Alejandro Alarcón

Arbuscular Mycorrhiza and Petroleum-Degrading Microorganisms Enhance Phytoremediation of Petroleum-Contaminated Soil

While plants can phytoremediate soils that are contaminated with petroleum hydrocarbons, adding microbes to remediate contaminated sites with petroleum-degrading microorganisms and arbuscular mycorrhizal fungi (AMF) is not well understood. The phytoremediation of Arabian medium crude oil (ACO) was done with a Lolium multiflorum system inoculated with an AMF (Glomus intraradices) and a mixture of petroleum-degrading microorganisms—the bacterium, Sphingomonas paucimobilis (Sp) and the filamentous fungus, Cunninghamella echinulata (Ce, SpCe)—or with a combination of microorganisms (AMF + SpCe). Based on an earlier study on screening plants for phytoremediation of ACO, L. multiflorum (Italian ryegrass) was selected for its tolerance and rapid growth response (Alarcón, 2006). The plants were exposed to ACO-contaminated soil (6000 mg kg−1) for 80 d under greenhouse conditions. A modified Long Ashton Nutrient Solution (LANS) was supplied to all treatments at 30 μg P mL−1, except for a second, higher P, control treatment at 44 μg P mL−1. Inoculation with AMF, SpCe, or AMF + SpCe resulted in significantly increased leaf area as well as leaf and pseudostem dry mass as compared to controls at 30 μg P mL−1. Populations of bacteria grown on a nitrogen-free medium and filamentous fungi increased with AMF + SpCe and SpCe treatments. The average total colonization and arbuscule formation of AMF-inoculated plants in ACO-contaminated soil were 25% and 8%, respectively. No adverse effects were caused by SpCe on AMFcolonization. Most importantly, ACOdegradation was significantly enhanced by the addition of petroleum-degrading microorganisms and higher fertility controls, as compared to plants at 30 μg P mL−1. The highest ACOdegradation (59%) was observed with AMF + SpCe. The phytoremediation of ACO was also enhanced by single inoculation of AMF or SpCe. The effect of AMF and petroleum-degrading microorganisms on plant growth and ACOdegradation was not attributable to differences in proline, total phenolics, nitrate reductase levels, or variation in plant–gas exchange.

Arbuscular mycorrhizal fungi on growth, nutrient status, and total antioxidant activity of Melilotus albus during phytoremediation of a diesel-contaminated substrate

This research evaluated the effects of arbuscular mycorrhizal fungi (AMF) on growth, nutritional status, total antioxidant activity (AOX), total soluble phenolics content (TPC), and total nitrate reductase activity (NRA) of leaves and roots of Melilotus albus Medik planted in diesel-contaminated sand (7500 mg kg(-1)). Seedlings of Melilotus either Non inoculated (Non-AMF) or pre-inoculated plants (AMF) with the AMF-inoculum Glomus Zac-19 were transplanted to non-contaminated or contaminated sand. After 60 days, diesel significantly reduced plant growth. AMF- plants had no significant greater (64% and 89%, respectively) shoot and leaf dry weight than Non-AMF plants, but AMF plants had lower specific leaf area. AMF-plants had significantly greater content of microelements than non-AMF plants. Regardless diesel contamination, the total AOX and TPC were significantly higher in leaves when compared to roots; in contrast, NRA was higher in roots than leaves. Diesel increased total AOX of leaves, but AMF-plants had significantly lower AOX than non-AMF plants. In contrast, roots of AMF-plants had significantly higher AOX but lower NRA than non-AMF plants. AMF-colonization in roots detected via the fungal alkaline phosphatase activity was significantly reduced by the presence of diesel. AMF-inoculation alleviated diesel toxicity on M. albus by enhancing plant biomass, nutrient content, and AOX activity. In addition, AMF-plants significantly contributed in higher degradation of total petroleum hydrocarbons when compared to non-AMF-plants.

Tolerance and growth of 11 Trichoderma strains to crude oil, naphthalene, phenanthrene and benzo(a)pyrene

Petroleum hydrocarbons (PHs) are major organic contaminants in soils, whose degradation process is mediated by microorganisms such as the filamentous fungi Cunninghamella elegans and Phanerochaete chrysosporium. However, little is known about the tolerance and the degradation capability of Trichoderma species when exposed to PH. This research evaluated the tolerance and growth of 11 Trichoderma strains to crude oil (COil), naphthalene (NAPH), phenanthrene (PHE) and benzo[a]pyrene (B[a]P) by using in vitro systems. Petri dishes containing solid mineral minimum medium were separately contaminated with COil, with seven doses of either NAPH or PHE (250, 500, 750, 1000, 2000, and 3000 mg L(-1)), and with six doses of B[a]P (10, 25, 50, 75, and 100 mg L(-1)). Non-contaminated plates were used as controls. Trichoderma strains were exposed to all the contaminants by triplicate, and the growth of each fungal colony was daily recorded. No significant differences were observed among Trichoderma strains when they were exposed to COil in which the maximum fungal growth was reached at 96 h. In contrast, Trichoderma strains showed variations to tolerate and grow under different doses of either NAPH, PHE or B[a]P. Increasing NAPH doses resulted on significant greater fungal growth inhibition than PHE doses. The exposure to B[a]P did not inhibited growth of some Trichoderma strains.

