Effect of Placement and Sequence of Inoculation with Fusarium oxysporum on its Interaction with Meloidogyne arenaria on Subterranean Clover

The effect of the placement of inoculum of Fusarium oxysporum at two soil depths, and the sequences of inoculations with Meloidogyne arenaria and Fusarium oxysporum on root growth and development of root disease in Trifolium subterraneum L. (subterranean clover) were investigated. The timing of infection and the proximity of root tips of the host root system to infection by M. arenaria and F. oxysporum appeared to be the major determining factors of root growth and of disease development in plants exposed to the pathogens. Immediate contact of roots with F. oxysporum (where the fungus was placed at seed level of 10 mm depth) appeared to result in more severe effects on roots in the presence of the nematode than later infection by the fungus placed at 30 mm depth. The production of galls by the nematode and early infection by F. oxysporum at 10 mm depth resulted in a severe inhibition of root growth, particularly of the lateral roots. But no such growth inhibition was evident when F. oxysporum and M. arenaria were introduced together at the lower depth of 30 mm. The lowest density of M. arenaria inoculum was sufficient to cause severe root rot if F. oxysporum was present at the host seed level. With the fungus at 30 mm depth, however, the expression of root rot appeared to be influenced by the inoculum level of the nematode. In sequential inoculation with F. oxysporum or M. arenaria, the organism added 2 weeks later had little or no effect on root development. The first organism (M. arenaria or F. oxysporum) to infect the germinated seedlings was the main cause of root growth inhibition. The organism that came into contact with the roots 2 weeks later had little or no effect on the roots. Concurrent infection by F. oxysporum and M. arenaria resulted in less M. arenaria gall production on the tap root system than those added with the nematode alone or in advance of the fungus.     

Fungi associated with boll and lint rot of cotton in Southern Khorasan province of Iran

During surveys of cotton fields since 2009 in the Southern Khorasan province of Iran, symptomatic samples with lint and boll rot disease signs were collected and cultured on potato dextrose, malt extract agar and PARPH media. Several fungal isolates were obtained and identified. Among them, we found Penicillium expansum which to our knowledge is reported as a new pathogen of cotton worldwide; Fusarium semitectum and Nigrospora oryzae (associated to mite Siteroptes sp.) were also recorded as lint or boll rot pathogens for the first time from Iran. Disease symptoms and fungal characteristics of Exserohilum rostratum, which has been reported briefly as a new causal agent of boll and lint rot disease, are illustrated and presented. In addition, Aspergillus niger and Rhizopus spp. were also isolated frequently from lint parts of cotton prior to harvesting. There was a positive correlation between disease occurrence and spiny cotton bollworm damage. The pathogenicity test was characterised by inoculating the attached and detached bolls of cotton plants.     

First Report of <i>Fusarium striatum</i> Causing Root Rot Disease of <i>Panax notoginseng</i> in Yunnan,China

Panax notoginseng is a traditional Chinese medicinal plant. Root rot of P. notoginseng is one of the most serious diseases affecting P. notoginseng growth and causes wilted leaves, fewer lateral roots and rotten roots. Root rot is a soil-borne disease, and mainly occurs from June to August in Yunnan Province when the temperatures are high and the air is humid. In this study, the endophytic fungal genus Fusarium isolate E-2018.1.22-#3.2 was obtained from a P. notoginseng embryo. Fusarium isolate E-2018.1.22-#3.2 was identified as Fusarium striatum based on morphological characteristics and molecular analysis. The fungus was found to have conidiophores and macroconidia, and its ITS, LSU and TEF-1α genes shared 100%, 99.2% and 99% identities with the homologous genes of Fusarium striatum, respectively. Isolate F. striatum E-2018.1.22-#3.2 can cause root rot symptoms, including black, soft roots, fewer lateral roots and leaf wilt, in 93% of the experimental P. notoginseng plants, and could be re-isolated, fulfilling Koch’s postulates. When the P. notoginseng plants were treated with the fungicide pyraclostrobin, isolate F. striatum E-2018.1.22-#3.2 was unable to cause root rot. We have therefore demonstrated that F. striatum E-2018.1.22-#3.2 is able to cause root rot disease in P. notoginseng. This is the first report of root rot disease caused by F. striatum on P. notoginseng in China.     

