The effects of Pythium sylvaticum, seed-borne pathogens and seed treatments on the emergence of brassica seedlots of high vigour

The emergence from brassica seedlots designated by seed merchants as being of high vigour was measured in sterilised soil and in sterilised soil re-inoculated with Pythium sylvaticum. The mean emergence level of 93·9% in the former compared favourably with the known mean total viability of the seedlots (97·7%). However, P. sylvaticum reduced emergence of 10 out of 16 untreated seedlots. The majority of seedlots gave > 75% emergence but in three cases emergence was reduced by 32·5–40·0%. The effects were not related directly to time taken to emerge, nor were the seedlots most affected those with high levels of infection. A compound seed treatment containing metalaxyl, fenpropimorph and gamma-HCH restored high emergence levels.     

Establishment and effectiveness of inoculated arbuscular mycorrhizal fungi in agricultural soils

Arbuscular mycorrhizal fungi (AMF) are promoted as biofertilizers for sustainable agriculture. So far, most researchers have investigated the effects of AMF on plant growth under highly controlled conditions with sterilized soil, soil substrates or soils with low available P or low inoculum potential. However, it is still poorly documented whether inoculated AMF can successfully establish in field soils with native AMF communities and enhance plant growth. We inoculated grassland microcosms planted with a grass–clover mixture (Lolium multiflorum and Trifolium pratense) with the arbuscular mycorrhizal fungus Rhizoglomus irregulare. The microcosms were filled with eight different unsterilized field soils that varied greatly in soil type and chemical characteristics and indigenous AMF communities. We tested whether inoculation with AMF enhanced plant biomass and R. irregulare abundance using a species specific qPCR. Inoculation increased the abundance of R. irregulare in all soils, irrespective of soil P availability, the initial abundance of R. irregulare or the abundance of native AM fungal communities. AMF inoculation had no effect on the grass but significantly enhanced clover yield in five out of eight field soils. The results demonstrate that AMF inoculation can be successful, even when soil P availability is high and native AMF communities are abundant.     

Effect of seed exudates ofPinus resinosa on the germination of sporangia and on the population ofPythium irregulare in the soil

Summary Exudates from germinating seed ofP. resinosa stimulated the germination of sporangia and increased thePythium populations in soil. Sporangia ofP. irregulare did not germinate in natural soil and needed exogenous nutrition for their germination. Different components of the exudate, particularly glucose and asparagine, effectively stimulated sporangial germination. This is in agreement with an earlier finding withPythium ultimum ³.     

Interaction of nutrients and four antifungal antibiotics in their effects on Pythium species in vitro and in soil

Summary Griseofulvin, streptomycin, aureomycin, and actidione, with and without nutrients, were tested againstPythium ultimum Trow (two strains),P. irregulare Buis., andP. rostratum Butler. Streptomycin was tolerated in the highest concentration, and actidione and aureomycin in the lowest.P. rostratum, a saprophytic species, was more resistant to the antibiotics than the pathogenic species, which were inhibited at low concentrations. The two strains ofP. ultimum behaved differently, the oogonial strain being more tolerant than the sporangial strain. Yeast extract was usually superior to sucrose or asparagine in counteracting the toxicity of antibiotics.Germination of sporangia ofPythium irregulare was studiedin vitro and in a natural soil in the presence of antibiotic and antibiotic-nutrient solutions. Very meager germination of sporangia occurred in soil amended with antibiotics. Actidione severely restricted germination in soil. Nutrients nullified the toxicity to varying degrees. Usually higher germination of sporangia occurredin vitro than in soil. Sub-lethal concentrations of antibiotics induced morphological abnormalities in germ tubes. Aureomycin caused a dendroid type of branching; actidione caused distortion, vacuolation, and vesiculation; griseofulvin caused lysis and septation; and streptomycin caused distortion.Germ tubes were lysed in a natural soil and in soils amended with antibiotics, but nutrients, in general, prevented early lysis at low concentrations of antibiotics. No resistant bodies were formed prior to the initiation of lysis of germ tubes.     

