Effects of Soil Temperatures and Inoculum Levels of Meloidogyne incognita and Rhizoctonia solani on Seedling Disease of Cotton

Soreshin of cotton was more severe from combined infections of Rhizoctonia solani and Meloidogyne incognita than from either organism alone, when both critical soil temperature and inoculum concentrations were present. Optimum soil temperatures for disease development from combined infections were 18 and 21 C. Either 2,500 or 5,000 M. incognita larvae per plant, combined with R. solani, increased seedling disease severity over that caused by R. solani alone. When 100 or 500 larvae per plant were added with R. solani, disease severity did not change. Disease severity increased with the highest level of R. solani inoculum either alone or combined with M. incognita.     

Identification and pathogenicity of Rhizoctonia species isolated from bean and soybean plants in Samsun,Turkey

A total of 434 isolates of Rhizoctonia belonging to 10 anastomosis groups were obtained from the roots and rhizosphere soils of bean and soybean plants grown in Samsun, Turkey. AG-4 was found to be the most common group on bean and soybean plants and AG-5, AG-6, binucleate AG-A, AG-B and R. zeae were other groups isolated from the both plant species. AG-1, AG-7 and AG-K from bean and AG-E from soybean were other groups obtained in the study. The pathogenicity tests on bean and soybean seedlings showed that the highest disease severities were caused by AG-4 isolates, whereas AG-1 and AG-6 isolates were moderately pathogenic. Binucleate Rhizoctonia AG-B isolates were also moderately pathogenic, while other binucleate Rhizoctonia were found to be weakly pathogenic. Rhizoctonia zeae isolates caused moderate disease symptoms on bean, but soybean plants were slightly affected by this group of isolates. This is the first reported observation of R. solani AG-6 and AG-7 and binucleate Rhizoctonia AG-B on bean, and R. solani AG-5 and AG-6 and binucleate Rhizoctonia AG-A, AG-B and AG-E on soybean, in Turkey.     

Scarlet-Rz1, an EMS-generated hexaploid wheat with tolerance to the soilborne necrotrophic pathogens <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG-8 and <Emphasis Type="Italic">R. oryzae</Emphasis>

The necrotrophic root pathogens Rhizoctonia solani AG-8 and R. oryzae cause Rhizoctonia root rot and damping-off, yield-limiting diseases that pose barriers to the adoption of conservation tillage in wheat production systems. Existing control practices are only partially effective, and natural genetic resistance to Rhizoctonia has not been identified in wheat or its close relatives. We report the first genetic resistance/tolerance to R. solani AG-8 and R. oryzae in wheat (Triticum aestivum L. em Thell) germplasm ‘Scarlet-Rz1’. Scarlet-Rz1 was derived from the allohexaploid spring wheat cultivar Scarlet using EMS mutagenesis. Tolerant seedlings displayed substantial root and shoot growth after 14 days in the presence of 100–400 propagules per gram soil of R. solani AG-8 and R. oryzae in greenhouse assays. BC₂F₄ individuals of Scarlet-Rz1 showed a high and consistent degree of tolerance. Seedling tolerance was transmissible and appeared to be dominant or co-dominant. Scarlet-Rz1 is a promising genetic resource for developing Rhizoctonia-tolerant wheat cultivars because the tolerance trait immediately can be deployed into wheat breeding germplasm through cross-hybridization, thereby avoiding difficulties with transfer from secondary or tertiary relatives as well as constraints associated with genetically modified plants. Our findings also demonstrate the utility of chemical mutagenesis for generating tolerance to necrotrophic pathogens in allohexaploid wheat. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. P. A. Okubara and C. M. Steber contributed equally to this work.     

