Physiological response of Cucurbita pepo var. pepo mycorrhized by Sonoran desert native arbuscular fungi to drought and salinity stresses

ABSTRACT Plants response to symbiosis with arbuscular mycorrhizal fungi (AMF) under water stress is important to agriculture. Under abiotic stress conditions native fungi are more effective than exotics in improving plant growth and water status. Mycorrhization efficiency is related to soil fungi development and energy cost-benefit ratio. In this study, we assessed the effect on growth, water status and energy metabolism of Cucurbita pepo var. pepo when inoculated with native AMF from the Sonoran desert Mexico (mixed isolate and field consortium), and compared with an exotic species from a temperate region, under drought, low and high salinity conditions. Dry weights, leaf water content, water and osmotic potentials, construction costs, photochemistry and mycorrhization features were quantified. Under drought and low salinity conditions, the mixed isolate increased plant growth and leaf water content. Leaf water potential was increased only by the field consortium under drought conditions (0.5-0.9 MPa). Under high salinity, the field consortium increased aerial dry weight (more than 1 g) and osmotic potential (0.54 MPa), as compared to non-mycorrhized controls. Plants inoculated with native AMF, which supposedly diminish the effects of stress, exhibited low construction costs, increased photochemical capacity, and grew larger external mycelia in comparison to the exotic inoculum.

Dark septate endophyte decreases stress on rice plants

Abstract Abiotic stress is one of the major limiting factors for plant development and productivity, which makes it important to identify microorganisms capable of increasing plant tolerance to stress. Dark septate endophytes can be symbionts of plants. In the present study, we evaluated the ability of dark septate endophytes isolates to reduce the effects of water stress in the rice varieties Nipponbare and Piauí. The experiments were performed under gnotobiotic conditions, and the water stress was induced with PEG. Four dark septate endophytes were isolated from the roots of wild rice (Oryza glumaepatula) collected from the Brazilian Amazon. Plant height as well as shoot and root fresh and dry matter were measured. Leaf protein concentrations and antioxidant enzyme activity were also estimated. The dark septate endophytes were grown in vitro in Petri dishes containing culture medium; they exhibited different levels of tolerance to salinity and water stress. The two rice varieties tested responded differently to inoculation with dark septate endophytes. Endophytes promoted rice plant growth both in the presence and in the absence of a water deficit. Decreased oxidative stress in plants in response to inoculation was observed in nearly all inoculated treatments, as indicated by the decrease in antioxidant enzyme activity. Dark septate endophytes fungi were shown to increase the tolerance of rice plants to stress caused by water deficiency.

Nitrogen-fixing bacteria and arbuscular mycorrhizal fungi in Piptadenia gonoacantha ( Mart. ) Macbr

Abstract The family Leguminosae comprises approximately 20,000 species that mostly form symbioses with arbuscular mycorrhizal fungi (AMF) and nitrogen-fixing bacteria (NFB). This study is aimed at investigating and confirming the dependence on nodulation and biological nitrogen fixation in the specie Piptadenia gonoacantha (Mart.) Macbr., which belongs to the Piptadenia group. Two consecutive experiments were performed in a greenhouse. The experiments were fully randomized with six replicates and a factorial scheme. For the treatments, the two AMF species and three NFB strains were combined to nodulate P. gonoacantha in addition to the control treatments. The results indicate this species’ capacity for nodulation without the AMF; however, the AMF + NFB combinations yielded a considerable gain in P. gonoacantha shoot weight compared with the treatments that only included inoculating with bacteria or AMF. The results also confirm that the treatment effects among the AMF + NFB combinations produced different shoot dry weight/root dry weight ratios. We conclude that AMF is not necessary for nodulation and that this dependence improves species development because plant growth increases upon co-inoculation.

Production of native arbuscular mycorrhizal fungi inoculum under different environmental conditions

Abstract In order to obtain an arbuscular mycorrhizal fungi (AMF) native inoculum from Sierra de Moa and determine the most appropriate conditions for its big scale production, four light and temperature combinations were tested in three plant species (Calophyllum antillanum, Talipariti elatum and Paspalum notatum). Growth and development parameters, as well as the mycorrhizal functioning of the seedlings were evaluated. The natural light treatment under high temperatures (L-H) was the most suitable for the growth and development of the three plant species, showing the highest total biomass values, mainly of root, and a positive root-shoot ratio balance. This treatment also promoted higher values of root mycorrhizal colonization, external mycelium and AMF spore density. A total of 38 AMF species were identified among the plants and environmental conditions tested. Archaeospora sp.1, Glomus sp.5, Glomus brohultii and G. glomerulatum were observed in all the treatments. The L-H condition can be recommended for native inoculum production, as it promotes a better expression of the AM symbiosis and an elevated production of mycorrhizal propagules.

