Factors affecting host range in a generalist seed pathogen of semi-arid shrublands

Generalist pathogens can exhibit differential success on different hosts, resulting in complex host range patterns. Several factors operate to reduce realized host range relative to potential host range, particularly under field conditions. We explored factors influencing host range of the naturally occurring generalist ascomycete grass seed pathogen Pyrenophora semeniperda. We measured potential host range in laboratory experiments at high inoculum loads with 26 grass species, including the primary host Bromus tectorum, and developed models to predict susceptibility and tolerance based on host traits, including germination speed, seed hardness, seed size, and phylogenetic relations. We also examined pathogen and host density effects on infection and mortality. All species tested were at least somewhat susceptible to the pathogen at high inoculum loads, but both infection and mortality varied widely. Species more closely related to the original host (B. tectorum) were more susceptible to infection, whereas species with slower germination were less tolerant and therefore more likely to suffer mortality. Infection and mortality were sharply reduced as inoculum load was reduced. Intermediate loads had major negative impacts on dormant B. tectorum seeds but generally minimal effects on native species. In addition, field seed bank studies determined that P. semeniperda rarely exploits native grass species as hosts. This marked reduction in realized host range relative to potential host range indicates that laboratory host range studies are potentially a poor predictor of either the current or possible future realized host range for wildland plant pathogens.     

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.     

Fungal Antagonists of Gerlachia nivalis

Trichoderma viride was strongly antagonistic to Gerlachia nivalis in agar plate culture and it controlled infection of barley seedlings from seed-borne inoculum when incorporated in soil as straw culture and from soil-borne inoculum when conidia were applied to the seed coat. Penicillium expansum and Papulaspora byssina were inhibitory towards G. nivalis in culture, although not as strongly as T. viride, and they did not control seedling infection as effectively.     

Effects of organomercury treatment of wheat seed on Fusarium seedling disease in inoculated soil

In growth room experiments, treatment of wheat seed with phenyl mercuric acetate significantly reduced seedling disease caused by soil-borne Fusarium culmorum, F. nivale and F. avenaceum in inoculated soil. Seed treatment improved the low germination caused by F. culmorum, but did not affect the reduced seedling vigour caused by F. nivale. Seed treatment was normally as effective in conditions favouring disease as it was in less favourable conditions. Conditions favouring disease in soil inoculated with F. culmorum were high inoculum level, deep sowing and dry soil.     

Strobilurins Seed Treatment Enhances Resistance of Common Bean Against Bean common mosaic virus

Commercial formulations of strobilurins (azoxystrobin, kresoxim‐methyl, trifloxystrobin and pyraclostrobin) were evaluated for their efficacy against Bean common mosaic virus (BCMV) in screenhouse and field conditions. Highest seed germination and seedling vigour were recorded with 20 μg/ml pyraclostrobin seed treatment in comparison with the control. In screenhouse studies, 76% protection against BCMV was recorded with pyraclostrobin seed treatment at 10 μg/ml. Under field conditions with natural BCMV inoculum, pyraclostrobin seed treatment resulted in 65% protection against BCMV. The protection offered by strobilurins against BCMV was evaluated by ELISA, with lowest immunoreactive values recorded in common bean seedlings raised from seeds treated with pyraclostrobin and kresoxim‐methyl. Strobilurins in addition to exerting a direct positive physiological effect on common bean plants also protect bean plants against BCMV infection in screen house and field conditions. Thus, it is proposed that these reduced‐risk pesticides are potential inducers against BCMV and growth enhancers and could be a beneficial component of integrated disease management of common bean.     

