Biochemical Systematics and Ecology 29 (2001) 305–311
Betaine distribution in the Bromeliaceae
Maricela Adrian-Romero
a,b, Gerald Blunden
a,*
aSchool of Pharmacy and Biomedical Sciences, University of Portsmouth, St Michael’s, Building, White Swan Road, Portsmouth PO1 2DT, UK
b
Faculty of Pharmacy, University of Los Andes, Me´rida ZP-5101-A, Venezuela
Received 23 November 1999; accepted 15 March 2000
Abstract
Aerial parts of 38 taxa, distributed in 21 genera and all three subfamilies of the Bromeliaceae have been examined for the presence of betaines. Glycinebetaine and trigonelline were isolated from all the plants studied, although the yields of both compounds were low. For glycinebetaine, values (based upon dry weight) varied from 0.0004% forPitcairnia corallinato 0.087% forTillandsia usneoides. Trigonelline levels varied from 0.0003 to 0.022%; the highest yields were recorded for Billbergia rubicunda (0.022%) and Vriesia splendens (0.020%).
# 2001 Elsevier Science Ltd. All rights reserved.
Keywords:Bromeliaceae; Betaines; Glycinebetaine; Trigonelline; Chemotaxonomy
1. Introduction
The taxonomic value of betaines in plants, particularly at the generic level, has been reported for the Gramineae (Wyn Jones, 1980), marine algae (Blunden et al., 1982, 1986, 1992), the Labiatae (Blunden et al., 1996) and the Capparaceae (McLean et al., 1996). However, systematic studies to determine the distribution of betaines in the plant kingdom have been restricted and concentrated primarily on herbaceous plants of saline and dry habitats in the temperate regions (Rhodes and Hanson, 1993).
The most studied role of betaines is that of compatible osmolytes aiding adaptation to saline and dry conditions. For this use, however, the concentrations of the compounds must be high, for example, as in the Chenopodiaceae (Adrian-Romero et al., 1998) and Amaranthaceae (Blunden et al., 1999), but in many betaine-containing plants, the levels of these compounds are low, as in the
*Corresponding author. Tel.: +44-23-92843593; fax: +44-23-92843565.
Polygonaceae (Adrian-Romero et al., 1998). Recently, it has been demonstrated that betaines have a role in aiding plants to resist attack by pathogens. Application of betaines in low amounts to plants has resulted in significant enhancement of the ability of the treated plants to resist attack by fungi (Kraska and Scho¨nbeck, 1992; Manninger et al., 1992) and root knot nematodes (Jenkins et al., 1998).
Continuing our studies of betaines in the plant kingdom, we report on the qualitative and quantitative distribution of these compounds in 38 taxa, representing 21 genera of Bromeliaceae in all three subfamilies (Smith and Downs, 1974, 1977, 1979). To our knowledge, this family has not been examined previously for betaines. Bromeliaceae was chosen because many of the species grow in regions with plentiful rainfall for part of the year followed by long periods of dryness; the presence of betaines in plants growing under such conditions of stress might be expected. The Bromeliaceae is a family of xerophytes and epiphytes containing in excess of 2000 species in 51 genera, originating from South and Central America and the southern states of the USA. The single exception isPitcairnia feliciana, which was discovered in West Africa (Brummitt, 1992).
2. Materials and methods
The species examined and their places of collection are given in Table 1. Voucher samples of most of the plants studied are maintained in the herbarium of either the Faculty of Pharmacy, University of Los Andes, Me´rida, Venezuela {(Tillandsia balbisiana, T. jenmani, T. schiedeana and T. spiculosa); Index Herbariorum code MERF} or the Hampshire County Council Museums Service, Winchester, Hampshire, UK (Index Herbariorum code HCMS; accession number Bi2000.16) Herbarium samples are not available for the species collected from the Royal Horticultural Society Gardens, Wisley, with the exception of Nidularium bill-bergioides.
Only leaves were used, which were oven-dried at 608C, powdered and extracted with 80% methanol for 24 h. The methods of extraction, purification of the extracts by passage through a column of cation exchange resin and two-way thin layer chromatographic (TLC) examination have been described previously (Blunden et al., 1981, 1985). The semi-purified extracts were examined by proton nuclear magnetic resonance (1H NMR) spectroscopy. The betaines were then isolated from the extracts by preparative TLC using air-dried silica gel G layers, 500mm, and methanol–water (50 : 50) as the development solvent. Zones corresponding to those of glycinebetaine and trigonelline (reference compounds run as bands at the edges of the plates) were removed, the compounds eluted with water, the extracts evaporated to dryness, the residues redissolved in deuterium oxide (D2O) and examined by 1H NMR spectroscopy (270 MHz).
