Occurrence and distribution of tomato seed-borne mycoflora in Saudi Arabia and its correlation with the climatic variables

One hundred samples of tomato seeds were collected in 2011 and 2012 from tomato-cultivated fields in Saudi Arabia and screened for their seed-borne mycoflora. A total of 30 genera and 57 species of fungi were recovered from the collected seed samples using agar plate and deep-freezing blotter methods. The two methods differed as regards the frequency of recovered seed-borne fungi. Seven fungi among those recovered from tomato seeds, which are known as plant pathogens, were tested for their pathogenicity and transmission on tomato seedlings. The recovery rate of these pathogens gradually decreased from root up to the upper stem, and did not reach to the stem apex. The distribution of tomato seed-borne fungi was also investigated throughout Saudi Arabia. In this concern, Al-Madena governorate recorded the highest incidence of fungal flora associated with tomato seeds. The impact of meteorological variables on the distribution of tomato seed-borne mycoflora was explored using the ordination technique (canonical correspondence analysis). Among all climatic factors, relative humidity was the most influential variable in this regard. Our findings may provide a valuable contribution to our understanding of future global disease change and may be used also to predict disease occurrence and fungal transfer to new uninfected areas.     

Treatment of lentil (Lens culinaris. L) seeds with various concentrations of salts to check their efficiency against seed-borne mycoflora during storage

Seed sample of lentil collected from the Swabi district were treated with NaCl and KCl at 0.01, 0.1 and 1.0% (w/w). Seed-borne mycoflora was observed at 0, 20, 40, 60 and 80 day intervals. Seed treatment with both the salts was found to be effective against storage fungi; however, KCl was more efficient against storage mycoflora such as Aspergillus species when compared with NaCl. With the passage of time, the incidence of deep-seated fungi was observed in salt-treated seed samples while untreated seed sample showed heavy infestation by the species of Aspergillus. Seed viability also remained unaffected in storage, except in seeds heavily infested with seed-borne mycoflora. Aspergillus spp. were the main cause of seed rot. Surface sterilisation of seeds with 1% Na(OCl)₂ reduced the fungal infestation of seeds. Among various concentrations of salts, 0.1% (w/w) of both salts were better in controlling seed-borne mycoflora.     

Hot Acidified Cupric Acetate Soaks for Eradication of Xanthomonas campestris from Crucifer Seeds

Acidified cupric acetate soaks were tested for eradication of Xanthomonas campestris from naturally infected crucifer seeds. The pathogen was eradicated from seeds by soaking in 0.5% cupric acetate dissolved in 0.005 N acetic acid for 20 min at 35, 40, 45, and 50°C but not 25°C. Moreover, normal bacterial flora of crucifer seeds and the seed-borne Phoma lingam and Alternaria spp. were reduced by 95, 92, and 81%, respectively, after the cupric acetate treatment at 40°C. The seed germination percentage was generally reduced, but the amount of reduction depended upon the treatment temperature and plant cultivar. At 50°C, less than 50% of the seed of all 12 cultivars tested germinated, whereas at 40°C more than 50% of the seeds of most cultivars germinated. Treating seeds in cupric acetate at 40°C should prove useful for eradicating X. campestris from seeds of breeding lines and stock seed used for hybrid seed production. Furthermore, a significant reduction in total bacterial flora and seed-borne fungi suggests the usefulness of the treatment for other microorganisms associated with other seeds or foodstuffs.     

Sowing seasons and drying methods during post-harvest influence the seed vigour of soybean (Glycine max (L.) Merr.)

