Sadhna Alström

Cyanide production by rhizobacteria as a possible mechanism of plant growth inhibition

SummaryVolatile metabolites from a number of rhizosphere pseudomonads prevented lettuce root growth in a seedling bioassay. One of these metabolites was identified as cyanide. Direct contact between rhizobacteria and plant roots produced, with one exception, similar responses. However, not all cyanogenic isolates were plant-growth-inhibitory rhizobacteria. When grown in liquid culture, cyanogenic strains produced an average of 37 nmol HCN ml−1 over a 36-h period and inhibition of root growth occurred at concentrations as low as 20 nmol ml−1. Cyanogenic strains introduced into sand or soil also produced HCN. Two cyanogenic strains ofPseudomonas fluorescens, one (5241) a plant-growth inhibitory rhizobacterium and the other (S97) a plant-growth-promotory rhizobacterium, were used to treat bean and lettuce seedlings prior to planting in soil. Lettuce dry weight was reduced by 49.2% (day 28) and 37.4% (day 49) when inoculated with S241 whereas S97 increased growth initially (+64.5% at day 28, no difference from control at day 49). Equivalent figures for inoculated bean plants were: −52.9% and −65.1% (5241); +40.7% and +23.3% (S97). A more detailed experiment using only bean plants confirmed these contrasting affects. Inhibition by S241 was related to consistently higher levels of rhizosphere cyanide in comparison with S97-treated plants and control soils. S241 also survived in the rhizosphere at higher densities and for a longer period of time than S97. The possible contribution of rhizobacterial cyanogenesis to plant growth inhibition is discussed.

A study on microbial diversity in different cultivars of Brassica napus in relation to its wilt pathogen, Verticillium longisporum.

Oilseed rape (Brassica napus) is one of the major oilseed crops in the world but is vulnerable to attack by many pathogens and insect pests. In addition to the host plant genotype, micro-organisms present in the rhizosphere and within plant tissues affect the susceptibility to plant pathogens. While rapid progress has been achieved concerning the concept of plant resistance genes, information on the role of the microbial community in plant protection is less apparent. We have studied the endophytic bacterial populations present in different tissues of oilseed rape and also analysed several cultivars (Express, Libraska, Maluka and Westar), which differ in their susceptibility to the wilt pathogen Verticillium longisporum. The population diversity was studied using agar plating assay, fatty acid methyl ester analysis and functional characterisation of isolated strains. Our work shows that already in the seeds there exists diversity in populations as well as in the total microbial load between two of the four tested cultivars. About 50% of the strains isolated from cultivars Express and Libraska showed moderate to strong direct inhibition of V. longisporum. The diversity of the endophytic flora isolated from oilseed rape and its implications in crop protection are discussed.

Different crop rotation systems as drivers of change in soil bacterial community structure and yield of rice, Oryza sativa

Intensive cropping, especially of rice, is considered to contribute to negative effects not only on soil chemical and biological properties but also on long-term grain yield. Appropriate crop rotation is often practiced as an alternative strategy to overcome the negative side effects of intensive cropping. Although soil microbial diversity and community structure have been shown to respond differently to altered agricultural management practices, little is known about possible links between crop rotation and grain yield on bacterial communities in rice paddy soil. In this study, we investigated the impact of specific rotational crops and compared it with intensive rice cultivation. The main crop rice (Oryza sativa) was rotated with maize (Zea mays) and mungbean (Phaseolus aureus) in different combinations in a system cultivating three crops per year. Soil bacterial communities were studied in two different cropping periods using pyrosequencing of the variable V4 region of the 16S rRNA. Our results showed that rotation with alternative crops increased rice yield by 24–46% depending on rotation structure and that bacterial community structure was altered in the presence of mungbean and/or maize compared to that in rice monoculture. In the crop rotation systems, composition, abundance, and diversity of soil bacterial communities were significantly different and higher than those in rice monoculture. Our results show that effects of crop rotation relate to changes in soil bacterial community structure suggesting that appropriate crop rotations provide a feasible practice to maintain the equilibrium in soil microbial environment for sustainable rice cultivation.

