Allelopathic growth inhibition of Heterosigma akashiwo by the three Ulva spcieces (Ulva Pertusa, Ulva Linza, Enteromorpha intestinalis) under laboratory conditions

Allelopathic effects of several concentrations of fresh tissue, dry powder and dry tissue of three bloom-forming green macroalgae Ulva pertusa, Ulva linza and Enteromopha intestinalis on the red tide microalga Heterosigma akashiwo were evaluated in microcosms systems. The effects of macroalgae culture medium filtrate were investigated on H. akashiwo using initial or semi-continuous filtrate addition. Preliminary studies on the algicidal effects of one aqueous and four organic solvent extracts from the macroalgae on the microalga were carried out to confirm the existence of allelochemicals in the tissue of these macroalgae. The dry powder of U. pertusa was extracted with methanol, and the methanol extracts were partitioned to petroleum ether phase, ethyl acetate phase, butanol phase and distilled phase by liquid-liquid fractionation. The bioassays of the activity of every fraction were carried out on H. akashiwo. The resultant microcosms assay showed that the growth of H. akashiwo was strongly inhibited by using fresh tissues, dry powder or dry tissue of these three macroalgae, while aqueous and methanol extracts of both macroalgae had strong inhibitory effects on the growth of H. akashiwo, and the EC50 values for methanol extract of U. pertusa, U. linza or E. intestinalis were 0.016, 0.028×10-12 or 0.033×10-12, respectively. While the other three organic solvent extracts (acetone, ether and chloroform) had no apparent effect on its growth, this suggests that the allelochemicals from these three macroalgae had relatively high polarities. The activity of petroleum ether phase, ethyl acetate phase, butanol phase and distilled phase of U. pertusa methanol extract was carried out on H. akashiwo indicating that petroleum ether phase and ethyl acetate phase had stronger algicidal effect on H. akashiwo. The inhibition effect of the ethyl acetate phase was not as strong as that of petroleum ether phase, and effective concentration of petroleum ether phase was 17 mg/L for H. akashiwo. However, no significant algicidal effects were observed on the butanol phase and distilled water phase. These three macroalgae's culture medium filtrate exhibited no apparent growth inhibitory effect on the microalga under initial filtrate addition whereas the growth of H. akashiwo was significantly inhibited under semi-continuous filtrate addition, which suggests that continuous release of small quantities of rapidly degradable allelochemicals from the fresh tissue of both macroalgae was effective in inhibiting the growth of H. akashiwo.