Cloning of Genes Involved in the Synthesis of Pyrrolnitrin from Pseudomonas fluorescens and Role of Pyrrolnitrin Synthesis in Biological Control of Plant Disease

A soil isolate of Pseudomonas fluorescens (BL915) was shown to be an effective antagonist of Rhizoctonia solani-induced damping-off of cotton. Investigation of the biological basis of this antagonism revealed that the strain produces pyrrolnitrin, a secondary metabolite known to inhibit R. solani and other fungi. Mutants of strain BL915 that did not produce pyrrolnitrin and did not suppress damping-off of cotton by R. solani were generated by exposure to N-methyl-N′ -nitro-N-nitrosoguanidine. A gene region that was capable of restoring pyrrolnitrin production to the non-pyrrolnitrin-producing mutants and of conferring this ability upon two other P. fluorescens strains not otherwise known to produce this compound or to be capable of suppressing damping-off caused by R. solani was isolated from strain BL915. The non-pyrrolnitrin-producing strains (mutants of BL915 and the other two P. fluorescens strains) which synthesized pyrrolnitrin after the introduction of the gene region from strain BL915 were also shown to be equal to strain BL915 in their ability to suppress R. solani-induced damping-off of cotton. These results indicate that we have isolated from P. fluorescens BL915 a gene(s) that has a role in the synthesis of pyrrolnitrin and that the production of this compound has a role in the ability of this strain to control damping-off of cotton by R. solani.     

ROS and trehalose regulate sclerotial development in Rhizoctonia solani AG-1 IA

Rhizoctonia solani AG-1 IA is the causal agent of rice sheath blight (RSB) and causes severe economic losses in rice-growing regions around the world. The sclerotia play an important role in the disease cycle of RSB. In this study, we report the effects of reactive oxygen species (ROS) and trehalose on the sclerotial development of R. solani AG-1 IA. Correlation was found between the level of ROS in R. solani AG-1 IA and sclerotial development. Moreover, we have shown the change of ROS-related enzymatic activities and oxidative burst occurs at the sclerotial initial stage. Six genes related to the ROS scavenging system were quantified in different sclerotial development stages by using quantitative RT-PCR technique, thereby confirming differential gene expression. Fluorescence microscopy analysis of ROS content in mycelia revealed that ROS were predominantly produced at the hyphal branches during the sclerotial initial stage. Furthermore, exogenous trehalose had a significant inhibitory effect on the activities of ROS-related enzymes and oxidative burst and led to a reduction in sclerotial dry weight. Taken together, the findings suggest that ROS has a promoting effect on the development of sclerotia, whereas trehalose serves as an inhibiting factor to sclerotial development in R. solani AG-1 IA.     

Timelapse scanning reveals spatial variation in tomato (Solanum lycopersicum L.) root elongation rates during partial waterlogging

Aims

Effects of soil amendments with crop residues on suppression of damping-off of sugar beet were examined by growing the seedlings in pasteurized, Rhizoctonia solani (AG2-2 IIIB)-infested soil at different temperatures. Dried residues of five dasheen or taro (Colocasia esculenta (L.) Schott) cultivars were compared with those of peanut (Arachys hypogaea L.) and Brassica rapa Olsson for their effects on disease suppression.

Methods and Results

When the seedlings were grown at 17/12 °C (day/night), all residues equally suppressed the disease when amended into the soil just before sowing. At 22/17 or 32/27 °C, damping-off developed in non-treated soil within 10 days, and differential suppressive effects by the residues became apparent by 21 days. When non-pasteurized, non-treated soil was infested with the pathogen, seedling survival was markedly better than in the same but pasteurized, infested soil. Yet, the effect was not different within the entire temperature ranges. Growth of both R. solani and the seedlings peaked near 25 °C and leveled off at higher temperatures.

Conclusions

These results suggest that damping-off was suppressed by antagonistic soil microorganisms, and individual residues elevated their effects differently. Under cool conditions, the antagonists dominated the pathogen to suppress the disease. Under warmer conditions, pathogenesis overcame antagonism depending on the residue, resulting in differential effects of disease suppression.     