Bioleaching of gold, copper and nickel from waste cellular phone PCBs and computer goldfinger motherboards by two Aspergillus nigerstrains

In an effort to develop alternate techniques to recover metals from waste electrical and electronic equipment (WEEE), this research evaluated the bioleaching efficiency of gold (Au), copper (Cu) and nickel (Ni) by two strains of Aspergillus niger in the presence of gold-plated finger integrated circuits found in computer motherboards (GFICMs) and cellular phone printed circuit boards (PCBs). These three metals were analyzed for their commercial value and their diverse applications in the industry. Au-bioleaching ranged from 42 to 1% for Aspergillus niger strain MXPE6; with the combination of Aspergillus niger MXPE6 + Aspergillus niger MX7, the Au-bioleaching was 87 and 28% for PCBs and GFICMs, respectively. In contrast, the bioleaching of Cu by Aspergillus niger MXPE6 was 24 and 5%; using the combination of both strains, the values were 0.2 and 29% for PCBs and GFICMs, respectively. Fungal Ni-leaching was only found for PCBs, but with no significant differences among treatments. Improvement of the metal recovery efficiency by means of fungal metabolism is also discussed.

Interactions between arbuscular mycorrhizal fungi and Meloidogyne incognita in the ornamental plant Impatiens balsamina

Biocontrol traits of arbuscular mycorrhizal fungi (AMF), in terms of single and mixed species inoculum, against the root knot nematode Meloidogyne incongita in Impatiens balsamina L., were examined with and without mineral fertilization in a greenhouse pot experiment. At harvest, 60 days after sowing, general plant growth parameters and plant defense response in terms of antioxidant activity and content of phenolic compounds in roots and leaves were measured. Also AMF root colonization and abundance ofnematode root-knots were determined. Mineral fertilization increased all plant growth parameters measured, which coincided with an increased disease development caused by M. incognita. Inoculation with AMF mitigated the observed plant growth reduction caused by M.incognita, though, higher abundance of M. incognita root knots was found in mycorrhizal plants. Plant defense responses in terms of antioxidant activity and content of phenolic compounds did not seem to be linked to the observed biocontrol traits of AMF against M. incognita. However, roots inoculated with a consortium of AMF, which presented less nematode root knots than roots with the single species inoculum, had the highest level of phenolic compounds. The results from the present study suggest that AMF induce tolerance in I. balsamina against the root knot nematode M. incognita.

Efectividad de siete consorcios nativos de hongos micorrízicos arbusculares en plantas de café en condiciones de invernadero y campo

Se probo el efecto de siete consorcios de hongos micorrizicos arbusculares (HMA) aislados de fincas cafetaleras con diferente nivel de tecnologia (bajo, medio y alto) del estado de Veracruz, en el crecimiento de plantas de cafe (Coffea arabica L.) var. Garnica en condiciones de invernadero y de campo. El grado de tecnificacion influyo en la composicion de especies de HMA, a mayor tecnificacion menor numero de especies de HMA encontradas. En condiciones de invernadero, los consorcios incrementaron la altura en un 91 % con respecto al testigo absoluto y al testigo fertilizado con fosforo (800 mg Ca(PO4)3 L-1). A los 130 dias despues de la inoculacion (DDI), el mejor consorcio fue La Estanzuela (ES). En condiciones de campo, a los 290 DDI, las plantas inoculadas con los consorcios ES, Miradores (MI), y Paso Grande (PG) tuvieron mayor supervivencia (> 80 %). Los consorcios mas efectivos en la promocion de la altura y supervivencia de las plantas en condiciones de campo, procedieron de agroecosistemas con nivel de tecnologia medio (MI y ES), mismos que tuvieron mayor numero de especies de HMA.

