Effects of temperature and humidity on infection of Fusarium oxysporum in seedlings of Salvia miltiorrhiza / 中国中药杂志

Wilt disease is a major disease of cultivated Salvia miltiorrhiza, which is caused by Fusarium oxysporum. Since the infection process of F. oxysporum in plants is affected by environment factors, this study was conducted to reveal the relationship between disease severity and concentration of the pathogen in plants in the infection process of F. oxysporum in seedlings of S. miltiorrhiza by pot experiments and to reveal the effects of temperature and humidity on the infection process. The results showed that, after inoculation of S. miltiorrhiza seedlings with F. oxysporum, the pathogen in different parts was detected at different time, and it was first detected in substrates. With the continuous propagation of the pathogen(4-5 d), it gradually infected the roots and stems of the seedlings, and the plants had yellowing leaves and withering. The number of the pathogen reached the maximum in each part after 7-8 d, and then gradually decreased in the later stage of the disease. The concentration of the pathogen in substrates, roots and stems of S. miltiorrhiza showed a trend of decreasing after increasing with the aggravation of the disease and reached the maximum in the samples of moderate morbidity, while the concentration in the samples of severe morbidity decreased. In addition, the infection of F. oxysporum in seedlings of S. miltiorrhiza was affected by temperature and humidity. The suitable temperature was 25-30 ℃ and the suitable humidity was 80%-90%. This study could provide guidance for the experiments on pathogenicity of F. oxysporum, screening of biocontrol bacteria and controlling of wilt.

The improvement of bioactive secondary metabolites accumulation in Rumex gmelini Turcz through co-culture with endophytic fungi

Abstract Aspergillus sp., Fusarium sp., and Ramularia sp. were endophytic fungi isolated from Rumex gmelini Turcz (RGT), all of these three strains could produce some similar bioactive secondary metabolites of their host. However the ability to produce active components degraded significantly after cultured these fungi alone for a long time, and were difficult to recover. In order to obtain more bioactive secondary metabolites, the co-culture of tissue culture seedlings of RGT and its endophytic fungi were established respectively, and RGT seedling was selected as producer. Among these fungi, Aspergillus sp. showed the most significant enhancement on bioactive components accumulation in RGT seedlings. When inoculated Aspergillus sp. spores into media of RGT seedlings that had taken root for 20 d, and made spore concentration in co-culture medium was 1 × 104 mL-1, after co-cultured for 12 d, the yield of chrysophaein, resveratrol, chrysophanol, emodin and physcion were 3.52-, 3.70-, 3.60-, 4.25-, 3.85-fold of the control group. The extreme value of musizin yield was 0.289 mg, which was not detected in the control groups. The results indicated that co-culture with endophytic fungi could significantly enhance bioactive secondary metabolites production of RGT seedlings.

Biocontrol potential of saline- or alkaline-tolerant Trichoderma asperellum mutants against three pathogenic fungi under saline or alkaline stress conditions

ABSTRACT Salinity and alkalinity are major abiotic stresses that limit growth and development of poplar. We investigated biocontrol potential of saline- and alkaline-tolerant mutants of Trichoderma asperellum to mediate the effects of salinity or alkalinity stresses on Populus davidiana × P. alba var. pyramidalis (PdPap poplar) seedlings. A T-DNA insertion mutant library of T. asperellum was constructed using an Agrobacterium tumefaciens mediated transformation system; this process yielded sixty five positive transformants (T1-T65). The salinity tolerant mutant, T59, grew in Potato Dextrose Agar (PDA) containing up to 10% (1709.40 mM) NaCl. Under NaCl-rich conditions, T59 was most effective in inhibiting Alternaria alternata (52.00%). The alkalinity tolerant mutants, T3 and T5, grew in PDA containing up to 0.4% (47.62 mM) NaHCO3. The ability of the T3 and T5 mutants to inhibit Fusarium oxysporum declined as NaHCO3 concentrations increased. NaHCO3 tolerance of the PdPap seedlings improved following treatment with the spores of the WT, T3, and T5 strains. The salinity tolerant mutant (T59) and two alkalinity tolerant mutants (T3 and T5) generated in this study can be applied to decrease the incidence of pathogenic fungi infection under saline or alkaline stress.

