Disease cycle involved in damping-off of groundnut caused byRhizoctonia solani

Summary By taking into consideration of the results of various studies on damping-off of groundnut caused byRhizoctonia solani, the mechanisms leading to ultimate collapse and death of the seedlings, are described.     

Effect of peat-bran inoculum ofTrichoderma species on biological control ofRhizoctonia solani in lettuce

A range of known biocontrol or plant growth-stimulating species ofTrichoderma orGliocladium were grown on peat-bran substrate to yield between 5×10⁷–3×10¹⁰ colony forming units (cfu's)g^–1 substrate after 14 days growth. Inocula were incorporated into peat:sand potting compost infested withRhizoctonia solani to give 7–8 × 10⁴ cfu's of antagonist g^–1 compost and assessed for biological control activity using lettuce seedlings. Six of the eight antagonists decreased daming-off and three of these consistently increased yield in comparison withR. solani treatment alone.Subsequently, peat-bran inoculum ofT. harzianum isolate TH1 was incorporated at 0.5% w/v intoR. solani infested potting compost. Both autoclaved and nonautoclaved inoculum ofT. harzianum TH1 decreased disease and increased yield. Incorporation of ethyl acetate-extracted autoclaved inoculum ofT. harzianum TH1 resulted in similar levels of biocontrol and improved plant growth as did incorporation of nonautoclaved and autoclavedT. harzianum TH1 inoculum. The need to standardize inocula and controls is emphasized.     

Control ofRhizoctonia solani in cotton by seed-coating withTrichoderma spp. spores

Summary Coating cotton seeds withTrichoderma spp. reduced the incidence of disease caused byR. solani by up to 83% in the greenhouse.T. hamatum was more efficient in disease control at 20°C, whileT. harzianum was superior at 27°C. Disease severity was reduced by 47–60% in two field experiments, showing no statistical difference from treatment with pentachloronitrobenzene.     

Enzyme Production by the Mycoparasite Verticillium biguttatum against Rhizoctonia solani

Verticillium biguttatum, a mycoparasite of the ubiquitous soil-borne plant pathogen Rhizoctonia solani, excreted chitinase and beta-1,3-glucanase into liquid medium when grown on laminarin and chitin, respectively. Neither chitinase nor beta-1,3-glucanase was produced by the mycoparasite when grown on cell walls of two isolates of R. solani representing anastomosis groups (AG)-3 and AG-8. Extracellular protease was induced by growth on cell walls of the pathogen, whereas beta-1,3-glucanase and chitinase were produced bound to the cell wall of V. biguttatum. This is the first report of chitinase, beta-1,3-glucanase and protease production by V. biguttatum. These enzymes may play a previously unforeseen role in dissolving and penetrating the cell walls of R. solani.     

Changes in Azospirillum brasilense motility and the effect of wheat seedling exudates

The rhizobacterium Azospirillum brasilense Sp245 swims, swarms (Swa⁺ phenotype) or, very rarely, migrates with the formation of granular macrocolonies (Gri⁺ phenotype). Our aims were (i) to identify Sp245 mutants that swarm faster than the parent strain or differ from it in the mode of spreading and (ii) to compare the mutants’ responses to wheat seedling exudates. In isotropic liquid media, the swimming speeds of all motile A. brasilense strains were not influenced by the exudates. However, the exudates significantly stimulated the swarming of Sp245. In several Sp245 mutants, the superswarming phenotype was insensitive to local colonial density and to the presence of wheat seedling exudates. An A. brasilense polar-flagellum-defective Gri⁺ mutant BK759.G gave rise to stable Swa⁺⁺ derivatives with restored flagellum production. This transition was concurrent with plasmid rearrangements and was stimulated in the presence of wheat seedling exudates. The swarming rate of the Swa⁺⁺ derivatives of BK759.G was affected by the local density of their colonies but not by the presence of the exudates.     

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.     

