Arbuscular mycorrhiza in landrace and modern wheat : its impact on plant performance and how it is influenced by plant breeding and modern cropping practices

Modern agriculture has a negative impact on climate and environment and is challenged by climate change. A transition to a more sustainable, secure and yet high food production is necessary. Landrace cereals attract attention for their ability to sustain nutrient deficiency, drought and harsh climate conditions. Arbuscular mycorrhiza is a common symbiosis with ancient origin between plant roots and fungi. The aim of this thesis is to review the current knowledge on arbuscular mycorrhiza regarding its impact on wheat landrace performance during cultivation, and to understand if arbuscular mycorrhiza in wheat has been affected by breeding and conventional cropping practices. Arbuscular mycorrhiza may contribute to uptake of phosphorous and other nutrients and enhance drought resistance in wheat. However, the response to arbuscular mycorrhiza in plants is highly variable and dependent on genetical and environmental factors and cropping practices. This thesis concludes that the ability of wheat landrace cereals to adapt to nutrient deficiency and drought stress are most likely not dependent on arbuscular mycorrhiza under Swedish conditions. In addition, input of fertilizers reduces the plant responsiveness to arbuscular mycorrhiza, the crop rotation system might favour or disfavour arbuscular mycorrhiza depending on which crops are included, tillage disfavours arbuscular mycorrhizal fungi and fungicides might reduce spore germination. Thus, conventional practices might have a negative impact on arbuscular mycorrhiza in wheat, even though the farmer’s choice of cultivation methods are more important for arbuscular mycorrhiza than the type of cropping system. Finally, it is concluded that more molecular genetic research is needed in order to understand if arbuscular mycorrhiza in wheat has been affected by breeding for conventional cropping systems

Development of arbuscular mycorrhizal fungi in the presence of different patterns of Trifolium repens shoot flavonoids

We tested the effects of the flavonoid 3-methoxi-5,6,7,8-hydroxy-4'hydroxy flavone (NMHTV) isolated from shoots of non arbuscular mycorrhizal (AM) inoculated clover, and of the flavonoids 5,6,7,8-hydroxy-3-methoxy flavone (MH-1); 5,6,7,8-hydroxy-4'- hydroxy flavone (MH-2); and 5,7-hydroxy-3,4'-methoxy flavone (MH-3); isolated from AM clover (Trifolium repens) shoots, on spore germination, hyphal length, hyphal branches and the number of cluster of auxiliary cells or the number of secondary spores (Presymbiotic stage) and on the number of entry points and the percentage of AM colonized root of tomato (Lycopersicum esculentum) by the AM fungi Gigaspora rosea, Giaspora margarita, Glomus mosseae and Glomus intraradices (Symbiotic stage). Non significant effects of the flavonoids isolated from the shoot of mycorrhizal colonized clover on the presymbiotic and symbiotic stages of Gigaspora and Glomus endophytes were found. The flavonoid NMHTV isolated from non AM clover shoot, did not affect the percentage of germination of spores but significantly increased (P < 0.05) the other steps of the presymbiotic stage of Gi. margarita spores when 2 μM concentration was used. The symbiotic stage of Gi. margarita was also significantly increased when 2 μM of the flavonoid NMHTV was applied. This flavonoid had no effect on the presymbiotic development of G. mosseae, G. intraradices and Gi. rosea except when 8 μM concentration was used, which inhibited the hyphal length of Gi. rosea. These results suggest the possible implication of the flavonoid NMHTV in the susceptibility of tomato roots to the AM formation by Gi. margarita. The absence of stimulation of the AM presymbiotic and symbiotic stages in tomato by exogenous application of the newly synthesized flavonoids MH-1, MH-2, and MH-3, in clover shoots after AM colonization, indicated that the autorregulation of the AM symbiosis can be, at least partially, due to the disappearance of flavonoids in AM colonized plants that stimulated the AM symbiosis.Fil: Scervino, Jose Martin. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; ArgentinaFil: Ponce, María Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Centro de Investigaciones en Hidratos de Carbono. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Centro de Investigaciones en Hidratos de Carbono; ArgentinaFil: Della Mónica, Ivana Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Centro de Investigaciones en Hidratos de Carbono. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Centro de Investigaciones en Hidratos de Carbono; ArgentinaFil: Vierheilig, Horst. Universitat Fur Bodenkultur Wien; Austria. Consejo Superior de Investigaciones Científicas. Estación Experimental del Zaidín; EspañaFil: Ocampo, Juan Antonio. Consejo Superior de Investigaciones Científicas. Estación Experimental del Zaidín; EspañaFil: Godeas, Alicia Margarita. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Influence of soil chemical properties on relative abundance of arbuscular mycorrhiza in forested soils in Malaysia

