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[Allelopathy of root exudates from two genotypes soybeans on root pathogenic fungi].

With biological simulation experiment and chemical analysis, this paper studied the allelopathy of carbohydrates, amino acids and organic acids in the root exudates from two genotypes soybeans (9536 and Jilin 30) on the pathogenic fungi of root rot. The results showed that the water soluble carbohydrates in the root exudates from test soybeans significantly promoted the growth of Fusarium oxysporium and Fusarium semitectum at low concentrations and inhibited their growth at high concentrations, but had no evident influence on Gliocladium roseum. The water soluble amino acids from the root exudates demonstrated different actions, i. e., at middle and high concentrations, those from 9536 significantly inhibited the growth of Fusarium oxysporium, Fusarium semitectum and Gliocladium roseum, while those from Jilin 30 mostly promoted their growth. The organic acids from the root exudates of 9536 and Jilin 30 significantly inhibited the growth of Fusarium oxysporium, Fusarium semitectum and Gliocladium roseum. It's suggested that there existed interactions between the root exudates of the two genotypes soybeans and the pathogenic fungi of root rot. Different genotypes of soybean may have different allelopathy on pathogenic fungi of root rot.

Fusarium↗

Analysis of variation in Alternaria alternata by pathogenicity and RAPD study.

Alternaria alternata were isolated and identified from root, foliage and soil of both wheat and rice crops and their aggressiveness was studied using aggressiveness analysis. Isolates of Alternaria alternata were genetically characterized using RAPD's. The investigations were based on surveys of wheat and rice crops in the rice-wheat cropping areas of Pakistan. The study showed that Alternaria alternata is root rot causing fungi and during root rot aggressiveness analysis the isolates showed higher aggressiveness on rice varieties than on wheat but in foliar aggressiveness the overall number of aggressive isolates was higher on wheat varieties than on rice. In genetic characterization Random Amplified Polymorphism DNA (RAPD) was used to study the polymorphism and genetic variation within the population of Alternaria alternata that established correlation between aggressiveness and genetical characters of fungi. Alternaria alternata tree is constructed based on the pattern of bands. This study highlighted the correlation between morphological, aggressiveness and genetic variations of Alternaria alternata.

Alternaria↗

Effects of fungal root pathogens on the population dynamics of biocontrol strains of fluorescent pseudomonads in the wheat rhizosphere.

The influences of Gaeumannomyces graminis var. tritici (which causes take-all of wheat), Rhizoctonia solani AG-8 (which causes rhizoctonia root rot of wheat), Pythium irregulare, P. aristosporum, and P. ultimum var. sporangiiferum (which cause pythium root rot of wheat) on the population dynamics of Pseudomonas fluorescens 2-79 and Q72a-80 (bicontrol strains active against take-all and pythium root rot of wheat, respectively) in the wheat rhizosphere were examined. Root infection by either G. graminis var. tritici or R. solani resulted in populations of both bacterial strains that were equal to or significantly larger than their respective populations maintained on roots in the absence of these pathogens. In contrast, the population of strain 2-79 was significantly smaller on roots in the presence of any of the three Pythium species than on noninfected roots and was often below the limits of detection (50 CFU/cm of root) on Pythium-infected roots after 40 days of plant growth. In the presence of either P. aristosporum or P. ultimum var. sporangiiferum, the decline in the population of Q72a-80 was similar to that observed on noninfected roots; however, the population of this strain declined more rapidly on roots infected by P. irregulare than on noninfected roots. Application of metalaxyl (which is selectively inhibitory to Pythium spp.) to soil naturally infestated with Pythium spp. resulted in significantly larger rhizosphere populations of the introduced bacteria over time than on plants grown in the same soil without metalaxyl. It is apparent that root infections by fungal pathogens may either enhance or depress the population of fluorescent pseudomonads introduced for their control, with different strains of pseudomonads reacting differentially to different genera and species of the root pathogens.

Journal Article↗

Importance of antibiotic production in antagonism of selected Streptomyces species to two soil-borne plant pathogens.

