Effects of Elevated Substrate-Ethylene on Colonization of Leek (\u3cem\u3eAllium porrum\u3c/em\u3e) by the Arbuscular Mycorrhizal Fungus \u3cem\u3eGlomus aggregatum\u3c/em\u3e

There are very few studies of hormonal regulation of arbuscular mycorrhiza formation that include the gaseous hormone ethylene. Ethylene is considered inhibitory to the formation of arbuscular mycorrhizae; however, very low concentrations may promote their formation. We used an improved method of exogenous ethylene application to determine whether ethylene concentration dependent changes in colonization occur in the leek (Allium porrum L. cv. Giant Musselburgh) – Glomus aggregatum Schenck & Smith emend. Koske system. This improved method allowed for a continuous flow of constant concentration of the gas to be applied to a substrate. The 0.6 μL/L substrate–ethylene treatment reduced both root and leaf length and resulted in significantly lower arbuscular colonization compared with controls, whereas the 0.3 μL/L treatment reduced root length only and did not significantly affect colonization levels. Despite continuous application of exogenous ethylene, the amount of ethylene detected in inoculated substrates was reduced to near zero 20 days after inoculation. This decrease may be either due to an increased capacity for ethylene oxidation by arbuscular mycorrhizal roots or because arbuscular mycorrhizal fungi (or other microbes in the pot-cultured inoculum) are capable of metabolizing ethylene. The present study highlights the need for investigations into arbuscular mycorrhizal fungal physiology and the mechanisms by which ethylene regulates arbuscular mycorrhiza formation

A Model for the Development of the Rhizobial and Arbuscular Mycorrhizal Symbioses in Legumes and Its Use to Understand the Roles of Ethylene in the Establishment of these two Symbioses

We propose a model depicting the development of nodulation and arbuscular mycorrhizae. Both processes are dissected into many steps, using Pisum sativum L. nodulation mutants as a guideline. For nodulation, we distinguish two main developmental programs, one epidermal and one cortical. Whereas Nod factors alone affect the cortical program, bacteria are required to trigger the epidermal events. We propose that the two programs of the rhizobial symbiosis evolved separately and that, over time, they came to function together. The distinction between these two programs does not exist for arbuscular mycorrhizae development despite events occurring in both root tissues. Mutations that affect both symbioses are restricted to the epidermal program. We propose here sites of action and potential roles for ethylene during the formation of the two symbioses with a specific hypothesis for nodule organogenesis. Assuming the epidermis does not make ethylene, the microsymbionts probably first encounter a regulatory level of ethylene at the epidermis–outermost cortical cell layer interface. Depending on the hormone concentrations there, infection will either progress or be blocked. In the former case, ethylene affects the cortex cytoskeleton, allowing reorganization that facilitates infection; in the latter case, ethylene acts on several enzymes that interfere with infection thread growth, causing it to abort. Throughout this review, the difficulty of generalizing the roles of ethylene is emphasized and numerous examples are given to demonstrate the diversity that exists in plants

Cytokinin accumulation and an altered ethylene response mediate the pleiotropic phenotype of the pea nodulation mutant R50 (sym16)

R50 (sym16), a pleiotropic mutant of Pisum sativum L., is short, has thickened internodes and roots, and has a reduced number of lateral roots and nodules. Its low nodule phenotype can be restored with the application of ethylene inhibitors; furthermore, it can be mimicked by applying cytokinins (CKs) to the roots of the parent line 'Sparkle'. Here, we report on the etiolation phenotypes of R50 and 'Sparkle', and on the interactive roles of ethylene and CKs in these lines. R50 displayed an altered etiolation phenotype, as it was shorter and thicker, and had more developed leaves than dark-grown 'Sparkle'. Shoot morphological differences induced by exogenous ethylene or CKs were found to be less severe for R50. Ethylene inhibitor application induced root and shoot elongation and encouraged apical hook opening in both etiolated lines. Liquid chromatography - tandem mass spectrometry analysis indicated that CK concentrations in R50 were higher than in 'Sparkle', particularly in mature shoots where the levels were maintained at elevated concentrations. These differences indicate a reduction in the CK catabolism of R50. The accumulation of CKs can be directly related to several traits of R50, with the reduced number of nodules and altered shoot ethylene response being likely indirect effects

Reduced arbuscular mycorrhizal colonization in tomato ethylene mutants Reduzida formação de micorrízas arbusculares em tomateiros mutantes em etileno

Plant hormones are likely key regulators of arbuscular mycorrhizae (AM) development. However, their roles in AM are not well known. Here mutants in five hormone classes introgressed in a single tomato (Lycopersicon esculentum Mill. Syn Solanum lycopersicum L.) background (cv. Micro-Tom) were used to determine their effects on AM development and the expression of defense-related genes (chitinases and b-1,3-glucanases) in roots. Under low P conditions, mutant epinastic (epi) and Never ripe (Nr), ethylene overproducer and low sensitivity, respectively, had the intraradical colonization by Glomus clarum highly inhibited, as compared to the control Micro-Tom (MT). No significant alterations in fungal colonization were observed in mutants affecting other hormone classes. Under low P conditions, the steady state levels of transcripts encoding a class I basic chitinase (chi9) were higher in mycorrhizal epi and Nr mutant roots as compared to MT controls. In contrast the steady state levels of a class III acidic b-1,3-glucanase (TomPR-Q'a) transcripts in mycorrhizal epi mutant roots were significantly lower than in mycorrhizal MT roots. Root colonization in epi mutants was accompanied by several alterations in fungal morphology, as compared to root colonization in MT controls. The data suggest that ethylene may play an important role in controlling intraradical arbuscular mycorrhizal fungal growth.<br>Os hormônios vegetais são possíveis reguladores chave do desenvolvimento de micorrizas arbusculares (MAS). Contudo, seus papéis em MA são pouco conhecidos. No presente estudo, foram utilizados mutantes em cinco classes hormonais introgredidos em uma única cultivar (cv. Micro-Tom) de tomateiro (Lycopersicon esculentum Mill. Syn Solanum lycopersicum L.) para determinar seus efeitos no desenvolvimento de MA e expressão de genes relacionados à defesa (quitinases e b-1,3-glucanases) em raízes. Sob condição de baixo P, os mutantes epinastic (epi) e Never ripe (Nr), os quais são super produtores e pouco sensíveis a etileno, respectivamente, tiveram a colonização intra-radicular por Glomus clarum inibida quando comparada com o controle Micro-Tom (MT). Não se observou alterações significativas na colonização fúngica nos mutantes afetando outras classes hormonais. Sob condição de baixo P, o nível de transcritos codificando uma quitinase básica de classe I (chi9) foi mais elevado em raízes micorrizadas dos mutantes epi e Nr, quando comparado com o controle MT. Em contraste, o nível de transcritos de uma b-1,3-glucanase ácida da classe III (TomPR-Q'a) em raízes micorrizadas do mutante epi foi significativamente menor que em raízes micorrizadas de MT. A colonização de raízes no mutante epi foi acompanhada por várias alterações na morfologia fúngica, quando comparada com o controle MT. Os resultados sugerem que o etileno pode desempenhar um importante papel controlando o crescimento fúngico intra-radicular nas MAS