Demonstration of the biofumigation activity of <Emphasis Type="Italic">Muscodor albus</Emphasis> against <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> in soil and potting mix

This research demonstrates the role of antimicrobial volatiles produced by Muscodor albus in disease control in soil and potting mix. The volatiles controlled damping-off of broccoli seedlings when pots containing soil or soilless potting mix infested with Rhizoctonia solani were placed in the presence of active M. albus culture without physical contact in closed containers. Conversely, plugs of R. solani on potato dextrose agar were inhibited when they were placed in the presence of M. albus incorporated into garden soil or soilless potting mix. Gas chromatographic analysis with solid-phase micro extraction showed that isobutyric acid and 2-methyl-1-butanol were released from the treated substrates. There was a significant relationship between the production of isobutyric acid in soil and damping-off control (P = 0.0415). Production of isobutyric acid was short-lived in treated substrates, peaking at 24 h in potting mix and 48 h in soil. Amounts of isobutyric acid released from soil were several times higher than those released from potting mix. Also, higher rates of M. albus rye grain culture were required to control damping-off in potting mix than in soil. This suggests that the soil used in this study is a better environment than soilless potting mix for the biological activity or viability of M. albus and components from the potting mix might bind the volatiles. The release of volatiles from soil during the biofumigation process suggests that containment measures such as tarping could be used to improve the control of soil-borne diseases and reduce use rate of the biocontrol agent.     

Biocontrol of Rhizoctonia solani damping-off and promotion of tomato plant growth by endophytic actinomycetes isolated from native plants of Algerian Sahara

Thirty-four endophytic actinomycetes were isolated from the roots of native plants of the Algerian Sahara. Morphological and chemical studies showed that twenty-nine isolates belonged to the Streptomyces genus and five were non-Streptomyces. All isolates were screened for their in vitro antifungal activity against Rhizoctonia solani. The six that had the greatest pathogen inhibitory capacities were subsequently tested for their in vivo biocontrol potential on R. solani damping-off in sterilized and non-sterilized soils, and for their plant-growth promoting activities on tomato seedlings. In both soils, coating tomato seeds with antagonistic isolates significantly reduced (P < 0.05) the severity of damping-off of tomato seedlings. Among the isolates tested, the strains CA-2 and AA-2 exhibited the same disease incidence reduction as thioperoxydicarbonic diamide, tetramethylthiram (TMTD) and no significant differences (P < 0.05) were observed. Furthermore, they resulted in a significant increase in the seedling fresh weight, the seedling length and the root length of the seed-treated seedlings compared to the control. The taxonomic position based on 16S rDNA sequence analysis and phylogenetic studies indicated that the strains CA-2 and AA-2 were related to Streptomyces mutabilis NBRC 12800^T (100% of similarity) and Streptomyces cyaneofuscatus JCM 4364^T (100% of similarity), respectively.     

Quantification of rice sheath blight progression caused by Rhizoctonia solani

Rhizoctonia solani has a wide host range, including almost all cultivated crops and its subgroup anastomosis group (AG)-1 IA causes sheath blight in rice. An accurate measurement of pathogen’s biomass is a convincing tool for enumeration of this disease. Mycological characteristics and molecular diagnosis simultaneously supported that all six strains in this study were R. solani AG-1 IA. Heterokaryons between strains Rs40104, Rs40105, and Rs45811 were stable and viable, whereas Rs40103 and Rs40106 did not form viable fused cells, except for the combination of Rs40106 and Rs40104. A primer pair was highly specific to RsAROM gene of R. solani strains and the amplified fragment exists as double copies within fungal genome. The relationship between crossing point (CP) values and the amount of fungal DNA was reliable (R ² >0.99). Based on these results, we determined R. solani’s proliferation within infected stems through real time PCR using a primer pair and a Taqman probe specific to the RsAROM gene. The amount of fungal DNA within the 250 ng of tissue DNA from rice cv. Dongjin infected with Rs40104, Rs40105, and Rs45811 were 7.436, 5.830, and 5.085 ng, respectively. In contrast, the fungal DNAs within the stems inoculated with Rs40103 and Rs40106 were 0.091 and 0.842 ng. The sheath blight symptom progression approximately coincided with the amount of fungal DNA within the symptoms. In summary, our quantitative evaluation method provided reliable and objective results reflecting the amount of fungal biomass within the infected tissues and would be useful for evaluation of resistance germplasm or fungicides and estimation of inoculum potential.     

