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The sensitivities of Selenastrum capricornutum and the toxic strain Microcystis aeruginosa to exudates from Potamogeton maackianus and P. malaianus were compared using exudation experiment, and the potential allelochemicals released by these two pondweeds into surrounding water were also analyzed. The growth of S. capricornutum and M. aeruginosa was inhibited by the exudates from the two macrophytes. Compared to the control, the cell densities of S. capricornutum decreased by 42.7% and 61.9% in 2.5 and 5 g・L⁻¹ FW P. maackianus treatments, and the cell densities of S. capricornutum also decreased by 65.8% and 73.5% in the two biomass density treatments of P. malaianus after three days of treatments. After 3 days’ incubation in 2.5 and 5 g FW・L⁻¹ P. maackianus exudates, the M. aeruginosa cell densities were higher in control than in treatment. As for P. malaianus treatments, the cell densities of M. aeruginosa were reduced by 16.5 and 65.8% of the control in 2.5 and 5 g・L⁻¹ FW marophytes at the end of incubation period, respectively. The allelochemicals exuded from the macophytes, which inhibited both S. capricornutum and M. aeruginosa, belonged to lipophilic and moderately lipophilic compounds according to the bioassay results of exudate fractionations. By multiple comparison statistics, the results showed that P. maackianus had stronger inhibitory effects on M. aeruginsa, while S. capricornutum was more sensitive to the allelochemicals of P. malaianus. The different sensitivities of the two algae were probably caused by three alcohol compounds (1-methoxy-2-methyl-2-Propanol, 2-methyl-2-Hexanol, and 4-ethyl-2,6-dimethyl-4-Heptanol) through the GC-MS analysis of the most active exudate fractions.
Culture solutions of the submerged freshwater macrophytes Hydrilla verticillata and Vallisneria spiralis, separately incubated at 10 g of fresh weight (FW) per liter for three days, were extracted by solid phase extraction (SPE) followed by liquid liquid extraction (LLE), and the acquired ethyl acetate fractions were analyzed by gas chromatograph-mass spectrometry (GC-MS) to identify potential allelochemicals exuded from these plant species. Freeze-dried plant tissues were separately methanol-extracted with a similar LLE-GC-MS procedure to compare allelochemical production and exudation. Four phenolic compounds were identified for both species: vanillic acid (VA), protocatechuic acid (PA), ferulic acid (FA), and caffeic acid (CA). H.verticillata produced 179 times the amount of phenolic compounds of V. spiralis. The phenolic contents in the culture solution were lower than 10% of those in the plant tissues for both submerged species in six of the eight cases. When evaluating the joint effects of the compounds using the Toxicity Index (TI) model, it was observed that the four phenolic compounds exerted additive and synergistic inhibition effects on the growth of Microcystis aeruginosa depending on the mixing ratios. These results indicate that H. verticillata and V. spiralis could release some phenolic allelochemicals to inhibit the growth of M. aeruginosa, and the joint action of multiple allelochemicals may be an important allelopathic pattern of submerged macrophytes to inhibit the growth of noxious cyanobacteria in natural aquatic ecosystems.
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