Experiments were conducted to study the influence of sowing seasons and drying methods on the seed vigour of two spring soybean (Glycine max (L.) Merr.) cultivars. Two cultivars, ‘Huachun18’ and ‘Huachun 14’, were sown in three seasons viz., spring, summer and autumn and the harvested seeds were dried using three different methods. The results showed that soybean sown in spring had a higher number of branches per plant, pods per branch and seed weight, and consequently resulted in higher seed yields than that of soybean sown in autumn or summer seasons. Seeds sown in the autumn season had the lowest values of electrical conductivity during seed imbibitions, higher peroxidase (POD) activity in germinated seedlings and lower contamination by the seed-borne fungi on the MS medium, which indirectly improved the seed vigour, which was followed by summer sown seeds. Seeds sown during the spring season resulted in poor seed vigour. In addition, the effect of drying methods on the seed vigour was also clarified. Seeds that hung for four days before threshing and then air-dried had the poorest seed vigour which was determined by germination, electrical conductivity, POD activity and seed borne fungal growth. There was no difference in seed vigour between other methods, i.e. seeds threshed directly at harvest and then air-dried on a bamboo sifter or concrete floor. These results indicated that autumn sowing soybean and the drying method in which seeds were threshed directly at harvest and then air-dried on a bamboo sifter resulted in higher seed vigour.     

Stomatal and chlorophyll fluorescence characteristics in European beech cultivars during leaf development

Changes in stomatal and chlorophyll fluorescence characteristics were analyzed in the course of leaf expansion in European beech (Fagus sylvatica L.) cultivars Aurea Pendula, Cristata, Rohanii, Rotundifolia and Viridivariegata. Stomatal length increased gradually from the second to the fifth phenological stage. Rotundifolia reached the highest mean stomatal length whereas Aurea Pendula and Cristata had the lowest values. Stomatal density for all cultivars decreased from the second to the fifth stage. Aurea Pendula reached the highest stomatal density in all phenological stages. The highest values of variable to maximum fluorescence ratio (F_v/F_m) were recorded in Rotundifolia, Rohanii, and the wild type, whereas Viridivariegata showed the lowest F_v/_Fm. Similar trend was found in maximum to initial fluorescence ratio (F_m/F₀), but extremely low F_m/F₀ values were recorded in Viridivariegata in the last phenological stage. The highest potential electron capacity was found in Rohanii, Viridivariegata and the wild type and lowest in Cristata. This parameter increased in the course of early leaf development.     

Fungi pathogenic to ryegrass seedlings

Summary Fungi borne on or in ryegrass (Lolium spp.) seeds or invading ryegrass seedlings grown on field soils were isolated and identified. Selected isolates were tested to determine their pathogenicity to ryegrass seedlings. Seed-borne fungi were generally weakly virulent or non-pathogenic to ryegrass seedlings. Pathogenic seed-borne fungi includedChaetomium globosum Kunze: Fr.,Curvularia trifolii (Kauffm.) Boedijn, and species ofPenicillium Link andAspergillus Mich. ex Link. Species of fungi isolated from seedlings grown on field soils de pended on soil and temperature. Soil-borne fungi pathogenic to seedlings includedFusarium avenaceum (Fr.) Sacc.,F. culmorum (W. G. Smith) Sacc.,F. equiseti (Corda) Sacc.,F. oxysporum Schlecht.: Fr.,F. solani (Mart.) Sacc.,Pythium afertile Kanouse and Humphrey,P. debaryanum auct. non Hesse,P. irregulare Buisman,P. ultimum Trow, a sphaerosporangiatePythium sp.,Chaetomium globosum, Thanatephorus cucumeris (Frank) Donk,Trichoderma koningii Oudem., and aPhomopsis sp. Individual isolates of fungi differed in virulence to ryegrass seedlings, and ryegrass cultivars differed in susceptibility to seedling pathogens.     

Culturable Endophytes Associated with Soybean Seeds and Their Potential for Suppressing Seed-Borne Pathogens