RFLP analysis of the PCR-amplified 28S rDNA inRhizoctonia solani

RFLP analyses of a portion of the 28S rDNA gene region were conducted by using four restriction endonucleases for 57 isolates of 13 intraspecific groups (ISGs) representing 7 anastomosis groups (AGs) ofRhizoctonia solani. Variations in the PCR-amplified rDNA products and the polymorphisms on digestion with restriction enzymes (BamHI,HaeIII,HhaI andHpaII) were observed among three AGs, AG 1, 2 and 4. These differences were also conserved among some ISGs of AG 1 and AG 2. Among ISGs of AG 1, the pattern of rDNA fragments of AG 1-IA obtained by digestion withHpaII was significantly different from those of AG 1-IB and IC. Such difference in the fragment pattern was also observed among AG 2-1, 2-2 IIIB and 2-2 IV by the digestion withHhaI andHpaII. A dendrogram derived from the restriction enzyme data showed that ISGs from AG 1 and AG 2 can each be subdivided into distinct groups, those are distantly related to the majority isolates of the other AGs.     

Co-utilization of Bacillus subtilis and Flutolanil in controlling damping-off of tomato caused by Rhizoctonia solani

Damping-off of tomato caused by Rhizoctonia solani was controlled in a pot test using the biological agent, Bacillus subtilis RB14-C, and the chemical pesticide, Flutolanil. The co-utilization of B. subtilis RB14-C, and Flutolanil decreased the amount of Flutolanil used from 375 g/pot when Flutolanil was used alone to 94 g/pot, while exerting the same effect of reducing disease occurrence.     

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.     

Effect of certain fungicides on the production of enzymes byRhizoctonia solani

The effect of fungicidesviz. Benomyl, Carbendazim, Chloroneb, Kitazin, Edifenphos, Thiabendazole and Wettable ceresan on the production of enzymes byR. solani was studied. The fungicides tested were inhibitory to the production and activity of amylase, invertase and cellulase. The fungicides inhibited the growth of the fungus to a considerable extent.     

Antagonism against Rhizoctonia solani and fungitoxic metabolite production by some Penicillium isolates

A number of Penicillium isolates were recovered in association to Rhizoctonia solani strains pathogenic on tobacco and from soil on plates pre-colonized by the pathogen itself. Their antagonism toward R. solaniAG-2-1 was evaluated in dual cultures in vitro. Inhibition of growth was evident to some extent in most pairings, while hyphal interactions referable to mycoparasitic relationships were not observed. However, the occurrence of plasmolysis and/or vacuolisation and the induction of monilioid cells were indicative of the release of bioactive compounds. Therefore, production of fungitoxic metabolites was tested by adding concentrated culture filtrates of each Penicillium isolate to the growth medium of R. solani. Complete and lasting inhibition was incited by culture filtrates of some isolates belonging to P. brevicompactum, P. expansum, and P. pinophilum. Three purified compounds, respectively mycophenolic acid, patulin and 3-O-methylfunicone, which were extracted from culture filtrates, were able to inhibit R. solani in vitro. Their production was also detected in dual cultures of the same Penicilliumstrains with R. solani prepared in sterilized soil and when the Penicilliumstrains were cultured directly on R. solani mycelium harvested from liquid cultures. The possible role of such metabolites in antagonism of the above-mentioned Penicilliumspecies against R. solani is discussed.     

Expression of Dm-AMP1 in rice confers resistance to Magnaporthe oryzae and Rhizoctonia solani

Magnaporthe oryzae and Rhizoctonia solani, are among the most important pathogens of rice, severely limiting its productivity. Dm-AMP1, an antifungal plant defensin from Dahlia merckii, was expressed in rice (Oryza sativa L. sp. indica cv. Pusa basmati 1) using Agrobacterium tumefaciens-mediated transformation. Expression levels of Dm-AMP1 ranged from 0.43% to 0.57% of total soluble protein in transgenic plants. It was observed that constitutive expression of Dm-AMP1 suppresses the growth of M. oryzae and R. solani by 84% and 72%, respectively. Transgenic expression of Dm-AMP1 was not accompanied by an induction of pathogenesis-related (PR) gene expression, indicating that the expression of DmAMP1 directly inhibits the pathogen. The results of in vitro, in planta and microscopic analyses suggest that Dm-AMP1 expression has the potential to provide broad-spectrum disease resistance in rice.     

