ABSTRACT
Here, we report a quick and low-cost method to improve plant transformation using Agrobacterium tumefaciens. This method involves the use of physical wounding, ultrasound, and an increase in exposure time to the bacteria. We show how the transformation rate increased from 0 to 14% when an ultrasound pulse of 10 s was used in conjunction with 96 h of bacterial exposure in Eclipta alba explants.
Subject(s)
Agrobacterium tumefaciens/genetics , DNA, Bacterial/genetics , Eclipta/genetics , Plant Stems/genetics , Plants, Genetically Modified , Transformation, Genetic , Agrobacterium tumefaciens/metabolism , Anti-Bacterial Agents/pharmacology , DNA, Bacterial/metabolism , Eclipta/drug effects , Eclipta/microbiology , Eclipta/radiation effects , Genetic Vectors/chemistry , Genetic Vectors/metabolism , Kanamycin/pharmacology , Kanamycin Resistance , Plant Stems/drug effects , Plant Stems/microbiology , Plant Stems/radiation effects , Ultrasonic WavesABSTRACT
In terrestrial ecosystems, plant growth, plant community structure, and ultimately the ecosystem services provided by plants are dependent on the presence and composition of below ground arbuscular mycorrhizal (AM) fungal communities. AM fungi form obligate symbioses with plants providing nutrients to their host plants in exchange for photosynthates. While AM have been found in most wetland ecosystems, the effects of urban contaminants on AM associations are largely unknown. Triclosan (5-chloro-2-[2,4-dichlorophenoxy]phenol; TCS) is a widespread contaminant found in surface waters throughout North America and in addition to antimicrobial properties is purported to have antifungal properties. To determine the effects of TCS on arbuscular mycorrhizal associations, we exposed AM inoculated wetland plant species (Eclipta prostrata, Hibiscus laevis, and Sesbania herbacea) to TCS at concentrations of 0.0, 0.4 and 4.0 µg/L in a continuous flow-through exposure system. TCS exposure caused significant reductions in hyphal and arbuscular colonization while no significant effect was detected for vesicular colonization. Across all species, hyphal colonization was significantly higher in controls (18.58 ± 1.84%) compared to 0.4 and 4.0 µg/L (10.20 ± 1.34% and 9.86 ± 1.32% respectively) TCS treatments. Similarly, arbuscular colonization was significantly higher in the controls (4.58 ± 0.75%) compared to 0.4 µg/L (2.20 ± 0.38%) and 4.0 µg/L (1.22 ± 0.24%) TCS exposures. Since our lowest effect concentration, 0.4 µg/L, lies within the range of concentrations found in North American streams it is plausible that AM colonization has been impacted in streams receiving WWTP effluent. Further studies are required to understand the mechanism of TCS inhibition of mycorrhizal colonization in wetland plant species as well as the potential ecological consequences that a decline in the AM colonization levels may represent.
Subject(s)
Eclipta/microbiology , Hibiscus/microbiology , Mycorrhizae/drug effects , Sesbania/microbiology , Triclosan/toxicity , Water Pollutants, Chemical/toxicity , Dose-Response Relationship, Drug , Hyphae/drug effects , Mycorrhizae/physiology , Plant Roots/drug effects , Plant Roots/microbiology , Toxicity Tests/methods , WetlandsABSTRACT
To identify the importance of arbuscular mycorrhizal fungi (AMF) colonizing wetland seedlings following flooding, we assessed the effects of AMF on seedling establishment of two pioneer species, Bidens frondosa and Eclipta prostrata grown under three levels of water availability and ask: (1) Do inoculated seedlings differ in growth and development from non-inoculated plants? (2) Are the effects of inoculation and degree of colonization dependent on water availability? (3) Do plant responses to inoculation differ between two closely related species? Inoculation had no detectable effects on shoot height, or plant biomass but did affect biomass partitioning and root morphology in a species-specific manner. Shoot/root ratios were significantly lower in non-inoculated E. prostrata plants compared with inoculated plants (0.381 ± 0.066 vs. 0.683 ± 0.132). Root length and surface area were greater in non-inoculated E. prostrata (259.55 ± 33.78 cm vs. 194.64 ± 27.45 cm and 54.91 ± 7.628 cm(2) vs. 46.26 ± 6.8 cm(2), respectively). Inoculation had no detectable effect on B. frondosa root length, volume, or surface area. AMF associations formed at all levels of water availability. Hyphal, arbuscular, and vesicular colonization levels were greater in dry compared with intermediate and flooded treatments. Measures of mycorrhizal responsiveness were significantly depressed in E. prostrata compared with B. frondosa for total fresh weight (-0.3 ± 0.18 g vs. 0.06 ± 0.06 g), root length (-0.78 ± 0.28 cm vs.-0.11 ± 0.07 cm), root volume (-0.49 ± 0.22 cm(3) vs. 0.06 ± 0.07 cm(3)), and surface area (-0.59 ± 0.23 cm(2) vs.-0.03 ± 0.08 cm(2)). Given the disparity in species response to AMF inoculation, events that alter AMF prevalence in wetlands could significantly alter plant community structure by directly affecting seedling growth and development.