Impact of two soil-applied herbicides on damping-off of cowpea caused byRhizoctonia solani

Summary Alachlor was more inhibitory toRhizoctonia solani growth than fluchloralin in potato dextrose broth (PDB). Infective capacity of the pathogen was not altered by growing it in a medium containing either of the herbicides. Cowpea seedlings grown in alachlor-treated soil were more susceptible toR. solani than those treated with fluchloralin and the untreated seedlings. Pre-sowing application of alachlor in soil (5 l a.i./kg) aggravated damping-off whereas fluchloralin decreased the disease incidence to nearly half of that in untreated soil in greenhouse pot tests (av. temperature 31±5°C). Both herbicides reduced damping-off in pots kept at constant temperature of 30°C and increased the disease incidence at 20°C. Fungus growth in culture was stimulated at 20° but was strongly inhibited at 30°C by both herbicides. Growth inhibition by herbicides was maximum in PDB of pH 8 and decreased steadily up to pH 5.Impact of fluchloralin and alachlor onR. solani damping-off of cowpea appears to be due to the predisposing effect by the herbicides on the susceptibility of the host and is influenced by atmospheric temperature.     

Phylogeography of the Solanaceae-infecting Basidiomycota fungus <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG-3 based on sequence analysis of two nuclear DNA loci

Background  

The soil fungus Rhizoctonia solani anastomosis group 3 (AG-3) is an important pathogen of cultivated plants in the family Solanaceae. Isolates of R. solani AG-3 are taxonomically related based on the composition of cellular fatty acids, phylogenetic analysis of nuclear ribosomal DNA (rDNA) and beta-tubulin gene sequences, and somatic hyphal interactions. Despite the close genetic relationship among isolates of R. solani AG-3, field populations from potato and tobacco exhibit comparative differences in their disease biology, dispersal ecology, host specialization, genetic diversity and population structure. However, little information is available on how field populations of R. solani AG-3 on potato and tobacco are shaped by population genetic processes. In this study, two field populations of R. solani AG-3 from potato in North Carolina (NC) and the Northern USA; and two field populations from tobacco in NC and Southern Brazil were examined using sequence analysis of two cloned regions of nuclear DNA (pP42F and pP89).     

Antifungal properties of essential oil and crude extracts of Hypericum linarioides Bosse

The chemical composition of the essential oil isolated from the aerial parts of Hypericum linarioides Bosse by hydrodistillation was analysed by GC–MS. It was determined that 74 compounds, which represent 84.1% of total oil, were present in the oil. The oil contains mainly δ-cadinene (6.9%), (Z)-β-farnesene (5.2%), γ-muurolene (5.5%), spathulenol (4.8%), hexahydrofarnesyl acetone (4.5%) and α-selinene (4.0%). The oil was also characterized by high content of sesquiterpenes (64.2% of total oil). The oil was tested for antifungal activity using mycelial growth inhibition assays (in vitro) against 11 agricultural pathogenic fungi, which consisted of six Fusarium species (Fusarium acuminatum, Fusarium culmorum, Fusarium equiseti, Fusarium oxysporum, Fusarium sambucinum and Fusarium solani) and three anastomosis groups of Rhizoctonia solani (AG-5, AG-9 and AG-11), Alternaria solani and Verticillium albo-atrum. The oil of H. linarioides showed antifungal activity against AG-9 and V. albo-atrum. In addition, petroleum ether, chloroform, acetone and methanol extracts of H. linarioides were tested against species of 11 fungi. The extracts showed moderate inhibition effects on the growth of A. solani, F. culmorum, F. equiseti and all anastomosis groups of R. solani.     