Occurrence of Pythium spp. and Phytophthora spp. in Norwegian Greenhouses and their Pathogenicity on Cucumber Seedlings

Samples of tomato, lettuce and cucumber submitted for diagnosis to the Plant Protection Centre at the Norwegian Crop Research Institute and samples of soil, water and cucumber collected from greenhouses employing hydroponic cultures were examined for the occurrence of Pythium spp. and Phytophthora spp. Two species of Phytophthora and 16 species of Pythium were identified. Phytophthora cryptogea was found on tomato and lettuce. Phytophthora nicotianae was found on tomato fruit. Phytophthora was not found on cucumbers. Pythium irregulare and Pythium group F were the two most commonly found Pythium species in hydroponically cultivated cucumbers. A pathogenicity test with 56 isolates was performed on cucumber seedlings. The most aggressive species were Pythium aphanidermatum, P. irregulare, Pythium paroecandrum and Pythium ultimum.     

Use of Brassicaceous seed meals to improve seedling emergence of tomato and pepper in Pythium ultimum infested soils

Growth room studies were conducted to determine the impact of Brassicaceae seed meals on the emergence of tomato and pepper seedlings in Pythium ultimum infested soils. Pasteurised Burch sandy loam soils were amended with intact and denatured seed meal of rape seed and mustard. Brassica juncea or Brassica napus intact seed meal increased the tomato and pepper seedling emergence. Interestingly, B. napus amended soils resulted in the same seedling emergence with B. juncea regardless of their relatively lower glucosinolate content compared to mustard-based seed meals. Seedling emergence in soils amended with intact Sinapis alba seed meal was significantly the lowest for both tomato and pepper seedlings. In contrast, seedling emergence was higher in soils amended with denatured than intact S. alba seed meals suggesting some glucosinolate-related inhibitory effect on seedling emergence of both crops. Glycine max seed meal amendment improved the seedling emergence better than the control but to a lower-level when compared to glucosinolate containing seed meals. This finding suggests that even though improvement of seedling emergence of tomato and pepper in P. ultimum infested soils can be achieved using Brassicaceae seed meals, it cannot be entirely attributed to glucosinolate-related processes. These studies demonstrate that intact B. napus and B. juncea seed meals can be used to improve tomato and pepper seedling emergence in P. ultimum infested soils.     

Effect ofAgrobacterium radiobacter and its polysaccharide on emergence and damping-off of tomato plants

Agrobacterium radiobacter B6 and agrobacteran (an exopolysaccharide of the succinoglycan group) stimulated seed germination and tomato plant emergence. The germination was most stimulated by dipping the seeds in 0.1 % agrobacteran for 30 min whereas plant emergence in garden soil was best with 0.4 % agrobacteran at 10–20°C. Treatment of the plants withA.radiobacter cells and agrobacteran solution at 30–35°C. had a lower effect.A.radiobacter cells applied on seed surface protected the plants against damping off in garden soil artificially inoculated with the fungiRhizoctonia solani andPythium ultimum; in soils contaminated withFusarium solani 0.1 to 0.2% agrobacteran had a higher protective effect than the bacterization. The difference can be attributed to the varying density ofA.radiobacter population in plant rhizosphere in the presence of different plantpathogenic fungi, different interactions of microorganisms in the rhizosphere and different mode of penetration of the pathogens into plant roots.     

Oomycetes associated with Prunus serotina persist in soil after tree harvest

Soil-borne pathogens can shape forest communities by lowering seedling survivorship. Many soil pathogens can persist long-term as survival spores, but how long pathogens outlive tree hosts in gap soils and whether they continue to affect seedling survival is uncertain. We studied the presence of oomycetes and evaluated seedling performance in soils near live Prunus serotina trees, and 0.5 and 1.5-y-old stumps. We isolated five species of oomycetes from soils, two of which were pathogenic (Pythium intermedium and Pythium irregulare) to Prunus serotina. There was a non-significant ~10.5% increase in conspecific seedling survival in stumps versus live trees, and pathogens were present in soils of all stump ages. The continued presence of pathogens of Prunus serotina in gap soils demonstrates the potential for impacts on conspecific regeneration after tree death, though the slight improvement in survival suggests that these effects may weaken with time.     