Th eeff ect of soil chemical properties on the diversity and colonization of arbuscular mycorrhiza (AM) varies among ecosystems. This study was conducted to assess and compare the abundance of AM in a rehabilitated forest and a logged-over forest soil using the most probable number and spore number methods. Glomus (71.7%-82.1%) and Acaulospora (17.4%-19.5%) were found to be abundant in both sites, while Gigaspora was found only in the loggedover forest. Th e abundance of AM in the rehabilitated forest based on the spore count was signifi cantly higher than in the logged-over forest by a 6-fold diff erence. Furthermore, root colonization in the rehabilitated forest was found to be almost 9-fold higher than in the logged-over forest. Such diff erences are linked to the soil chemical properties. The addition of organic matter during forest rehabilitation activity had created favorable conditions for AM sporulation. Soil P in both forests was positively related to the spore count (r > 0.68, P < 0.001) while the most probable number (MPN)was negatively infl uenced by soil K (r = –0.632, P <0.01). In conclusion, this study showed that soil chemical properties have a direct eff ect on the abundance of AM

Excluding arbuscular mycorrhiza lowers variability in soil respiration but slows down recovery from perturbations

The role of mutualisms in mediating temporal stability in an ecosystem has been debated extensively. Here, we focus on how a ubiquitous mutualism, arbuscular mycorrhiza, influences temporal stability of a key ecosystem process, ecosystem respiration. We discriminated between two forms of temporal stability, temporal variability and resilience, and hypothesized that excluding arbuscular mycorrhiza would be detrimental for both of them. We analyzed a set of 10 parallel manipulation experiments to assess how excluding arbuscular mycorrhiza modulates temporal stability compared to other common experimental factors. We quantified the temporal variability of ecosystem respiration and the resilience to experimental perturbations (i.e., pulses, stresses, and a disturbance) following manipulations of mycorrhizal state. We observed lower temporal variability in the absence of arbuscular mycorrhiza in discord to our main hypothesis. Manipulating arbuscular mycorrhiza had a stronger impact on temporal variability than the pulse (application of urea), the stress (addition of salt), and a disturbance (experimental defoliation) but weaker than excluding primary producers or comparing across different plant species. Resilience to experimental perturbations declined in non‐mycorrhizal microcosms. We present an empirical study on how mutualisms impact temporal stability. Arbuscular mycorrhiza differentially alters temporal variability and resilience, highlighting that generalizing across different forms of temporal stability could be misleading

SymGRASS: a database of sugarcane orthologous genes involved in arbuscular mycorrhiza and root nodule symbiosis : from Seventh International Meeting on Computational Intelligence Methods for Bioinformatics and Biostatistics, (CIBB 2010), Palermo, Italy, 16 - 18 September 2010