Compatibility of Azospirillum brasilense and Pseudomonas fluorescens in growth promotion of groundnut ( Arachis hypogea L)

ABSTRACT We attempted to study the compatibility among plant beneficial bacteria in the culture level by growing them near in the nutrient agar plates. Among all the bacteria tested, Rhizobium was found to inhibit the growth of other bacteria. From the compatible group of PGPR, we have selected one biofertilizer (Azospirillum brasilense strain TNAU) and one biocontrol agent (Pseudomonas fluorescens strain PF1) for further studies in the pot culture. We have also developed a bioformulation which is talc powder based, for individual bacteria and mixed culture. This formulation was used as seed treatment, soil application, seedling root dip and foliar spray in groundnut crop in vitro germination conditions. A. brasilense was found to enhance the tap root growth and P. fluorescens, the lateral root growth. The other growth parameters like shoot growth, number of leaves were enhanced by the combination of both of the bacteria than their individual formulations. Among the method of application tested in our study, soil application was found to be the best in yielding better results of plant growth promotion.

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.

Establishment of Mimosa biuncifera (Fabaceae) inoculated with arbuscular mycorrhizal fungi in greenhouse and field drought conditions / Establecimiento de Mimosa biuncifera (Fabaceae) inoculada con hongos micorrízicos arbusculares en invernadero y campo, en condiciones de sequía

AbstractMexico is dominated by arid or semi-arid ecosystems, predominantly characterized as xeric shrublands. These areas are frequently deteriorated due to agriculture or over-grazing by livestock (sheep and goats). The vegetation type mainly consists of thorny plant species, and among these, the dominant one in overgrazed areas is catclaw (Mimosa biuncifera). This is a nurse plant that facilitates establishment of other vegetation and promotes plant succession. Catclaw plants form a mutualistic association with arbuscular mycorrhizal fungi (AMF), which improves uptake of nutrients and water. The objective of this study was to determine the effect of inoculating catclaw plants with native AMF and starting their growth under a low water availability treatment in a greenhouse, and later transplanting them to field conditions of drought and deterioration. Field plants were evaluated according to their survivorship and growth. The seeds of catclaw plants and soil with AMF spores were collected in the Mezquital Valley of Hidalgo State, in Central Mexico. Seedlings were grown in individual pots in a greenhouse. The experimental design consisted of two levels of pot irrigation, wet (W) and dry (D), as well as the presence (M+) or absence (M-) of AMF inoculum, with 20 replicates for each treatment. The following plant parameters were recorded every week: height, number of leaves and pinnae, and mean diameter of coverage. After 20 weeks in the greenhouse, determination was made of fresh and dry biomass, relative growth rate (RGR), root/shoot ratio, real evapotranspiration (RET), water-use efficiency (WUE), and percentage of mycorrhizal colonization. The remaining plants growing under the dry treatment (M+ and M-) were then transplanted to a semi-arid locality in the Mezquital Valley. During one year, monthly records were kept of their height, number of leaves, mean diameter of coverage and survival. Results showed that compared to greenhouse plants under other treatments, those under the wet mycorrhizal (WM+) treatment were taller, had more pinnae, and were characterized by greater coverage, faster RGR, and greater fresh and dry biomass. Moreover, inoculated plants (WM+ and DM+) showed higher WUE than those uninoculated (WM- and DM-, respectively). After one year in field conditions, there was a higher survival rate for previously inoculated versus uninoculated plants. Hence, mycorrhization of M. biuncifera with native AMF inoculum increased plant efficiency in biomass production, thus favoring establishment and survival in field conditions. We concluded that inoculation of catclaw plants is recommendable for revegetation programs in deteriorated semi-arid zones. Rev. Biol. Trop. 64 (2): 791-803. Epub 2016 June 01.