An Evaluation of Soil Colonisation Potential of Selected Fungi and their Production of Ligninolytic Enzymes for Use in Soil Bioremediation Applications

Initially sixteen fungi were screened for potential ligninolytic activity using decolourisation of a polymeric dye Poly R-478. From this, four fungi were selected, Trametes versicolor, Pleurotus ostreatus, Collybia sp., and an isolate (identified as Rhizoctonia solani) isolated from a grassland soil. Differences in the ligninolytic enzyme profiles of each of the fungi were observed. All of the four fungi tested produced MnP and laccase while the Collybia sp. and R. solani produced LiP in addition. Enzyme activity levels also varied greatly over the 21 days of testing with T. versicolor producing levels of MnP and laccase three to four times greater than the other fungi. The four fungi were then tested for their ability to colonise sand, peat (forest) and basalt and marl mixed till (field) soils through visual measurement and biomass detection in soil microcosms. Trametes versicolor and the Collybia sp. failed to grow in any of the non-sterilised soils whereas the R. solani and P. ostreatus isolates grew satisfactorily. Primers were␣designed to detect MnP and laccase genes in P.␣ostreatus and RTPCR was used to detect that these genes are expressed in forest and field soils.     

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.     

First Report of Pythium indigoferae and P. irregulare Associated to Apple Trees Decline in Tunisia

Pythium indigoferae and Pythium irregulare, identified based on morphological and physiological characteristics, were isolated from necrotic roots, crown tissues and the rhizosphere of apple trees in Tunisia from 23 apple orchards in spring and autumn 2007–2009. The virulence assays on excised twigs, using different Pythium species isolated demonstrated that these oomycetes were pathogenic on the Anna, Lorka and Meski varieties and the MM106 rootstock. However, the biggest lesion area was noted on MM106 rootstock. Thus, it appeared that this rootstock is more susceptible to Pythium infections than Anna, Meski and Lorka apple varieties. Furthermore, it is important to note that in vitro tests showed that P. indigoferae seems to be more virulent than P. irregulare.     

Regenerative role of seed banks following an intense soil disturbance

Our main aim was to determine the contribution of the seed bank to vegetation regeneration following a disturbance consisting in a deep ploughing and a thorough homogenisation of a perennial grassland. In the seed bank prior to disturbance, seed distribution through the vertical soil profile was evaluated to determine the initial seed species structure. Then, several characteristics of the shallow seed bank and the extant vegetation were evaluated prior and following field disturbance: seed species composition and abundance, and species composition of the aboveground vegetation. The contribution of seed rain versus seed bank was evaluated by means of the comparison of the vegetation developed in plots filled with sterilised soil (seed bank removal) and the vegetation developed in non-sterilised plots in the field. The distribution of seeds through the profile indicated a sharp decline in abundance with depth, and it was probably linked to propagule morphology, with small and rounded seeds proner to being buried deeper than larger seeds. In the grassland prior to disturbance, the aboveground vegetation and seed bank species composition showed very low similarity index, most likely because during the 5 years following field abandonment, sheep pressure had caused a faster change in aboveground vegetation species composition than in seed bank species composition. Ploughing and homogenisation of the grassland led to low seed abundance in the shallow soil layer caused by dilution of the seed bank. Regardless of impoverishment in seed abundance and species richness, comparison between sterilised and non-sterilised plots showed that the seed bank acted as an effective source of colonising species and determined the aboveground species composition. To summarise, this study outlines the importance of considering several characteristics of the seed bank, such as species composition and seed abundance, in the understanding of the function of seed bank and dynamics of the vegetation following a deep ploughing and homogenisation treatment.     