Antagonism of ten Streptomyces spp., five of which produce antibiotics, to the plant pathogens Rhizoctonia solani and Phytophthora megasperma var. sojae was studied. Antibiotic activity was detected in culture for the five antibiotic producers. S. griseus, S. hygroscopicus var. geldanus, and S. noursei produced wide zones of inhibition to R. solani and P. megasperma var. sojae. Similar activity was found for S. reticuli var. protomycicus to P. megasperma var. sojae. S. cellulosae reduced Rhizoctonia root rot on pea when sterile soil was infested simultaneously with the antagonist and R. solani. S. hygroscopicus gave almost complete disease control when the streptomycete was added 7 days prior to infesting the soil with R. solani. Several of the Streptomyces spp. reduced Phytophthora root rot on soybean when the streptomycetes were added to soil at the same time as P. megasperma var. sojae or 7 days prior to adding the pathogen. S. herbaricolor and S. coeruleofuscus gave the most consistent control. No relationship was found between reported antibiotic activity or antagonism on agar media and reduction in disease severity. Only S. hygroscopicus var. geldanus gave both control of Rhizoctonia root rot and large zones of inhibition on agar media when the streptomycetes were preincubated in soil for 7 days.

Anti-Bacterial Agents↗

Introduction of the phzH gene of Pseudomonas chlororaphis PCL1391 extends the range of biocontrol ability of phenazine-1-carboxylic acid-producing Pseudomonas spp. strains.

Pseudomonas chlororaphis PCL1391 controls tomato foot and root rot caused by Fusarium oxysporum f. sp. radicis-lycopersici. Its biocontrol activity is mediated by the production of phenazine-1-carboxamide (PCN). In contrast, the take-all biocontrol strains P. fluorescens 2-79 and P. aureofaciens 30-84, which produce phenazine-1-carboxylic acid (PCA), do not control this disease. To determine the role of the amide group in biocontrol, the PCN biosynthetic genes of strain PCL1391 were identified and characterized. Downstream of phzA through phzG, the novel phenazine biosynthetic gene phzH was identified and shown to be required for the presence of the 1-carboxamide group of PCN because a phzH mutant of strain PCL1391 accumulated PCA. The deduced PhzH protein shows homology with asparagine synthetases that belong to the class II glutamine amidotransferases, indicating that the conversion of PCA to PCN occurs via a transamidase reaction catalyzed by PhzH. Mutation of phzH caused loss of biocontrol activity, showing that the 1-carboxamide group of PCN is crucial for control of tomato foot and root rot. PCN production and biocontrol activity of the mutant were restored by complementing the phzH gene in trans. Moreover, transfer of phzH under control of the tac promoter to the PCA-producing biocontrol strains P. fluorescens 2-79 and P. aureofaciens 30-84 enabled these strains to produce PCN instead of PCA and suppress tomato foot and root rot. Thus, we have shown, for what we believe is the first time, that the introduction of a single gene can efficiently extend the range of the biocontrol ability of bacterial strains.

Amino Acid Sequence↗

Phellinus Weirii and Other Native Root Pathogens as Determinants of Forest Structure and Process in Western North America.

The population structure and ecological roles of the indigenous pathogen Phellinus weirii, cause of laminated root rot in conifer forests of western North America, are examined. This pathogen kills trees in slowly expanding mortality centers, creating gaps in the forest canopy. It is widespread, locally abundant, and very long-lived. It is among the most important disturbance agents in the long intervals between stand-replacing events such as wildfire or harvest in these ecosystems and shapes the structure and composition of both wild and managed forests. Trees are infected and killed regardless of individual vigor. Management of public lands is changing dramatically, with renewed emphasis on natural forest structures and processes but pathogens, especially root rot fungi, remain a significant challenge to "ecosystem management."

Douglas-fir↗

Phylogenetic analyses of DNA sequences reveal species partitions amongst isolates of Armillaria from Africa.