Background: The rationale for gathering information from plants procuring nitrogen through symbiotic interactions controlled by a common genetic program for a sustainable biofuel production is the high energy demanding application of synthetic nitrogen fertilizers. We curated sequence information publicly available for the biofuel plant sugarcane, performed an analysis of the common SYM pathway known to control symbiosis in other plants, and provide results, sequences and literature links as an online database. Methods: Sugarcane sequences and informations were downloaded from the nucEST database, cleaned and trimmed with seqclean, assembled with TGICL plus translating mapping method, and annotated. The annotation is based on BLAST searches against a local formatted plant Uniprot90 generated with CD-HIT for functional assignment, rpsBLAST to CDD database for conserved domain analysis, and BLAST search to sorghum's for Gene Ontology (GO) assignment. Gene expression was normalized according the Unigene standard, presented as ESTs/100 kb. Protein sequences known in the SYM pathway were used as queries to search the SymGRASS sequence database. Additionally, antimicrobial peptides described in the PhytAMP database served as queries to retrieve and generate expression profiles of these defense genes in the libraries compared to the libraries obtained under symbiotic interactions. Results: We describe the SymGRASS, a database of sugarcane orthologous genes involved in arbuscular mycorrhiza (AM) and root nodule (RN) symbiosis. The database aggregates knowledge about sequences, tissues, organ, developmental stages and experimental conditions, and provides annotation and level of gene expression for sugarcane transcripts and SYM orthologous genes in sugarcane through a web interface. Several candidate genes were found for all nodes in the pathway, and interestingly a set of symbiosis specific genes was found. Conclusions: The knowledge integrated in SymGRASS may guide studies on molecular, cellular and physiological mechanisms by which sugarcane controls the establishment and efficiency of endophytic associations. We believe that the candidate sequences for the SYM pathway together with the pool of exclusively expressed tentative consensus (TC) sequences are crucial for the design of molecular studies to unravel the mechanisms controlling the establishment of symbioses in sugarcane, ultimately serving as a basis for the improvement of grass crops

Effect of tillage and crop on arbuscular mycorrhiza

Large-scale inoculation with arbuscular mycorrhizal fungi (AMF) is generally impractical in most regions and we have little understanding of the factors that determine inoculation success. Nevertheless, the ability to take full advantage of indigenous AMF for sustainable production needs to be developed within cropping systems. We used part of a long-term field experiment to understand the influence of tillage and the preceding crop on AMF colonization over the growing season. Arbuscular mycorrhiza colonization rate was more affected by treatment (tillage or the combination of crop and preceding crop) than by the total number of AMF spores in the soil. Conventional tillage (CT) had a statistically significant negative effect (P £ 0.05) on spore numbers isolated from the soil, but only in the first year of study. However, the AMF colonization rate was significantly reduced by CT, and the roots of wheat, Triticum aestivum, L, cv. Coa after sunflower, Helianthus annuus L., were less well colonized than were those of triticale, X Triticosecale Wittmack, cv. Alter after wheat, but the affect of tillage was more pronounced than was the effect of crop combination. Under no-till there was a significant increase in AMF colonization rate throughout the sampling period in both wheat and triticale,indicating that the extraradical mycelium previously produced acted as a source of inoculum. In general, triticale showed greater AMF colonization than wheat, despite the preceding crop being less mycotrophic. Under these experimental conditions, typical of Mediterranean agricultural systems, AMF colonization responded more strongly to tillage practices than to the combination of crop and preceding crop

Arbuscular mycorrhizas in phosphate-polluted soil: interrelations between root colonization and nitrogen

To investigate whether arbuscular mycorrhizal fungi (AMF) – abundant in a phosphate-polluted but nitrogen-poor field site – improve plant N nutrition, we carried out a two-factorial experiment, including N fertilization and fungicide treatment. Percentage of root length colonized (% RLC) by AMF and tissue element concentrations were determined for four resident plant species. Furthermore, soil nutrient levels and N effects on aboveground biomass of individual species were measured. Nitrogen fertilization lowered % RLC by AMF of Artemisia vulgaris L., Picris hieracioides L. and Poa compressa L., but not of Bromus japonicus Thunb. This – together with positive N addition effects on N status, N:P-ratio and aboveground biomass of most species – suggested that plants are mycorrhizal because of N deficiency. Fungicide treatment, which reduced % RLC in all species, resulted in lower N concentrations in A. vulgaris and P. hieracioides, a higher N concentration in P. compressa, and did not consistently affect N status of B. japonicus. Evidently, AMF had an influence on the N nutrition of plants in this P-rich soil; however – potentially due to differences in their mycorrhizal responsiveness – not all species seemed to benefit froma mycorrhiza-mediated N uptake and accordingly, N distribution

