In vitro propagation of a cuban Arbuscular Mycorrhizal Fungal strain and factible bacteria's association / Propagación in vitro de una cepa cubana de Hongos Micorrízicos Arbusculares y su factible asociación a bacterias

ABSTRACT In vitro root cultivation techniques based on modified root systems are often used in studies on Arbuscular Mycorrhizal Fungi (AMF). It is a simplified but powerful tool to investigate AMF root colonization and development of the extraradical mycelium. The aim of this study was to establish and characterize the in vitro culture of a Cuban strain of Rhizophagus irregularis (INCAM 11) by using transformed chicory roots. For that, superficially disinfected propagules of R. irregularis were co-culture with the hairy transformed chicory roots on Modified Strullu and Romand (MSR) medium during five months. Spore germination was observed 3-5 days after surface disinfection. The first contact between AMF hyphae and roots occurred 1 - 3 days after germination and a significant production of extensive extraradical mycelium was observed. New spore formation started within 21 - 25 days. After 5 months, 2000 spores could be observed per plate which were able to germinate, colonize, establish and reproduce again spores when associated to young transformed roots of chicory. The most frequent associated microorganism to the in vitro culture of INCAM 11 was isolated and identified as Paenibacillus sp.

RESUMEN Las técnicas de cultivo de raíces in vitro basadas en sistemas de raíces modificadas se utilizan a menudo en los estudios sobre hongos micorrízicos arbusculares (HMA). Es una herramienta simplificada pero poderosa para investigar la colonización de las raíces de los HMA y el desarrollo del micelio extrarradical. El objetivo de este estudio fue establecer y caracterizar el cultivo in vitro de una cepa cubana de Rhizophagus irregularis (INCAM 11) utilizando raíces transformadas de achicoria. Para ello, propágulos de R. irregularis desinfectados superficialmente fueron co-cultivados con las raíces transformadas de achicoria en medio Strullu y Romand modificado (SRM) durante cinco meses. La germinación de las esporas se observó 3-5 días después de la desinfección superficial. El primer contacto entre las hifas y las raíces se produjo entre 1 y 3 días después de la germinación y se observó una producción significativa de micelio extrarradical. La formación de nuevas esporas comenzó entre 21 - 25 días. Después de 5 meses, se pudieron observar 2000 esporas por placa que fueron capaces de germinar, colonizar, establecer y reproducir nuevas esporas cuando se asociaron a raíces jóvenes transformadas de achicoria. El microorganismo asociado frecuentemente al cultivo in vitro de INCAM 11 fue aislado e identificado como Paenibacillus sp.

Yield increase of corn inoculated with a commercial arbuscular mycorrhizal inoculant in Brazil / Rendimento de grãos da cultura do milho com inoculante micorrízico arbuscular comercial no Brasil

ABSTRACT: Arbuscular mycorrhizal fungi (AMF) play an important role in plant growth. However, there are no reports of legally commercialized AMF-based inoculants for agricultural crops in Brazil. The objective of this research was to evaluate the agronomic efficiency of a Rhizophagus intraradices inoculant in combination with phosphate fertilization in grain yield of corn under different edaphoclimatic conditions in Brazil. Experiments were conducted in five Brazilian states (Goiás, Mato Grosso, Minas Gerais, Rio Grande do Sul and Santa Catarina) in a 2 x 3 factorial scheme, with two inoculation treatments (inoculated and non-inoculated seeds) and three doses of phosphate fertilization (0, 50 and 100% of the recommended P). At the end of the crop cycle (stages R4-R5), inoculation provided increases in biomass (average of 48%) regardless of the applied dose of P, higher P absorption, and 54% average increase in grain yield. In conclusion, the mycorrhizal inoculant increases biomass yield, P uptake and corn grain yield under different edaphoclimatic conditions in Brazil, especially in soils that originally had low or medium levels of available P.