Seed-borne pathogens in crops reduce the seed germination rate and hamper seedling growth, leading to significant yield loss. Due to the growing concerns about environmental damage and the development of resistance to agrochemicals among pathogen populations, there is a strong demand for eco-friendly alternatives to synthetic chemicals in agriculture. It has been well established during the last few decades that plant seeds harbor diverse microbes, some of which are vertically transmitted and important for plant health and productivity. In this study, we isolated culturable endophytic bacteria and fungi from soybean seeds and evaluated their antagonistic activities against common bacterial and fungal seed-borne pathogens of soybean. A total of 87 bacterial isolates and 66 fungal isolates were obtained. Sequencing of 16S rDNA and internal transcribed spacer amplicon showed that these isolates correspond to 30 and 15 different species of bacteria and fungi, respectively. Our antibacterial and antifungal activity assay showed that four fungal species and nine bacterial species have the potential to suppress the growth of at least one seed-borne pathogen tested in the study. Among them, Pseudomonas koreensis appears to have strong antagonistic activities across all the pathogens. Our collection of soybean seed endophytes would be a valuable resource not only for studying biology and ecology of seed endophytes but also for practical deployment of seed endophytes toward crop protection.     

Growth promotion activity and biological control for the management of leaf blight incited by Alternaria helianthi

The Pseudomonas fluorescens (Pf1), Bacillus subtilis (Bs1) are the major potential biocontrol agents against foliar pathogens. MPf1 and MBs1 were found to be the most effective in inhibiting the mycelial growth of Alternaria helianthi. These biocontrol agents have the maximum capacity in controlling the spore germination, and data showing the growth-promoting effect of biocontrol agents and inhibition of seed-borne fungi are available. Seed-borne infections of A. helianthi are controlled by seed treatment with P. fluorescens, which showed least seed infection. The root length and shoot length has also been increased.     

Pathogenic response of seed mycoflora associated with cowpea,Vigna unguiculata

Cowpea, Vigna unguiculata (L.) Walp. (Fabaceae), has always been an important grain legume in tropical countries and a veritable source of dietary protein for the teeming population of human and livestock. The crop is often attacked and deteriorated by a number of pathogens; many of these are seed-borne fungi, which produce an adverse effect on the growth and development of plants. In this study, the effects of fungi on germinability and seedling health were determined, and the pathogenicity was established. Eleven fungal species were isolated from cowpea seeds which were used as inocula on cowpea. All the tested fungi show a reduction in the germination rate of the cowpea plants, and different types of disease symptoms including reduction in shoot and root lengths were noted for different pathogenic fungi. The symptoms included seed rot, seedling rot, yellowing of leaves, deformation of leaves, etc. Many phytopathogenic fungi penetrate deep into the seeds. These fungi produce enzymes, toxins and metabolites which adversely affect the plant growth and produce characteristic symptoms and cause deterioration of this important protein-rich crop.     

Application of beneficial rhizospheric microbes for the mitigation of seed-borne mycotoxigenic fungal infection and mycotoxins in maize

In the present investigation, seven rhizobacteria and nine Trichoderma spp. were evaluated to suppress seed-borne mycotoxigenic fungi (Aspergillus flavus and Fusarium verticillioides) and mycotoxin (aflatoxin and fumonisin) and to improve planting value of maize. Under in vitro conditions, these beneficial microorganisms suppressed the growth of A. flavus and F. verticillioides to various extents. Bacillus sp. (Bsp 3/aM), Pseudomonas putida (Has 1/c), Trichoderma asperellum (M5) and T. asperellum (T2) exhibited the greatest antagonistic effect on seed-borne mycotoxigenic fungi, and subsequently reduced mycotoxin concentrations in seeds. Under greenhouse conditions, these four biocontrol strains were also found to increase root length, shoot length, % germination, vigour index, fresh weight and dry weight of seedlings. Considering their overall performances, strains Bsp 3/aM, Has 1/c, M5 and T2 were selected for field studies as microbial talcum formulations. Among the tested microbial formulations, strain Bsp 3/aM significantly increased yield by 9.4% and 6.2% over the control in two maize cultivars Hema and Pearl, respectively. Increased plant growth and yield was also correlated with nutrient uptake in both the tested cultivars. All microbial formulation recorded significantly (p ≤ 0.05) reduced A. flavus infection and aflatoxin contamination in harvested seeds. But, none of the microbial formulations were found significant (p ≤ 0.05) in reducing F. verticilliodes incidence and fumonisin contamination. Our findings indicate that these microbial antagonists indirectly improve host health by suppressing seed-borne incidence of mycotoxigenic fungi and directly by facilitating nutrient uptake, thereby revealing their potential as both biofertilisers and biopesticides for maize production.     

