Genome wide association mapping for agronomic, fruit quality, and root architectural traits in tomato under organic farming conditions.

BACKGROUND: Opportunity and challenges of the agriculture scenario of the next decades will face increasing demand for secure food through approaches able to minimize the input to cultivations. Large panels of tomato varieties represent a valuable resource of traits of interest under sustainable cultivation systems and for genome-wide association studies (GWAS). For mapping loci controlling the variation of agronomic, fruit quality, and root architecture traits, we used a heterogeneous set of 244 traditional and improved tomato accessions grown under organic field trials. Here we report comprehensive phenotyping and GWAS using over 37,300 SNPs obtained through double digest restriction-site associated DNA (dd-RADseq). RESULTS: A wide range of phenotypic diversity was observed in the studied collection, with highly significant differences encountered for most traits. A variable level of heritability was observed with values up to 69% for morphological traits while, among agronomic ones, fruit weight showed values above 80%. Genotype by environment analysis highlighted the strongest genotypic effect for aboveground traits compared to root architecture, suggesting that the hypogeal part of tomato plants has been a minor objective for breeding activities. GWAS was performed by a compressed mixed linear model leading to 59 significantly associated loci, allowing the identification of novel genes related to flower and fruit characteristics. Most genomic associations fell into the region surrounding SUN, OVATE, and MYB gene families. Six flower and fruit traits were associated with a single member of the SUN family (SLSUN31) on chromosome 11, in a region involved in the increase of fruit weight, locules number, and fruit fasciation. Furthermore, additional candidate genes for soluble solids content, fruit colour and shape were found near previously reported chromosomal regions, indicating the presence of synergic and multiple linked genes underlying the variation of these traits. CONCLUSIONS: Results of this study give new hints on the genetic basis of traits in underexplored germplasm grown under organic conditions, providing a framework for the development of markers linked to candidate genes of interest to be used in genomics-assisted breeding in tomato, in particular under low-input and organic cultivation conditions.

Effects of organic farming and genotype on alimentary and nutraceutical parameters in tomato fruits.

BACKGROUND: In recent years there has been growing interest in the influence of sustainable cultivation systems on the biochemical quality of vegetables. In this study, two genotypes of tomato, Giulianova and Perbruzzo, were grown in both organic (Or) and conventional (Conv) systems for three years and harvested at commercial ripening in each year. The Conv system was established according to traditional techniques and the Or system according to current EU regulations. Samples were evaluated for volatile substances, sugars, organic acids, dry matter, pH and lycopene. RESULTS: During the three years, volatile substances increased in Or samples of Giulianova, while this response was less evident in Perbruzzo. Other parameters of both genotypes were not influenced by Or cultivation. Lycopene content in both Conv-cultivated genotypes was constant during the whole experimental period. In Or samples, lycopene content was lower than in Conv samples during the first and second years. In the last year, lycopene content in Or samples of Giulianova was similar to that in Conv samples, while lycopene content in Or samples of Perbruzzo was higher than that in Conv samples. CONCLUSION: Changes in biochemical parameters of tomato fruits could be affected by both cultivation system and genotype, with a significant increase in both volatile substances and lycopene in the 2010 crop.

Influence of year, genotype and cultivation system on nutritional values and bioactive compounds in tomato (Solanum lycopersicum L.).

Two tomato genotypes were grown in open field by three cultivation systems (one conventional and two distinct organic for mulching) in three years, 2015, 2016 and 2017. Yields, sugars, organic acids, amino acids, ascorbic acid, biothiols, carotenoids and phenols were measured. Weather conditions largely differed among harvest years, with summer 2016 rainier and less warm, and an opposite summer 2017. Organic systems had lower yields than conventional one but also, interestingly, lower waste percentages. Furthermore, tilled and no-tilled organic systems provided comparable yields. With respect to 3-year average, sugars were higher in 2017, acids in 2016 and in organic fruits, and amino acids increased in 2015 and in conventional samples. A higher glutathione content was found in organic samples, and higher carotenoids in 2017. Phenols increased in 2016, with a higher chlorogenic acid content in organic tomatoes. Some differences between genotypes were observed, highlighting their different adaptability to growing systems.