ResumenMás de la mitad del territorio mexicano está dominado por ecosistemas áridos y semiáridos. El principal tipo de vegetación de estos ambientes son matorrales xerófilos y normalmente están deteriorados por agricultura y sobrepastoreo de ovejas y cabras inducido por la actividad humana. Las plantas espinosas dominan en estas zonas, como el gatuño (Mimosa biuncifera Benth., Fabaceae), que es una planta nodriza que promueve la sucesión vegetal. Las plantas de gatuño forman junto con hongos micorrizógenos arbusculares (HMA), una asociación mutualista llamada micorriza arbuscular que mejora la captación vegetal de nutrimentos y agua. El objetivo de este trabajo fue determinar el efecto de la inoculación con HMA nativos en plantas de gatuño, para evaluar su crecimiento y establecimiento bajo un tratamiento de baja disponibilidad hídrica, tanto en condiciones de invernadero como de campo. Para esto, se recolectaron semillas de Mimosa biuncifera y suelo con esporas de HMA en el Valle del Mezquital, estado de Hidalgo, en el Centro de México. Las plántulas fueron cultivadas en macetas individuales en condiciones de invernadero bajo un diseño experimental que consistió en dos tratamientos de riego de las macetas, húmedo (W) y seco (D), y dos tratamientos de inoculación: con y sin inóculo de HMA (M+, M-). Cada tratamiento tuvo 20 repeticiones. Semanalmente se registró altura, diámetro medio de la cobertura y número de hojas y pinnas de las plantas. Después de 20 semanas, se determinaron biomasa húmeda y seca, tasa relativa de crecimiento (TRC), proporción raíz vástago, evapotranspiración real, eficiencia en el uso del agua (WUE) y porcentaje de colonización micorrícica. Posteriormente, los individuos del tratamiento seco (M+ y M-) fueron trasplantados a una localidad semiárida en el Valle del Mezquital, donde la altura, número de hojas y supervivencia fueron registrados mensualmente durante un año. Los resultados muestran que las plantas en el tratamiento húmedo y micorrizado (WM+) tuvieron una mayor altura, cobertura vegetal, biomasa húmeda y seca, TRC y más pinnas que las plantas de los otros tratamientos; también los tratamientos micorrizados (WM+ y DM+) tuvieron mayor WUE que la plantas sin inóculo (M-). En condiciones de campo, después de un año, la supervivencia de las plantas M+ fue mayor que en los testigos. Se concluyó que la micorrización de M. biuncifera con inóculo nativo incrementa su eficiencia en la producción de biomasa y favorece el establecimiento y supervivencia en condiciones de campo. Finalmente, se recomienda la inoculación de plantas de gatuño con HMA en programas de revegetación de zonas semiáridas deterioradas.

Caracterización de rizobacterias promotoras de crecimiento en plántulas de Eucalyptus nitens / Characterization of growth-promoting rhizobacteria in Eucalyptus nitens seedlings

Se aislaron bacterias rizosféricas y endófitas a partir de rizósfera y tejidos de raíz de árboles de Eucalyptus nitens con el objetivo de evaluar su capacidad de promover el crecimiento en plántulas de la misma especie en condiciones de invernadero. Los aislamientos que incrementaron el crecimiento de las plántulas fueron identificados y caracterizados por su capacidad de producir ácido indolacético (AIA), solubilizar fosfato y expresar la 1-aminociclopropano-1-carboxilato (ACC) desaminasa. Los 105 aislamientos obtenidos fueron morfológicamente diferentes y solo 15 promovieron significativamente el crecimiento de plántulas de E. nitens. Los máximos incrementos observados fueron en el peso seco aéreo (142 %) y de la raíz (135 %); también aumentaron la altura de las plantas (50 %) y el largo de raíces (45 %) de las mismas. Las rizobacterias pertenecieron a los géneros Arthrobacter, Lysinibacillus, Rahnella y Bacillus. Los aislados identificados como A. phenanthrenivorans 21 y B. cereus 113 incrementaron la emergencia de E. nitens a los 12 días en un valor promedio de 3,15 veces con relación al control. R. aquatilis aislado 78 presentó la mayor producción de AIA (97,5 ± 2,87 μg/ml) en presencia de triptófano y el mayor índice de solubilización de fósforo (2,4). B. amyloliquefaciens aislado 60 fue positivo para la actividad ACC desaminasa. Los resultados obtenidos indican el potencial de las rizobacterias estudiadas como promotoras de emergencia y crecimiento de plántulas de E. nitens y su posible uso como inoculantes, ya que presentan más de un mecanismo de acción asociado a la promoción del crecimiento.