Research on the aflatoxin problem in groundnut at ICRISAT

Summary Aflatoxin contamination of groundnut is a serious problem in most groundnut producing countries and as such is given high research priority by the Groundnut Improvement Program of ICRISAT. Since 1979 we have concentrated on selecting cultivars resistant to seed invasion and colonization by toxigenicAspergillus flavus, and/or to aflatoxin production following invasion by the fungus. Resistance to invasion and colonization byA. flavus of rehydrated, mature seed has been found, and confirmed, in some cultivars. We have also screened several groundnut cultivars for seed resistance in the field, both under natural conditions and with the inoculum of the fungus added to the soil in the pod zone. Some cultivars with resistance to seed colonization also showed resistance to seed invasion byA. flavus. None of the cultivars tested has shown complete resistance to aflatoxin production but significant cultivar differences occurred in the amounts of aflatoxin produced in seeds inoculated with a toxigenic strain ofA. flavus.ICRISAT Journal Article No. JA-316     

Dispersal of rice sheath blight fungus,Rhizoctonia solani AG-1(IA), and subsequent disease development in paddy fields, from survey of vegetative compatibility groups

The rice sheath blight fungus,Rhizoctonia solani AG-1(IA), was isolated from surface soil and stubble in the pretrans-planting season (early March) and from sheath blight lesions on plants at the maturing stage (late September) in two paddy fields (TW and TM:8 a). Isolates were classified into vegetative compatibility groups (VCG) based on the formation of a barrage zone (or interaction zone) at the colony interface between isolates on paired cultures. In field TW surveyed in 1991 and 1992, 128–142 isolates obtained in the pretransplanting season and 82–85 isolates from lesions on mature plants were classified into 27–29 and 28–30 VCGs, respectively. Seven to 10 VCGs (26–35%) were common between both samples, and of 30 VCGs from mature plants 12 (40%) survived the winter 1991/1992 and recovered from stubble and/or soil. In field TM in 1992, 111 isolates in the pretransplanting season and 99 from mature plants were divided into 18 and 14 VCGs, respectively. Seven (39%) out of 18 VCGs in the pretransplanting season were recovered again from mature plants. Occurence of common VCGs throughout the year indicated that ca. one third of VCGs found in the pretransplanting season caused sheath blight on rice plants about 6 mo later in the same field. Further, it was shown that about the same rate of VCGs which had caused disease overwintered on stubble and/or surface soil until the following pretransplanting season.     

Studies of 13C in the foliage reveal interactions between nutrients and water in forest fertilization experiments

The severity of a root rot disease of cereals, caused by Rhizoctonia solani Kühn AG8, was inversely correlated to the Zn status of plants in field studies in 1989 and 1990. In 1989, a preliminary survey was conducted in a farmer's field in South Australia where Zn deficiency and disease were both widespread. Zn concentration in Spear wheat plants at the 3-leaf to early tillering stage was negatively correlated with severity of the disease. For the elevent elements analysed, a correlation matrix showed that Zn had the highest, and only significant (R²=0.52^**) association with disease. The effect of Zn applications and their residual value on disease severity was further studied in a long-term field experiment in 1989 and 1990 to which Zn had been applied in 1986. There was a decrease in the area of Rhizoctonia bare patch as Zn rate was increased, a result consistent with the field survey results; the recommended rate of 2.5 kg Zn ha^–1 reduced the area affected by bare patch from 42% to 21% of the total crop area compared with no Zn application, overcame Zn deficiency and increased grain yield from 1.1 to 2.8 t ha^–1. In 1990, fresh Zn application treatments were applied to trial plots designed for this purpose, in order to compare the response with the older Zn treatments applied in 1986. The areas of bare patch in the older Zn treatments were approximately 5% greater than those in the fresh Zn treatments. The results are consistent with the hypothesis that Zn deficient plants are more susceptible to root rot caused by R. solani. Testing this hypothesis is the subject of a companion paper.     

Development of vesicular-arbuscular mycorrhiza in groundnut and other hosts

Summary A vesicular-arbuscular mycorrhizal fungus, identified asGlomus mosseae Gerdemann and Trappe, was found to occur in groundnut and some other hosts. In groundnut roots under experimental conditions, the fungus showed three phases of development-a lag phase of 3–4 weeks by the end of which formation of vesicles was noticed, a phase of gradual development upto 12 weeks, by which time an average of 6 vesicles per centimeter of root developed and a constant phase where there was no further increase of the fungus. Pigeon-pea, black gram, green gram, angular gourd, onion, maize, sorghum and pearl millet also formed mycorrhizae with this fungus, but tomato and egg-plant did not. The lag phase was longer and the average number of vesicles developed per unit root length was less in the non-leguminous hosts.     