Comparison of pathogenicity of Clavibacter michiganensis subsp. insidiosus and Pseudomonas viridiflava strains on alfalfa

Hamedan province of Iran is a suitable niche for alfalfa growth but many diseases including alfalfa bacterial wilt, bacterial crown and root rot diseases cause economic crop losses. Bacterial wilt is caused by Clavibacter michiganensis subsp. insidiosus, and bacterial crown and root rot diseases are caused by Pseudomonas viridiflava. In this study, we investigated the pathogenicity of C. michiganensis subsp. insidiosus and P. viridiflava strains collected from the main alfalfa growing areas of Hamedan province. Pathogenicity of the virulent strains was tested on alfalfa and the bacterial strains caused symptoms, and data were collected about stem length, root length, wet weight and dry weight of the infected plantlets. The data about the pathogenicity of C. michiganensis subsp. insidiosus and P. viridiflava on alfalfa were compared with each other and were analysed by SAS software and Dunkan's test. Resulted data showed more pathogenicity of C. michiganensis subsp. insidiosus than P. viridiflava on alfalfa. These data also showed that both of these bacteria produced the most losses on wet weight and dry weight of alfalfa plantlets.     

Minor root rot pathogens of cassava (Manihot esculenta Crantz) in Nigeria

Many different species of fungi are often isolated from rotted cassava root tubers and pathogenicity studies have often implicated Botryodiplodia theobromae and Fusarium solani as the major causal pathogens. Consequently, more attention has often been focused on Botryodiplodia theobromae and Fusarium solani with little or no attention on the other minor pathogens. Considering the increasing importance of cassava to the Nigerian economy and the fact that minor root rot pathogens of cassava today could become major tomorrow, the aim of this research is to determine the incidence, pathogenicity and symptoms of the minor root rot pathogens in cassava from cassava fields within the derived savanna and the humid forest of Nigeria. Isolation of associated fungi was done on rotted root samples and the pathogenicity of these isolates were established by inoculating them into healthy cassava tuberous roots and subsequently reisolating them from resulting rotted tissue. The less frequently isolated fungi where Macrophomina sp., Trichoderma sp., Aspergillus niger, Aspergillus flavus, Sclerotium rolfsii and Fungus ‘A’ (a yet to be identified fungus). Repeated experiments confirmed a constant relationship between inoculated fungus and the resulting rotted tissue colour. The root rot tissue colours associated with inoculated pathogens in the laboratory were identical with the pathogens colony colour on potato dextrose agar.     

Efficiency of different application methods of biocontrol agents and biocides in control of Fusarium root rot on some citrus rootstocks

Abstract

The efficiency of two biocontrol agents (Trichoderma harzianum NB and Bacillus subtilis NB) and two commercial biocides (Plant Guard and Rhizo-N) in controlling Fusarium root rot disease on some citrus rootstocks was evaluated under artificially infested soil in green house.

Fusrium root rot on citrus rootstocks seedlings i.e. sour orange (SO), volkamer lime (VL), rangpur lime (RP) and cleopatra mandarin (CL) was successfully controlled by dipping the root system of such seedlings in water suspensions of each biological treatment i.e. Trichoderma harzianum (spore suspension 5×10⁶ spore/ml), Bacillus subtilis (cell suspension 8×10⁷ cell/ml), Plant Guard (3 g/l) and Rhizo-N (4 g/l), then transplanted into artificially infested soil with Fusarium solani and drenched with enough water suspension of such biological treatments. Plant Guard (3 g/l) and Rhizo-N (4 g/l) were highly effective treatments in decreasing infection and severity of the disease, Fusarium density in rhizosphere soil and colonization of Fusarium solani in the roots of all tested seedlings.

Meanwhile, root dipping or soil drenching with the same treatments individually gave the least effect in reducing root rot incidenceon all tested rootstocks compared with application of the two methods together.

