Optimizing Chemically Induced Resistance in Tomato Against Botrytis cinerea.

Resistance-inducing chemicals can offer broad-spectrum disease protection in crops, but can also affect plant growth and interactions with plant-beneficial microbes. We have evaluated different application methods of ß-aminobutyric acid (BABA) and jasmonic acid (JA) for long-lasting induced resistance in tomato against Botrytis cinerea. In addition, we have studied nontarget effects on plant growth and root colonization by arbuscular mycorrhizal fungi (AMF). Germinating seeds for 1 week in BABA- or JA-containing solutions promoted seed germination efficiency, did not affect plant growth, and induced resistance in 4-week-old plants. When formulating BABA and JA in carboxymethyl cellulose seed coating, only BABA was able to induce resistance in 4-week-old plants. Root treatment of 1-week-old seedlings with BABA or JA also induced resistance in 4-week-old plants. However, this seedling treatment repressed plant growth at higher concentrations of the chemicals, which was particularly pronounced in hydroponically grown plants after BABA treatment. Both seed coating with BABA, and seedling treatments with BABA or JA, did not affect AMF root colonization in soil-grown tomato. Our study has identified commercially feasible application methods of BABA and JA, which induce durable disease resistance in tomato without concurrent impacts on plant growth or colonization by plant-beneficial AMF.

Interactions between biochar, arbuscular mycorrhizal fungi and photosynthetic processes in potato (Solanum tuberosum L.).

Pyrolyzed biomass, generating biochar for use as soil amendment, is recognized as a promising strategy for carbon sequestration. Current understanding of the interactions between biochar, arbuscular mycorrhizal (AM), and plant photosynthesis, in terms of biochemical processes and CO2 uptake, is fragmentary. The aim of this study was to investigate the effects on photosynthesis in potato including maximum rate of carboxylation by Rubisco (Vcmax), maximum rate of electron transport rate for RuBP-regeneration (Jmax), mesophyll conductance (gm) and other plant traits. Four types of biochar (wheat or miscanthus straw pellets pyrolyzed at temperatures of either 550 °C or 700 °C) were amended into low phosphorus soil. Potato plants were inoculated with the AM fungus Rhizophagus irregularis (M+) or not (M-). The results showed that four types of biochar generally decreased nitrogen and phosphorus content of potato, especially the biochars pyrolyzed at high temperature. This negative effect of biochar on nutrient content was alleviated by AM. It was found that Vcmax was limited by low plant nitrogen content as well as leaf area and phosphorus content. Plant phosphorus content also limited Jmax, which was mutually constrained by Vcmax of leaves. Low gm was an additional limiting factor for photosynthesis. The gm was positively correlated to nitrogen content, which influenced the leaf anatomical structure by alteration of leaf mass per area. In conclusion, the influence of interactions between quality of biochar and AM symbiosis on photosynthesis of potato seems to relate to effects on plant nutrient content and leaf structures. Accordingly, a model for the dependence of Vcmax on nitrogen and phosphorus content and their interactive effect exhibited a high correlation coefficient. As potato plants form AM symbiosis under natural field conditions, the extent and interaction with the quality of amended biochar can be a determining factor for plant nutrient content, growth and yield.

Influence of DNA extraction and PCR amplification on studies of soil fungal communities based on amplicon sequencing.

Most studies involving next-generation amplicon sequencing of microbial communities from environmental studies lack replicates. DNA extraction and PCR effects on the variation of read abundances of operational taxonomic units generated from deep amplicon 454 pyrosequencing was investigated using soil samples from an agricultural field with diseased pea. One sample was extracted four times, and one of these samples was PCR amplified four times to obtain eight replicates in total. Results showed that species richness was consistent among replicates. Variation among dominant taxa was low across replicates, whereas rare operational taxonomic units showed higher variation among replicates. The results indicate that pooling of several extractions and PCR amplicons will decrease variation among samples.

Activity in the Arbuscular Mycorrhizal Hyphosphere Warning Neighbouring Plants.

Pathogen infections of the phyllosphere have been investigated in detail, however, the changes induced by these infections on the arbuscular mycorrhizal hyphosphere, and the consequent signalling to the neighbouring plants have been scarcely investigated. Here, our objectives were to document that B.fabae infection of connected Vicia faba plants resulted in changes in the metabolism and microbial community of the hyphosphere, confirming the induction of plant defence in connected plants through gene-expression evaluations. Infected plants were challenged with B. fabae for 72 h. Changes in gene-expression of pathogenesis-related proteins 1,2, and 5 (PR1, PR2, PR5) of both infected- and non-infected plants were analysed, to confirm signalling through the hyphosphere. The primary metabolic profiles and changes in the level of microbiota in the hyphosphere were assessed. Changes in expression of PR1, PR2, and PR5 genes occurred in the neighbouring plants 24 hours after infection. Mannitol levels decreased in presence of AMF. A decrease in the level of actinobacteria in the hyphosphere of infected plants was detected. We conclude that B.fabae infection induced a signalling event through the AM hyphosphere, confirmed by changes in defence gene-expression in non-infected neighbouring plants, influenced primary metabolic activity of-, and affected the microbial composition within-, the AM hyphosphere.

Discussion paper: Sustainable increase of crop production through improved technical strategies, breeding and adapted management - A European perspective.