紫花苜蓿种子对逆境贮藏条件的反应

以陇东紫花苜蓿 (MedicagosativaL .cv .“Longdong”)种子为材料 ,在室温、35℃和 35℃ +10 %的种子含水量 (SMC) 3种贮藏、接种或不接种燕麦镰刀菌 (Fusariumavenaceum (Fr.)Sacc .)的条件下 ,1年贮藏期内对各逆境处理的种子每隔 6 0d进行 1次标准发芽试验 ,2 0℃恒温、第 10d统计种子的发芽率和死亡率 ,试验结束时计测种子幼苗的长度和感病率 ;在大田条件下观测各处理种子的出苗率 ,确定催腐 (CD)与各种贮藏条件下的苜蓿种带真菌种类和检出率 .结果表明 ,随着贮藏温度和种子含水量等逆境贮藏条件胁迫的加剧 ,苜蓿种带真菌检出率逐渐增高 ,从室温、35℃条件下的 10 %上升到CD +35℃ +10 %SMC条件下的 2 9% ;抗病性逐渐减弱 ,35℃ +10 %SMC条件下幼苗的感病率和种子死亡率显著 (P <0 .0 5 )高于室温和 35℃下的感病率和种子死亡率 ;室内种子发芽率和田间出苗率逐渐下降 ,35℃ +10 %SMC条件下的种子的发芽率和田间出苗率显著 (P <0 .0 5 )低于在室温和 35℃下的发芽率和田间出苗率 ;幼苗生长受到抑制 ,35℃ +10 %SMC条件下的苗长和根长显著 (P <0 .0 5 )低于在室温和 35℃下的幼苗长度 .随着贮藏时间的延长 ,种子真菌检出率和田间出苗率下降 ,幼苗感病率增加 .与未接种的对照相比 ,接种燕麦镰刀菌的种子     

Prevalence and control of wheat (Triticum aestivum L.) seed borne fungi in farmer-saved seeds

High incidence of diseases associated with the use of seeds saved from previous harvests as desire of maintaining local varieties with special attributes is of increased concern in wheat industry worldwide. Prevalent of seed-borne fungi in farmer-saved seeds and seed dressing fungicides to prevent infection from seeds to seedlings was studied in Northern Tanzania. One hundred and thirty five untreated farmer-saved seed lots were collected randomly from farmers. Alternaria alternata, Bipolaris sorokiniana, Drechslera tritici, Fusarium graminearum, Fusarium moniliforme, Aspergillus flavus, Cladosporium sphaerospermum, Epicoccum purpurascens, Pyricularia oryzae and Penicillium corylophilum were fungi isolated in farmer-saved seeds. Mean seed infection was 29% causing average grain yield loss of 1.2 mt/ha^?1. Seed dressing with Metalaxy plus (Methyl carboxaitide), Mancozeb (Manganase-zinc salt) and Baytan (Chlorophenoxy ethanol) increased seed germination by 14, 13 and 17%, respectively, and grain yield by 28, 20 and 18%, respectively. Farmer-saved seeds were heavily infected by fungi with low grain yield performance.     

The effect of iprodione on the seed-borne phase of Alternaria brassicicola

Alternaria brassicicola infection of Brassica oleracea seeds was effectively controlled by a dust application of iprodione (Rovral 50% w.P.). At 2.5 g a.i./kg the seed-borne fungus was usually eliminated from samples with up to 61.5% affected seeds (35.5% internally diseased) but higher levels of infection required increased doses for complete eradication of the fungus. The germination of healthy seeds, including samples from 7–yr-old stocks, on filter paper was unaffected by the treatment. However, the germination of diseased samples, particularly those internally infected with A. brassicicola, was improved. More seedlings emerged from iprodione treated than from untreated seeds in glasshouse soil but the differences were not significant. The application of gamma-hexachlorocyclohexane to iprodione treated seeds sown in soil did not adversely affect subsequent emergence or disease control. Disease control was maintained and germination was not affected by the treatment when treated infected seeds were stored for 2 yr at 10 °C, 50% r.h. In a field trial iprodione seed treatment reduced seedling infection in a cabbage crop grown from naturally diseased seeds (100% contaminated, 45.5% internally infected) from 5.6 to 0.04%.     

Transmission of Alternaria macrospora in Cotton Seeds

Alternaria macrospora was isolated from seeds only after the natural opening of the bolls and exposure of seeds to an environment in which the fungus was present. The fungus lacks the ability to penetrate the boll wall and reach the seed site. Attempts to isolate the pathogen from seeds of immature bolls at different developmental stages failed. Internal infection by slow injection resulted in seed infection and partial shedding of the injected plant parts which was high in buds and decreased with the ripening to mature bolls. Severity of seed infection was not dependent on either inoculum level, boll physiological age or even if the boll itself was not diseased. Infection of flowers under field conditions caused flower shedding. Naturally infected seeds or inoculated seeds with inoculum levels of 100 spores/ml and above resulted in diseased cotyledons, the incidence of which was, for inoculated seeds, positively correlated with inoculum level. A small difference was observed between cultivars in susceptibility to artificial inoculation at the cotyledon stage. A. macrospora survived on commercial cotton seeds and on post-season plants left growing at the field edges. Survival in plant debris under field conditions was minimal and may only have a minor effect on field reinfestation.     