RESUMO: Os fungos micorrízicos arbusculares (FMA) desempenham um papel importante no crescimento das plantas, no entanto, não há relatos de inoculantes legais baseados em AMF para culturas agrícolas no Brasil. O objetivo deste trabalho foi avaliar a eficiência agronômica de um inoculante a base de Rhizophagus intraradices em combinação com a adubação fosfatada no rendimento de grãos de milho sob diferentes condições edafoclimáticas no Brasil. Os experimentos foram conduzidos em cinco estados brasileiros (Goiás, Mato Grosso, Minas Gerais, Rio Grande do Sul e Santa Catarina) em esquema fatorial 2 x 3, com dois tratamentos de inoculação (sementes inoculadas e não inoculadas) e três doses de adubação fosfatada (0, 50 e 100% do P recomendado). No final do ciclo da cultura (estágios R4-R5), a inoculação proporcionou aumentos na biomassa (média de 48%), independentemente da dose aplicada de P, maior absorção de P e aumento médio de 54% no rendimento de grãos. Conclui-se que o inoculante micorrízico arbuscular aumenta a produção de biomassa, a absorção de P e a produtividade de grãos de milho em diferentes condições edafoclimáticas no Brasil, especialmente em solos que originalmente apresentavam níveis baixos ou médios de P.

Arbuscular mycorrhizal fungal communities in the rhizosphere of a continuous cropping soybean system at the seedling stage

Abstract Arbuscular mycorrhizae (AM) fungi play a crucial role in the growth of soybean; however, the planting system employed is thought to have an effect on AM fungal communities in the rhizosphere. This study was performed to explore the influence of continuous soybean cropping on the diversity of Arbuscular mycorrhizal (AM) fungi, and to identify the dominant AM fungus during the seedling stage. Three soybean cultivars were planted under two and three years continuous cropping, respectively. The diversity of AM fungi in the rhizosphere soil at the seedling stage was subsequently analyzed using polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE). The results showed that an increase in cropping years improved the colonization rate of AM in all three soybean cultivars. Moreover, the dominant species were found to be Funneliformis mosseae and Glomus species. The results of cluster analysis further confirmed that the number of years of continuous cropping significantly affected the composition of rhizospheric AM fungal communities in different soybean cultivars.

Abundancia y riqueza de hongos micorrizógenos arbusculares en cafetales de Soconusco, Chiapas, México / Abundance and richness of arbuscular mycorrhizal fungi in coffee plantations from Soconusco, Chiapas, Mexico

Resumen El cultivo del café, en la economía mexicana y en la región del Soconusco en Chiapas, aporta importantes divisas; sin embargo es susceptible a plagas y enfermedades, por lo que se requiere reforzar su producción a través de un manejo más sustentable. Este estudio analizó la abundancia, riqueza y composición de hongos micorrizógenos arbusculares (HMA) nativos que pudieran emplearse en un futuro como biofertilizantes en el cultivo del café. En 21 muestras de suelo recolectadas en noviembre 2015 de siete sitios en la región del Soconusco, Chiapas, México, cultivados con café "robusta" (Coffea canephora), se cuantificó la abundancia de esporas, la riqueza y composición de morfoespecies de HMA y se exploró su relación con las propiedades del suelo. Se obtuvo un total de 20 morfoespecies y cinco nuevos registros de HMA para Chiapas, las morfoespecies más frecuentes pertenecen a los géneros Acaulospora y Glomus. Los sitios Toluca y Victoria tuvieron mayor riqueza de especies (17 spp. c/u) que San Agustín, 20 de noviembre y San Luis Nexapa (4 - 7 spp. c/u); mientras que Providencia y Platanar registraron una riqueza intermedia y los mayores valores de abundancia de esporas. Los sitios Victoria y Toluca presentaron un 97 % de disimilitud en la composición de especies de HMA con respecto a los demás sitios; la cual fue explicada por la baja concentración de PO4 -3en el suelo de Victoria y Toluca. La disponibilidad de P, ligada a la acidez del suelo, fueron los factores que podrían estar regulando las comunidades de HMA en el suelo de la rizosfera de los cafetos en la región del Soconusco. Se propone que podrían haber consorcios de HMA específicos para los niveles de P y acidez presentes en diferentes sitios con cafetos, o bien consorcios de especies de Acaulospora y Glomusacordes con las condiciones ambientales en las que se desarrolla el café, cuya compatibilidad y funcionalidad debe examinarse antes de emplearlos como biofertilizantes nativos en los cafetos de esta región tropical de México.

