Citation:

s10811-015-0563-z

Always quote citation above when using data! You can download the citation in several formats below.

RIS Citation BibTeX Citation Text Citation Facebook TwitterShow Map Google Earth

Abstract:

In search of a meaningful stress indicator for Fucus vesiculosus we found that the often used quantitative determination procedures for the polysaccharide laminarin (beta-1,3-glucan) result in different kind of problems, uncertainties and limitations. This chemical long-term storage form of carbon enables perennial brown algae in seasonally fluctuating ecosystems to uncouple growth from photosynthesis. Because of this high ecological relevance a reliable and precise method for determination and quantification of laminarin is needed. Therefore, a simple, cold water extraction method coupled to a new quantitative liquid chromatography-mass spectrometrical method (LC-MS) was developed. Laminarin was determined in nine out of twelve brown algal species, and its expected typical molar mass distribution of 2000-7000 Da was confirmed. Furthermore, laminarin consisted of a complex mixture of different chemical forms, since fifteen chemical laminarin species with distinct molecular weights were measured in nine species of brown algae. Laminarin concentrations in the algal tissues ranged from 0.03 to 0.86% dry weight (DW). The direct chemical characterization and quantification of laminarin by LC-MS represents a powerful method to verify the biochemical and ecological importance of laminarin for brown algae.

Single individuals of Laminaria hyperborea, L. digitata, Saccharina latissima, F. serratus, F. vesiculosus, F. spiralis, Himanthalia elongata, Cystoseira tamariscifolia, Pelvetia canaliculata, Ascophyllum nodosum, Halidrys siliquosa and Dictyota dichotoma were collected in fall (18.11.2013) during spring low tide from the shore of Finavarra, Co. Clare, west coast of Ireland (53° 09' 25'' N, 09° 06' 58'' W). After sampling, the different algae were immediately transported to the lab, lyophilized and sent to the University of Rostock.

Laminarin was extracted with cold ultrapure water from the algal samples. Before extraction they were ground to < 1 mm grain size with an analytical mill (Ika MF 10 Basic). The algal material (approx. 1.5 g DW) was extracted in ultrapure water (8 mL) on a shaker (250 rpm) for 5 h. After the addition of surplus ultrapure water (4 mL) and shaking manually, 1 mL of the sample was filter centrifuged (45 µm) at 14,000 rpm (Hettich Mikro 22 R). The slightly viscous supernatant was free of suspended material and converted into a microvial (300 µL) for further analysis. The extracts were analyzed using liquid chromatography-mass spectrometry (LC-MS) analysis (LTQ Velos Pro ion trap spectrometer with Accela HPLC, Thermo Scientific). Laminarin species were separated on a KinetexTM column (2.6 µm C18, 150 x 3 mm). The mobile phase was 90 % ultrapure water and 10 % acetonitrile, run isocratically at a flow rate of 0.2 mL min-1. MS was working in ESI negative ion mode in a mass range of 100 - 4000 amu.

Glucose contents were determined after extraction using high-performance liquid chromatography (HPLC). Extracted samples were analyzed in an HPLC (SmartLine, Knauer GmbH) equipped with a SUPELCOGELTM Ca column (30 x 7,8 mm without preColumn) and RI-detector (S2300 PDA S2800). Water was used as eluent at a flow rate of 0.8 mL min-1 at 75 °C. Glucose was quantified by comparison of the retention time and peak area with standard solutions using ChromGate software.

Mannitol was extracted from three subsamples of 10-20 mg powdered alga material (L. hyperborea, L. digitata, S. latissima, F. serratus, F. vesiculosus, F. spiralis, H. elongata, P. canaliculata, A. nodosum, H. siliquosa) and quantified, following the HPLC method described by Karsten et al. (1991).

For analyzing carbon and nitrogen contents, dried algal material was ground to powder and three subsamples of 2 mg from each alga thalli were loaded and packed into tin cartridges (6×6×12 mm). The packages were combusted at 950 °C and the absolute contents of C and N were automatically quantified in an elemental analyzer (Elementar Vario EL III, Germany) using acetanilide as standard according to Verardo et al. (1990).