Genotypic and Environmental Effects on Morpho-Physiological and Agronomic Performances of a Tomato Diversity Panel in Relation to Nitrogen and Water Stress Under Organic Farming.

The agricultural scenario of the upcoming decades will face major challenges for the increased and sustainable agricultural production and the optimization of the efficiency of water and fertilizer inputs. Considering the current and foreseen water scarcity in several marginal and arid areas and the need for a more sustainable farming production, the selection and development of cultivars suitable to grow under low-input conditions is an urgent need. In this study, we assayed 42 tomato genotypes for thirty-two morpho-physiological and agronomic traits related to plant, fruit, and root characteristics under standard (control) and no-nitrogen fertilization or water deficit (30% of the amount given to non-stressed trials) treatments in two sites (environments), which corresponded to organic farms located in Italy and Spain. A broad range of variation was found for all traits, with significant differences between the applied treatments and the cultivation sites. Dissection of genotypic (G), environmental (E), and treatment (T) factors revealed that the three main factors were highly significant for many traits, although G was the main source of variation in most cases. G × E interactions were also important, while G × T and E × T were less relevant. Only fruit weight and blossom end rot were highly significant for the triple interaction (G × E × T). Reduction of water supply significantly increased the soluble solid content in both locations, whereas both nitrogen and water stress led to a general decrease in fruit weight and total yield. Despite so, several accessions exhibited better performances than the control when cultivated under stress. Among the accessions evaluated, hybrids were promising in terms of yield performance, while overall landraces and heirlooms exhibited a better quality. This suggests the possibility of exploiting both the variation within ancient varieties and the heterosis for yield of hybrids to select and breed new varieties with better adaptation to organic farming conditions, both under optimal and suboptimal conditions. The results shed light on the strategies to develop novel varieties for organic farming, giving hints into the management of inputs to adopt for a more sustainable tomato cultivation.

The ground beetle Pseudoophonus rufipes gut microbiome is influenced by the farm management system.

Intensive conventional farm management, characterized by high agrochemicals input, could alter the composition of microbial communities with potential negative effects on both functional traits and the ecosystem services provided. In this study, we investigated the gut microbial composition of a high ecological relevance carabid Pseudoophonus rufipes, sampled in two fields subjected to conventional and organic management practices. Carabids' gut microbiota was analyzed via qPCR and NGS. Profound differences between the microbial composition of organic and conventional samples were detected: the abundance of Tenericutes and Proteobacteria was significant higher in organic and conventional samples, respectively. Spiroplasmataceae and Bifidobacteriaceae families were significantly more abundant in samples from organic management, while Enterococcaceae, Morganellaceae and Yersiniaceae were more abundant in samples from conventional management. The diverse gut microbial composition of insects between the two management systems is related to the pressure of environmental stressors and it may representing an important bioindication of ecological functions and services provided by a carabid species.

Can multi-cropping affect soil microbial stoichiometry and functional diversity, decreasing potential soil-borne pathogens? A study on European organic vegetable cropping systems.

Crop diversification in spatial and temporal patterns can optimize the synchronization of nutrients plant demand and availability in soils, as plant diversity and soil microbial communities are the main drivers of biogeochemical C and nutrient cycling. The introduction of multi-cropping in organic vegetable production can represent a key strategy to ensure efficient complementation mediated by soil microbiota, including beneficial mycorrhizal fungi. This study shows the effect of the introduction of multi-cropping in five European organic vegetable systems (South-West: Italy; North-West: Denmark and Belgium; North-East: Finland and Latvia) on: (i) soil physicochemical parameters; (ii) soil microbial biomass stoichiometry; (iii) crop root mycorrhization; (iv) bacterial and fungal diversity and composition in crop rhizosphere; (v) relative abundance of selected fungal pathogens species. In each site, three cropping systems were considered: (1) crop 1-monocropping; (2) crop 2-monocropping; (3) crop 1-crop 2-intercropping or strip cropping. Results showed that, just before harvest, multi-cropping can increase soil microbial biomass amount and shape microbial community toward a predominance of some bacteria or fungi phyla, in the function of soil nutrient availability. We mainly observed a selection effect of crop type on rhizosphere microbiota. Particularly, Bacteroidetes and Mortierellomycota relative abundances in rhizosphere soil resulted in suitable ecological indicators of the positive effect of plant diversity in field, the first ones attesting an improved C and P cycles in soil and the second ones a reduced soil pathogens' pressure. Plant diversity also increased the root mycorrhizal colonization between the intercropped crops that, when properly selected, can also reduce the relative abundance of potential soil-borne pathogens, with a positive effect on crop productivity in long term.