The basidiomycete genus Armillaria causes root rot and death to woody plants in boreal, temperate and tropical regions of the world. Armillaria root rot has been described from various parts of Africa on many different hosts. However, very little is known regarding the evolutionary relationships among Armillaria species in Africa. The aim of this study was to determine the phylogenetic relationships between isolates originating from different regions in Africa using nDNA sequences from two non-coding gene regions. The ITS and the IGS-1 regions of the ribosomal DNA operon were sequenced and analysed using different phylogenetic tree searching methods. Phylogenetic trees grouped the African taxa in two strongly supported clades. One of these represented A. fuscipes and the other an undescribed but distinct species.

Africa↗

[Presearch on preventing the medicinal plant diseases with Trichoderma harzianum preparation].

OBJECTIVE: To control the medicinal plant diseases with the preparation of Trichoderma harzianum. METHOD: Antagonistic action of the preparation to the pathogens of the medicinal plants in vitro, and controling effects of the preparation on these diseases in greenhouse and in the field were tested. RESULT: The test in vitro showed that Trichoderma harzianum, used as a biocontrol factor, had stronger antagonistic action to Fusarium equiseti, Sclerotinia sp. and Rhizoctonia solani which were the medicinal plant pathogens of Astragalus membranaceus, Glehnia littoralis and Panax quinquefolium respectively. Biological controling effects on sclerotium root rot of Glehnia littoralis were 83.6% and 72.5% respectively in greenhouse and in the field with the preparation of Trichoderma harzianum. And controling effects on root rot of Astragalus membranaceus and seedling damping-off of Panax quinquefolium were 80% and 60% respectively in the field. The dosage of the preparation used in the field was 10 g.m-2. CONCLUSION: The preparation of Trichoderma harzianum can be used as a substitute for such chemicals as Carbendazim. Using the preparation to control medicinal plant diseases provides a technical safeguard for the good agricultural practice of medicinal plants.

Apiaceae↗

Relationships between Fusarium population structure, soil nutrient status and disease incidence in field-grown asparagus.

Fusarium species cause important diseases in many crops. Lack of knowledge on how Fusarium species and strains interact with their environment hampers growth management strategies to control root diseases. A field experiment involving asparagus as host plant and three phosphorus fertilization levels was designed to examine the seasonal changes and ecological relationships between Fusarium populations and their soil and plant environments. Fusarium taxa were identified and assessed using PCR-denaturing gradient electrophoresis of the EF1-alpha gene. Resulting profiles were analyzed with respect to 17 ecological parameters measured during the three main asparagus phenological phases across a growing season. Multivariate statistical analysis showed that Fusarium population structure was strongly influenced by soil P level while seasonal variation was less important. A significant relationship between Fusarium population composition and Fusarium crown and root rot incidence was also found in September. Canonical analysis further revealed significant relationships between Fusarium population structure, and plant manganese and iron contents, soil dehydrogenase activity and soil calcium concentration. If higher Fusarium crown and root rot incidence is related to the Fusarium community structure, strategies to reduce the incidence in asparagus plantations may be found through manipulation of the soil fertility.

Asparagus Plant↗

Combined effects of Brassica napus seed meal and Trichoderma harzianum on two soilborne plant pathogens.

The effects of soil amendment with rapeseed meal from Brassica napus cv. 'Dwarf Essex' (high glucosinolate concentrations) and 'Stonewall' (low glucosinolate concentrations) on the biological control activity of Trichoderma harzianum towards Sclerotinia sclerotiorum and Aphanomyces euteiches were evaluated. Trichoderma harzianum added to soil reduced myceliogenic germination of S. sclerotiorum by 94%, but did not affect carpogenic germination. In contrast, 100% reduction in carpogenic germination was observed in soil amended with Dwarf Essex meal, along with a 33% reduction in myceliogenic germination. With Stonewall meal as soil amendment, carpogenic germination was reduced by 44% and myceliogenic germination was not affected. Both Dwarf Essex and Stonewall meals inhibited colonization of sclerotia in soil by T. harzianum, from 100% to 0% and 8%, respectively, so that biocontrol activity of T. harzianum was reduced in the presence of either meal. Aphanomyces euteiches root rot of pea was significantly reduced by T. harzianum alone (100%), by amendment with Dwarf Essex meal alone (77%), and by T. harzianum in combination with Dwarf Essex meal (100%). Amendment with Stonewall meal alone did not control root rot, and combination of Stonewall meal with T. harzianum reduced the biocontrol efficacy of T. harzianum.

