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.     

Mixed PGPR consortium: an effective modulator of antioxidant network for management of collar rot in cauliflower

Abstract

Sclerotinia sclerotiorum is a cosmopolitan fungal pathogen causing stem and collar watery soft rot of cauliflower. Previous management of this disease with chemical pesticides caused hazardous results which lead to use of more eco-friendly microbial approaches. In the present study, consortia of Trichoderma harzianum TNHU27 and Pseudomonas aeruginosa PJHU15 were assessed for their ability in controlling Sclerotinia rot in cauliflower. The seedlings of cauliflower were challenged singly and in consortium with these two compatible microbes. The plants were evaluated upon challenge inoculation with S. sclerotiorum for changes in total phenolic content, the activity of defense and antioxidative enzymes. The microbial consortium comprising of T. harzianum and P. aeruginosa significantly enhanced the defense responses of the plant in comparison to pathogen challenged and unchallenged control. The study elucidates that plant beneficial microbes in the consortium may provide superior protection by induction of faster and enhanced defense responses in comparison to unchallenged and single microbe challenged plants under pathogen challenged conditions.     

Heterothallism revealed in the root rot fungi Berkeleyomyces basicola and B. rouxiae

Berkeleyomyces basicola and Berkeleyomyces rouxiae, two sister species previously treated collectively as Thielaviopsis basicola, reside in the Ceratocystidaceae (Microascales, Ascomycota). Both species are important root pathogens of many important agricultural crops and ornamental plants. Although T. basicola has been known for more than 150y, a sexual state has never been found and it has been assumed to be an asexual pathogen. The aim of this study was to determine the mating strategy of the two Berkeleyomyces species. Investigation of the genome sequences of two B. basicola isolates allowed for the complete characterization of the MATlocus, revealing that it has a typical heterothallic mating system with the MAT1-1andMAT1-2 idiomorphs occurring in different isolates. PCR amplification using mating type primers developed in this study, showed that the MAT1-1-1andMAT1-2-1 genes were also present in different isolates of B. rouxiae. Pairing of isolates representing the two mating types of both species,using a variety of techniques failed to produce sexual structures. Although we have found no direct evidence that they reproduce sexually, these fungi are clearly heterothallic with both mating types occurring in some countries suggesting that a cryptic sexual cycle could exist for them.     

中国木本植物病原木材腐朽菌研究

简要论述了中国木本植物致病木材腐朽菌,报道危害活立木的木腐菌152种,其中49种（占总数的34%）在中国首次报道为林木病原菌。对每个种的寄主、侵染部位、腐朽类型、发生频率及分布进行了报道。它们中有135种（占总数的89%）造成木材白色腐朽,17种导致褐色腐朽;67种为常见种,33种为偶见种,52种为少见种。     

Biological Control of Phytophthora Root Rot of Pepper Using Trichoderma harzianum and Streptomyces rochei in Combination

A combination of two compatible micro‐organisms, Trichoderma harzianum and Streptomyces rochei, both antagonistic to the pathogen Phytophthora capsici, was used to control root rot in pepper. The population of the pathogen in soil was reduced by 75% as a result. Vegetative growth of the mycelium of P. capsici was inhibited in vitro on the second day after P. capsici and T. harzianum were placed on the opposite sides of the same Petri plate. Trichoderma harzianum was capable of not only arresting the spread of the pathogen from a distance, but also after invading the whole surface of the pathogen colony, sporulating over it. Scanning electron microscopy showed the hyphae of P. capsici surrounded by those of T. harzianum, their subsequent disintegration, and the eventual suppression of the pathogen's growth. Streptomyces rochei produced a zone of inhibition, from which was obtained a compound with antioomycete property secreted by the bacteria. When purified by high‐pressure liquid chromatography, this compound was identified as 1‐propanone, 1‐(4‐chlorophenyl), which seems to be one of the principal compounds involved in the antagonism. A formulation was prepared that maintained the compound's capacity to inhibit growth of the pathogen for up to 2 years when stored at room temperature in the laboratory on a mixture of plantation soil and vermiculite. The two antagonists, added as a compound formulation, were effective at pH from 3.5 to 5.6 at 23–30°C. The optimal dose of the antagonists in the compound formulation was 3.5 × 10⁸ spores/ml of T. harzianum and 1.0 × 10⁹ FCU/ml of S. rochei. This is the first report of a compound biocontrol formulation of these two antagonists with a potential to control root rot caused by P. capsici.     

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.     

