Abstract
Macroalgae-associated bacteria play an important role in their algal hosts’ biological processes. They are localized on surfaces of the host thalli, as well as between and even within algal cells. To examine the differences in community structures and functions between epi- and endo- bacteria, an effective approach for maximizing epiphyte removal from delicate seaweeds while retaining endophyte fidelity must be developed. In this study, a variety of surface sterilization methods for Ulva prolifera were compared, including mechanical, chemical, and enzymatical treatments. According to the results of scanning electron microscope (SEM) and denaturing gradient gel electrophoresis (DGGE) analysis, almost complete removal of epiphytic bacteria on Ulva was obtained simply by co-vortex of seaweeds with silica sands, causing minimal disturbance to endosymbionts when compared to previous published methods. In addition, the adaptability was also confirmed in additional U. prolifera strains and Ulva species with blade-like or narrow tubular thallus shapes. This easy mechanical method would enable the analysis of community composition and host specificity for Ulva-associated epi- and endo-bacteria separately.
Highlights
Macroalgal-bacterial associations were demonstrated to be widely distributed in marine habitats [1]
Bacterial communities have been shown to play a key role in terms of algal growth [2], nutrition acquisition [3], resistance to biofouling [4], adaptation to environmental stresses [5], spore germination, and colonization of seaweeds [6], and may even act as an indispensable factor in algal morphogenesis process [7,8,9,10,11,12]
Macroalgae-associated bacteria were found on surfaces of host thalli, as well as between or even within algal cells [13,14]
Summary
Macroalgal-bacterial associations were demonstrated to be widely distributed in marine habitats [1]. Among reported enzymatic and chemical methods applied to macroalgae, the UNSET buffer (urea and SDS as principal ingredients) and 3MTM Rapid Multi-Enzyme Cleaner (Sydney, NSW, Australia) were designed for selective collection of epiphytic bacteria [19,20] Both of them were successful in obtaining representative samples for epiphytes, the staining with fluorescent dye 40 ,6-diamidino-2-phenylindole (DAPI) still showed that some bacteria remained on the outer surface of algae, which made the algal thalli not suitable for the extraction of endophytes [21]. We attempted to develop an efficient protocol to maximize the removal of the epiphytic bacteria while maintaining the native endophytic community for the blooming-causative green seaweed U. prolifera O.F. Müller [31,32], facilitating the analysis of community composition and host specificity for Ulva-associated epi- and endo-bacteria separately
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