Online supplement
Can targeted defense elicitation improve seaweed aquaculture?
Gaoge Wang1,2, Lirong Chang3, Rui Zhang1,2, Shasha Wang4, Xiaojiao Wei1, Esther Rickert4, Peter Krost5, Luyang Xiao3, Florian Weinberger4
1 College of Marine Life Sciences, Ocean University of China, Yushan Road No. 5, Qingdao 266003, China
2 Institute of Evolution & Marine Biodiversity, Ocean University of China, Qingdao 266003, China
3 Weihai Changqing Ocean Science & TechnologyCo., Ltd, Rongcheng 264316, China
4 Department of Benthic Ecology, GEOMAR Helmholtz-Zentrum für Ozeanforschung Kiel, Düsternbrooker Weg 20, D-24105 Kiel, Germany
5 Coastal Research and Management (CRM), Tiessenkai 12, Kiel, Germany Corresponding author：
Florian Weinberger fweinberger@geomar.de
Figure S1. Dose-effect curves of oxidative burst responses after elicitation of Saccharina spp. with oligoalginate. Top: Responses of S. japonica 30 min after elicitation with different concentrations of oligoalginate B. Bottom: Responses of S. latissima 15 min after elicitation with different concentrations of oligoalginate A. Lines represent best fitting logistic dose-response functions.
Figure S2. Resistance of adult S. latissima sporophytes to 2-Propanol. Thalli were dipped into the solvent for (A.) 0 s, (B.) 1 s, (C.) 5 s, (D.) 10 s and (E.) 30 s. They were immediately afterwards rinsed in seawater for 30 s.
Damaged cells were then stained in seawater containing 0.05 % of the lethal stain Evans Blue. Exposure to 2- Propanol for 10 s visibly damaged cells of some thallus areas (D.), while an exposure time of 30 s resulted in 100 % damage (E.). Exposure times of 5 s or less caused no visible damage.
Fig.S3. Effects of oligoalginate concentration on the density and the length of S. japonica germlings after weekly treatments. Oligoalginate was applied between mid of September and mid of October 2012. Different letters or capitals indicate treatments and treatment frequencies that were significantly different (ANOVA & Tukey-test, p
< 0.05, see also table S1 below).
Table S1. (A) Density and (B) length of Saccharina japonica germlings after nursery cultivation with weekly application of oligoalginate at three different concentrations. Effect of concentration in a one-way-ANOVA.
(A)
Source DF SS MS F-Ratio p-value
Concentration 2 458.1 229.1 20.09 < 0.001
Error 12 136.8 11.4
(B)
Source DF SS MS F-Ratio p-value
Concentration 2 5.357 2.679 2.397 0.133
Error 12 13.41 1.118
Figure S4. Effect of oligoalginate elicitation on the density of bacterial cells on the surface of sea-cultivated S.
latissima sporophytes. Thallus parts of treated and control sporophytes were treated weekly over four weeks and conserved 24 h after an exposure to oligoalginate by freezing (-20℃). The protocol for quantification of associated bacteria followed Stratil (2013) with little modification. Approximately 0.5 cm2 of the algal material were cut out, applied on a microscope slide and directly stained with 10 µl DAPI (4,6-diamidino-2-phenylindole) containing mounting medium (Roti®-Mount FluorCare DAPI, Roth, Germany). Stacked images of five visual fields per replicate were taken and bacteria present were counted. n = 10.
Stratil, S.B. (2013) Ecology of microbes associated with a macroalgal host: Effects of environmental conditions on the composition of epibacterial communities and repellant effects of microbial epibionts on barnacle larvae.
PhD-thesis, Christian-Albrechts-Universität Kiel, Germany, 95 pp.