Ascomycota↗

Soil suppressiveness to Rhizoctonia solani and microbial diversity.

Rhizoctonia solani anastomosis group 2-2IIIB causes damping-off, black root rot and crown rot in sugar beet (Beta vulgaris). Based on experiences of growers and field experiments, soils can become suppressive to R. solani. The fungus may be present in the soil, but the plant does not show symptoms. Understanding the mechanisms causing soil suppressiveness to R. solani is essential for the development of environmentally friendly control strategies of rhizoctonia root rot in sugar beet. A bioassay that discriminates soils in their level of disease suppressiveness was developed. Results of bioassays were in accordance with field observations. Preliminary results indicate an active role of microbial communities. Our research is focused on the disentanglement of biological mechanisms causing soil suppressiveness to R. solani in sugar beet. Therefore, we are handling a multidisciplinary approach through experimental fields, bioassays, several in vitro techniques and molecular techniques (PCR-DGGE).

Antibiosis↗

Characterization of the hcnABC gene cluster encoding hydrogen cyanide synthase and anaerobic regulation by ANR in the strictly aerobic biocontrol agent Pseudomonas fluorescens CHA0.

The secondary metabolite hydrogen cyanide (HCN) is produced by Pseudomonas fluorescens from glycine, essentially under microaerophilic conditions. The genetic basis of HCN synthesis in P. fluorescens CHA0 was investigated. The contiguous structural genes hcnABC encoding HCN synthase were expressed from the T7 promoter in Escherichia coli, resulting in HCN production in this bacterium. Analysis of the nucleotide sequence of the hcnABC genes showed that each HCN synthase subunit was similar to known enzymes involved in hydrogen transfer, i.e., to formate dehydrogenase (for HcnA) or amino acid oxidases (for HcnB and HcnC). These similarities and the presence of flavin adenine dinucleotide- or NAD(P)-binding motifs in HcnB and HcnC suggest that HCN synthase may act as a dehydrogenase in the reaction leading from glycine to HCN and CO2. The hcnA promoter was mapped by primer extension; the -40 sequence (TTGGC ... ATCAA) resembled the consensus FNR (fumarate and nitrate reductase regulator) binding sequence (TTGAT ... ATCAA). The gene encoding the FNR-like protein ANR (anaerobic regulator) was cloned from P. fluorescens CHA0 and sequenced. ANR of strain CHA0 was most similar to ANR of P. aeruginosa and CydR of Azotobacter vinelandii. An anr mutant of P. fluorescens (CHA21) produced little HCN and was unable to express an hcnA-lacZ translational fusion, whereas in wild-type strain CHA0, microaerophilic conditions strongly favored the expression of the hcnA-lacZ fusion. Mutant CHA21 as well as an hcn deletion mutant were impaired in their capacity to suppress black root rot of tobacco, a disease caused by Thielaviopsis basicola, under gnotobiotic conditions. This effect was most pronounced in water-saturated artificial soil, where the anr mutant had lost about 30% of disease suppression ability, compared with wild-type strain CHA0. These results show that the anaerobic regulator ANR is required for cyanide synthesis in the strictly aerobic strain CHA0 and suggest that ANR-mediated cyanogenesis contributes to the suppression of black root rot.

Aerobiosis↗

Associations between DNA markers and resistance to diseases in sugarcane and effects of population substructure.