Integration of Pythium nunn and Trichoderma harzianum isolate T-95 for the biological control of Pythium damping-off of cucumber

Two biological control agents, Pythium nunn and Trichoderma harzianum isolate T-95, were combined to reduce Pythium damping-off of cucumber in greenhouse experiments lasting 3–4 weeks. T. harzianum T-95, a rhizosphere competent mutant, was applied to seeds and P. nunn was applied to pasteurized and raw soils naturally and artificially infested with Pythium ultimum. Some treatments were also amended with bean leaves to enhance the activity of P. nunn. The biological control of Pythium damping-off was evaluated in a Colorado soil (Nunn sandy loam) and an Oregon soil mix, which were replanted twice after 2 and 3 months. Interactions between P. nunn and T-95 were detected in the Colorado but not the Oregon soil. No consistent evidence of antagonism between P. nunn and T. harzianum was seen, and significant interactions were detected in the Colorado, but not the Oregon soil. In the first planting of some treatments, the combination of P. nunn and T. harzianum gave greater control of damping-off than either applied alone. P. nunn was most effective in soils that were pasteurized or amended with bean leaves. T. harzianum controlled Pythium damping-off in the Colorado, but not the Oregon soil. In both soils, disease declined over time in treatments amended with bean leaves but without P. nunn or T. harzianum added. This suppression was greater in the Colorado soil, which contained an indigenous population of P. nunn. This work demonstrates that two compatible biological control agents can be combined to give additional control of a soil-borne plant pathogen.     

Growth of Trifolium alpinum: Effects of soil properties, symbionts and pathogens

The lowland cultivation of Trifolium alpinum, a clover species found on acid soils in the Alps and suitable for the restoration of erosion areas at high altitudes, failed repeatedly in previous experiments. Three experiments were carried out in a controlled environment to elucidate the reasons for the failure and to develop possible cultivation strategies. In experiment I, T. alpinum was grown in an autochthonous soil from the Alps (high elevation) and in two allochthonous soils, a grassland soil from the Hercynian mountains (medium elevation), and an arable soil (low elevation), in which the seed propagation of T. alpinum had failed previously. The two allochthonous soils had lower contents of soil organic C and ergosterol, an indicator for fungal biomass, than the autochthonous high-elevation soil, but higher levels of exchangeable Ca and extractable P. Plants grown in the allochthonous soils achieved higher biomass and total N amounts per plant than those from the high elevation soil if inoculated with this autochthonous material to establish rhizobial infection. In the allochthonous high elevation soil, the growth of T. alpinum was P-limited as shown in experiment II. In experiment I, plants grown in the low elevation soil had a lower biomass and smaller number of active leaves at 120 days after emergence than those grown on the medium elevation soil. This difference can be explained by strong colonization with the phytophagous nematode Pratylenchus sp., as demonstrated in experiment III by comparing plant growth either in untreated or in autoclaved low-elevation soil. Successful propagation of T. alpinum at low elevation may be achieved through suitable inoculation with autochthonous soil biota, especially Rhizobia, and avoidance of soils infested by Pratylenchus species by choosing sites with acidic soil and ensuring adequate P-availability.     

The influence ofPythium on the growth of barley seedlings as affected by soil water and inoculum density

Summary Pythium spp. were more abundant in the southerly and more temperature regions of the barley growing region of South Australia than in the drier and hotter north. Populations were more abundant in the top 10 cm than in the 10 to 20 cm soil zone. Eleven species ofPythium were identified from barley crops.P. irregulare appeared to be the most abundant and was one of the most pathogenic species on barley.P. volutum was also highly pathogenic; it had not been recorded in South Australia before. A factorial experiment using ninePythium spp. and four levels of soil water indicated that unlike other species,P. irregulare, P. volutum andP. graminicolum were most pathogenic in soils with a water content close to field capacity. A factorial experiment usingP. irregulare at four levels of soil water and six inoculum levels showed that inhibition of growth in barley seedlings byP. irregulare increased as the level of water in the soil increased. The experiments support the hypothesis that inhibition of growth of barley seedlings byPythium spp. is most severe in the southerly parts of the barley growing area of South Australia particularly where there is a combination of high soil water and high population densities.     