Project(s):

Biological Impacts of Ocean Acidification (BIOACID)

Coverage:

Latitude: 54.156944 * Longitude: -9.116110

Date/Time Start: 2013-11-18T00:00:00 * Date/Time End: 2013-11-18T00:00:00

Minimum Elevation: -5.0 m * Maximum Elevation: -5.0 m

Event(s):

Finavarra (West Coast Ireland) * Latitude: 54.156944 * Longitude: -9.116110 * Date/Time: 2013-11-18T00:00:00 * Elevation: -5.0 m * Method/Device: Sampling by hand (HAND)

Parameter(s):

#	Name	Short Name	Unit	Principal Investigator	Comment
1	Species	Species		Graiff, Angelika
2	Tissues	Tissues		Graiff, Angelika
3	Replicate	Repl		Graiff, Angelika
4	Laminarin	Laminarin	mg/g	Graiff, Angelika
5	Glucose	Glc	µg/g	Graiff, Angelika	dry weight
6	Mannitol	Mannitol	mg/g	Graiff, Angelika
7	Carbon	C	%	Graiff, Angelika
8	Nitrogen, total	TN	%	Graiff, Angelika
9	Carbon/Nitrogen ratio	C/N		Graiff, Angelika

License:

Creative Commons Attribution 3.0 Unported (CC-BY-3.0)

Size:

276 data points

Data

Download dataset as tab-delimited text — use the following character encoding:

1 Species	2 Tissues	3 Repl	4 Laminarin [mg/g]	5 Glc [µg/g]	6 Mannitol [mg/g]	7 C [%]	8 TN [%]	9 C/N
Laminaria hyperborea	meristematic blade	1	8.60	0	374.26	35.71	1.45	24.62
Laminaria hyperborea	meristematic blade	2	8.67		325.02	35.52	1.46	24.35
Laminaria hyperborea	meristematic blade	3	8.49		330.94	35.25	1.43	24.58
Laminaria digitata	meristematic blade	1	7.52	213	240.82	29.91	2.17	13.79
Laminaria digitata	meristematic blade	2	7.58		239.25	29.85	2.29	13.03
Laminaria digitata	meristematic blade	3	7.44		245.36	29.79	2.16	13.81
Saccharina latissima	whole blade	1	0.83	0	285.06	29.51	1.89	15.62
Saccharina latissima	whole blade	2	1.05		254.91	29.28	1.99	14.70
Saccharina latissima	whole blade	3	0.85		271.92	29.36	2.00	14.71
Fucus serratus	blade incl. meristematic tips	1	3.81	0	134.88	34.51	1.13	30.63
Fucus serratus	blade incl. meristematic tips	2	4.75		137.30	32.54	1.30	24.96
Fucus serratus	blade incl. meristematic tips	3	2.89		126.05	35.11	1.07	32.93
Fucus vesiculosus	blade incl. meristematic tips	1	2.51	0	204.68	35.34	0.97	36.51
Fucus vesiculosus	blade incl. meristematic tips	2	2.77		205.21	34.87	0.83	42.09
Fucus vesiculosus	blade incl. meristematic tips	3	2.40		191.93	34.41	1.19	28.87
Fucus spiralis	blade incl. meristematic tips	1	0.03	111	100.68	31.90	1.33	23.94
Fucus spiralis	blade incl. meristematic tips	2	0.04		86.00	32.99	1.28	25.81
Fucus spiralis	blade incl. meristematic tips	3	0.03		88.64	32.28	1.14	28.21
Himanthalia elongata	reproductive sporophyte	1	0.50	0	42.53	31.23	1.20	26.03
Himanthalia elongata	reproductive sporophyte	2	0.37		51.83	30.38	1.11	27.44
Himanthalia elongata	reproductive sporophyte	3	0.49		42.74	30.47	1.18	25.80
Cystoseira tamariscifolia	whole	1	0.00	468
Cystoseira tamariscifolia	whole	2	0.00
Cystoseira tamariscifolia	whole	3	0.00
Pelvetia canaliculata	whole	1	0.30	0	47.56	33.82	1.04	32.66
Pelvetia canaliculata	whole	2	0.30		46.56	34.46	1.05	32.85
Pelvetia canaliculata	whole	3	0.30		49.55	34.25	1.08	31.61
Ascophyllum nodosum	tips and middle part	1	0.00	0	90.76	34.95	0.96	36.37
Ascophyllum nodosum	tips and middle part	2	0.00		93.31	35.01	0.95	36.79
Ascophyllum nodosum	tips and middle part	3	0.00		100.05	35.53	0.87	40.79
Halidrys siliquosa	whole	1	0.00	0	127.66	36.79	1.38	26.58
Halidrys siliquosa	whole	2	0.00		118.05	35.18	1.18	29.90
Halidrys siliquosa	whole	3	0.00		100.32	37.25	1.09	34.21
Dictyota dichotoma	whole	1	0.65	1280
Dictyota dichotoma	whole	2	0.68
Dictyota dichotoma	whole	3	0.65