Incidence of seed-borne fungi and aflatoxins in Sudanese lentil seeds

Thirteen seed samples of lentil (Lens esculenta) were incubated on agar plate and moist filter papers (Moist Chambers) at 28 ± 2 °C for determination of the incidence of seed-borne fungi. Aflatoxins content of the seeds was measured using the bright greenish-yellow fluorescence test (BGYF) and thin-layer chromatography (TLC). Sixty-nine species and seven varieties, which belong to 24 genera of fungi, were isolated from this crop. Of these fungi, 51 species and two varieties are considered new for this crop, whereas seven genera and 13 species are new to the mycoflora of the Sudan. The genus Aspergillus (13 species and 6 varieties) which comprising 44% of the total colony count was the most prevalent genus followed by Rhizopus (2 species, 19%), Penicillium (6 species) and Fusarium (8 species) (12%), Chaetomium (3 species) and Cladosporium (5 species) (6%), where the 18 genera (1–4 species) showed very low level of incidence (19%). Of the possible pathogens of lentil plants, F. oxysporum the main cause of vascular wilt was recovered from seeds of this crop. Thin layer chromatographic analysis of chloroform extracts of 13 seed samples showed that only one samples was naturally contaminated with aflatoxins B₁, B₂, G₁ and G₂ (14.3 μg/kg). This revised version was published online in June 2006 with corrections to the Cover Date.     

The endophytic fungi from wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

In order to study the species composition of endophytes from wheat healthy plants in Buenos Aires Province (Argentina) and to determine their infection frequencies from leaves, stems, glumes and grains, wheat plants were collected from five cultivars at five growth stages from crop emergence to harvest. A total of 1,750 plant segments (leaves, stems, glumes and grains) were processed from the five wheat cultivars at five growth stages, and 722 isolates of endophytic fungi recovered were identified as 30 fungal genera. Alternaria alternata, Cladosporium herbarum, Epicoccum nigrum, Cryptococcus sp., Rhodotorula rubra, Penicillium sp. and Fusarium graminearum were the fungi that showed the highest colonization frequency (CF%) in all the tissues and organs analysed. The number of taxa isolated was greater in the leaves than those in the other organs analysed.     

Cultivar response against root-infecting fungi and efficacy of Pseudomonas aeruginosa in controlling soybean root rot

Abstract

A study was conducted in the greenhouse to examine the resistance of three soybean cultivars against root-infecting fungi, and to determine the role of five strains of Pseudomonas aeruginosa in protecting the roots from these fungal pathogens. In this study soybean cv RAWAL was found to be less susceptible against charcoal rot fungus Macrophomina phaseolina than cvs PARC and BRAGG. Most of the strains of P. aeruginosa used as seed dressing significantly reduced M. phaseolina and Rhizoctonia solani infection on all three cvs PARC, BRAGG and RAWAL (p < 0.05). Most of the strains of P. aeruginosa were effective on cv PARC against Fusarium solani infection, while on cv BRAGG P. aeruginosa strain Pa₃, and on cv RAWAL strain Pa₅ were effective. Both strains Pa₃ and Pa₂₂ gave maximum plant height and fresh weight of shoots, respectively on cvs PARC and BRAGG than other strains. These characteristics make these P. aeruginosa strains good candidates for use as biocontrol agents against soil-borne plant pathogens.     

Seed-borne fungi of five cruciferous vegetables and relative efficacy of aqueous seed extracts against some associated fungi

A total of 121 seed samples of cabbage, cauliflower, cress, radish and turnip collected from five localities in Upper Egypt were assayed for their fungal flora. The highest count of fungi was recorded on cabbage seeds (75%), whereas the lowest count was observed on turnip seeds (33%). Thirty-five fungal species and two varieties belonging to 16 genera were identified. The broadest number of species (22 species + 1 variety) were isolated from cress seeds. However, only five species,viz. Aspergillus flavus, A. niger, Penicillium chrysogenum, P. funiculosum andRhizopus stolonifer, were found to be associated with seeds of the five plants. The relative efficacy of aqueous seed extracts against these five fungal species was tested. The antifungal drug Trosyd (tioconazole) was taken as a standard inhibitor. The aqueous extract of cabbage seeds inhibited three fungal species, cress extract two and cauliflower and turnip one species; radish seed extract was ineffective.     