During the next decade it will be necessary to develop novel combinations of management strategies to sustainably increase crop production and soil resilience. Improving agricultural productivity, while conserving and enhancing biotic and abiotic resources, is an essential requirement to increase global food production on a sustainable basis. The role of farmers in increasing agricultural productivity growth sustainably will be crucial. Farmers are at the center of any process of change involving natural resources and for this reason they need to be encouraged and guided, through appropriate incentives and governance practices, to conserve natural ecosystems and their biodiversity, and minimize the negative impact agriculture can have on the environment. Farmers and stakeholders need to revise traditional approaches not as productive as the modern approaches but more friendly with natural and environmental ecosystems values as well as emerging novel tools and approaches addressing precise farming, organic amendments, lowered water consumption, integrated pest control and beneficial plant-microbe interactions. While practical solutions are developing, science based recommendations for crop rotations, breeding and harvest/postharvest strategies leading to environmentally sound and pollinator friendly production and better life in rural areas have to be provided.

Aphid infestation in the phyllosphere affects primary metabolic profiles in the arbuscular mycorrhizal hyphosphere.

While effects of (a)biotic stress events in the phyllosphere have been studied intensively, possible influences of stress on the arbuscular mycorrhizal hyphosphere has scarcely been investigated. We hypothesised that stress challenge in the phyllosphere could alter primary metabolite profiles of the hyphosphere - the mycelial network connecting plants. Donor plants, connected to receiver plants by mycelial networks, were aphid-challenged during 84 h. Primary metabolite profiles in the hyphosphere were investigated. Gene-expression of plant defence gene PR1 was measured in one of the receiver plants during the challenge. Hexose levels in the hyphosphere increased when donor plants were aphid-challenged. This change in metabolic profile was influenced by leaf sampling from receiver plant. PR1 expression increased in donor plants 48 h after challenge, and consequently 60 h after, in receiver plants. We conclude that aphid infestation of donor plants modified primary carbon metabolism in the hyphosphere. Plant defence response in receiver plants, occurred 12 h after detection of response in the aphid-challenged donor plants. While this work is the first to reveal primary metabolic profiles of the AM hyphosphere, more work is needed to elucidate the possible role of transient changes of hexose metabolism in stress response and signalling processes in the hyphosphere of connected plants.

Arbuscular mycorrhizal fungi differentially affect expression of genes coding for sucrose synthases in maize roots.

• The effects of three arbuscular mycorrhizal (AM) fungal isolates on the expression of genes coding for sucrose synthase and on the nonstructural carbohydrate (CHO) status of maize roots were evaluated. Gene expression and CHOs were compared with their status in nonmycorrhizal plants grown at three soil phosphorus (P) concentrations. • The AM fungi and soil P supply influenced expression of genes coding for sucrose synthase in maize roots. In general, up to 18 d after plant emergence, AM colonization increased the expression of genes coding for sucrose synthase in maize roots, whereas increasing soil P decreased this gene expression. The responses in gene expression were detected earlier than other effects of AM fungal colonization, such as increased leaf P status and plant growth response under limiting P supply. • Higher sucrose-synthase gene expression was not related to the concentration of sucrose, reducing sugars or starch in the root tissue. • Higher gene expression in AM roots confirms that there is greater allocation of sucrose from nonstructural CHO pools in roots for the AM fungus during the earliest phase of colonization than in nonmycorrhizal roots.

Linking fungal communities in roots, rhizosphere, and soil to the health status of Pisum sativum.

Changes in fungal communities associated with healthy and diseased pea roots were investigated using deep amplicon pyrosequencing in three spatial compartments: roots, rhizosphere, and surrounding soil. Thirty root systems were collected from three fields, half of which showing clear symptoms of root rot. In total, 500,461 internal transcribed spacer-1 sequences were obtained that were clustered into 123 (roots), 271 (rhizosphere), and 440 (bulk soil) nonsingleton operational taxonomic units (OTUs). Species richness was highest in bulk soils and lowest in roots; however, no notable differences in richness were observed between samples associated with diseased and healthy roots. Health status and field both had significant effects on fungal community structures in roots, whereas only field had significant effects on communities in rhizosphere and bulk soils. Indicator species analysis across the three fields identified a number of OTUs that were more abundant in healthy roots. Pathogens such as Fusarium oxysporum were abundant in diseased roots in some fields. Patterns of disease and causal agents of root rot were different among the three fields, which were also reflected in fungal communities. In conclusion, health status of roots was only vaguely reflected in rhizosphere and bulk soil fungal communities, whereas health status was more important for shaping root communities.

Biotransformation of 2-benzoxazolinone to 2-amino-(3H)-phenoxazin-3-one and 2-acetylamino-(3H)-phenoxazin-3-one in soil.

An alternative to the use of synthetic pesticides is to exploit the natural defense chemicals produced by cereals. An important class of allelochemicals is cyclic hydroxamic acids and related benzoxazolinones. A prolonged degradation experiment of the allelochemical compound from rye 2-benzoxazolinone (BOA) was carried out for up to 90 d at 15 degrees C at three different concentration levels, 3, 3000, and 30,000 nmol BOA g soil(-1), respectively, in a sandy loam soil. Two main degradation products, 2-amino-(3H)-phenoxazin-3-one (APO) and 2-acetylamino-(3H)-phenoxazin-3-one (AAPO), were identified and quantified by LC-ESI-MS-MS. The half-life of BOA increased with higher levels of BOA added to the soil. Half-lives of BOA, APO, and AAPO were determined by fitting a single first-order model to the degradation data. Half-life of BOA was determined to be 0.6 d in the 3 nmol BOA g soil(-1) treatment. Half-lives of BOA, APO, and AAPO were 3.1, 2.7, and 2.1 d, respectively, in the 3000 nmol BOA g soil(-1) treatment. In the 30,000 nmol BOA g soil(-1) treatment, the half-lives were 31 d for BOA and 45 d for APO. The microbial community structure was not affected by addition of BOA to the soil as investigated by analysis of signature fatty acids. The results suggest that the exploitability of BOA for crop protection is dependent on the existing concentration of BOA in the soil and the timing of incorporation of hydroxamic acid synthesizing crops into the soil.