Extracellular Enzyme Activities and Carbon/Nitrogen Utilization in Mycorrhizal Fungi Isolated From Epiphytic and Terrestrial Orchids

Fungi employ extracellular enzymes to initiate the degradation of organic macromolecules into smaller units and to acquire the nutrients for their growth. As such, these enzymes represent important functional components in terrestrial ecosystems. While it is well-known that the regulation and efficiency of extracellular enzymes to degrade organic macromolecules and nutrient-acquisition patterns strongly differ between major fungal groups, less is known about variation in enzymatic activity and carbon/nitrogen preference in mycorrhizal fungi. In this research, we investigated variation in extracellular enzyme activities and carbon/nitrogen preferences in orchid mycorrhizal fungi (OMF). Previous research has shown that the mycorrhizal fungi associating with terrestrial orchids often differ from those associating with epiphytic orchids, but whether extracellular enzyme activities and carbon/nitrogen preference differ between growth forms remains largely unknown. To fill this gap, we compared the activities of five extracellular enzymes [cellulase, xylanase, lignin peroxidase, laccase, and superoxide dismutase (SOD)] between fungi isolated from epiphytic and terrestrial orchids. In total, 24 fungal strains belonging to Tulasnellaceae were investigated. Cellulase and xylanase activities were significantly higher in fungi isolated from terrestrial orchids (0.050 ± 0.006 U/ml and 0.531 ± 0.071 U/ml, respectively) than those from epiphytic orchids (0.043 ± 0.003 U/ml and 0.295 ± 0.067 U/ml, respectively), while SOD activity was significantly higher in OMF from epiphytic orchids (5.663 ± 0.164 U/ml) than those from terrestrial orchids (3.780 ± 0.180 U/ml). Carboxymethyl cellulose was more efficiently used by fungi from terrestrial orchids, while starch and arginine were more suitable for fungi from epiphytic orchids. Overall, the results of this study show that extracellular enzyme activities and to a lesser extent carbon/nitrogen preferences differ between fungi isolated from terrestrial and epiphytic orchids and may indicate functional differentiation and ecological adaptation of OMF to local growth conditions.

Plant and Soil Enzyme Activities Regulate CO2 Efflux in Alpine Peatlands After 5 Years of Simulated Extreme Drought

Increasing attention has been given to the impact of extreme drought stress on ecosystem ecological processes. Ecosystem respiration (Re) and soil respiration (Rs) play a significant role in the regulation of the carbon (C) balance because they are two of the largest terrestrial C fluxes in the atmosphere. However, the responses of Re and Rs to extreme drought in alpine regions are still unclear, particularly with respect to the driver mechanism in plant and soil extracellular enzyme activities. In this study, we imposed three periods of extreme drought events based on field experiments on an alpine peatland: (1) early drought, in which the early stage of plant growth occurred from June 18 to July 20; (2) midterm drought, in which the peak growth period occurred from July 20 to August 23; and (3) late drought, in which the wilting period of plants occurred from August 23 to September 25. After 5 years of continuous extreme drought events, Re exhibited a consistent decreasing trend under the three periods of extreme drought, while Rs exhibited a non-significant decreasing trend in the early and midterm drought but increased significantly by 58.48% (p < 0.05) during the late drought compared with the ambient control. Plant coverage significantly increased by 79.3% (p < 0.05) in the early drought, and standing biomass significantly decreased by 18.33% (p < 0.05) in the midterm drought. Alkaline phosphatase, polyphenol oxidase, and peroxidase increased significantly by 76.46, 77.66, and 109.60% (p < 0.05), respectively, under late drought. Structural equation models demonstrated that soil water content (SWC), pH, plant coverage, plant standing biomass, soil β-D-cellobiosidase, and β-1,4-N-acetyl-glucosaminidase were crucial impact factors that eventually led to a decreasing trend in Re, and SWC, pH, β-1,4-glucosidase (BG), β-1,4-xylosidase (BX), polyphenol oxidase, soil organic carbon, microbial biomass carbon, and dissolved organic carbon were crucial impact factors that resulted in changes in Rs. Our results emphasize the key roles of plant and soil extracellular enzyme activities in regulating the different responses of Re and Rs under extreme drought events occurring at different plant growth stages.

Differential Organic Carbon Mineralization Responses to Soil Moisture in Three Different Soil Orders Under Mixed Forested System

Soil microbial respiration is one of the largest sources of carbon (C) emissions to the atmosphere in terrestrial ecosystems, which is strongly dependent on multiple environmental variables including soil moisture. Soil moisture content is strongly dependent on soil texture, and the combined effects of texture and moisture on microbial respiration are complex and less explored. Therefore, this study examines the effects of soil moisture on the mineralization of soil organic C Soil organic carbon in three different soils, Ultisol, Alfisol and Vertisol, collected from mixed forests of Georgia, Missouri, and Texas, United States , respectively. A laboratory microcosm experiment was conducted for 90 days under different moisture regimes. Soil respiration was measured weekly, and destructive harvests were conducted at 1, 15, 60, and 90 days after incubation to determine extractable organic C (EOC), phospholipid fatty acid based microbial community, and C-acquiring hydrolytic extracellular enzyme activities (EEA). The highest cumulative respiration in Ultisol was observed at 50% water holding capacity (WHC), in Alfisol at 100% water holding capacity, and in Vertisol at 175% WHC. The trends in Extractable Organic Carbon were opposite to that of cumulative microbial respiration as the moisture levels showing the highest respiration showed the lowest EOC concentration in all soil types. Also, extracellular enzyme activities increased with increase in soil moisture in all soils, however, respiration and EEA showed a decoupled relationship in Ultisol and Alfisol soils. Soil moisture differences did not influence microbial community composition.

ISOLATE ENDOPHYTIC FUNGI FROM LOCAL CATHARANTHUS ROSEUS AND ANALYZE THEIR EXTRACELLULAR ENZYME ACTIVITIES

Endophytic microbia are known as natural sources for producing valuable enzymes. In this study, four endophytic fungi were isolated from roots of local Catharanthus roseus (L.) G. Don var. roseus (purple flower) and C. roseus var. ocellatus (red stamens white flower) widely grown in Nha Trang. They were identified as Fusarium solani RN1, Chaetomium funicola RN3, Penicillium rugulosum RN4 and Chaetomium homopilatum WN1 based on morphologies colonies and spores. The activity analysis of their extracellular enzymes indicted all isolated endophytic fungi are able to produce protease, cellulose, xylanase as well as amylase. This is the first report on the endophytic fungi inhabited in C. roseus plant growing in the coastal regions of Vietnam, which could provide an attractive source for bioactive enzyme exploitation.