Agroecological Service Crops Drive Plant Mycorrhization in Organic Horticultural Systems.

Mycorrhizal symbiosis represents a valuable tool for increasing plant nutrient uptake, affecting system biodiversity, ecosystem services and productivity. Introduction of agroecological service crops (ASCs) in cropping systems may determine changes in weed community, that can affect the development of the mycorrhizal mycelial network in the rhizosphere, favoring or depressing the cash crop mycorrhization. Two no-till Mediterranean organic horticultural systems were considered: one located in central Italy, where organic melon was transplanted on four winter-cereals mulches (rye, spelt, barley, wheat), one located in southern Italy (Sicily), where barley (as catch crop) was intercropped in an organic young orange orchard, with the no tilled, unweeded systems taken as controls. Weed "Supporting Arbuscular Mycorrhiza" (SAM) trait, weed density and biodiversity indexes, mycorrhization of coexistent plants in the field, the external mycelial network on roots were analyzed by scanning electron microscopy, crop P uptake, yield and quality were evaluated. We verified that cereals, used as green mulches or intercropped, may drive the weed selection in favor of the SAM species, and promote the mycelial network, thus significantly increasing the mycorrhization, the P uptake, the yield and quality traits of the cash crop. This is a relevant economic factor when introducing sustainable cropping practices and assessing the overall functionality of the agroecosystem.

Weed Functional Diversity as Affected by Agroecological Service Crops and No-Till in a Mediterranean Organic Vegetable System.

This paper explores the effect of agroecological service crops (ASCs), i.e., crops included in the crop rotation for their ecosystem services, terminated with an in-line tillage roller crimper (ILRC) on weed community composition and their functional traits in comparison to a tilled control without ASC. A two-year study was performed in a long-term experiment with vegetables under organic management. Four different cereal crops were introduced as ASCs. Weed abundance and richness and the functional traits were assessed at three different stages, i.e., before and after ASC termination and before harvest of the following crop, melon. All the ASCs showed strong weed suppression, with few differences between the cereals tested. Weed communities with ASCs had later flowering onset and wider flowering span compared to the control, which positively affects weed dispersal and attraction of beneficial insects. However, weed communities with ASCs had higher values for traits related to competition (specific leaf area, seed weight and more perennials). A trade-off between weed suppression and selection of more competitive weed communities by the introduction of ASCs managed with the ILRC should be evaluated in the long-run. The use of the ILRC alternating with other soil management practices seems the more effective strategy to benefit from the minimal soil tillage while avoiding the selection of disservice-related traits in the weed community.

ddRAD sequencing-based genotyping for population structure analysis in cultivated tomato provides new insights into the genomic diversity of Mediterranean 'da serbo' type long shelf-life germplasm.

