seed population for resistance toRhizoctonia solani

Field and laboratory studies were conducted in 1985–1986 to evaluate the relative severity ofRhizoctonia solani damage among 26 clones and five potato (Solanum tuberosum) cultivars. A test with true potato seeds showed that resistance toR. solani could be identified in two years. One highly resistant line was identified by this method. Moderate degrees of resistance were found in lines from the USDA Potato Breeding Program (BARC) with russet types producing a higher percentage of highly resistant lines than round whites. A laboratory test revealed that apical sprout damage (nipping) was variable within and between lines and that it was an important phase of disease development to measure when determining resistance because of its effects on the resulting plant growth and production of marketable tubers. It was also determined that to accurately define resistance toR. solani in potato, all phases of the disease must be evaluated to insure against resistance “breakdown” at some point in plant development.     

Biocontrol of Rhizoctonia solani and Sclerotium rolfsii on tomato by delivering antagonistic bacteria through a drip irrigation system

In a 2-year assessment carried out on tomato crops of Central and Southern Italy, a high incidence of Rhizoctonia solani and Sclerotium rolfsii was found. These fungal pathogens attack horticultural crops and are responsible for severe crown and stem rot. Because of technical, economical and environmental issues their chemical control is an arduous task. To find alternative and eco-compatible control methods, the effectiveness of two new antagonistic bacterial isolates (Burkholderia cepacia, T1A-2B, and Pseudomonas sp., T4B-2A), previously selected from suppressive organic amendments, were tested on tomato plants grown under both growth chamber and field conditions. The potential antagonists were compared with two commercial biofungicides, based on Bacillus subtilis (BSF4) and Trichoderma asperellum (TV1), and four synthetic fungicides (tolclofos-methyl, azoxystrobin, fosetyl-Al and fosetyl-Al + propamocarb). In 2-year field experiments carried out on tomato plants, the biocontrol bacteria as well as the other treatments were applied to the soil, proximal to the plant crowns and main roots, by means of an effective and specific system of drip irrigation. In all the experiments the novel selected biocontrol bacteria significantly reduced both incidence and severity of the diseases caused by S. rolfsii or R. solani, with results demonstrating effectiveness equal to TV1, better than BSF4 and comparable with the synthetic fungicides, except for tolclofos-methyl which was the most effective treatment. In field experiments, carried out for two consecutive years, isolate T1A-2B reduced up to 58.33% and up to 63.8% the severity of the diseases caused by S. rolfsii and R. solani respectively; whereas isolate T4B-2A gave reduction of S. rolfsii and R. solani diseases severity up to 73.2% and up to 62.7%, respectively.     

Disease variation and chemical control of Ramularia leaf spot in sugar beet

Data from 1999 to 2009 on Ramularia leaf spot caused by Ramularia beticola in sugar beet showed that it was a serious disease in sugar beet in 5 out of 11 seasons. The severity and significance of the disease was found to vary depending on events with precipitation, particularly in two specific weeks in July and September. Several fungicides were found to give effective control, and positive net yield responses were found in 9 out of 11 seasons. The average sugar yield response varied in individual years between 0.7 and 2.2 t ha⁻¹. High levels of control of Ramularia leaf spot was obtained in field trials, a semi-field trial and an in vitro test using the compounds pyraclostrobin, epoxiconazole, difenoconazole and propiconazole. Dose response trials with epoxiconazole from two seasons showed both reduced efficacy and yield responses from low doses. They also proved that the optimal input of fungicides varies significantly between seasons depending on disease severity. A sensitivity test of R. beticola to different fungicides showed a normal distribution of sensitivity with no sign of resistance development to either strobilurins or triazoles. Results from a semi-field trial showed both good preventive and curative effects with 84–100% disease control from epoxiconazole, difenoconazole and pyraclostrobin. In order to optimize an IPM control strategy better forecasting systems are needed along with cultivars providing higher levels of resistance to the disease.     

Achievements and prospects in breeding for rhizomania resistance in sugar beet

Economic viability of a sugar beet crop largely depends on its successful protection against rhizomania, a most devastating disease that causes severe losses in root yield, sucrose content and quality. Rhizomania disease is caused by Beet necrotic yellow vein virus (BNYVV), a virus present in most sugar beet growing regions being vectored by the widely spread soil borne protoctist Polymyxa betae Keskin. The only practical means to control the disease is the use of genetically resistant varieties and, to date, such resistance is mainly based on a dominant gene (Rz1) that when present confers a sufficiently high level of protection against BNYVV. However, the emergence of virus strains capable of compromising the resistance employed in commercial varieties as well as a possible spread of more pathogenic isolates threatens crop's protection efficiency in the future. All these point to the necessity for exploiting new and more effective genetic sources of rhizomania resistance, both by classical and molecular breeding approaches, a practice that is being pursued by the relevant breeding firms. This article critically reviews the various issues related to the disease and its management and particularly to the ones pertaining to pathogen genetic diversity, types of genetic resistance currently employed, as well as to novel biotechnological approaches aiming at the development of better resisting cultivars.     

Cultivar Effects on the Interaction between Free-Living Plant-Parasitic Nematodes and the Fungal Pathogen <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> in Potato

Crop damage is associated with infection by plant pathogens but can also arise through abiotic factors. However, the plant pathogens are involved in biotic interactions with other plant pathogens, and these interactions may differ depending of the cultivar of the crop. Here, the interaction between the fungus Rhizoctonia solani (AG3) and free-living plant-parasitic nematodes was investigated in a pot experiment with different potato cultivars. No synergistic interaction between R. solani and plant-parasitic nematodes was found, instead there was an effect of treatment with lower tuber yield when nematodes occurred alone. There were differences among the cultivars regarding incidence of black scurf, dry weight of stems and tubers, and there was interactive effects between treatment and cultivar regarding dry weight of stolons and roots. Therefore, results concerning incidence and damage of R. solani and/or plant-parasitic nematodes found for one cultivar may not be applicable to other cultivars.