A seed treatment for the control of Pythium damping-off diseases and Peronospora parasitica on brassicas

In soil inoculated with Pythium ultimum or Pythium irregulare, seed treatment with either Apron 70 (=1 g metalaxyl and 1 g captan/kg seed) or thiram gave control of pre-emergence damping-off of Brussels sprout and cabbage seedlings. On cauliflower, Apron 70 was significantly more effective than thiram. No post-emergence damping-off occurred in either of these crops or in oil-seed rape following seed treatment with Apron 70 whilst post-emergence losses from untreated seed ranged from 10·2–19·4% and from thiram treated seed from 5·7-7·4%. Apron 70 gave complete control of Peronospora parasitica on cauliflower inoculated 10 days after sowing; thiram was ineffective. Following seed treatment with Apron 70, metalaxyl was detected in the cotyledons, true leaves and roots of cabbage seedlings up to 4 wk from sowing.     

Soil seed bank and community dynamics in an annual-dominated Mediterranean salt-marsh

Abstract. The seed bank in the soil and litter of an annual grassland in the upper salt marsh of the Guadalquivir delta (SW Spain) contained on average 49110 seeds/m²; 29 species were represented. Field censuses of the grassland, in two consecutive springs, yielded 7 410 and 9 520 plants/m² respectively. A combined Correspondence Analysis of the seed bank samples and field censuses showed differences in grassland composition between wet and dry years. The environmental changes induced different expressions of the soil seed bank, mediated by demographic processes. The effect of salinity (25 dS/m of NaCl) was studied on intact soil blocks transplanted to a greenhouse. Salinity did not affect the emergence of seedlings, but reduced the growth of the dominant species, Trisetaria panicea. The importance of the role of seed banks in community dynamics and their use for conservation management are discussed.     

Effect of different soil types,pH and phosphorus levels on the wheat seedling and the incidence ofPythium graminicolum

Summary In the present study, the effect of different types of soil, pH and phosphorus levels on root, shoot development and browning extent on shoot of wheat seedlings growth in soil inoculated withPythium graminicolum; was studied. Out of the four soil types the development of root and shoot was better in Domatta soil where as the disease development was more in black clay soil. The pH values below 5.5 and above 8.0 are favourable for the root and shoot development whereas these values are unfavourable for the development of the Pythium and consequently for infection by it. The percentage of browning of the collar region was more in soils near neutral (pH 6.5–7.1) and it was less in acidic (pH 5.0) or alkaline (pH 8.6) soils. The development of wheat roots and shoot in soil with 30 lb/acre phosphorus was good as compared to that in soil with P₀, P_15,45 and P₆₀ lb/acre in inoculated soil. The percentage of extent of browning on collar region was less at 15 lb and 30 lb/acre P, due to less development of the disease.     

Inter-relationship between shoot weight,severity of root rot and survival rate of subterranean clover inoculated with certain pathogenic fungi

Summary Plant survival rate, root disease index and fresh shoot weight of subterranean clover seedlings inoculated with fungal pathogens (Fusarium avenaceum, F. oxysporum, Phoma medicaginis, Pythium irregulare andRhizoctonia solani, both singly and in combinations) were generally inter-correlated over a wide range of temperature (10, 15, 20 and 25°C) and moisture conditions (45, 65% water holding capacity and flooding). There was a negative correlation between root disease index and shoot weight for all treatments exceptF. avenaceum + P. irregulare. Root disease index and seedling survival rate were negatively correlated except forF. oxygsporum, Phoma medicaginis, P. irregulare andF. oxysporum + F. avenaceum. However, a good positive correlation was found between the survival rate and shoot weight for all treatments with the exception ofPhoma medicaginis.