It should be noted that using biocontrol agents and commercial biocides could be successfully used in controlling root rot pathogens on citrus in commercial greenhouses or under field conditions before transplanting in new reclaimed lands in the desert.     

一种由粉红粘帚霉引起的青稞根腐病

[背景]根腐病在青稞生产中的危害日趋严重,阻碍了青稞根腐病的有效防控及青海省青稞产业的发展。然而人们对青稞根腐病的研究甚少且病原菌不详。[目的]明确青稞根腐病发生的危害、病原及致病性,为青稞根腐病的防控提供理论依据。[方法]采用常规的组织分离法分离青稞根腐病病原,通过形态鉴定与分子鉴定结合的方法对病原进行鉴定,并采用烧杯水琼脂法测定其致病性。[结果]共分离得到4株青稞根腐病病原菌,鉴定为Clonostachys rosea,有较强的致病性且致病性差异显著,经柯赫氏法则验证为青稞根腐病病原菌,并且是一种新的青稞根腐病病原,该类根腐病也是一种新的根腐类病害,在国内外属首次发现。[结论]Clonostachys rosea可引起青稞根腐病且致病性强。     

Green manures and crop sequences influence alfalfa root rot and pathogen inhibitory activity among soil-borne streptomycetes

A two-year trial was conducted to determine the effects of green manures and crop sequences on plant disease, streptomycete and bacterial densities, and inhibitory activity of indigenous streptomycetes against four target pathogens. Green manure treatments, buckwheat (Fagopyrum esculentum L.), canola (Brassica napus L.), sorghum-sudangrass (Sorghum bicolor) (L.) Moench × Sorghum sudanense (Piper) Stapf.), and fallow control were tested in conjunction with three crop sequences in a Phytophthora-infested soil placed in containers. Alfalfa (Medicago sativa L.), potato (Solanum tubersoum L.), or corn (Zea mays L.) was grown in the first year, and alfalfa was grown in all containers in the second year. Compared to fallow controls, alfalfa grown in sorghum-sudangrass- or buckwheat-treated soil had significantly greater stand counts and total biomass, respectively. In addition, alfalfa grown in fallow-treated soils had the greatest Phytophthora root rot as a function of stand count. Crop rotation also had a significant effect on alfalfa root rot and yield. Potato scab disease intensity was greatest on tubers grown in fallow-treated soils, while tubers grown in canola-treated soils had the highest yields (total tuber weight). Green-manure-treated soils tended to have greater streptomycete and bacterial densities than fallow-treated soils. In addition, buckwheat- or sorghum-sudangrass-treated soils had greater proportions of streptomycetes that were antagonistic against the target pathogens than fallow-treated soils. The proportion of antagonists in soil was negatively correlated with alfalfa root rot, and positively correlated with alfalfa stand counts. Inhibitory activity of the streptomycetes was also negatively correlated with potato scab and positively correlated with potato yield. These data suggest that green manures may provide a strategy for increasing pathogen inhibitory activity within the streptomycete community in soil, and, in conjunction with crop rotation, may contribute to the control of a diverse collection of soil-borne plant pathogens on multiple crop species.     

Endophytic colonisation of Bacillus subtilis in the roots of Robinia pseudoacacia L

The endophytic colonisation of Bacillus subtilis strain GXJM08, isolated from roots of Podocarpus imbricatus B1. Enum. P1. Jav., in roots of the leguminous plant Robinia pseudoacacia L. was investigated. Ultrastructure observations showed that B. subtilis caused morphological changes in the root hair and colonised the plant through infected root hairs. The structure of the infection thread was similar to that of rhizobia, but the structure of infected cells was different. B. subtilis is also different from rhizobia and plant pathogens in terms of the formation of a peribacteroid membrane and the mode of penetration through the host cell wall. Our results provide a basis for studying development of the mutualistic symbiotic relationship between B. subtilis and plants, and a basis for studying the mechanism of the B. subtilis–plant interaction.     