Evaluation of Some Fungi and Bacteria for Potential Control of Safflower Rust

Some fungal and bacterial isolates applied as soil and seed treatments in greenhouse trials gave a good protection against Puccinia carthami Cda. on safflower, significantly reducing the infection from rust-infested seeds. Trichoderma viride and T. harzianum added alone and in combination as air-dried inoculum to the soil were the most effective fungal isolates. However T. viride at two higher rates reduced the rate of emergence of safflower seedlings. Among bacteria, Bacillus subtilis, B. cereus, isolates of Pseudomonas fluorescens and B. thuringiensis reduced rust infection when added both as soil drench and as seed treatment. Good results in the biocontrol of P. carthami were also obtained with some combination antagonist treatments such as T. viride + B. cereus, T. viride + P. fluorescens (16), T. viride + T. harzianum + B. cereus and T. viride +, T. harzianum + P. fluorescens (16). Fungal isolates added as soil treatment increased seedling length, whereas no similar effects were observed when these isolates were applied as seed treatment.     

Seed biopriming with novel strain of Trichoderma harzianum for the control of toxigenic Fusarium verticillioides and fumonisins in maize

Fusarium verticillioides is one of the most important fungal pathogens in maize causing both pre- and post-harvest losses and also capable of producing Fumonisins. In the present study attempts have been made for screening potential T. harzianum from native rhizosphere and to study its effect on Fusarium ear rot disease, fumonisin accumulation in different maize cultivars grown in India. Eight isolates of T. harzianum were isolated and T. harzianum isolate Th-8 exhibited better antifungal activity than carbendizim. Th-8 was formulated in different solid substrates like wheat bran, paddy husk, talcum powder and cornstarch. Maize seeds of kanchan (moderately resistant), pioneer (resistant) and sweet corn (susceptible) were selected for laboratory and field studies and these seeds were treated with a conidial suspension of T. harzianum at the rate of 1 × 10⁸ spore/ml and formulation at the rate of 10 g/kg. Treated seeds were subjected to evaluate F. verticillioides incidence, seed germination, seedling vigour and field emergence, yield, thousand seed weight and fumonisin production. It was found that the pure culture of T. harzianum was more effective in reducing the F. verticillioides and fumonisin incidence followed by Talc formulation than the carbendizim treated and untreated control. Formulations of T. harzianum were effective at reducing the F. verticillioides and Fumonisin infection and also increasing the seed germination, vigour index, field emergence, yield, and thousand seed weight in comparison with the control.     

The Seed-borne Inoculum of Peronospora manshurica, Causal Agent of Soybean Downy Mildew

The seed-borne inoculum of P. manshurica is described using various microscopic techniques and staining reactions. The inoculum, in addition to the oospores, consists of a thin walled mycelium under the hourglass cell layer of the seed coat and a thick walled resting mycelium in the oospore crust on the seed surface. By a modified Feulgen staining technique it was possible to demonstrate that the thin walled and the thick walled resting mycelium as well as thin and thick walled oogonia contain nuclei which are Feulgen positive suggesting that portions of the mycelium of P. manshurica may survive even on the surface of dry, mature seeds.     

Effect of harvest seasons on the incidence of seed-borne mycoflora of three Amaranthus cultivars

Fifteen fungal species were found to be associated with seeds of three cultivars ofAmaranthus cruentus L. There was a seasonal variability in the frequency of isolation, distribution and quantity of these seed-borne fungi among cultivars. The wetness, high temperature and air humidity that accompanied harvests of the first and second seasons greatly increased the number and amount of seed-borne fungi. Least number and mean percentage seed infection were recorded for NHAc₁₀₀ while NHAc₃₀ and NHAc₃₃ carried highest percentage inoculum of these pathogens. The seed testa of all cultivars was found to harbour the highest number and amount of some of these pathogens also in the second season. These seed-borne fungi,Choanephora cucurbitarum, Rhizoctonia solani andPythium aphanidermatum were highly pathogenic on seedlings of NHAc₃₃ and NHAc₃₀. Five of these seed-borne mycoflora have not been recorded onAmaranthus in Nigeria.     