The role of rhizosphere bacteria in herbicide-mediated increase in Rhizoctonia solani-induced cotton seedling damping-off

A study was carried out to determine the role of rhizosphere-residing bacteria in the observed pendimethalin- and prometryn-mediated increase in cotton seedling damping-off incidence caused by Rhizoctonia solani. Judging from the results, pendimethalin-mediated increase in disease incidence may be due to a decrease in the populations of indigenous cotton root colonizing bacteria in the presence of pendimethalin. Prometryn-mediated increase in disease may be due to a decrease in the populations of indigenous root colonizing bacteria as well as increased susceptibility of cotton seedlings to R. solani infection in the presence of prometryn. Pendimethalin and prometryn neither affected the growth of R. solani nor caused any visible change in seedling physical characteristics.     

Pectic zymogram variation and pathogenicity of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG-4 to bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis>) isolates in Isfahn,Iran

Rhizoctonia disease, caused by Rhizoctonia solani is one of the most important fungal diseases in bean fields in Isfahan, Iran. Bean plants showing stem and root cankers were collected and Rhizoctonia-like fungi obtained from the samples were identified by anastomosis. Pure cultures of bean isolates of R. solani were identified as AG-4. There were also AG-4 isolates from tomato, potato, cucumber, alfalfa and sugar beet in the areas sampled. A total of 163 isolates of R. solani AG-4 originating from stem and root cankers of beans were examined using pectic zymogram electrophoresis. Polygalacturonase (PG) and pectin estrase isozymes were observed in all AG-4 isolates tested. One (PG) and one pectic esterase (PE) band was found in common between all isolates examined. The electrophoretic patterns were grouped into seven zymogram groups (ZGs) according to the diagnostic PG and PE bands. One ZG occurred in a high frequency throughout the areas sampled. A pathogenicity test was conducted and representative isolates of each ZG were used to inoculate healthy bean plants. The results showed that each ZG caused different symptoms with varying severity. Isolates belonging to two ZGs were highly pathogenic causing root, stem and hypocotyl cankers whereas isolates of the other ZGs produced weak or no symptoms.     

Phylogenetics and histology provide insight into damping-off infections of ‘Poblano’ pepper seedlings caused by <Emphasis Type="Italic">Fusarium</Emphasis> wilt in greenhouses

The ‘Poblano’ pepper crop is economically important in Mexico and throughout the world as it is used as a hot spice in food. The cultivated area of the ‘Poblano’ pepper crop is decreasing yearly for many reasons, among them a wilt disease commonly associated with Fusarium spp. This disease is a problem of field and greenhouse production plants. Moreover, it is not clear whether the pathogens that cause wilt in mature plants are the same as those involved in the damping-off symptoms and death of pepper seedlings in greenhouses. For this reason, the aim of the present study was to identify the causal agent of damping-off in pepper during seedling production, establish its relationship with the causal agent of wilting in mature plants, and determine whether histological damage in seedlings occurs. Isolates were recovered from the crown rot and stem base of 4-month-old infected ‘Poblano’ mature pepper plants and were identified using morphological and phylogenetic approaches. Fusarium oxysporum and F. solani were isolated from the crown rot and base stem, respectively. A pathogenicity test showed that both species caused damping-off in pepper seedlings. Histological studies with inoculated seedlings of both isolates showed several changes in the external cortex, epidermal cells, endodermis, Casparian strips, cell size, and xylem wall. Casparian strip rupture resulted in permeability loss and regulatory activity to maintain the cellular equilibrium inside the vascular bundles. Hence, according to these findings, producers should avoid seedling contamination by infected mature plants because the aggressiveness of Fusarium isolates can cause rapid seedling mortality.     

Fungitoxicity of mineral oils against Rhizoctonia solani causing damping-off of mung bean (Phaseolus aureus)

Three mineral oils, BSO, EWOS and E9267 and one vegetable oil (mustard oil), did not appreciably inhibit the mycelial growth of Rhizoctonia solani. However, treatment of 100 g seeds of mung bean with 2 ml EWOS and E9267 oils controlled more than 90% of the pre- and post-emergence damping-off and protected seedlings in soil inoculated with R. solani 5 days after sowing. Soaking seeds in solutions of these oils or drenching the soil did not control damping-off. Mustard oil controlled only pre-emergence damping-off.     