Studies on seed-borne fungi of wheat in seed health testing programme

Seed mycoflora associated with wheat was studied on different media with a particular reference to Blotter and potato dextrose agar (PDA) procedures of ISTA. Seed-borne fungi, viz. Fusarium moniliforme, Rhizopus spp., Mucor spp., Alternaria alternata, Aspergillus niger, Aspergillus flavus, Curvularia lunata, Drechslera spp, Alternaria spp. and Penicillium spp., were isolated from the variety HD264. Blotter method was found to be the best media for the isolation of mycoflora whether borne externally or internally. Total number and frequency of occurrence of fungi were recorded. The effect of seed treatment with different chemicals and eco-friendly botanicals was analysed on germination, and growth, better percentage of seed germination and reduction in fungal pathogen were due to biochemical seed treatment.     

Characterization of agronomic traits and markers of recombinant inbred lines from intra- and interracial populations of Phaseolus vulgaris L.

The value of intra- and interracial populations in common bean (Phaseolus vulgaris L.) needs to be determined in order to create useful genetic variation for maximizing gains from selection, broadening the genetic base of commercial cultivars, and making efficient use of available resources. Five large-seeded parents of race Nueva Granada (N), two small-seeded race Mesoamerica (M), and one medium-seeded race Durango (D) were hybridized to produce one intraracial (N x N) and three interracial (two N x M and one N x D) populations. Seventy-nine F₂-derived F₆ lines randomly taken from each population along with their parents were evaluated for agronomic traits and markers at Palmira and Popayán, Colombia, in 1990 and 1991. Variation for agronomic traits and for morphological, protein, and isozyme markers was larger in interracial populations than in the intraracial population. Mean seed yield of all lines as well as yield of the highest yielding line from two interracial populations were significantly higher than that of the intraracial population. The highest ( 0.80±0.15) heritability was recorded for 100-seed weight. Values for seed yield varied from 0.19±0.17 to 0.50±0.16. Gains from selection (at 20% selection pressure) for seed yield ranged from 3.9% to 11.4%. Seed yield was positively associated with biomass yield, pods/m², and days to maturity, but harvest index showed negative correlations with these traits and a positive value with 100-seed weight. Polymorphism was recorded for phaseolin, lectins, protein Group-1 and protein Group-2 fractions, and six isozyme loci. Lines with indeterminate growth habit had significantly (P < 0.01) higher seed yield than lines with determinate growth habit in a Redkloud x MAM 4 population. Also, 23 other associations of markers with agronomic traits other than seed yield were recorded. Of these associations, lines with T phaseolin, the Diap1 ² allele, and lilac flower color tended to possess greater seed weight.     

The effect of leaf exudates on the spore germination of phylloplane mycoflora of chilli (Capsicum annuum L.) cultivars

Summary Ninetten aminoacids, twelve sugars, eleven organic acids and ten phenols were detected in the leaf exudates of three cultivars of chilli. The number of aminoacids, sugars, organic acids and phenols increased as the plants grew older. More aminoacids and sugars were detected in the exudate from the susceptible cultivar (Malwa). More organic acids and phenols were detected from the resistant cultivar (Simla). The leaf exudate of the resistant cultivar (Simla) inhibited spore germination of the pathogen (Alternaria solani) while that of susceptible (Malwa) stimulated spore germination. The cultivar ‘Patna’ which is moderately resistant, occupied an intermediate position. Spore germination of the isolated fungi was enhanced in leaf exudate of susceptible cultivar (Malwa), while leaf exudates of the moderately resistant (Patna) and resistant (Simla) inhibited spore germination of the majority of fungi isolated. Most of the antagonistic fungi were not isolated from the susceptible cultivar and the percentage spore germination of these fungi was less in leaf exudate of the susceptible cultivar, while leaf exudates of resistant cultivars enhanced the percentage spore germination of antagonistic fungi,viz Aspergillus flavus, A. fumigatus, A. versicolor, Penicillium citrinum, P. restrictum andTrichoderma viride.     