针叶树病原菌异担子菌属的物种多样性及相关检疫建议

异担子菌属Heterobasidion种类是北半球针叶树最严重的森林病原菌,在世界范围内能侵染27种针叶树,对欧洲和北美洲的经营林己造成重大损失.基于传统形态学研究,认为异担子菌属有2个种,即多年异担子菌H.annosum和岛生异担子菌H.insulare,然而单孢交配实验研究证明2个种为复合种.分子系统发育分析研究表...     

Resistance Induction by Salicylic Acid Formulation in Cassava Plant against Fusarium solani

Fusarium root rot caused by the soil-borne fungus Fusarium solani is one of the most important fungal diseases of cassava in Thailand, resulting in high yield losses of more than 80%. This study aimed to investigate if the exogenous application of salicylic acid formulations (Zacha) can induce resistance in cassava against Fusarium root rot and observe the biochemical changes in induced cassava leaf tissues through synchrotron radiation based on Fourier-transform infrared (SR-FTIR) microspectroscopy. We demonstrated that the application of Zacha11 prototype formulations could induce resistance against Fusarium root rot in cassava. The in vitro experimental results showed that Zacha11 prototype formulations inhibited the growth of F. solani at approximately 34.83%. Furthermore, a significant reduction in the disease severity of Fusarium root rot disease at 60 days after challenge inoculation was observed in cassava plants treated with Zacha11 at a concentration of 500 ppm (9.0%). Population densities of F. solani were determined at 7 days after inoculation. Treatment of the Zacha11 at a concentration of 500 ppm resulted in reduced populations compared with the distilled water control and differences among treatment means at each assay date. Moreover, the SR-FTIR spectral changes of Zacha11-treated epidermal tissues of leaves had higher integral areas of lipids, lignins, and pectins (1,770–1,700/cm), amide I (1,700–1,600/cm), amide II (1,600–1,500/cm), hemicellulose, lignin (1,300–1,200/cm), and cellulose (1,155/cm). Therefore, alteration in defensive carbohydrates, lipids, and proteins contributed to generate barriers against Fusarium invasion in cassava roots, leading to lower the root rot disease severity.     

Rhizoctonia solani injuries in oilseed crops in Finland and impacts of different crop management practices on disease incidence and severity

A disease survey in Finnish oilseed Brassica (OSR) fields in 2007–09 revealed the widespread occurrence and several fold increase of necrotic stem base lesions and severely injured blackened roots in comparison to a corresponding survey carried out in 1984–89. Rhizoctonia solani was the predominant fungi detected in the isolations and was followed by several species of Fusarium and Thielaviopsis basicola. In 60% of the samples all three species were detected together. Only the R. solani AG 2–1 strains isolated from OSR and other cruciferous hosts caused damping off or stem base symptoms on turnip rape in a greenhouse experiment. Therefore R. solani AG 2–1 was considered the main pathogen associated with the observed symptoms in OSR crops. Cultural practices changed significantly between the 1980s and 2000s. In the 2007–09 survey there was an increase in the cultivation of oilseed rape instead of turnip rape, increase in the use of no soil or reduced soil tillage and of chemical control of weeds, but a reduction in macronutrient fertilization, especially P and K, when compared to the 1980s survey. The risk for high incidence of stem base lesions and blackened roots was affected by different cultural practices. No tillage and maintaining sufficient soil pH and NPK fertilisation decreased the risk for both types of R. solani induced symptoms. Late sowing date increased the risk for high incidence of stem base lesions, while application of fungicides against Sclerotinia reduced it. The incidence of R. solani damages in many fields was very high in spite of relatively long crop rotations and therefore the average effect of crop rotation in the disease was insignificant. Current turnip rape cultivars are vulnerable to root rot while oilseed rape is vulnerable to stem base symptoms. The higher incidence of R. solani induced diseases could be associated with the decline in productivity of OSR crops in Finland. This study showed that cultural practices such as reduced or no soil tillage, adequate levels of pH and of NPK fertilization could reduce the severity of the symptoms in OSR fields.     