Association between markers and sugarcane diseases were investigated in a collection of 154 sugarcane clones, consisting of important ancestors or parents, and cultivars. 1,068 polymorphic AFLP and 141 SRR markers were scored across all clones. Data on the four most important diseases in the Australian sugarcane industry were obtained; these diseases being pachymetra root rot (Pachymetra chaunorhiza B.J. Croft & M.W. Dick), leaf scald (Xanthomonas albilineans Dowson), Fiji leaf gall (Fiji disease virus), and smut (Ustilago scitaminea H. & P. Sydow). By a simple regression analysis, association between markers and diseases could be readily detected. However, many of these associations were due to the effects of embedded population structure and random effects. After taking population structure into account, we found that 59% of the phenotypic variation in smut resistance ratings could be accounted for by 11 markers, 32% of variation for leaf scald and pachymetra root rot rating by 4 markers, and 26% of Fiji leaf gall by 5 markers. The results suggest that marker-trait associations can be readily detected in populations generated from modern sugarcane breeding programs. This may be due to special features of past sugarcane breeding programs leading to persistent linkage disequilibrium in modern parental populations.

Genetic Linkage↗

Molecular profiling of rhizosphere microbial communities associated with healthy and diseased black spruce (Picea mariana) seedlings grown in a nursery.

Bacterial and fungal populations associated with the rhizosphere of healthy black spruce (Picea mariana) seedlings and seedlings with symptoms of root rot were characterized by cloned rRNA gene sequence analysis. Triplicate bacterial and fungal rRNA gene libraries were constructed, and 600 clones were analyzed by amplified ribosomal DNA restriction analysis and grouped into operational taxonomical units (OTUs). A total of 84 different bacterial and 31 different fungal OTUs were obtained and sequenced. Phylogenetic analyses indicated that the different OTUs belonged to a wide range of bacterial and fungal taxa. For both groups, pairwise comparisons revealed that there was greater similarity between replicate libraries from each treatment than between libraries from different treatments. Significant differences between pooled triplicate samples from libraries of genes from healthy seedlings and pooled triplicate samples from libraries of genes from diseased seedlings were also obtained for both bacteria and fungi, clearly indicating that the rhizosphere-associated bacterial and fungal communities of healthy and diseased P. mariana seedlings were different. The communities associated with healthy and diseased seedlings also showed distinct ecological parameters as indicated by the calculated diversity, dominance, and evenness indices. Among the main differences observed at the community level, there was a higher proportion of Acidobacteria, Gammaproteobacteria, and Homobasidiomycetes clones associated with healthy seedlings, while the diseased-seedling rhizosphere harbored a higher proportion of Actinobacteria, Sordariomycetes, and environmental clones. The methodological approach described in this study appears promising for targeting potential rhizosphere-competent biological control agents against root rot diseases occurring in conifer nurseries.

Bacteria↗

Soil microbial populations, community composition, and activity as affected by repeated applications of hog and cattle manure in eastern Saskatchewan.

A field site near Humboldt, Saskatchewan, was annually treated with hog or cattle manure and cropped to canola, spring wheat, barley, and canola from 1997 to 2000. During each growing season, soil was analyzed for microbial populations in terms of activity and community structure, and crops were assessed for root rot and foliar diseases. Microbial activity in soils treated with cattle manure was higher than in soils treated with hog manure or urea. Similarly, nitrous oxide emissions from soil increased with increasing rates of hog and cattle manure. Potential human pathogens, including Rahnella, Serratia, Proteus, Leclercia, and Salmonella species, were identified in soils that received cattle manure, whereas pseudomonads were the dominant species in the hog-manure-treated soil. Fecal coliforms were confirmed in soils that received hog or cattle manure. However, Enterobacteriaceae populations were 10-fold higher in soils receiving cattle manure than in soils receiving the other treatments. Increasing cattle manure rates increased fecal coliform population, but there was no indication that increased hog manure rates increased fecal coliform populations. Addition of urea, hog manure, or cattle manure to the soil did not increase foliar disease in wheat, barley, and canola and had variable effects on root rot incidence in cereals.