Chemical,physical and biological control of carrot seedling diseases

In naturally infested soil containingPythium ultimum, P. acanthicum andPhytophthora megasperma, onlyP. ultimum was associated with root rot and damped-off seedlings. Damping-off was promoted by low soil temperatures and by flooding. Seedling stands were markedly reduced when seed was pre-incubated in soil at 12°C but not at 25°C or 35°C. Dusting carrot seed with metalaxyl significantly increased seedling stands in the field at rates from 1.5–6 g kg⁻¹ seed and in both flooded and unflooded, naturally infested soil at 3.15 g kg⁻¹. In greenhouse experiments using artifically infested soil,P. ultimum andP. paroecandrum caused damping-off of carrot seedlings andRhizoctonia solani reduced root and shoot weights.R. solani caused damping-off in nutrient-enriched soil.P. acanthicum andP. megasperma were not pathogenic to seedlings, although both fungi colonized roots. Soil populations of allPythium spp., particularlyP. ultimum, increased during growth of seedlings and population growth ofP. megasperma was promoted by periodic flooding. Infestation of soil withP. acanthicum did not reduce damping-off of carrot seedlings byP. ultimum orP. paroecandrum, but significantly increased root and shoot weights and decreased root colonization byR. solani P. acanthicum has potential as a biocontrol agent againstR. solani.     

Isolation and characterization of rhizosphere bacteria for the biocontrol of the damping-off disease of tomatoes in Tunisia

Fluorescent Pseudomonas spp., isolated from tomato and pepper plants rhizosphere soil, was evaluated in vitro as a potential antagonist of fungal pathogens. Pseudomonas strains were tested against the causal agents of tomatoes damping-off (Sclerotinia sclerotiorum), root rot (Fusarium solani), and causal agents of stem canker and leaf blight (Alternaria alternata). For this purpose, dual culture antagonism assays were carried out on 25% tryptic soy agar, King B medium and potato dextrose agar to determine the effect of the strains on mycelial growth of the pathogens. In addition, strains were screened for their ability to produce exoenzymes and siderophores. All the strains significantly inhibited Alternaria alternata, particularly in 25% TSA medium. Antagonistic effect on Sclerotinia sclerotiorum and Fusarium solani was greater on King B medium. Protease was produced by 30% of the strains, but no strain produced cellulase or chitinase. Finally, the selected Pseudomonas strain, Psf5, was evaluated on tomato seedling development and as a potential candidate for controlling tomato damping-off caused by Sclerotinia sclerotiorum, under growth chamber conditions. In vivo studies resulted in significant increases in plant stand as well as in root dry weight. Psf5 was able to establish and survive in tomato plants rhizosphere after 40 days following the planting of bacterized seeds.     

Suppression of sugar beet damping-off caused by Rhizoctonia solani using bacterial and fungal antagonists

Rhizoctonia damping-off caused by Rhizoctonia solani Kühn, is one of the most damaging sugar beet diseases. It causes serious economic damage wherever sugar beets are grown. Biological control is an efficient and environmentally friendly way to prevent damping-off disease. Suppression of damping-off disease caused by R. solani was carried out by four isolates of Bacillus subtilis (Ehrenberg) Cohn as well as three isolates of each of Trichoderma harzianum Rifai and Trichoderma hamatum (Bonord.) Bainier. The effect of Bacillus and Trichoderma isolates against R. solani was investigated in vitro and tested on sugar beet plants under greenhouse conditions. Isolates of Bacillus and Trichoderma were able to inhibit the growth of R. solani in dual culture. Furthermore, Trichoderma isolates gave high antagonistic effect than isolates of B. subtilis. Under greenhouse conditions, coating seeds by T. harzianum and B. subtilis separately, reduced seedling damping-off significantly. However, applications of T. harzianum increased the percentage of surviving plants more than B. subtilis in comparison to control. The obtained results indicate that T. harzianum and B. subtilis are very effective biocontrol agents that offer potential benefit in sugar beet damping-off and should be harnessed for further biocontrol applications.