Double digest restriction-site associated sequencing (ddRAD-seq) is a flexible and cost-effective strategy for providing in-depth insights into the genetic architecture of germplasm collections. Using this methodology, we investigated the genomic diversity of a panel of 288 diverse tomato (Solanum lycopersicum L.) accessions enriched in 'da serbo' (called 'de penjar' in Spain) long shelf life (LSL) materials (152 accessions) mostly originating from Italy and Spain. The rest of the materials originate from different countries and include landraces for fresh consumption, elite cultivars, heirlooms, and breeding lines. Apart from their LSL trait, 'da serbo' landraces are of remarkable interest for their resilience. We identified 32,799 high-quality SNPs, which were used for model ancestry population structure and non-parametric hierarchical clustering. Six genetic subgroups were revealed, clearly separating most 'da serbo' landraces, but also the Spanish germplasm, suggesting a subdivision of the population based on type and geographical provenance. Linkage disequilibrium (LD) in the collection decayed very rapidly within <5 kb. We then investigated SNPs showing contrasted minor frequency allele (MAF) in 'da serbo' materials, resulting in the identification of high frequencies in this germplasm of several mutations in genes related to stress tolerance and fruit maturation such as CTR1 and JAR1. Finally, a mini-core collection of 58 accessions encompassing most of the diversity was selected for further exploitation of key traits. Our findings suggest the presence of a genetic footprint of the 'da serbo' germplasm selected in the Mediterranean basin. Moreover, we provide novel insights on LSL 'da serbo' germplasm as a promising source of alleles for tolerance to stresses.

Mycorrhiza-mediated interference between cover crop and weed in organic winter cereal agroecosystems: The mycorrhizal colonization intensity indicator.

The mycorrhizal fungi are symbiotic organisms able to provide many benefits to crop production by supplying a set of ecosystem functions. A recent ecological approach based on the ability of the fungi community to influence plant-plant interactions by extraradical mycelium development may be applied to diversified, herbaceous agroecosystems. Our hypothesis is that the introduction of a winter cereal cover crop (CC) as arbuscular mycorrhizal fungi (AMF)-host plant in an organic rotation can boosts the AMF colonization of the other plants, influencing crop-weed interference. In a 4-years organic rotation, the effect of two winter cereal CC, rye and spelt, on weed density and AMF colonization was evaluated. The AMF extraradical mycelium on CC and weeds roots was observed by scanning electron microscopy analysis. By joining data of plant density and mycorrhization, we built the mycorrhizal colonization intensity of the Agroecosystem indicator (MA%). Both the CC were colonized by soil AMF, being the mycorrhizal colonization intensity (M%) affected by environmental conditions. Under CC, the weed density was reduced, due to the increase of the reciprocal competition in favor of CC, which benefited from mycorrhizal colonization and promoted the development of AMF extraradical mycelium. Even though non-host plants, some weed species showed an increased mycorrhizal colonization in presence of CC respect to the control. Under intense rainfall, the MA% was less sensitive to the CC introduction. On the opposite, under highly competitive conditions, both the CC boosted significantly the mycorrhization of coexistent plants in the agroecosystem. The proposed indicator measured the agroecological service provided by the considered CCs in promoting or inhibiting the overall AMF colonization of the studied agroecosystems, as affected by weed selection and growth: It informs about agroecosystem resilience and may be profitably applied to indicate the extent of the linkage of specific crop traits to agroecosystem services, contributing to further develop the functional biodiversity theory.

Impact of drying techniques, seasonal variation and organic growing on flavor compounds profiles in two Italian tomato varieties.

The industrial transformation of tomato (Lycopersicon esculentum Mill.) produces processed foods, such as dried tomatoes. In this study two varieties (SaAb and PerBruzzo), grown in three cropping systems (one conventional and two organic ones), were processed by two types of small-scale drying (oven or sun drying), over two years of production. The dried samples were analyzed for their non-volatile and volatile composition, relating the results with sensory analysis. The multivariate analysis performed on collected data allowed a detailed comparison of the effects of processing, year-to year variation and cropping systems. Results indicated that drying methods mainly influenced the composition and flavor profile, also affected by the production year. The cropping system significantly influenced some quality indices, such as the acid and sugar amounts, and the aldehydes, respectively higher and lower in organic samples. The comprehensive PCA analysis allowed discrimination of drying methods and, to a lesser extent, cropping systems.

Effect of tomato variety, cultivation, climate and processing on Sola l 4, an allergen from Solanum lycopersicum.