Lack of Host Specialization on Winter Annual Grasses in the Fungal Seed Bank Pathogen Pyrenophora semeniperda

Generalist plant pathogens may have wide host ranges, but many exhibit varying degrees of host specialization, with multiple pathogen races that have narrower host ranges. These races are often genetically distinct, with each race causing highest disease incidence on its host of origin. We examined host specialization in the seed pathogen Pyrenophora semeniperda by reciprocally inoculating pathogen strains from Bromus tectorum and from four other winter annual grass weeds (Bromus diandrus, Bromus rubens, Bromus arvensis and Taeniatherum caput-medusae) onto dormant seeds of B. tectorum and each alternate host. We found that host species varied in resistance and pathogen strains varied in aggressiveness, but there was no evidence for host specialization. Most variation in aggressiveness was among strains within populations and was expressed similarly on both hosts, resulting in a positive correlation between strain-level disease incidence on B. tectorum and on the alternate host. In spite of this lack of host specialization, we detected weak but significant population genetic structure as a function of host species using two neutral marker systems that yielded similar results. This genetic structure is most likely due to founder effects, as the pathogen is known to be dispersed with host seeds. All host species were highly susceptible to their own pathogen races. Tolerance to infection (i.e., the ability to germinate even when infected and thereby avoid seed mortality) increased as a function of seed germination rate, which in turn increased as dormancy was lost. Pyrenophora semeniperda apparently does not require host specialization to fully exploit these winter annual grass species, which share many life history features that make them ideal hosts for this pathogen.     

Fungal volatiles associated with moldy grain in ventilated and non-ventilated bin-stored wheat

The fungal odor compounds 3-methyl-l-butanol, l-octen-3-ol and 3-octanone were monitored in nine experimental bins in Winnipeg, Manitoba containing a hard red spring wheat during the autumn, winter and summer seasons of 1984–85. Quality changes were associated with seed-borne microflora and moisture content in both ventilated and non-ventilated bins containing wheat of 15.6 and 18.2% initial moisture content. All three odor compounds occurred in considerably greater amounts in bulk wheat in non-ventilated than in ventilated bins, particularly in those with wheat having 18.2% moisture content. The presence of these compounds usually coincided with infection of the seeds by the fungi Alternaria alternata (Fr.) Keissler, Aspergillus repens DeBarry, A. versicolor (Vuill.) Tiraboschi, Penicillium crustosum Thom, P. oxalicum Currie and Thom, P. aurantiogriseum Dierckx, and P. citrinum Thom. High production of all three odor compounds in damp wheat stored in non-ventilated bins was associated with heavy fungal infection of the seeds and reduction in seed germinability. High initial moisture content of the harvested grain accelerated the production of all three fungal volatiles in non-ventilated bins.     

Biological Control of Bacterial Wilt of Bean Using a Bacterial Endophyte, Pantoea agglomerans

Indoor studies were conducted to determine the potential use of Pantoea agglomerans isolate LRC 8311 as a biocontrol agent for control of bacterial wilt of bean caused by Curtobacterium flaccumfaciens pv. flaccumfaciens. Soaking seeds of great northern bean cv. US1140 in a suspension of 3 × 10⁸ cfu/ml P. agglomerans resulted in thorough endophytic colonization of the entire bean seedling from root to apical stem after 7 days, regardless of whether the inoculated seeds were hilum injured or not. Colonization of seedlings by P. agglomerans increased seedling height after 10 days, and had no negative effect on seedling emergence. Treatment of hilum‐injured bean seeds of great northern bean cv. US1140 or navy bean cv. Morden003 with a mixture of P. agglomerans + C. flaccumfaciens pv. flaccumfaciens resulted in a high rate of colonization of seedlings by P. agglomerans, reduced frequency of infection by C. flaccumfaciens pv. flaccumfaciens, improved seedling emergence and height, and reduced disease severity, compared with seeds treated with the wilt pathogen alone. Application of P. agglomerans as a soil drench 24 h after planting was also effective in suppressing bacterial wilt in some instances, but was generally not as effective as seed treatment. The study suggests that seed treatment with P. agglomerans may be an effective and practical method for control of bacterial wilt of bean.     