Agriculture↗

Hydroponic chrysanthemum production: cultural and pathological issues.

Hydroponic culture has not replaced soil culture in greenhouse production of chrysanthemum (commonly known as 'mum'). This study examines cultural or pathological conditions that might have affected the conversion from soil to hydroponic production. Cultural factors investigated included hydroponic container size and shape and oxygenation of the nutrient solution. Disorders encountered during the studies included salt wicking during rooting of the cuttings and severe Pythium infection in the third and successive crops in a hydroponic system. Mums did not appear to respond to various shapes and sizes of containers and troughs in which they were grown. Also, increased oxygenation had little effect on pythium root rot and plant growth. Rooting of cuttings in a polyethylene covered hydroponic system reduced wilting, whereas salt wicking was severe without a plastic covering. Pythium disease was severe in the third and successive crops in the same hydroponic system. The disease could be overcome by raising the cuttings in a peat-based growing medium in perforated plastic cells and then transplanting the seedlings along with the peat moss cubes into a hydroponic system.

Chrysanthemum↗

Loss of pectin is an early event during infection of cocoyam roots by Pythium myriotylum.

Cocoyam (Xanthosoma sagittifolium) is an important tuber crop in most tropical zones of Africa and America. In Cameroon, its cultivation is hampered by a soil-borne fungus Pythium myriotylum which is responsible for root rot disease. The mechanism of root colonisation by the fungus has yet to be elucidated. In this study, using microscopical and immunocytochemical methods, we provide a new evidence regarding the mode of action of the fungus and we describe the reaction of the plant to the early stages of fungal invasion. We show that the fungal attack begins with the colonisation of the peripheral and epidermal cells of the root apex. These cells are rapidly lost upon infection, while cortical and stele cells are not. Labelling with the cationic gold, which binds to negatively charged wall polymers such as pectins, is absent in cortical cells and in the interfacial zone of the infected roots while it is abundant in the cell walls of stele cells. A similar pattern of labelling is also found when using the anti-pectin monoclonal antibody JIM5, but not with anti-xyloglucan antibodies. This suggests that early during infection, the fungus causes a significant loss of pectin probably via degradation by hydrolytic enzymes that diffuse and act away from the site of attack. Additional support for pectin loss is the demonstration, via sugar analysis, that a significant decrease in galacturonic acid content occurred in infected root cell walls. In addition, we demonstrate that one of the early reactions of X. sagittifolium to the fungal invasion is the formation of wall appositions that are rich in callose and cellulose.

Cell Wall↗

A role for jasmonate in pathogen defense of Arabidopsis.

To investigate the role of jasmonate in the defense of plants against fungal pathogens, we have studied a mutant of Arabidopsis, fad3-2 fad7-2 fad8, that cannot accumulate jasmonate. Mutant plants were extremely susceptible to root rot caused by the fungal root pathogen Pythium mastophorum (Drechs.), even though neighboring wild-type plants were largely unaffected by this fungus. Application of exogenous methyl jasmonate substantially protected mutant plants, reducing the incidence of disease to a level close to that of wild-type controls. A similar treatment with methyl jasmonate did not protect the jasmonate-insensitive mutant coi1 from infection, showing that protective action of applied jasmonate against P. mastophorum was mediated by the induction of plant defense mechanisms rather than by a direct antifungal action. Transcripts of three jasmonate-responsive defense genes are induced by Pythium challenge in the wild-type but not in the jasmonate-deficient mutant. Pythium species are ubiquitous in soil and root habitats world-wide, but most (including P. mastophorum) are considered to be minor pathogens. Our results indicate that jasmonate is essential for plant defense against Pythium and, because of the high exposure of plant roots to Pythium inoculum in soil, may well be fundamental to survival of plants in nature. Our results further indicate that the fad3-2 fad7-2 fad8 mutant is an appropriate genetic model for studying the role of this important signaling molecule in pathogen defense.

Journal Article↗