Research on arbuscular mycorrhizae in Mexico: an historical synthesis and future prospects

This review analyzes the historical development and advances of the research on arbuscular mycorrhizal fungi (AMF) in Mexico, as well as the prospects for future research. AMF-research has been focused on studying both diversity and functionality in several ecosystems of Mexico, but mainly in the tropical dry and rainy ecosystems, and the agricultural systems. In Mexico, 95 species of AMF have been recorded, representing 41% of the known species worldwide. The functional effects of AMF colonization have been examined in approximately 10% of the known host plants, but greenhouse studies continue to dominate over those conducted under field conditions. Even though research to date has been at the organismic level, further effort is needed due to the high plant diversity in Mexico. Studies on AMF biomass under field conditions and more taxonomic determination are required based on morphological features, biochemical determinations (fatty acids) and molecular tools. In addition, ecophysiological and ecological in situ studies would help in understanding the relationships among AMF, soil fauna, nutrients, and host plants. The contribution of AMF to ecosystemic processes is a priority line of research that requires an integrated approach (inter- and multidisciplinary) in order to define the role of AM symbioses for biogeochemical models. The creation of a Mexican mycorrhizal research network has and will help to identify the main challenges. Generating similar research protocols, and sharing databases and experience will assist mycorrhizologists working under the diverse financial and ecological contexts that is to be found in Mexico and Latin America.

Controlled Release Fertilizer Increased Phytoremediation of Petroleum-Contaminated Sandy Soil

A greenhouse experiment was conducted to determine the effect of the application of controlled release fertilizer [(CRF) 0, 4, 6, or 8 kg m–3] on Lolium multiflorum Lam. survival and potential biodegradation of petroleum hydrocarbons (0, 3000, 6000, or 15000 mg kg–1) in sandy soil. Plant adaptation, growth, photosynthesis, total chlorophyll, and proline content as well as rhizosphere microbial population (culturable heterotrophic fungal and bacterial populations) and total petroleum hydrocarbon (TPH)-degradation were determined. Petroleum induced-toxicity resulted in reduced plant growth, photosynthesis, and nutrient status. Plant adaptation, growth, photosynthesis, and chlorophyll content were enhanced by the application of CRF in contaminated soil. Proline content showed limited use as a physiological indicator of petroleum induced-stress in plants. Bacterial and filamentous fungi populations were stimulated by the petroleum concentrations. Bacterial populations were stimulated by CRF application. At low petroleum contamination, CRF did not enhance TPH-degradation. However, petroleum degradation in the rhizosphere was enhanced by the application of medium rates of CRF, especially when plants were exposed to intermediate and high petroleum contamination. Application of CRF allowed plants to overcome the growth impairment induced by the presence of petroleum hydrocarbons in soils.

Enhanced Pb Absorption by Hordeum vulgare L. and Helianthus annuus L. Plants Inoculated with an Arbuscular Mycorrhizal Fungi Consortium

The effect of an arbuscular mycorrhizal fungi (AMF) consortium conformed by (Glomus intraradices, Glomus albidum, Glomus diaphanum, and Glomus claroideum) on plant growth and absorption of Pb, Fe, Na, Ca, and 32P in barley (Hordeum vulgare L.) and sunflower (Helianthus annuus L.) plants was evaluated. AMF-plants and controls were grown in a substrate amended with powdered Pb slag at proportions of 0, 10, 20, and 30% v/v equivalent to total Pb contents of 117; 5,337; 13,659, and 19,913 mg Pb kg−1 substrate, respectively. Mycorrhizal root colonization values were 70, 94, 98, and 90%, for barley and 91, 97, 95, and 97%, for sunflower. AMF inoculum had positive repercussions on plant development of both crops. Mycorrhizal barley absorbed more Pb (40.4 mg Pb kg−1) shoot dry weight than non-colonized controls (26.5 mg Pb kg−1) when treated with a high Pb slag dosage. This increase was higher in roots than shoots (650.0 and 511.5 mg Pb kg−1 root dry weight, respectively). A similar pattern was found in sunflower. Plants with AMF absorbed equal or lower amounts of Fe, Na and Ca than controls. H. vulgare absorbed more total P (1.0%) than H. annuus (0.9%). The arbuscular mycorrizal consortium enhanced Pb extraction by plants.

The reduction in proline buildup in mycorrhizal plants affected by nematodes

Abstract Plants stressed by pathogens activate a variety of defense mechanisms to survive. The osmoprotector amino acids, including proline, are among these defense mecha -nisms. In this work, the effects of arbuscular mycorrhizal fungi on plants infested by root-knot nematodes were evaluated with regard to the accumulation of the os-moprotectant proline. A 2x3factorial design was established with 8 treatments – with and without nematodes, with and without mycorrhizae, and with and without fertilizer application – with 4 replicates. Two weeks after inoculation with arbuscu-lar mycorrhizal fungi, the plants were infested with 4 nematode egg masses, and 8 weeks later, the plants were harvested. The inoculation with the arbuscular mycor-rhizal fungi significantly reduced the proline content, with the non-inoculated plants exhibiting a higher concentration. Neither the infestation of the nematodes nor the addition of fertilizer significantly affected the proline content. Plant height, stem diameter, leaf area, number of leaves, and fresh weight were significantly improved by the presence of the arbuscular mycorrhizal fungi. The interaction of the fungi and the fertilizer did have a significant effect for height and leaf area. The nematode infestation and the fertilization did not affect mycorrhizal colonization.