A comparison ofin vitro andin vivo effect of clay minerals,humic acid and micronutrients on the activity of fungicides againstRhizoctonia solani

Activity of nine fungicides against mycelial growth ofRhizoctonia solani in potato dextrose broth and in pot tests as seed treatment against cowpea seedling rot in infested soil was differentially in fluenced by clay minerals, humic acid and micronutrients. Humic acid, extracted from farmyard manure, considerably lowered the activity, bothin vitro andin vivo, of all fungicides except chloroneb. Montmorillonite caused substantial decrease in disease control by fungicides but enhanced the toxicity of 2-methoxyethyl mercury chloride (MEMC), and quintozene in culture. Kaolinite inactivated carbendazim, benomyl and thiophanatemethyl in nutrient broth but had little effect on disease control by these fugicides. The six micronutrients altered the activity of fungicides to varying levels often without definite correlation betweenin vitro andin vivo results. The results ofin vitro growth inhibition tests are largely inapplicable to dieseas control tests in infested soil mainly due to the differences in the ambient conditions of the two systems.     

The role of cuticle and epidermal cell wall in resistance of rapeseed and mustard to Rhizoctonia solani

A study was conducted to determine whether the cuticles in two genera of the family Cruciferae are effective barriers to infection by Rhizoctonia solani, and whether differences in cuticle and epidermal cell wall thickness and morphology of epicuticular wax exist between resistant and susceptible cultivars. As Canola/rapeseed (Brassica napus) and mustard (Sinapis alba) plants develop from 1 to 3 weeks of age, they become increasingly resistant to R. solani AG2-1 seedling root rot. Seven-day-old seedlings of S. alba cultivars are invariably more resistant than B. napus cultivars. Brassica napus cultivars do not show an obvious cuticle layer at 1 week but at 3 weeks the presence of a cuticle is seen through autofluorescence with a concomitant increase in resistance to R. solani. Removal of the cuticle from 3-week-old hypocotyls by chloroform treatment results in a decrease in cuticular autofluorescence and a significant increase in disease severity in both resistant and susceptible cultivars. Three-week-old plants of S. alba have a much lower percent disease rating and a significantly (p=0.05) thicker cuticle layer than similar-age plants of B. napus. The results suggest that the cuticle plays an important role in the resistance of S. alba and older plants of B. napus to infection by R. solani.     

Mycetoma caused by Fusarium solani with osteolytic lesions on the hand: case report

Eumycetoma is a mycotic disease caused by saprophytic soil fungi that are usually inoculated through minor injuries. A case of mycetoma in a Brazilian farmer aged71 years is reported. This patient presented erythema and edema on the dorsal surface of the left hand with multiple crusted and cicatricial lesions. No macroscopic grains were observed. The histopathological findings showed grains consisted of numerous hyphae which stained well with Gomori-Grocott method. This material obtained by cutaneous biopsy was submitted to culture on Sabouraud’s medium and the colonies were identified as Fusarium solani. The radiological studies revealed bone osteolytic lesions and the ultrasound showed pseudocysts and fistulae at the site of this infection. The patient was treated with oral ketoconazole with a good clinical response. This revised version was published online in June 2006 with corrections to the Cover Date.     

Fusarium solani invader of the eggs of the insectPanstrongylus geniculatus in a vivarium

A strain ofFusarium solani sensu Snyder & Hansen invaded the eggs of the insectPanstrongylus geniculatus in a vivarium. None of the invaded eggs hatched. To establish experimentally the pathogenicity of thisFusarium species against the eggs ofP. geniculatus, the fungus and the eggs were incubated together under different relative humidities and temperatures. At 64% relative humidity and 26 °C, the fungus grew well colonizing and penetrating all of the chorions.Three embryos died and were also colonized byF. solani. Only 4 nymphs hatched and survived to day 20. It is concluded that the isolate ofF. solani was capable of colonizing and invading the chorion of the eggs under certain humidity and temperature conditions and cause the death of the embryos.