Abstract In the Mexican economy, and especially in the Soconusco region of Chiapas, coffee is economically important, and sustainable management most cover pests and diseases. In this study, we searched for native arbuscular mycorrhizal fungi (AMF) that could be used as biofertilizers in coffee cultivation. We collected 21 soil samples from seven coffee plantations (Coffea canephora) in November 2015. We used wet sieving and decanting to quantify abundance, richness and composition of morphospecies, as well as their relationships with soil properties. A total of 20 morphospecies and five new records of AMF were obtained, and the most frequent genera were Acaulospora and Glomus. The Toluca and Victoria sites had more morphospecies richness (17 spp. each) than San Agustín, November 20 and San Luis Nexapa (4-7 spp. each). Providencia and Platanar were intermediate but had the highest spore abundance. The dissimilarity of Victoria and Toluca may reflect a low concentration of PO4 -3in the soil. P availability, linked to soil acidity, sems to be regulating these mycorrhizae communities at Soconusco. AMF consortia specific for soil P-levels and acidity, i.e. Acaulospora and Glomusconsortia, may occur and be common in the environmental conditions of Mexican coffee plantations, Strains should be tested for compatibility and functionality before using them as native biofertilizers. Rev. Biol. Trop. 66(1): 91-105. Epub 2018 March 01.

MANEJO INTEGRADO DE NEMATODOS FITOPARÁSITOS EN ALMÁCIGOS DE PLÁTANO DOMINICO HARTÓN (Musa AAB SIMMONDS) / INTEGRATED MANAGEMENT OF PHYTOPARASITIC NEMATODES IN DOMINICO HARTÓN PLANTAIN (Musa AAB SIMMONDS) SEEDLINGS

Como consecuencia de la alimentación de Radopholus similis, se generan daños en las raíces y el cormo que conllevan a un crecimiento deficiente de las plantas y reducción en rendimiento hasta 80%. Por tal motivo, el objetivo de esta investigación fue determinar el efecto de la integración de prácticas de manejo como limpieza sanitaria de cormos y biológicas como Micorrizas Arbusculares (MA), Purpureocillium lilacinum y Bacillus subtilis sobre el crecimiento de las plántulas de Dominico Hartón y la población de nematodos fitoparásitos. El experimento se realizó en la granja Montelindo de la Universidad de Caldas, a 1010 msnm, 23ºC de temperatura promedio y 1800 mm de precipitación. Los tratamientos fueron asignados en un diseño experimental completamente aleatorio, cada uno conformado por 20 cormos (unidades experimentales) sembrados en bolsas negras de 5 kg de capacidad, así: cormos con limpieza sanitaria sin aplicación de productos (T1), cormos con limpieza sanitaria + MA (T2), cormos con limpieza sanitaria + P. lilacinum (T3), cormos con limpieza sanitaria + B. subtilis (T4), y cormos con limpieza sanitaria + MA + P. lilacinum + B. subtilis (T5). El anterior procedimiento también se realizó a cormos tradicionales sembrados en suelo sin esterilizar (T6 a T10, respectivamente). En los tratamientos que fueron establecidos en suelo esterilizado y cormos con limpieza sanitaria, no hubo presencia de individuos de los principales géneros de nematodos fitoparásitos del plátano como Radopholus, Meloidogyne y Helicotylenchus. Los tratamientos en los cuales se utilizó suelo infestado y cormos con limpieza sanitaria, presentaron una disminución de hasta un 64% en la población de nematodos fitoparásitos en comparación con la inicial, aunque mostraron también una cantidad de nematodos menor al testigo. De estos tratamientos el que menor población presentó fue el T10 (Suelo infestado + cormos con limpieza sanitaria + MA + P. lilacinum + B. subtilis) con una población promedio de 472 nematodos fitoparásitos en 100 g de suelo. La integración de todas las medidas de manejo resultó más eficiente en el control de los nematodos fitoparásitos en plátano, con o sin la esterilización del suelo; y la limpieza sanitaria de cormos no afectó negativamente el desarrollo de las raíces ni de la parte aérea de las plantas, pues no se presentaron diferencias significativas respecto a los tratamientos donde se usaron cormos tradicionales.