Factors associated with detrimental effects of rhizobacteria on plant growth

Some of the factors interfering with the specific response of young common bean plants to two rhizosphere fluorescent pseudomonads were studied. These two bacterial strains produced symptoms in foliar plant parts and reduced yield in beans and several other plant species when inoculated on roots. Sensitivity in the plants subjected to bacterial application was highest at early growth stages (up to ten days old plants) giving typical symptoms for each strain in first expanding leaves and typically stunted plant growth. Symptoms to some extent also appeared on plants treated at the age of 2–3 weeks and the fresh shoot weight was affected also in such plants. Deleterious effects of the tested bacteria were found to be related to the level of inoculum (cfu/ml) used and presence or absence of certain nutrients in the bacterial suspensions used for inoculation. The two tested strains differed in nutritional requirements for affecting plant growth. One strain needed sucrose only. The other tested strain required peptone or yeast extract in addition to sucrose to induce typical symptoms and significant yield reductions. Supply of peptone as the only nutrient source to the bacterial suspensions eliminated the deleterious effects of both bacterial strains as tested on beans under nonsterile conditions. The two strains are assumed to affect test plants by different modes of action.

Root bacteria from nematicidal plants and their biocontrol potential against trichodorid nematodes in potato

Trichodorid nematodes (Nematoda: Trichodoridae) are vectors of tobacco rattle virus (TRV), one of the causal agents of spraing disease in potato. Root bacteria from nematicidal plants and their control potential against Trichodoridae were the focus of this study. Bacteria isolated from the roots of 12 nematicidal plants and potato were characterized for their production of hydrolytic enzymes, hydrogen cyanide, phenol oxidation ability and antifungal activity towards the potato pathogen Rhizoctonia solani. Based on these functional traits, bacteria isolates were selected and tested in greenhouse conditions on potato (cv. Saturna) for their effect on plant growth, and screened for nematicidal activity against Paratrichodorus pachydermus and Trichodorus primitivus in naturally infested soil. Sixteen bacteria isolates out of 44 reduced nematode densities by 50–100%. Nine selected isolated were further tested by bacterizing potato tubers (cv. King Edward) which were planted in a trichodorid and TRV-infested soil. Four bacterial isolates consistently reduced nematode densities (by 56.7–74.4%) with no visual negative effect on plant growth. These isolates were tentatively identified, partly by fatty acid methyl ester (FAME) analysis as: Stenotrophomonas maltophilia, Bacillus mycoides, Pseudomonas sp., and one unidentified bacterium. The isolates originated from potato, Plantago major, Thymus vulgaris and Asparagus officinalis, respectively. Two Pseudomonas isolates obtained from Zinnia elegans and selected for their strong nematicidal activity in soil screening tests, did not reduce the nematode population when tested on potato. It is concluded that plants releasing nematicidal compounds may harbour nematode-antagonistic bacteria as well.

Interactions among Glomus irregulare, arbuscular mycorrhizal spore-associated bacteria, and plant pathogens under in vitro conditions

Arbuscular mycorrhizal (AM) fungi interact with bacteria (AM fungi-associated bacteria, AMB) in the mycorrhizosphere. We previously identified a set of AMB that enhance AM fungal colonization, plant growth, and inhibit pathogens. Here, we used transformed carrot root cultures in a two-compartment plate system for further in vitro studies on interactions taking place among Glomus irregulare (syn.Glomus intraradices), AMB, and plant pathogens. We found that exudates of G. irregulare stimulated growth of all ten AMB isolates tested in multi-well plates. AMB growth stimulation was observed also during co-cultivation of three of these AMB with G. irregulare in the hyphal compartment. In addition, co-cultivation stimulated growth of G. irregulare hyphae and spore production, as well as G. irregulare root colonization. GC/MS analysis in a preliminary screening of metabolites revealed differences in concentrations of several identified but also unidentified compounds in G. irregulare hyphal exudates. Exudates in presence of three different AMB isolates co-cultivated with G. irregulare contained several additional compounds that differed in amount compared with G. irregulare alone. The results indicate that G. irregulare exudates contain carbohydrates, amino acids, and unidentified compounds that could serve as a substrate to stimulate AMB growth. With regard to effects on plant pathogens, growth inhibition of Rhizoctonia solani, Verticillium dahliae, and Pectobacterium carotovorum ssp. carotovorum was evident in the presence of the AMB isolates tested together with the G. irregulare exudates. These in vitro studies suggest that G. irregulare and AMB stimulate growth of each other and that they together seem to provide an additive effect against growth of both fungal and bacterial pathogens.