Molecular diversity of arbuscular mycorrhizal fungi in onion roots from organic and conventional farming systems in the Netherlands

Diversity and colonization levels of naturally occurring arbuscular mycorrhizal fungi (AMF) in onion roots were studied to compare organic and conventional farming systems in the Netherlands. In 2004, 20 onion fields were sampled in a balanced survey between farming systems and between two regions, namely, Zeeland and Flevoland. In 2005, nine conventional and ten organic fields were additionally surveyed in Flevoland. AMF phylotypes were identified by rDNA sequencing. All plants were colonized, with 60% for arbuscular colonization and 84% for hyphal colonization as grand means. In Zeeland, onion roots from organic fields had higher fractional colonization levels than those from conventional fields. Onion yields in conventional farming were positively correlated with colonization level. Overall, 14 AMF phylotypes were identified. The number of phylotypes per field ranged from one to six. Two phylotypes associated with the Glomus mosseae-coronatum and the G. caledonium-geosporum species complexes were the most abundant, whereas other phylotypes were infrequently found. Organic and conventional farming systems had similar number of phylotypes per field and Shannon diversity indices. A few organic and conventional fields had larger number of phylotypes, including phylotypes associated with the genera Glomus-B, Archaeospora, and Paraglomus. This suggests that farming systems as such did not influence AMF diversity, but rather specific environmental conditions or agricultural practice

NO-TILL PROVIDES THE OPPORTUNITY TO MANAGE UNDERGROUND INTERACTIONS BETWEEN ARBUSCULAR MYCORRHIZAL FUNGI, WEEDS AND CROP PLANTS UNDER MEDITERRANEAN CONDITIONS

Early colonization of crop roots by arbuscular mycorrhiza (AM) is considered beneficial but its importance likely depends on the possible stresses faced by the host plant. Manganese toxicity is one such stress that AM can alleviate. Colonization initiated by extraradical mycelium (ERM) is faster than other sources of inoculum. No-till creates the possibility of encouraging inoculation via this source. At seeding time the ERM available for colonizing plants under no-till would have developed in association with previous crops or those weeds that germinated after the first autumn rain. However, the long, hot and dry summer under Mediterranean conditions might reduce the effectiveness of the ERM to colonize the new crop. The hypothesis that an intact ERM developed by weeds can affect the earlier AM colonization of wheat and alleviate Mn toxicity was tested in a pot experiment. Two mycotrophic (Ornithopus compressus L., Lolium rigidum Gaudin), and one non-mycotrophic (Silene galica L) weed species were grown for 7 weeks before being controlled with Glyphosate (the ERM remaining intact) or by mechanical disturbance (which also disrupted the ERM). Wheat was then planted and allowed to grow for 21 days. AM colonization, plant dry weight and shoot nutrient content were evaluated for both weeds and wheat. When an intact mycelium was present at the seeding of wheat (treatments with Ornithopus compressus and Lolium rigidum controlled by Glyphosate) there was a threefold increase in the AM colonization rate and growth of the crop compared with results for all the other treatments. The enhanced growth of wheat was associated with an alleviation of Mn toxicity, consistent with the hypothesis. However, there was a significant difference of the wheat growth after Ornithopus and Lolium (1.9 times), suggesting functional diversity within mycotrophic weeds and crops

