Prasun Ray

Correlation between organic acid exudation and metal uptake by ectomycorrhizal fungi grown on pond ash in vitro

Experiments were conducted to investigate the effect of coal ash on organic acid exudation and subsequent metal uptake by ectomycorrhizal fungi. Four isolates of ectomycorrhizal fungi namely, Pisolithus tinctorius (EM-1293 and EM-1299), Scleroderma verucosum (EM-1283) and Scleroderma cepa (EM-1233) were grown on pond ash moistened with Modified Melin-Norkans medium in vitro. Exudation of formic acid, malic acid and succinic acid by these fungi were detected by HPLC. Mycelial accumulation of Al, As, Cd, Cr, Ni and Pb by these fungi was assayed by atomic absorption spectrophotometer. Relationship between organic acid exudation and metal uptake was determined using classical multivariate linear regression model. Correlation between organic acid exudation and metal uptake could be substantiated when several metals are considered collectively. The finding supports the widespread role of low molecular weight organic acid as a function of tolerance, when exposed to metals in vitro.

A Novel Delivery System for the Root Symbiotic Fungus, Sebacina vermifera, and Consequent Biomass Enhancement of Low Lignin COMT Switchgrass Lines

Sebacina vermifera (MAFF-305830) is a mycorrhizal fungus originally isolated from the roots of orchids that we have previously shown to be tremendously beneficial in enhancing biomass yield and drought tolerance in switchgrass, an important bioenergy crop for cellulosic ethanol production in the United States. Towards this end, we have developed a bentonite clay particle-based delivery system for mass production and dissemination of S. vermifera for large-scale field trials. A greenhouse-based experiment was conducted to evaluate this novel delivery method for biomass enhancement of wild type and transgenic, low lignin (COMT down-regulated) switchgrass lines compared to an efficient in vitro colonization method. S. vermifera colonization enhanced plant biomass regardless of delivery method, although the percentage of fungal biomass in planta increased with the clay-based delivery system. Further, we found that release of some clay minerals in solution was enhanced in the presence of S. vermifera, while others were seemingly reduced. Intriguingly, the presence of S. vermifera has little or no impact on cell wall composition, including lignification. This research is the first report documenting the development of a bentonite clay particle-based delivery system for mass production of any symbiotic microbe and suggests that S. vermifera can be packaged with a mineral composite and effectively delivered to a target host plant.

Sebacina vermifera: a unique root symbiont with vast agronomic potential.

The Sebacinales belong to a taxonomically, ecologically, and physiologically diverse group of fungi in the Basidiomycota. While historically recognized as orchid mycorrhizae, recent DNA studies have brought to light both their pandemic distribution and the broad spectrum of mycorrhizal types they form. Indeed, ecological studies using molecular-based methods of detection have found Sebacinales fungi in field specimens of bryophytes (moss), pteridophytes (fern) and all families of herbaceous angiosperms (flowering plants) from temperate, subtropical and tropical regions. These natural host plants include, among others, liverworts, wheat, maize and Arabidopsisthaliana, the model plant traditionally viewed as non-mycorrhizal. The orchid mycorrhizal fungus Sebacinavermifera (MAFF 305830) was first isolated from the Australian orchid Cyrtostylisreniformis. Research performed with this strain clearly indicates its plant growth promoting abilities in a variety of plants, while demonstrating a lack of specificity that rivals or even surpasses that of arbuscular mycorrhizae. Indeed, these traits thus far appear to characterize a majority of strains belonging to the so-called “clade B” within the Sebacinales (recently re-classified as the Serendipitaceae), raising numerous basic research questions regarding plant–microbe signaling and the evolution of mycorrhizal symbioses. Further, given their proven beneficial impact on plant growth and their apparent but cryptic ubiquity, sebacinoid fungi should be considered as a previously hidden, but amenable and effective microbial tool for enhancing plant productivity and stress tolerance.

Non-targeted Colonization by the Endomycorrhizal Fungus, Serendipita vermifera, in Three Weeds Typically Co-occurring with Switchgrass

Serendipita vermifera (=Sebacina vermifera; isolate MAFF305830) is a mycorrhizal fungus originally isolated from the roots of an Australian orchid that we have previously shown to be beneficial in enhancing biomass yield and drought tolerance in switchgrass, an important bioenergy crop for cellulosic ethanol production in the United States. However, almost nothing is known about how this root-associated fungus proliferates and grows through the soil matrix. Such information is critical to evaluate the possibility of non-target effects, such as unintended spread to weedy plants growing near a colonized switchgrass plant in a field environment. A microcosm experiment was conducted to study movement of vegetative mycelia of S. vermifera between intentionally inoculated switchgrass (Panicum virgatum L.) and nearby weeds. We constructed size-exclusion microcosms to test three different common weeds, large crabgrass (Digitaria sanguinalis L.), Texas panicum (Panicum texanum L.), and Broadleaf signalgrass (Brachiaria platyphylla L.), all species that typically co-occur in Southern Oklahoma and potentially compete with switchgrass. We report that such colonization of non-target plants by S. vermifera can indeed occur, seemingly via co-mingled root systems. As a consequence of colonization, significant enhancement of growth was noted in signalgrass, while a mild increase (albeit not significant) was evident in crabgrass. Migration of the fungus seems unlikely in root-free bulk soil, as we failed to see transmission when the roots were kept separate. This research is the first documentation of non-targeted colonization of this unique root symbiotic fungus and highlights the need for such assessments prior to deployment of biological organisms in the field.

Comparative Analysis of Metal Uptake Potential of Hyphal Fusion Progenies of AMF and Their Parents

The main objective of the present study was to compare 15 arbuscular mycorrhizal fungal isolates (8 parents representing 3 Glomus species under the same genera Glomaceae and their 7 stable hyphal fusion progenies) for their heavy metal uptake potential so that they can be recommended for soil reclamation, environmental protection, and enhanced agricultural production. In vitro studies were conducted with AMF cultures grown in jars with minimal media amended with or without coal ash containing heavy metals. The experimental design was completely randomized with three replicates per treatment. Uptake of ten metals namely Al, Cd, Co, Cr, Cu, Mn, Ni, Pb, Si and Zn was analyzed by atomic absorption spectrophotometer. Study results showed that heavy metal uptake by hyphal fusion progenies and their parental isolate differs considerably among themselves and also for different metals. Hyphal fusion progeny 14 and parental isolate P46 showed high tolerance for a wide range of metals.