Rhizospheric and endophytic bacteria were isolated from the rizosphere and root tissue of Eucalyptus nitens. The objective of this work was to evaluate their capacity to promote growth in seedlings of the same species under greenhouse conditions. The isolates that improved seedling growth were identified and characterized by their capacity to produce indoleacetic acid (IAA), solubilize phosphates and increase 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity. One hundred and five morphologically different strains were isolated, 15 of which promoted E. nitens seedling growth, significantly increasing the height (50%), root length (45%) as well as the aerial and root dry weight (142% and 135% respectively) of the plants. Bacteria belonged to the genus Arthrobacter, Lysinibacillus, Rahnella and Bacillus. Isolates A. phenanthrenivorans 21 and B. cereus 113 improved 3.15 times the emergence of E. nitens after 12 days, compared to control samples. Among isolated R. aquatilis, 78 showed the highest production of IAA (97.5±2.87 μg/ml) in the presence of tryptophan and the highest solubilizer index (2.4) for phosphorus, while B. amyloliquefaciens 60 isolate was positive for ACC deaminase activity. Our results reveal the potential of the studied rhizobacteria as promoters of emergence and seedling growth of E. nitens, and their possible use as PGPR inoculants, since they have more than one mechanism associated with plant growth promotion.

Eucalyptus grandis Hill ex Maiden inoculated with Pisolithus microcarpus (UFSC-Pt116) in land subject to the sandy process in southern Brazil

Eucalypts is one of the main species used for commercial reforestation in the state of Rio Grande do Sul, Brazil. This study aimed to evaluate the survival and early growth of eucalyptus trees in an area subject to sandy process after three years of growth. The Eucalyptus grandis seedlings were grown in a greenhouse, innoculated or not with the isolated ectomycorrhizal Pisolithus microcarpus (UFSC-Pt116), produced in peat or Entisol. After 120 days, the seedlings were transplanted to an area subject to the sandy process, in the city of São Francisco de Assis, RS. The plants have been evaluated regarding survival, height, stem diameter, nitrogen, phosphorus and potassium levels and total phosphorus, inorganic phosphorus, organic phosphorus and wood production on different days after planting. The seedlings grown on the Entisol which was inoculated with the isolated UFSC-Pt116 presented higher survival rates, height, stem diameter, nitrogen concentration and wood production compared to the non-inoculated seedlings. Inoculation with ectomycorrhizal fungi enhanced the production of E. grandis seedlings in survival rates, height, stem diameter.

Microorganisms associated to tomato seedlings growing in saline culture act as osmoprotectant

Less than 0.5% of total water in the world is available for human consumption and agriculture. The major part of the world's water is saline and salinity in soils interferes in germination of seeds and the posterior development of the plant. In order to increase the osmotolerance of tomato, seedlings were associated with Azospirillum brasilense Cd, Azospirillum brasilense Cd transformed bacteria with a plasmid harboring a trehalose biosynthesis gene-fusion or Chlorella vulgaris. Two plant culture media: Hydroponic and Murashige and Skoog were tested. In the first set of studies seedlings were associated to single free cells meanwhile in a second set single and combined free cells were studied. A positive interaction between transformed Azospirillum and Chlorella vulagris and tomato plants was observed. Seedlings showed a salt concentration tolerance, as sodium chloride, up to 200 mM. According to our results, the association of plants with A. brasilense Cd-BIF and C. vulgaris is a viable approach to increase their salt tolerance and biomass, as consequence the possible use of sea water to irrigate horticultural plants.