Tomatoes (Solanum lycopersicum) are one of the most consumed vegetables worldwide. However, tomato allergies in patients suffering from birch pollen allergy occur frequently. Due to highly similar protein structures of the tomato allergen Sola l 4 and the major birch pollen allergen Bet v 1, patients cross-react with allergenic proteins from tomato as well as other fruits or vegetables. The aim of this study was to quantify Sola l 4 in various tomatoes differing in color, size and shape for identification of varieties with a reduced allergen level. Therefore, an indirect competitive ELISA using a specific polyclonal Sola l 4 antibody was developed. In addition, two varieties, both cultivated either conventionally or organically and furthermore dried with different methods, were analyzed to investigate the influence of the cultivation method and processing techniques on Sola l 4 level. Within 23 varieties, Sola l 4 content varied significantly between 0.24 and 1.71 µg Sola l 4/g FW. The tomato cultivars Rugantino and Rhianna showed the significantly lowest level, whereas in cultivars Farbini and Bambello the significantly highest concentration was determined. Drying of tomatoes in the oven and by sun resulted in a significant decrease. The thermal instability was verified for the recombinant Sola l 4 emphasizing the results for the native protein in dried tomato samples. Overall, the Sola l 4 content is cultivar-dependent and no correlation between color and Sola l 4 amount was found. During the drying process of tomatoes Sola l 4 level was significantly reduced due to thermal instability. Growing conditions have a minor effect whereas seasonal effects show a more pronounced impact. These findings could extend the knowledge about the allergen level of different tomato varieties and may help to improve food safety to potentially increase the life quality of patients suffering from birch pollen allergy.

Long-term impact of farm management and crops on soil microorganisms assessed by combined DGGE and PLFA analyses.

In the present study, long-term organic and conventional managements were compared at the experimental field of Monsampolo del Tronto (Marche region, Italy) with the aim of investigating soil chemical fertility and microbial community structure. A polyphasic approach, combining soil fertility indicators with microbiological analyses (plate counts, PCR-denaturing gradient gel electrophoresis [DGGE] and phospholipid fatty acid analysis [PLFA]) was applied. Organic matter, N as well as some important macro and micronutrients (K, P, Mg, Mn, Cu, and Zn) for crop growth, were more available under organic management. Bacterial counts were higher in organic management. A significant influence of management system and management x crop interaction was observed for total mesophilic bacteria, nitrogen fixing bacteria and actinobacteria. Interestingly, cultivable fungi were not detected in all analyzed samples. PLFA biomass was higher in the organic and Gram positive bacteria dominated the microbial community in both systems. Even if fungal biomass was higher in organic management, fungal PCR-DGGE fingerprinting revealed that the two systems were very similar in terms of fungal species suggesting that 10 years were not enough to establish a new dynamic equilibrium among ecosystem components. A better knowledge of soil biota and in particular of fungal community structure will be useful for the development of sustainable management strategies.

Variations in the phytochemical contents and antioxidant capacity of organically and conventionally grown Italian cauliflower (Brassica oleracea L. subsp. botrytis): results from a three-year field study.

A three-year field study (2009-2011) was performed to evaluate phytochemicals and antioxidant capacities of two genotypes (HF1 Emeraude and the local variety, Velox) of green cauliflower grown under organic and conventional management. The conventional system increased yield, but had little effect on the dry matter, whereas the organic system increased the soluble solids. Phytochemicals and antioxidant capacity showed significant year-to-year variability. During the third year, the scarce rainfall determined a significant increase of total glucosinolates and a general decrease of antioxidants in all samples. Interestingly, in the same year organic plants were less affected by the unfavorable climatic conditions, as they increased ascorbic acid, polyphenols, and carotenoids with respect to conventional ones. The overall results for the three years showed that the two genotypes responded differently. Compared to the conventional system, Velox showed 24, 21, 13, 48, and 44% higher content of ascorbic acid, polyphenols, carotenoids, volatiles, and antioxidant capacity, respectively. In contrast, no significant increase in the phytochemicals or the antioxidant potential was found in organic Emeraude, with the exception of total volatiles (+41%). These findings suggest that organic cultivation may be highly effective for particular cauliflower genotypes.