Functionality of arbuscular mycorrhizal fungi in three plant communities in the Managed Floristic Reserve San Ubaldo-Sabanalamar, Cuba / Funcionalidad de hongos micorrízicos arbusculares en tres comunidades vegetales de la Reserva Florística Manejada de San Ubaldo-Sabanalamar, Cuba

Despite the ubiquity and importance of indigenous arbuscular mycorrhizal fungi (AMF) for plant ecosystems; functioning of indigenus mycorrhizal symbiosis (IMS) and related environmental factors at coastal Caribbean ecosystems remains still scarce. In order to determine functionality of IMS under contrasting land uses and wet seasons from Cuba, the influence of the water stress on some AMF functionality parameters from a semi-natural savannah (NS), a recovered savannah (RS) and an agro-ecosystem (AG) from the Managed Floristic Reserve San Ubaldo-Sabanalamar, Pinar del Rio, Cuba were assessed during two-years. Soil and root samples were collected in April and October, during the dry and wet seasons, respectively, in 2008 and 2010. Four plots in each ecosystem were selected, and five soil sub-samples were randomly collected, bulked, mixed homogeneously and used as the composite sample per plot. The host plant root biomass, arbuscular mycorrhizal colonization of the host plant, density of the intraradical and extraradical AMF mycelia, fungal endophyte biomass and AMF spore density were assessed. The host plant root biomass increased in the NS environment during the dry season, and approximately 12.85g root/dm³ dry soil was recorded. The colonization degree were significantly higher in all environments during the wet season of the second year, with means ranging from 79% to 89%. The extraradical mycelia were significantly more abundant in the dry season of the second year in all environments, with a maximum of 279mg/dm³ in the RS ecosystem. The density of AMF spores was highest in the dry season of the second year for the three studied ecosystems. The RS ecosystem hosted 5 670 spores/100g dry soil. In general, the influence of rainfall seasonality on the function of AMF was stronger than the influence of ecosystem management. The root biomass and extraradical mycelia were high in the dry seasons, suggesting strategies to increase the volume of soil for the mutual benefit of the symbionts. The increase in spore density during the dry seasons appears as an adaptation allowing AMF to survive period of water shortage. This study improves our understanding of the adaptative responses of arbuscular mycorrhizal symbiosis to seasonal variations in soil water availability. Rev. Biol. Trop. 63 (2): 341-356. Epub 2015 June 01.

En el presente estudio se evaluó, durante dos años, la influencia de los períodos seco y lluvioso sobre el funcionamiento de hongos formadores de micorrizas arbusculares (HFMA) simbiontes de plantas nativas de una sabana semi natural, una sabana recuperada de la actividad agrícola y un agroecosistema, ubicados en la Reserva Florística Manejada San Ubaldo-Sabanalamar, Pinar del Río, Cuba. Se recolectaron muestras de suelo en abril y octubre (período seco y húmedo de 2008 y 2010, respectivamente). Dentro de cada ecosistema se seleccionaron cuatro parcelas y se recolectaron cinco submuestras al azar, las que fueron homogeneizadas para formar una muestra compuesta por parcela. Se cuantificó la biomasa de raicillas, la colonización micorrízica de las plantas hospedadoras en el campo, el micelio extrarradical, el micelio endófito y la densidad de esporas. La biomasa de raicillas en la sabana semi natural incrementó durante los períodos secos, la cual alcanzó hasta 12.85 g/dm3 de suelo. En todos los ecosistemas, la mayor colonización micorrízica ocurrió en la época lluviosa del segundo año con valores que oscilaron entre 79 y 89 %. Las mayores biomasas de micelio externo fueron registradas en los tres ecosistemas durante el período seco del segundo año, con un valor máximo de 279 mg/dm3 de suelo en la sabana recuperada. La mayor densidad de esporas de HFMA fue determinada en el periodo seco del segundo año para los tres ecosistemas estudiados, con el valor más alto en la sabana recuperada con 5 670 esporas/100 g de suelo seco. De manera general, se evidenció un efecto de la estacionalidad de la lluvia más que del manejo de ecosistema sobre el funcionamiento de los HFMA. La mayor actividad colonizadora se evidenció en períodos húmedos, mientras que mayor biomasa de raicillas y micelio externo durante los períodos secos, lo que sugiere estrategias de incremento del volumen del suelo explorado por parte de los HFMA como de las plantas hospedadoras. La proliferación de esporas evidenció la formación de estructuras de resistencia de los HFMA ante condiciones adversas. Los resultados obtenidos pondrían en evidencia la plasticidad de la simbiosis micorrízica ante variaciones en la disponibilidad de agua.