Production of Tsr factor byRhizobium meliloti

The root exudates of alfalfa (Medicago sativa) and mungbean (Vigna radiata) induced the Tsr (thick and short roots) factor production inRhizobium meliloti. The factor caused a 30–40% reduction of root length in alfalfa seedlings. Pea root exudate had no Tsr induction activity. The flavonoid naringenin could replace the roots in inducing Tsr production. Naringenin-induced Tsr factor caused 70% shortening of main roots. The Tsr inducing property of naringenin was specific since quercetin and syringaldehyde had no such effect.     

Phytohormones and Antibiotics Produced by Bacillus subtilis and their Effects on Seed Pathogenic Fungi and on Soybean Root Development

Summary Bacteria belonging to the genus Bacillus were isolated from soil samples of Paraná State, Brazil, with the aim of evaluating their potential biological control of soybean seed pathogens. Strain PRBS-1 was selected, showing similar effectiveness to that of the strain AP-3, used as a reference due to its known antibiotic potential. The sequencing of the ribosomal 16S rRNA gene confirmed that both strains belong to the species B. subtilis, although showing high genetic diversity in relation to this species. Both strains inhibited five soybean seed pathogenic fungi in vitro, Rhizoctonia solani, Colletotrichum truncatum, Sclerotinia sclerotiorum, Macrophomina phaseolina and Phomopsis sp. Furthermore, the metabolites of AP-3 increased production of root hairs, while the metabolites of PRBS-1 stimulated outgrowth of lateral roots in soybean. The antibiotic effect of both strains seemed to be related to compounds of the iturin group, while the root growth promotion by PRBS-1 was at least partially related to the production of indoleacetic acid. The results have shown the potential of using selected strains of B. subtilis in the biological control of seed pathogens, as well as in promoting soybean growth.     

Root rot of onion caused byPythium irregulare andPythium coloratum

Pythium irregulare andPythium coloratum were isolated consistently from roots of onion plants exhibiting root rot and associated foliar symptoms in fields located in Organe County, NY.P. coloratum predominated following extremely wet weather in 1984 and 1986, whileP. irregulare was prevalent following moderately wet weather in 1985. Both species produced root rot symptoms similar to field symptoms when 12 week old onion plants (cv Downing Yellow Globe) were inoculated and incubated in a growth chamber at 14°C for 10 days. This is the first report ofP. irregulare andP. coloratum as causes of root rot of onion.     

The effect of mode of application of Bacillus subtilis CA32r on control of Sclerotium rolfsii on Capsicum annuum

Abstract

Sclerotium rolfsii Sacc. the causal agent of collar and root rot of chili (Capsicum annuum L.), is one of the most important soil-borne pathogens in Sri Lanka. Bacillus subtilis CA32r, a stable spontaneous kanamycin resistant isolate, showing antagonism in a Petri plate assay, was selected for greenhouse pot experiments to control S. rolfsii. An ethyl acetate extract of the culture filtrate of B. subtilis CA32r inhibited radial colony growth as well as germination of sclerotia of the pathogen in vitro, indicating the presence of antifungal compound(s) in the culture extract. B. subtilis CA32r was investigated for its effectiveness as a biological control agent against S. rolfsii infecting transplanted chilli seedlings in greenhouse pot experiments. CA32r significantly decreased the disease incidence in terms of lesion development on stem base and roots depending on the mode of the bacterial application. Seed bacterization and soil application alone did not protect chilli plants, but root bacterization prior to the transplanting of seedlings significantly decreased the disease incidence caused by S. rolfsii. However, even in the combined treatment, seed bacterization and soil application, did not protect chilli plants. The best protection was achieved by combination of root bacterization prior to transplant and soil application of CA32r. Root bacterization resulted in maintaining higher numbers of bacteria at the collar region of chilli plants and may have shielded the most vulnerable area from the pathogen, resulting enhanced protection. Since the application of CA32r resulted in a significant reduction of the number of viable S. rolfsii propagules in the soil indicates that B. subtilis CA32r possesses not only protective but also eradicative potential.     