Agronomic Management of Indigenous Mycorrhizas

Many of the advantages conferred to plants by arbuscular mycorrhiza (AM) are associated to the ability of AM plants to explore a greater volume of soil through the extraradical mycelium. Sieverding (1991) estimates that for each centimetre of colonized root there is an increase of 15 cm3 on the volume of soil explored, this value can increase to 200 cm3 depending on the circumstances. Due to the enhancement of the volume of soil explored and the ability of the extraradical mycelium to absorb and translocate nutrients to the plant, one of the most obvious and important advantages resulting from mycorrhization is the uptake of nutrients. Among of which the ones that have immobilized forms in soil, such as P, assume particular significance. Besides this, many other benefits are recognized for AM plants (Gupta et al, 2000): water stress alleviation (Augé, 2004; Cho et al, 2006), protection from root pathogens (Graham, 2001), tolerance to toxic heavy metals and phytoremediation (Audet and Charest, 2006; Göhre and Paszkowski, 2006), tolerance to adverse conditions such as very high or low temperature, high salinity (Sannazzaro et al, 2006), high or low pH (Yano and Takaki, 2005) or better performance during transplantation shock (Subhan et al, 1998). The extraradical hyphae also stabilize soil aggregates by both enmeshing soil particles (Miller e Jastrow, 1992) and producing a glycoprotein, golmalin, which may act as a glue-like substance to adhere soil particles together (Wright and Upadhyaya, 1998). Despite the ubiquous distribution of mycorrhizal fungi (Smith and Read, 2000) and only a relative specificity between host plants and fungal isolates (McGonigle and Fitter, 1990), the obligate nature of the symbiosis implies the establishment of a plant propagation system, either under greenhouse conditions or in vitro laboratory propagation. These techniques result in high inoculum production costs, which still remains a serious problem since they are not competitive with production costs of phosphorus fertilizer. Even if farmers understand the significance of sustainable agricultural systems, the reduction of phosphorus inputs by using AM fungal inocula alone cannot be justified except, perhaps, in the case of high value crops (Saioto and Marumoto, 2002). Nurseries, high income horticulture farmers and no-agricultural application such as rehabilitation of degraded or devegetated landscapes are examples of areas where the use of commercial inoculum is current. Another serious problem is quality of commercial available products concerning guarantee of phatogene free content, storage conditions, most effective application methods and what types to use. Besides the information provided by suppliers about its inoculum can be deceiving, as from the usually referred total counts, only a fraction may be effective for a particular plant or in specific soil conditions. Gianinazzi and Vosátka (2004) assume that progress should be made towards registration procedures that stimulate the development of the mycorrhizal industry. Some on-farm inoculum production and application methods have been studied, allowing farmers to produce locally adapted isolates and generate a taxonomically diverse inoculum (Mohandas et al, 2004; Douds et al, 2005). However the inocula produced this way are not readily processed for mechanical application to the fields, being an obstacle to the utilization in large scale agriculture, especially row crops, moreover it would represent an additional mechanical operation with the corresponding economic and soil compaction costs. It is well recognized that inoculation of AM fungi has a potential significance in not only sustainable crop production, but also environmental conservation. However, the status quo of inoculation is far from practical technology that can be widely used in the field. Together a further basic understanding of the biology and diversity of AM fungi is needed (Abbott at al, 1995; Saito and Marumoto, 2002). Advances in ecology during the past decade have led to a much more detailed understanding of the potential negative consequences of species introductions and the potential for negative ecological consequences of invasions by mycorrhizal fungi is poorly understood. Schwartz et al, (2006) recommend that a careful assessment documenting the need for inoculation, and the likelihood of success, should be conducted prior to inoculation because inoculations are not universally beneficial. Agricultural practices such as crop rotation, tillage, weed control and fertilizer apllication all produce changes in the chemical, physical and biological soil variables and affect the ecological niches available for occupancy by the soil biota, influencing in different ways the symbiosis performance and consequently the inoculum development, shaping changes and upset balance of native populations. The molecular biology tools developed in the latest years have been very important for our perception of these changes, ensuing awareness of management choice implications in AM development. In this context, for extensive farming systems and regarding environmental and economic costs, the identification of agronomic management practices that allow controlled manipulation of the fungal community and capitalization of AM mutualistic effect making use of local inoculum, seem to be a wise option for mycorrhiza promotion and development of sustainable crop production