The effects of thermotherapy and sodium hypochlorite treatments on pepino seed germination, a crucial step in breeding programmes

We evaluated the effects of treatments with thermotherapy (80°C for 24 h) and dipping in sodium hypochlorite (NaOCl; 0.8% for 10 min) on the germination percentage, germination rate, vigour index and ToMV inactivation of seeds from three pepino (Solarium muricatum) accessions (96–5, B–2 and OV–8) and two wild relatives (S. tabanoense EC–26 and S. caripense EC–40). Thermotherapy decreased the germination percentage (especially in the wild species), germination rate in wild species but not in cultivated pepino, and vigour index in all cases. Sodium hypochlorite increased germination, except for nonthermotreated seeds of OV–8, improved the germination rate in wild species and the vigour index in all cases except for non thermotreated OV–8. Seed coats were altered by both treatments, especially by NaOCl. Thermotherapy was totally effective in the inactivation of seed-borne ToMV, while NaOCl was not. Thermotherapy combined with NaOCl allows effective seed disinfection of S. muricatum seeds without negatively affecting seed germination. For the wild species, although thermotherapy reduces germination, the higher number of seeds per fruit in these species reduces the magnitude of the problem. Even when no disinfection is necessary, treatments with NaOCl are always advisable as they improve germination.     

Influence of Seed‐borne Cochliobolus sativus (Anamorph Bipolaris sorokiniana) on Crown Rot and Root Rot of Barley and Wheat

The effect of seed‐borne pathogens of wheat and barley on crown and root rot diseases of seven barley cultivars (Jimah‐6, Jimah‐51, Jimah‐54, Jimah‐58, Omani, Beecher and Duraqi) and three wheat cultivars (Cooley, Maissani and Shawarir) was investigated. Bipolaris sorokiniana and Alternaria alternata were detected in seeds of at least eight cultivars, but Fusarium species in seeds of only two barley cultivars (Jimah‐54 and Jimah‐58). Crown rot and root rot symptoms developed on barley and wheat cultivars following germination of infected seeds in sterilized growing media. Bipolaris sorokiniana was the only pathogen consistently isolated from crowns and roots of the emerging seedlings. In addition, crown rot and root rot diseases of non‐inoculated barley cultivars correlated significantly with B. sorokiniana inoculum in seeds (P = 0.0019), but not with Fusarium or Alternaria (P > 0.05). These results indicate the role of seed‐borne inoculum of B. sorokiniana in development of crown rot and root rot diseases. Pathogenicity tests of B. sorokiniana isolates confirmed its role in inducing crown rot and root rot, with two wheat cultivars being more resistant to crown and root rots than most barley cultivars (P < 0.05). Barley cultivars also exhibited significant differences in resistance to crown rot (P < 0.05). In addition, black point disease symptoms were observed on seeds of three barley cultivars and were found to significantly affect seed germination and growth of some of these cultivars. This study confirms the role of seed‐borne inoculum of B. sorokiniana in crown and root rots of wheat and barley and is the first report in Oman of the association of B. sorokiniana with black point disease of barley.     

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.     

Incidence, field spread, seed transmission and effects of broad bean stain virus and Echtes Ackerbohnenmosaik-Virus in Vicia faba in eastern Scotland

In eastern Scotland seed-borne infection with broad bean stain virus (BBSV) and/or Echtes Ackerbohnenmosaik-Virus (EAMV) was detected in five of 39 seed lots of field bean in 1975 and in four of 21 commercial crops of field bean or broad bean sampled in 1975 or 1976. Tests failed to detect the main weevil vector of these viruses, Apion vorax, in 1975 and 1976 but Sitona weevils were found in most crops and were numerous in many, reaching maximum numbers in August. No spread of BBSV and EAMV was detected in commercial crops containing seed-borne infection. In experimental field bean crops containing plants manually inoculated with virus, no virus spread was detected in 1975, and only 0–015% uninoculated plants became infected with EAMV in 1976. Sitona, therefore, was an inefficient vector. The percentage of virus infection in seed harvested from field bean plants manually inoculated 3, 5, 7 and 11 wk after emergence in the field was 1–5, 2–7, 0–4 and 0–06 for BBSV and 0–5, 2-1, 0–6 and 0 for EAMV respectively. Seed harvested from unrogued and rogued plots of field bean grown from seed containing 3–4% seed-borne infection produced 0–05% and no infected plants, respectively. Yield losses in field bean plants manually inoculated with virus before flowering were up to 20% but were much greater in plants infected through the seed. Loss in yield was largely caused by a decrease in number of seeds per pod. The absence of A. vorax, the late arrival of Sitona weevils in the crop and their inefficiency as vectors, and the smaller effects of BBSV and EAMV on crop yield than in southern England appear to make eastern Scotland very suitable for the production of bean seed free from BBSV and EAMV.