Estimation of the content of each betaine present in the extracts, after passage through the ion-exchange column, was achieved using the1H NMR spectroscopic assay method of Blunden et al. (1986).
Table 1
Betaines in the Bromeliaceae
Species Place of collection Date of collection Glycinebetaine yield
(%, dry wt)
Trigonelline yield (%, dry wt)
Abromeitiella Croston Cactus, February,1999 0.011 0.012
brevifolia(Griseb) Eccleston, Chorley,
Castell Lancs, UK
A. brevifolia Croston Cactus, February, 0.006 0.003
subsp.chlorantha(Haum.) Eccleston, Chorley, 1999
Schultze-Motel Lancs, UK
Acanthostachys Royal Botanic October, 0.010 0.002
strobilacea Gardens, Kew, UK 1999
(Schultes f.) Klotzsch.
Aechmea Royal Horticultural February, 0.009 0.004
coelestis(K. Koch) E. Morr. Society Gardens, Wisley, Surrey, UK 1999
A. fasciata Rumsey Gardens, December, 0.001 0.002
(Lindley) Baker Clanfield, Hants, UK 1998
A. tonduziiMez and Pittier Royal Horticultural Society Gardens, Wisley, Surrey, UK
February, 1999 0.008 0.003
Ananas comosus(L.) Merrill Purchased in UK; origin Costa Rica November, 1998 0.025 0.002
Billbergia horridaRegel Royal Horticultural Society Gardens, Wisley, Surrey, UK
February, 1999 0.003 0.005
B. nutansH. Wendl. ex Regel Architectural Plants, Nuthurst, W. Sussex, UK
January, 1999 0.001 0.002
B. rubicundaMez Royal Horticultural Society Gardens, Wisley, Surrey, UK
February, 1999 0.005 0.022
B. xwindiiHort. ex Baker Rumsey Gardens, Clanfield, Hants, UK
January, 1999 0.002 0.004
Bromelia plumieri(E. Morr.) L. B. Smith Royal Botanic Gardens, Kew, UK October, 1999 0.005 0.004
Cryptanthus bivittatus(Hook.) Regel Cambridge University Botanic Garden, UK
June, 1999 0.007 0.003
}continued on next page
Table 1 (continued)
Species Place of collection Date of collection Glycinebetaine yield
(%, dry wt)
Trigonelline yield (%, dry wt)
Dyckia feroxMez Greenway Gardens Nursery, Churston Ferrers, Devon, UK
January, 1999 0.001 0.0007
Fascicularia bicolor(Ruiz and Pav.) Mez subsp.bicolorNelson and Zizkaa
Greenway Gardens Nursery, Churston Ferrers, Devon, UK
January, 1999 0.002 0.003
F. bicolorsubsp.canaliculataNelson and Zizkaa
Greenway Gardens Nursery, Churston Ferrers, Devon, UK
January, 1999 0.010 0.002
Fosterella rojasi(L. B. Smith) L. B. Smith
Royal Botanic Gardens, Kew, UK October, 1999 0.011 0.008
Guzmania lingulata(L.) Mez Keydell Nurseries, Horndean, Hants, UK
November, 1998 0.001 0.003
G. ‘Morado’ Purchased in supermarket, Havant,
Hants, UK
November, 1998 0.001 0.006
Hechtia argenteaBaker Brian Hiley, Wallington, Surrey, UK February, 1999 0.004 0.004
Hohenbergia blanchetii(Baker) E. Morr. Royal Botanic Gardens, Kew, UK October, 1999 0.001 0.0003
Neoregelia carolinae(Beer) L. B. Smith formatricolor(M. B. Foster)
M. B. Foster ex L. B. Smith
Royal Horticultural Society Gardens, Wisley, Surrey, UK
February, 1999 0.031 0.015
N. chlorosticta(Baker) L. B. Smith Royal Horticultural Society Gardens, Wisley, Surrey, UK
Ochagavia carnea(Beer) L. B. Smith and Looser
Hardy Exotics, Penzance, Cornwall, UK
February, 1999 0.004 0.004
Pitcairnia corallinaLind. and Andre´ Cambridge University Botanic Garden, UK
June, 1999 0.0004 0.004
Puya berteronianaMez Greenway Gardens Nursery, Churston Ferrers, Devon, UK
January, 1999 0.008 0.001
P. coeruleaLindley Greenway Gardens Nursery, Churston Ferrers, Devon, UK
January, 1999 0.004 0.004
Table 1 (continued)
Species Place of collection Date of collection Glycinebetaine yield
(%, dry wt)
Trigonelline yield (%, dry wt)
P. chilensisMolina Architectural Plants, Nuthurst, W. Sussex, UK
January, 1999 0.004 0.005
P. venustaPhilippi Greenway Gardens Nursery, Churston Ferrers, Devon, UK
January,1999 0.002 0.007