Extruded Granular Formulation with Biomass of Biocontrol Gliocladium virens and Trichoderma spp. to Reduce Damping-off of Eggplant Caused by Rhizoctonia solani and Saprophytic Growth of the Pathogen in Soil-less Mix

Extruded granular formulations containing rice flour, gluten, Pyrax, vermiculite, canola oil, and fermentor-produced biomass of isolates of Gliocladium virens (Gl-3, Gl-21 and Gl-32), Trichoderma hamatum (TRI-4 and 31-3), T. harzianum (Th-32 and Th-87) and T. viride (Tv-101) were evaluated for their effect on the reduction of eggplant damping-off caused by Rhizoctonia solani, reduction of pathogen inoculum and proliferation of the isolates in a soil-less mix. Granules with all isolates except 31-3 significantly (P < 0.01) reduced damping-off, and granules with Gl-3, Gl-21, Gl-32, TRI-4 and Th-87 yielded stands comparable to that (90%) of the non-infested control. Granules with isolates Gl-21 and TRI-4 were the most effective in the reduction of saprophytic growth of R. solani, and there was a significant inverse correlation (r ² = - 0.82) between eggplant stand and saprophytic growth of the pathogen over all treatments. Isolate propagules proliferated to about 10⁷ colony-forming units (CFU) g^?1 of soil-less mix after a 6-week incubation, but there was no correlation between the number of CFU and eggplant stand or saprophytic growth reduction of the pathogen. Granules with Gl-21 and TRI-4 amended to pathogen-infested soil-less mix at a rate as low as 0.06% significantly (P < 0.05) reduced damping-off and pathogen saprophytic growth, and a rate of 0.25% of Gl-21 granules resulted in an eggplant stand comparable to that of the non-infested control. There was no significant correlation between the rate of granule amendment and the proliferation of Gl-21 and TRI-4. Granules of Gl-21 and TRI-4 also significantly prevented the spread of R. solani in flats of eggplant seedlings when the biocontrol granules were applied to the soil-less mix 1 day before the pathogen inoculum.     

Fungicidal control of damping-off and seedling root rot in Brassica species caused by Rhizoctonia solani in the growth chamber*

Eight fungicides (benodanil, carboxin, cyproconazole, fenpropimorph, fur-mecyclox, iprodione, pencycuron and tolclofos-methyl) were evaluated, under growth chamber conditions, as seed treatments against pre-emergence damping-off and post-emergence seedling root rot in six Brassica species. Five cultivars of B. rapa, four cultivars of B. juncea, four cultivars of B. napus and one cultivar/ strain from each of B. carinata, B. nigra and B. oleracea were grown in soilless mix infested with an isolate of Rhizoctonia solani AG-2-1. B. nigra and B. juncea were considerably less susceptible to R. solani than the four other species. Cyproconazole at 0.05-0.1 g a.i./kg seed and the other fungicides at 2–4 g a.i./ kg seed provided almost complete control of pre-emergence damping-off in most Brassica species and their cultivars. Their efficacy varied against the post-emergence seedling root rot. Furmecyclox, iprodione, tolclofos-methyl and pencycuron consistently gave good control of seedling root rot in all Brassica species and their cultivars. Benodanil and fenpropimorph provided moderate control, and carboxin and cyproconazole gave poor control against root rot. Efficacy of carboxin, cyproconazole, benodanil and fenopropimorph against seedling root rot varied significantly (P ≤ 0.05) among cultivars within a Brassica species.     

Epidemiology,identification and disease management of grape black rot and potentially useful metabolites of black rot pathogens for industrial applications – a review

Phyllosticta ampelicida (teleomorph: Guignardia bidwellii) is the fungal plant pathogen that causes black rot on grapevine. It is able to infect all green, expanding grape tissues. Black rot poses a threat to both yield and wine quality; a severe attack can virtually cause a complete crop loss. The fungus is native to North America, and was spread to Europe at the end of the 19th century. With the beginning of the 21st century, an increasing importance of the disease was observed in several European winegrowing regions. Successful black rot control strategies combine sanitary measures, cultural techniques, growing cultivars with reduced susceptibility and the use of effective fungicides. Berries are most susceptible to infections between flowering and bunch closure and consequently, fungicide applications against black rot need to focus on this period. In this paper, forecast models and decision support systems that help to achieve satisfactory control with a minimum of fungicide input are presented. Black rot in grapevine was reported to be well controlled in field experiments by a broad range of fungicide classes including quinone outside inhibitors (98 ± 3% efficacy), demethylation inhibitors (98 ± 3% efficacy) and dithiocarbamates (92 ± 7% efficacy). Average efficacies of other fungicide classes tested so far ranged from 32 to 69%; meta data on the efficacy of fungicides in numerous field trials are given. At present, black rot causes major problems primarily if vineyard management and fungicide use were reduced or abandoned. Organically managed vineyards are highly affected. Hence, site and cultivar selection as well as cultural measures are of the highest importance especially in organic viticulture. Recent studies showed that some strains formerly classified as G. bidwellii in fact belong to a distinct species (P. parthenocissi). Black rot pathogens produce some phytotoxic secondary metabolites, such as phenguignardic acid, guignardic acid, alaguignardic acid and the guignardianones A, E and F, which could stimulate the development of new herbicides of natural origin.     