Phenotypic, molecular phylogenetic, and pathogenetic characterization of Fusarium crassistipitatum sp. nov., a novel soybean sudden death syndrome pathogen from Argentina and Brazil

A novel soybean sudden death syndrome (SDS) pathogen from Argentina and Brazil is formally described herein as Fusarium crassistipitatum based on detailed phenotypic analyses of macro- and microscopic characters and phylogenetic analyses of multilocus DNA sequence data. Fusarium crassistipitatum can be distinguished from the other soybean SDS and bean (Phaseolus/Vigna) root rot pathogens (BRR) phenotypically by the production of yellowish colonies on PDA; and tall, stout, and mostly unbranched conidiophores with a thick-walled base, which form multiseptate conidia apically. Phylogenetic species recognition based on genealogical concordance of a six-gene dataset strongly supported the reciprocal monophyly of F. crassistipitatum with respect to the other SDS and BRR pathogens. Isolates of F. crassistipitatum were able to induce typical SDS foliar and root rot symptoms on soybean that were indistinguishable from those caused by three other SDS pathogens (i.e., F. virguliforme, F. brasiliense, and F. tucumaniae) on susceptible cultivars A-6445RG and N-4613RG in a pathogenicity experiment.     

First report of Plectosphaerella plurivora causing root rot disease in Panax notoginseng in China

Panax notoginseng is a highly valuable herb, but root rot disease severely impairs its development and decreases the yield and quality of roots. In this study, a fungal isolate (3A-2-2) was obtained from P. notoginseng seedlings with root rot symptoms and was identified as Plectosphaerella plurivora based on morphological characteristics and molecular analysis. The fungal isolate 3A-2-2 could cause root rot disease and be re-isolated, fulfilling Koch's postulates. This is the first study to report on root rot disease caused by P. plurivora on P. notoginseng in China.     

Characterization of blackleg and soft rot from potato in northern Thailand

Blackleg and soft rot of potato cause economic loss through reduced yield and quality. The causal agents of bacterial blackleg and soft rot of potato were identified based on biological data and sequence analyses of the 16S rDNA gene. Between 2016 and 2018, diseased potato stems and tubers were collected in Chai Prakan District, Chiang Mai Province, and Chiang Khum District, Pa Yao Province. The symptoms included black stem lesions, soft rot on tubers, wilting, break down of the stem vascular ring and foliar yellowing. Of 13 bacterial isolates, five were identified as Pectobacterium carotovorum subsp. brasiliense, four‐Dickeya dadantii, two‐Pseudomonas putida and two‐Bacillus altitudinis. Pathogenicity tests of P. carotovorum subsp. brasiliense and D. dadantii resulted in lower leaves turning yellow and wilting followed by blackleg symptoms on lower stems and maceration of tuber tissue. Symptoms caused by P. putida were yellowing and wilting of leaves. B. altitudinis caused yellowing of the lower leaves and wilting followed by drying of leaf tissue. This is a first report of these bacterial pathogens causing blackleg and soft rot of potato in Thailand.     

Colonization of barley roots by Fusarium culmorum and influence of Pseudomonas fluorescens on the process

The stages of barley root colonization by Fusarium culmorum were studied in sterile vermiculite by the method of fluorescent antibodies. The influence of the antagonistic bacterium Pseudomonas fluorescens on the process of root colonization by F. culmorum was demonstrated. In vermiculite inoculated with F. culmorum, the fungus density on the roots increased gradually. In the case of joint inoculation of vermiculite with the fungus and the bacterium, the F. culmorum density on the roots changed abruptly. It was shown that the site of primary colonization of the roots by the fungus was mainly the zone of root hairs. When Pseudomonas fluorescens was present on the roots, F. culmorum colonized not only root hairs, but also the elongation zone, during the first two days. Introduction of Pseudomonas fluorescens into vermiculite resulted in lower intensity of barley root rot.     

Evaluation of isolates of Trichoderma,Pseudomonas and Bacillus species as treatment for the control of post-harvest fungal rot disease of yam (Dioscorea spp.)