As a result of feeding of Radopholus similis, both roots and corms are damaged and this results in plant growth deficiency and yield losses up to 80%. Therefore, the objective of this study was to determine the effect of the integration of management practices such as corms sanitary cleaning and biological cleaning such as Arbuscular mycorrhizae (AM), Purpureocillium lilacinum and Bacillus subtilis on the development of Dominico Hartón plantain seedlings and phytoparasitic nematods populations. The experiment was conducted at the Universidad de Caldas Montelindo farm located at 1,010 m.a.s.l.., average temperature 23ºC and 1,800 mm rainfall. The treatments were assigned in a completely randomized block design, each one with 20 corms (experimental units) planted in 5 kg black plastic bags as follows: corms with sanitary seed cleaning without application of products (T1); corms with sanitary cleaning + AM (T2); corms with sanitary cleaning + P. lilacinum (T3); corms with sanitary cleaning + B. subtilis (T4); and corms with sanitary cleaning + AM + P. lilacinum + B. subtilis (T5). The described procedure was also applied to traditional corms planted in soil without sterilization (T6 to T10, respectively). In the treatments established in sterilized soil and cleaned corms, there was not presence of individuals of the main plant parasitic nematodes of plantain, such as, Radopholus, Meloidogyne and Helicotylenchus. Treatments with infested soil with nematodes and cleaned corms, showed a reduction in plant parasitic nematodes up to 64% compared with the initial population, although they also showed lower amount of nematodes than the control population. From these treatments, the one that had the lowest population of nematodes was the T10 (Infested soil + cleaned corms + AM + P. lilacinum + B. subtilis) with an average population of 472 phytoparasitic nematodes/100 g of soil). The integration of all management practices resulted in the most efficient way to control the plant phytoparasitic nematodes of plantain, with or without soil sterilization, and the corms sanitary cleanness did not affect either the root development or the aerial parts of the seedlings, since there were not significant differences with respect to the treatments using traditional corms.

The Distribution of Cytoplasm and Nuclei within the Extra-radical Mycelia in Glomus intraradices, a Species of Arbuscular Mycorrhizal Fungi

Nuclear distribution within the extra-radical fungal structures and during spore production in the arbuscular mycorrhizae fungus Glomus intraradices was examined using an in vitro monoxenic culture system. A di-compartmental monoxenic culture system was modified using a nitrocellulose membrane and a coverglass slip for detailed observations. Nuclear distribution was observed using the fluorescent DNA binding probes SYBR Green I and DAPI. Both septate and non-septate mycelial regions were observed, but cytoplasmic contents were only found within non-septate mycelia. Nuclear fluorescent staining revealed that the non-septate hyphal region contained nuclei only with cytoplasm, and that nuclear distribution was limited by septa. Swollen hyphal bodies were often associated with septate and empty-looking hyphae. Cytoplasmic contents filled the swollen hyphal body from the non-septate hyphal region following removal of the septa. As a consequence, the swollen body developed into a new spore. These observations provide understanding about the distribution of AM fungal nuclei within extra-radical mycelia and during spore formation. The results suggest a mechanism by which the development of a cytoplasm-containing mycelium is controlled by the formation or removal of septa to efficiently maintain and proliferate essential contents. This mechanism may provide a survival strategy to the fungus.