qRT-PCR quantification of the biological control agent Trichoderma harzianum in peat and compost-based growing media

To ensure proper use of Trichoderma harzianum in agriculture, accurate data must be obtained in population monitoring. The effectiveness of qRT-PCR to quantify T. harzianum in different growing media was compared to the commonly used techniques of colony counting and qPCR. Results showed that plate counting and qPCR offered similar T. harzianum quantification patterns of an initial rapid increase in fungal population that decreased over time. However, data from qRT-PCR showed a population curve of active T. harzianum with a delayed onset of initial growth which then increased throughout the experiment. Results demonstrated that T. harzianum can successfully grow in these media and that qRT-PCR can offer a more distinct representation of active T. harzianum populations. Additionally, compost amended with T. harzianum exhibited a lower Fusarium oxysporum infection rate (67%) and lower percentage of fresh weight loss (11%) in comparison to amended peat (90% infection rate, 23% fresh weight loss).     

Root rot disease incidence and severity on some legume species grown in Samsun and the fungi isolated from roots and soils

Abstract

Root rot disease is very common in the bean, soybean, faba bean and pea plants growing areas in Samsun province. Disease incidence and severity were detected the highest at 93.8% and 55.4% in the bean growing area, and the lowest at 64.0% and 24.3% in the faba bean growing area respectively. In this study, a total of 2714 fungal isolates were obtained from some legume plants and soil samples. The most common fungi isolated from root and soil samples were Fusarium spp., multinucleate Rhizoctonia (MNR), binucleate Rhizoctonia (BNR) and Pythium spp. respectively. Fusarium spp. were isolated at high rates from all the examined areas. MN Rhizoctonia and BN Rhizoctonia were isolated both from inner and coastal areas of the province, whereas Pythium spp. were isolated in costal areas, except for the Vezirköprü district which is situated in the inner area. When looking at the interactions among pathogens causing root rot, it was found the great majority of the samples (30.4%) isolated both Fusarium spp. and MNR-BNR group fungi, whereas Fusarium spp. and Pythium spp. were isolated together from 10.9% of the samples and MNR-BNR and Pythium spp. from only 1.5% of the samples.     

Potential of endophytic Streptomyces spp. for biocontrol of Fusarium root rot disease and growth promotion of tomato seedlings

Sixteen endophytic actinobacteria isolated from roots of native plants were evaluated for their antagonistic potential against soil-borne phytopathogenic fungi. Among them, three strong antagonistic isolates were selected and characterised for in vitro plant-growth-promoting and biocontrol traits, including production of hydrogen cyanide, indole-3-acetic acid and siderophores, chitinase and β-1,3-glucanase activities, and inorganic phosphate solubilisation. In all trials, the strain Streptomyces sp. SNL2 revealed promising features. The selected actinobacteria were investigated for the biocontrol of Fusarium oxysporum f. sp. radicis lycopersici and for growth promotion of tomato (Solanum lycopersicum L. cv. Aïcha) seedlings in autoclaved and non-autoclaved soils. All seed-bacterisation treatments significantly reduced the root rot incidence compared to a positive control (with infested soil), and the isolate SNL2 exhibiting the highest protective activity. It reduced the disease incidence from 88.5% to 13.2%, whereas chemical seed treatment with Thiram® provided 14.6% disease incidence. Furthermore, isolate SNL2 resulted in significant increases in the dry weight, shoot and root length of seedlings. 16S rDNA sequence analysis showed that isolate SNL2 was related to Streptomyces asterosporus NRRL B-24328^T (99.52% of similarity). Its interesting biocontrol potential and growth enhancement of tomato seedlings open up attractive uses of the strain SNL2 in crop improvement.     