The efficacy of four biological control agents (BCAs): Trichoderma asperellum strain NGT158, T. longibrachiatum strain NGT167, Bacillus subtilis and Pseudomonas fluorescens for the management of post-harvest tuber rot among four yam species, Dioscorea rotundata, D. cayenensis, D. alata, and D. dumetorum was evaluated. Rotted yam tubers were collected across three agroecological zones in Nigeria to isolate six infecting fungal pathogens: Aspergillus niger, Lasiodiplodia theobromae, Rhizoctonia solani, Penicillium oxalicum, Fusarium oxysporum and Sclerotium rolfsii. The BCAs were isolated by serial dilution and rot inhibition of treated tubers was evaluated using destructive sampling method in vivo after six months of storage. Bacillus subtilis was generally most effective, especially when applied 24 h before the inoculation of test pathogens across the four yam species, with percent inhibition that ranged between 47.8 and 81.2%. However, the four BCAs showed good potential in the control of the fungal pathogens causing post-harvest yam rot.     

Characterization of Iranian Pectobacterium carotovorum Strains from Sugar Beet by Phenotypic Tests and Whole-cell Proteins Profile

Thirty isolates of Pectobacterium carotovorum from soft rot‐affected sugar beet plants in the Fars province of Iran were characterized phenotypically and by analysis of whole‐cell protein electrophoresis patterns. The isolates were found to be heterogeneous based on the results of physiological and biochemical tests and protein profiles. The results of numerical analysis of phenotypic characteristics and protein patterns showed that only 27% of the collected isolates (phenon 4) could be identified as P. betavasculorum when compared with reference strains. Strains of the first, second, third and fifth phenon shared similar characters with those of P. carotovorum subsp. carotovorum, P. betavasculorum and P. carotovorum subsp. odoriferum, but were distinct from these subspecies. Inoculation of phenon 4 isolates into wounded sugar beet petioles led to black streaking, root rot and vascular necrosis. Other isolates were incapable of causing systemic symptoms in inoculated plants.     

Fine root demography in alfalfa (Medicago sativa L.)

In perennial forages like alfalfa (Medicago sativa L.), repeated herbage removal may alter root production and mortality which, in turn, could affect deposition of fixed N in soil. Our objective was to determine the extent and patterns of fine-diameter root production and loss during the year of alfalfa stand establishment. The experiment was conducted on a loamy sand soil (Udorthentic Haploboroll) in Minnesota, USA, using horizontally installed minirhizotrons placed directly under the seeded rows at 10, 20, and 40 cm depths in four replicate blocks. We seeded four alfalfa germplasms that differed in N₂ fixation capacity and root system architecture: Agate alfalfa, a winter hardy commercially-available cultivar; Ineffective Agate, which is a non-N₂-fixing near isoline of Agate; a new germplasm that has few fibrous roots and strong tap-rooted traits; and a new germplasm that has many fibrous roots and a strongly branched root system architecture. Video images collected biweekly throughout the initial growing season were processed using C-MAP-ROOTS software.More than one-half of all fine roots in the upper 20 cm were produced during the first 7 weeks of growth. Root production was similar among germplasms, except that the highly fibrous, branch-rooted germplasm produced 29% more fine roots at 20 cm than other germplasms. In all germplasms, about 7% of the fine roots at each depth developed into secondarily thickened roots. By the end of the first growing season, greatest fine root mortality had occurred in the uppermost depth (48%), and least occurred at 40 cm (36%). Survival of contemporaneous root cohorts was not related to soil depth in a simple fashion, although all survivorship curves could be described using only five rates of exponential decline. There was a significant reduction in fine root mortality before the first herbage harvest, followed by a pronounced loss (average 22%) of fine roots at the 10- and 20-cm depths in the 2-week period following herbage removal. Median life spans of these early-season cohorts ranged from 58 to 131 days, based on fitted exponential equations. At all depths, fine roots produced in the 4 weeks before harvest (early- to mid-August) tended to have shorter median life spans than early-season cohorts. Similar patterns of fine root mortality did not occur at the second harvest. Germplasms differed in the pattern, but not the ultimate extent, of fine root mortality. Fine root turnover during the first year of alfalfa establishment in this experiment released an estimated 830 kg C ha^–1 and 60 kg N ha^–1, with no differences due to N₂ fixation capacity or root system architecture.