Establishment of Desmoncus orthacanthos Martius (Arecaceae): effect of inoculation with arbuscular mycorrhizae

Inoculation with arbuscular mycorrhizal (AM) fungi has often promoted increased growth of plants but very little work has been done in the tropics to evaluate the effects of inoculation on the establishment and development of seedlings in forests. Desmoncus orthacanthos Martius is a scandent palm present both in early and late succession, and consequently can be used in restoration processes. A test was conducted to determine the effect of AM on the establishment of Desmoncus orthacanthos in tropical forest in the Yucatan Peninsula, Mexico. Thirty inoculated and 30 non-inoculated seedlings were introduced in two sites of different successional age, a mature forest and an eight-year old abandoned cornfield (acahual).Survival and growth parameters were evaluated after 12 months. Leaf area and phosphorus, but not height, were greater in inoculated than non-inoculated plants in the forest but not in the acahual. However, mycorrhizae had a clear effect on plant survival in both sites, with a threefold increase in survival of inoculated compared with non-inoculated plants bassed on an odds ratio. The results suggest that inoculation will be important to increase the establishment of this commercially important palm.

La inoculación con hongos micorrizógenos arbusculares (AM)promueve un incremento en el crecimiento de las plantas, sin embargo poco trabajo se ha realizado en los trópicos para evaluar el efecto de la inoculación en el proceso de establecimiento de plántulas. Desmoncus orthacanthos Martius es una palmera trepadora que se distribuye tanto en etapas tempranas y tardías de la sucesión, por lo tanto puede ser empleada en procesos de restauración. Se realizó una prueba de establecimiento de D. orthacanthos en una selva tropical de la península de Yucatán, México. Treinta plántulas inoculadas y 30 no inoculadas se establecieron en dos sitios con diferentes etapas sucesionales: selva madura y un campo abandonado de maíz (acahual). Después de 12 meses el valor de área foliar y contenido de fósforo fue mayor en el tratamiento con micorrizas en la selva pero no en el acahual, sin embargo, el tratamiento con micorrizas mostró un claro efecto en la supervivencia de las palmeras en ambos sitios con un incremento tres veces mayor en la probabilidad de supervivencia de palmeras inoculadas comparadas con las no inoculadas de acuerdo al análisis de razón de disparidad (odds ratio). La inoculación podría ser importante para incrementar el establecimiento de esta palmera.

Contribution to the Checklist of Soil-inhabiting Fungi in Korea

A total of 82 genera and 271 species of soil-inhabiting fungi including saprobic, nematode-trapping, and arbuscular mycorrhizal but plant pathogenic fungi published hitherto in South Korea are listed with the information on geographic location, habitat, vegetation when available, and relevant literatures.

Multiple Symbiotic Associations Found in the Roots of Botrychium ternatum

Two types of mycorrhizae, orchid (OM) and arbuscular mycorrhizae (AM), were observed in the cortical cells of Botrychium ternatum roots. The vesicles or arbuscules of AM fungi were examined and the fresh or digestive pelotons by other species of basidiomycetes were also observed in the roots under light microscope. These symbioses were, as the genomic DNAs extracted from roots of B. ternatum reacted with the specific primers, confirmed with PCR technique, being added to more strong evidences. These discoveries were rarely happened in the roots, especially a fern in nature. OM was observed in the roots of B. ternatum collected from the nationwide areas, whereas AM was only in the roots of B. ternatum collected from Chung-Buk areas. It is speculated that OM are associated with the nitrogen cycle in Islands and the growth of B. ternatum in the inland of Central Korea is related to both the phosphate and nitrogen cycle in the nature. The results suggest that B. ternatum is a typical species with two types of mycorrhizae under various growing conditions.

Study on Arbuscular Mycorrhizae of Panax notoginseng / 微生物学通报

Arbuscular mycorrhizal fungi can infect the roots of Cercidiphyllaceae and form arbuscular mycorrhizae.Infection rate is from 12% to 30%,but infection strength is weak.4 species of Acaulospora and 11 species of Glomus were isolated and identified,including Acaulospora tuberculata,A.spinosa,A.foveata,A.denticulate,Glomus geosporum,G.clarum,G.constrictum,G.monosporum,G.laroideum,G.versifome,G.microcarpum,G.mosseae,G.hoi,G.halonatum and G.reticulatum.Among them,G.cercidiphyllorum is advantage species.So AMF may be a potent resource of biology which can stimulate the growth of Panax notoginseng.