Fungos micorrízicos arbusculares em sistema de aléias no Estado do Maranhão, Brasil / Arbuscular mycorrhizal fungi in an alley cropping system in the state of Maranhão, Brazil

Sistemas em aléias podem consistir numa solução para o uso da terra em regiões do Trópico Úmido. A relação dessa forma de manejo com a dinâmica dos fungos micorrízicos arbusculares (FMA) ainda é pouco compreendida. O objetivo desse estudo foi verificar a influência de leguminosas arbóreas em um sistema em aléias na capacidade infectiva e diversidade de FMA nativos em São Luís, Maranhão. Amostras de solo coletadas do sistema em aléias no campus experimental da Universidade Estadual do Maranhão (UEMA) - São Luís, em duas épocas do ano (Julho/2006 e Abril/2007), a duas distâncias (0 m e 0,5 m) do tronco de três leguminosas (Leucaena leucocephala, Clitoria fairchildiana e Acacia mangium) e em área testemunha (sem leguminosas) na profundidade de 0 - 20 cm. O solo coletado foi utilizado para avaliar a capacidade infectiva dos FMA nativos, densidade e identificação de glomerosporos. O sistema em aléias aumenta o potencial de infectividade dos FMA nativos dependendo da espécie de leguminosa arbórea associada, estação de coleta e proximidades das árvores. Dezesseis espécies de FMA foram encontradas na área distribuídas em cinco gêneros, sendo Scutellospora o mais representativo.

Alley cropping systems may be a solution for land use in tropical regions. How land use is connected to mycorrhizal arbuscular fungi (AMF) is poorly understood, especially in the tropics. The aim of this study was to evaluate the influence of leguminous trees in an alley cropping system in regard to the infectivity and diversity of native AMF species in São Luís, Maranhão, Brazil. Soil samples were collected in an experimental area of the Universidade Estadual do Maranhão (UEMA) - São Luís, in two seasons (July/2006 and April/2007), two distances (0 m and 0.5 m) from the trunk of three leguminous trees (Leucaena leucocephala, Clitoria fairchildiana e Acacia mangium), and in a control area 0-20 cm deep. Soil samples were used to evaluate the infectivity of AMF, density and diversity of glomerospores. The alley cropping systems increased the infectivity of AMF which varied according to the leguminous tree species, raining season and distance from trees. Sixteen AMF species of five genera were identified in the area, and Scutellospora was the most representative genus.

Dominance of Paris-type morphology on mycothallus of Lunularia cruciata colonised by Glomus proliferum / Predomínio da morfologia tipo-Paris em talo de Lunularia cruciata colonizado por Glomus proliferum

Microscopic evidence confirms that L. cruciata hosting G. proliferum shows major anatomical traits (arbuscules, coils, arbusculate coils and vesicles) generally associated arbuscular mycorrhizal roots and the anatomical morphology of intra-thalli mycelium is predominantly of the Paris-type. Colonised L. cruciata showed a reduction of biomass when compared with axenic plants suggesting a drain of resources towards the fungus and depletion of nutrients required for optimum plant growth. The behaviour of mycothalli regarding available KH2PO4 indicates that the nutritional stress threshold for phosphorus (P) is above the residual amount of P already present in PhytagelTM and in plant inoculum. These raise the possibility that in certain circumstances the relationship between L. cruciata and G. proliferum be parasitic rather than symbiotic and open the door for future studies to ascertain the nature of liverwort-AM fungi relationships.

Observações de microscopia ótica confirmam que L. cruciata colonizada por G. proliferum apresenta caracteres anatomicos (arbúsculos, hifas novelas, arbúsculos enovelados e vesículas) geralmente associadas a raízes micorrízicas arbusculares em que o micélio intra-tálico apresenta uma anatomia predominantemente do tipo Paris. L. cruciata colonizada apresentou redução de biomassa quando comparada com plantas axenicas, sugerindo dreno de recursos para o fungo e consequente redução de nutrientes necessários para o ótimo crescimento da planta. O comportamento do talo-colonizado em relação à disponibilidade de KH2PO4 no meio indica que o limiar de stress nutricional para fósforo se encontra acima do somatório das quantidades residuais deste elemento presentes no PhytagelTM e no inóculo. Os resultados aqui discutidos sugerem a possibilidade de, em certas circunstâncias, a relação entre L. cruciata e G. proliferum ter características de parasitismo e não de simbiose, abrindo novas perspectivas para futuros estudos na determinação da natureza da relação hepática-fungo arbuscular.

Elisa de captura com IgY para quantificação de acetato de lupeol em Vernonia scorpioides lam. Pers (asteraceae) / Capture IgY-elisa to quantify lupeol acetate in Vernonia scorpioides lam. Pers (asteraceae)

O presente experimento descreve, pela primeira vez, a elaboração de um ensaio imunoenzimático (ELISA de captura) com anticorpos IgY para a detecção e quantificação de acetato de lupeol (LAc) em Vernonia scorpioides Lam. Pers (Asteraceae). Anticorpos anti-LAc, obtidos partir do soro das aves e gemas de seus ovos, após imunização com o conjugado acetato de lupeol-cBSA, foram concentrados em coluna contendo LAc-BSA acoplado a sepharose. A eficácia da metodologia de detecção imunológica foi de 97 por cento, com índices de sensibilidade e especificidade de 99 por cento e 95 por cento, respectivamente com limites de detecção do ensaio entre 0,02µg.g-1 (inferior) e 10 µg.g-1 (superior). A robustez do método foi atestada pela sua elevada reprodutibilidade (entre 94,75 por cento e 96,81 por cento), e pelo baixo coeficiente de variação interna (4,22 ± 1,03 por cento) nas condições de execução descritas.