Growth response of maize plantlets inoculated with Enterobacter spp., as a model for alternative agriculture / Respuesta de plántulas de maíz inoculadas con Enterobacter spp. como un modelo de agricultura alternativa

A maize rhizosphere isolate was phenotypically and genotypically characterized and identifed as Enterobacter spp. bacterium. Germinated seeds were inoculated, the plantlets were sown in vermiculite and in soil and grown under laboratory and feld conditions, respectively. The adherence, colonization and plant growth promotion capability of Enterobacter sp. UAPS03001 was evaluated in "Rojo-Criollo" maize under laboratory conditions. Twenty days after inoculation, the treated plantlets showed larger biomass than non-inoculated ones. In feld grown plants, the kernel biomass was also greater in inoculated than in non-inoculated plants. The inoculation of maize sprouts with plant growth- promoting bacteria before their sowing in the feld would be an alternative practice for achieving successful yield in temporal agriculture.

En este trabajo se aisló una bacteria de la rizósfera de maíz, que fue caracterizada mediante métodos fenotípicos y genotípicos e identifcada como Enterobacter sp. UAPS03001. La bacteria fue inoculada en semillas de maíz "Rojo-Criollo" germinadas en forma axénica. Las semillas germinadas e inoculadas se plantaron en vermiculita y posteriormente las plántulas fueron cultivadas en vermiculita o en suelo, para evaluar el efecto promotor del crecimiento vegetal de dicha bacteria, bajo condiciones de laboratorio y de campo. Bajo condiciones de laboratorio, también se evaluó la capacidad de esta cepa para adherirse a las plantas de maíz y colonizarlas. Veinte días después de la inoculación, las plántulas inoculadas mostraron una biomasa mayor con referencia a las no inoculadas. En campo, la biomasa de la mazorca fue también mayor en las plantas inoculadas respecto de las plantas no inoculadas. La inoculación de germinados de maíz con una bacteria promotora del crecimiento vegetal y su posterior transferencia a campo podría ser una práctica alternativa para llevar a cabo una producción exitosa en agricultura de temporal.

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.

IP3 production in the hypersensitive response of lemon seedlings against Alternaria alternata involves active protein tyrosine kinases but not a G-protein

IP3 increase and de novo synthesis of scoparone are produced in the hypersensitive response (HR) of lemon seedlings against the fungus Alternaria alternata. To elucidate whether a G-protein and/or a protein tyrosine kinase (PTK) are involved in signal transduction leading to the production of such a defensive response, we studied the HR in this plant system after treatment with G-protein activators alone and PTK inhibitors in the presence of fungal conidia. No changes in the level of IP3 were detected in response to the treatment with the G-protein activators cholera toxin or mastoparan, although the HR was observed in response to these compounds as determined by the scoparone synthesis. On the contrary, the PTK inhibitors lavendustin A and 2,5-dihidroxy methyl cinnamate (DHMC) not only prevented the IP3 changes observed in response to the fungal inoculation of lemon seedlings but also blocked the development of the HR. These results suggest that the IP3 changes observed in response to A. alternata require a PTK activity and are the result of a G-protein independent Phospholipase C activity, even though the activation of a G-protein can also lead to the development of a HR. Therefore, it appears that more than one signaling pathway may be activated for the development of HR in lemon seedlings: one involving a G-protein and the other involving a PTK-dependent PLC.

Heavy metal toxicity on dehydrogenase activity on rhizospheric soil of ectomycorrhizal pine seedlings in field condition.

The present investigation was carried out to study the toxicity of heavy metals (Zn, Cd, Cu, Ni, Pb and Al) on the dehydrogenase activity of ectomycorrhizal (Suillus leutus, Scleroderma aurantium, Cenococcum graniforme and Boletus spp.) and non-mycorrhizal rhizospheric soil of pine seedlings. The treatment of heavy metals adversely affected the dehydrogenase activity. Inoculation of mixed ectomycorrhizal fungi harbored increased activity of dehydrogenase. It was observed that in absence of ectomycorrhizae, heavy metals drastically reduced the enzyme activity at higher concentration of metals.