Characterisation of bacteria in soils under barley monoculture and crop rotation

Abstract Rhizobacterial populations on barley roots, originating from experimental fields with barley monoculture (MC) and crop rotation (CR), were analyzed for their fatty profiles. In the first part of the study, the profiles of 1188 isolates were statistically analyzed to identify clusters of bacteria with a possible high prevalence in MC or CR soil. One such cluster was found, termed Ps4-C4, with characteristically high contents of C12:1 3OH (9.8%) and an unidentified fatty acid with the equivalent chain length (ECL) of 12.35, which was provisionally named ECL12.35 (4.5%). Bacteria in Ps4-C4 were also rich in C10:0 3OH (8.3%) and C12:0 3OH (8.1%). The cluster consisted of 109 isolates, 86 from MC populations and 23 from CR, none of which could be identified by means of fatty acid analysis. In the second part of the study, fatty acid profiles of 240 microbial populations from the same fields were analyzed. Results showed higher relative contents of C12:1 3OH, ECL12.35, C10:0 3OH and C12:0 3OH in MC populations, which also had a high proportion of C12:0 2OH and C16:0. A detailed statistical analysis of the correlations between the fatty acids indicated that Ps4-C4 alone explained the higher portions of C12:1 3OH and ECL12.35 in populations from MC soil, while other bacterial groups are seem to have contributed to the elevated contents of C10:0 3OH and C12:0 3OH. Some common functional characteristics of bacteria in the Ps4-C4 cluster are also described.

Evidence for specificity of cultivable bacteria associated with arbuscular mycorrhizal fungal spores.

Bacteria associated with arbuscular mycorrhizal (AM) fungal spores may play functional roles in interactions between AM fungi, plant hosts and defence against plant pathogens. To study AM fungal spore-associated bacteria (AMB) with regard to diversity, source effects (AM fungal species, plant host) and antagonistic properties, we isolated AMB from surface-decontaminated spores of Glomus intraradices and Glomus mosseae extracted from field rhizospheres of Festuca ovina and Leucanthemum vulgare. Analysis of 385 AMB was carried out by fatty acid methyl ester (FAME) profile analysis, and some also identified using 16S rRNA gene sequence analysis. The AMB were tested for capacity to inhibit growth in vitro of Rhizoctonia solani and production of fluorescent siderophores. Half of the AMB isolates could be identified to species (similarity index 0.6) within 16 genera and 36 species. AMB were most abundant in the genera Arthrobacter and Pseudomonas and in a cluster of unidentified isolates related to Stenotrophomonas. The AMB composition was affected by AM fungal species and to some extent by plant species. The occurrence of antagonistic isolates depended on AM fungal species, but not plant host, and originated from G. intraradices spores. AM fungal spores appear to host certain sets of AMB, of which some can contribute to resistance by AM fungi against plant pathogens.

Soil, But Not Cultivar, Shapes the Structure of Arbuscular Mycorrhizal Fungal Assemblages Associated with Strawberry

Arbuscular mycorrhizal fungi are widespread plant symbionts occurring in most agricultural crops, where they can play key roles in the growth and health of their plant hosts. Plant benefits can depend on the identity of the associated arbuscular mycorrhizal fungi (AMF), but little is known about the identity of the fungal partners in most agricultural systems. In this study, we describe the AMF assemblages associated with four cultivars of strawberry in an outdoor experiment using two field soils with different origin and management history. Assemblages were characterised by clone library sequencing of 18S rRNA gene fragments. Soil dramatically influenced the degree of mycorrhizal colonisation and AMF assemblage structure in the roots. No differences were observed between cultivars. Fungi belonging to the genus Acaulospora dominated the AMF assemblages in one soil, but they were not detected in the other. These results suggest that physicochemical soil characteristics and management can play a role in determining the identity and structure of microbial communities associated with particular hosts in agricultural systems.

Occurrence of Glomeromycota spores and some arbuscular mycorrhiza fungal species in arable fields in Sweden

Fungi within the phylum Glomeromycota were investigated in arable fields throughout Sweden. Sweden is located between 55° and 69° North. The fungi within this phylum form arbuscular mycorrhizal symbiosis with plant roots. Sampling of soil was carried out to a depth of 30 cm in the rhizosphere. Arbuscular mycorrhizal fungi were found at all 45 sampling sites, at densities between 3 and 44 spores per g dry weight of soil. No significant differences in spore densities were found between different agro-climatic zones or between semi-natural grassland and ploughed fields. Our study revealed that the upper half (0–15 cm) of the soil profiles had significantly more spores than the lower half (15–30 cm). Spores from eight sampling sites were identified from the indigenous soils. Almost 90% were shown to belong to the genus Glomus. The other genera found were Gigaspora and Scutellospora.