Isolation and characterization of mutants of <Emphasis Type="Italic">Pseudomonas maltophila</Emphasis> PM-4 altered in chitinolytic activity and antagonistic activity against root rot pathogens of clusterbean (<Emphasis Type="Italic">Cyamopsis tetragonoloba</Emphasis>)

Pseudomonas maltophila PM-4, an antagonist of pathogenic fungi including Rhizoctonia bataticola, R. solani, Fusarium oxysporum and Sclerotinia sclerotiorum associated with root rot of clusterbean (Cyamopsis tetragonoloba) was mutagenized with Tn5. Hyperchitinase producing mutants showing large zone of colloidal chitin dissolution were identified on medium containing calcoflor dye as an indicator. A mutant P-48 producing 137% higher chitinase activity than the parent strain PM-4 was identified. Seed bacterization of clusterbean (Cyamopsis tetragonoloba) with P-48 controlled the root rot upto 40.8% in the presence of conglomerate of all the four fungal pathogens Rhizoctonia bataticola, R. solani, F. oxysporum and Sclerotinia sclerotiorum.     

Biological control of grey mould in strawberry fruits by halophilic bacteria

Aims:  Grey mould caused by Botrytis cinerea is an economically important disease of strawberries in Tunisia and worldwide. The aim of this study was to select effective halophilic bacteria from hypersaline ecosystems and evaluate the abilities of antifungal bacteria to secrete extracellular hydrolytic enzymes, anti- Botrytis metabolites and volatiles.
Methods and Results:  Grey mould was reduced in strawberry fruits treated with halophilic antagonists and artificially inoculated with B. cinerea . Thirty strains (20·2%) were active against the pathogen and reduced the percentage of fruits infected after 3 days of storage at 20°C, from 50% to 91·66%. The antagonists were characterized by phenotypic tests and 16S rDNA sequencing. They were identified as belonging to one of the species: Virgibacillus marismortui , B. subtilis , B. pumilus , B. licheniformis , Terribacillus halophilus , Halomonas elongata , Planococcus rifietoensis , Staphylococcus equorum and Staphylococcus sp. The effective isolates were tested for antifungal secondary metabolites.
Conclusions:  Moderately halophilic bacteria may be useful in biological control against this pathogen during postharvest storage of strawberries.
Significance and Impact of the study:  The use of such bacteria may constitute an important alternative to synthetic fungicides. These moderate halophiles can be exploited in commercial production and application of the effective strains under storage and greenhouse conditions.     

Differential accumulation of Xanthomonas campestris pv. campestris proteins during the interaction with the host plant: Contributions of an in vivo system

Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot, a highly destructive disease that affects all brassicas. This work aimed to study the interaction Xcc–Brassica oleracea using an in vivo system in an attempt to identify proteins involved in pathogenicity. We used label‐free shotgun 2D‐nanoUPLC/MS^E to analyze Xcc proteins in three conditions: in the interaction with susceptible (REK) and resistant (REU) plants and in culture medium (control condition). A model of Xcc–susceptible host interaction is proposed and shows that Xcc increases the abundance of several crucial proteins for infection and cell protection. In this study, we also confirmed the differential expression by qPCR analysis of selected genes. This is the first report showing a large‐scale identification of proteins in an in vivo host plant condition. Considering that most studies involving phytopathogens are in vitro (growth in culture medium or in plant extract), this work contributes with relevant information related to the plant–pathogen interaction in planta.     

当归根腐病主要病原菌生物学特性及室内毒力测定

为探究当归(Angelica sinensis)根腐病的病原菌种类、生物学特性并筛选其防治药剂。采用常规组织法分离培养病原菌,利用柯赫氏法则验证其致病性,通过形态学和分子生物学相结合的方法进行病原菌的鉴定、生物学特性分析;采用菌丝生长速率法对病原菌进行室内毒力测定。结果表明,当归根腐病菌菌落毛毡状,在三种培养基上大型分生孢子呈镰刀形,大小为(27.5～47.5)μm×(3.8～7.5)μm(n=50),隔膜数1～5;小型分生孢子在PDA和SNA培养基上呈椭圆形、肾型、纺锤形,在CLA培养基上肾型,(3.8～15.0)μm×(2.5～7.5)μm(n=50),隔膜数0～3;厚垣孢子在CLA培养基上呈串生,PDA培养基上呈不规则形,SNA培养基上厚垣孢子着生方式多样,串生、圆形、不规则形,大小为(2.5～3.8)μm×(2.5～5.0)μm(n=50)。EF-1α、RPB1和RPB2联合序列系统发育树表明,该菌为锐顶镰刀菌(Fusarium acuminatum),其最适生长温度为25℃,最适生长初始pH为9,光照对其生长无显著影响。室内毒力测定显示10%苯醚甲环唑(WG)对其抑制效果最好...