Isolation and analysis of endophytic microorganisms in wheat (Triticum aestivum L.) leaves

The present investigation was undertaken in order to select the surface-sterilization technique most efficient for eliminating epiphytes, to document the spectrum of endophytes of healthy leaves from three wheat cultivars in Buenos Aires Province (Argentina) and to determine their infection frequencies at three growth stages. Surface-sterilization with undiluted commercial solution of sodium hypochlorite was reaffirmed as adequate for removing epiphytes on wheat leaves. From the 450 wheat leaf segments incubated, three bacterial isolates and 130 fungal isolates were obtained. From all the isolates, 19 fungal species were identified. Bacterial isolates were characterized as Bacillus sp. There were significant differences between microorganisms, stages of growth, and stages × microorganisms interaction. Differences between cultivars, stages × cultivars, microorganisms × cultivars and for the triple interaction were not significant. Frequency of microorganisms isolated increased with crop age, but it was statistically similar for the three wheat cultivars tested (Klein Centauro, Klein Dragón and Buck Ombú). Rhodotorula rubra, Alternaria alternata, Cladosporium herbarum and Epicoccum nigrum were isolated in the highest frequency. The other microorganisms were present at intermediate or low values. The species isolated may be assigned to three groups: (a) well-known and economically important pathogens of wheat, (b) commonly abundant phylloplane fungi considered to be primary saprobic and minor pathogens and (c) species occasionally present in wheat.     

Isolation and Characterization of Trichoderma spp. for Antagonistic Activity Against Root Rot and Foliar Pathogens

Trichoderma, soil-borne filamentous fungi, are capable of parasitising several plant pathogenic fungi. Twelve isolates of Trichoderma spp. isolated from different locations of South Andaman were characterized for their cultural, morphological and antagonistic activity against soil borne and foliar borne pathogens. The sequencing of these isolates showed seven different species. The isolates revealed differential reaction patterns against the test pathogens viz., Sclerotium rolfsii, Colletotrichum gloeosporioides and C. capsici. However, the isolates, TND1, TWN1, TWC1, TGD1 and TSD1 were most effective in percentage inhibition of mycelial growth of test pathogens. Significant chitinase and β-1,3-glucanase activities of all Trichoderma isolates has been recorded in growth medium. T. viride was found with highest chitinase whereas T. harzianum was recorded with highest β-1,3-glucanase activities.     

Characterization and screening of plant probiotic traits of bacteria isolated from rice seeds cultivated in Argentina

Many seeds carry endophytes, which ensure good chances of seedling colonization. In this work, we have studied the seed-borne bacterial flora of rice varieties cultivated in the northeast of Argentina. Surface-sterilized husked seeds of the rice cultivars CT6919, El Paso 144, CAMBA, and IRGA 417 contained an average of 5×10⁶ CFU/g of mesophilic and copiotrophic bacteria. Microbiological, physiological, and molecular characterization of a set of 39 fast-growing isolates from the CT6919 seeds revealed an important diversity of seed-borne mesophiles and potential plant probiotic activities, including diazotrophy and antagonism of fungal pathogens. In fact, the seed-borne bacterial flora protected the rice seedlings against Curvularia sp. infection. The root colonization pattern of 2 Pantoea isolates from the seeds was studied by fluorescence microscopy of the inoculated axenic rice seedlings. Both isolates strongly colonized the site of emergence of the lateral roots and lenticels, which may represent the entry sites for endophytic spreading. These findings suggest that rice plants allow grain colonization by bacterial species that may act as natural biofertilizers and bioprotectives early from seed germination.