Tillandsia balbisianaSchult. f Laguna de Caparu´, Lagunillas, Me´rida State, Venezuela
March, 1996 0.035 0.004
T. jenmaniBaker La Lagunita, District of Valera, Trujillo State, Venezuela
March, 1996 0.021 0.002
T. schiedeanaSteud. Laguna de Caparu´, Lagunillas, Me´rida State, Venezuela
March, 1996 0.030 0.002
T. spiculosaGriseb. La Lagunita, District of Valera, Trujillo State, Venezuela
March, 1996 0.073 0.010
T. usneoides(L.) L. Brazos State Park, Texas, USA April, 1999 0.087 0.002
Vriesea splendens(Brongn.) Lem. Cambridge University Botanic Garden, UK
June, 1999 0.011 0.020
Wittrockia amazonica (Baker) L. B. Smith
Royal Botanic Gardens, Kew, UK October, 1999 0.012 0.009
aSee Zizka et al. (1999).
M.
Adrian-Romero,
G.
Blunden
/
Biochemic
al
Systematics
and
Ecology
29
(2001)
305–311
3. Results and discussion
Aerial parts of 38 taxa, representing 21 genera of the Bromeliaceae were extracted and the extracts examined for the presence of betaines. These were not detected by TLC in most of the extracts, even after partial purification by passage through a column of ion-exchange resin. However, the extracts, when examined by 1H NMR spectroscopy indicated the presence, in low concentrations, of glycinebetaine {N+–(CH3)3 resonance at d 3.37} and trigonelline (N+–CH3 resonance at d 4.33). The extracts were thus subjected to preparative TLC and the bands corresponding to the regions of glycinebetaine and trigonelline were removed, dried and the compounds eluted and examined by1H NMR spectroscopy and TLC.
By both TLC and 1H NMR spectroscopy, glycinebetaine and trigonelline were detected in extracts of all the species examined. The yields of both betaines were consistently low. The highest contents of glycinebetaine were found in theTillandsia
species (0.021–0.087%), but the contents found in other genera were much lower, usually being less than 0.01%. The highest yield of trigonelline was 0.022% from
Billbergia rubicunda, but usually the values recorded were less than 0.01%, but, with yields as low as these, the1H NMR spectroscopic assay procedure used is subject to considerable error.
Rhodes and Hanson (1993) classified the betaine-yielding families and genera into those which accumulate betaines and those which do not. The data recorded here show that the Bromeliaceae should be included in the non-betaine accumulating group.
Unlike some families examined, for example, the Labiatae (Blunden et al., 1996) and Capparaceae (McLean et al., 1996), in which the species in different genera show significantly different betaine compositions, in the Bromeliaceae the species tested in all the genera, including examples in all three subfamilies (Smith and Downs, 1974, 1977, 1979), contained the same two betaines, indicating the homogeneity of the family.
Acknowledgements
We thank the following for the supply of plant materials: the Director, University of Cambridge Botanic Garden; the Curator and Mr. M. Marsh of the Royal Botanic Gardens, Kew, Richmond, Surrey; Mr. N. Morgan, The Royal Horticultural Society, Wisley, Surrey; Mr. J.L. Henshaw, Croston Cactus, Eccleston, Chorley, Lancashire; Mr. R. Clark, Greenway Gardens Nursery, Churston Ferrers, Devon; Ms. J. Rojas-Vera and Mr. G. Medina-Ramirez, Faculty of Pharmacy, University of Los Andes, Me´rida, Venezuela; and Mr. and Mrs. R. Knights. We are grateful to Mr. D. Gullick and Mr. N. Armstrong for the1H NMR spectra and Mr. S. Sumner for technical assistance. This work and earlier studies (Blunden et al., 1999; Adrian-Romero et al., 1999) were partially funded by the CDCHT, University of Los Andes, Venezuela (project code no. FA-138-94-09-B).
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