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Lipid MARKERS in CHEMOTAXONOMY of TROPICAL FRUITS: PRELIMINARY STUDIES with CARAMBOLA and LOQUAT H

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Lipid MARKERS in CHEMOTAXONOMY of TROPICAL FRUITS: PRELIMINARY STUDIES with CARAMBOLA and LOQUAT H 5. Hoffmeister, John E. 1974. Land from the Sea. Univ. of Miami 11. Popenoe, Wilson. 1920. Manual of Tropical and Subtropical Fruits. Press, Miami, FL. 143p. Hafner Press. 474p. 6. Ingram, Martha H. 1976. Crysophylum cainito—Star apple. The 12. Ruehle, G. D. and P. J. Westgate. 1946. Annual Rept., Fla. Agr. Propagation of Tropical Fruit Trees, (ed.) R. J. Garner et. al. Exp. Sta., Homestead, FL. Hort. Rev. No. 4, Commonwealth Agr. Bureaux, Farnham Royal, 13. Small, John K. 1933. Manual of Southeastern Florida. Hafner Pub. England, p. 314-320. Co., New York. 1554p. 7. Lendine, R. Bruce. 1952. The Naranjilla (Solanum quitoense 14. Stover, L. H. 1960. Progress in the Development of Grape Varieties Lam.) Proc. Fla. State Hort. Soc. p. 187-190. for Florida. Proc. Fla. State Hort. Soc. 73:320-323. 8. Long, Robert W. 1974. Origin of the vascular flora of South 15. Tomlinson, P. B. and F. C. Craighead, Sr. Growth-ring studies on Florida. In: Environments of South Florida (present and past), the native trees of sub-tropical Florida, p. 39-51. Research Trends in (ed.) Patrick J. Gleason, Miami Geological Soc, Miami, FL. p. 28- Plant Anatomy, K. A. Chowdhury Commemoration Volume 1972. 36. (eds.) A. K. M. Ghouse and Mohd. Yunus, New Delhi, India. 9. and O. Lakela. 1972. A Flora of Tropical Florida. Univ. 16. Winters, H. F. and Robert J. Knight, Jr. 1975. Selecting and breed of Miami Press, Miami, FL. 962p. ing hardy passion flowers. Am. Hort. 54(5):22-27. 10. Menzel, Margaret Young and F. O. Wilson. 1963. Allododecaploid 17. Zimmerman, G. A. 1941. Hybrids of the American pawpaw. J. of hybrid of Hibiscus diversifolius and some related J?1 Hybrids. /. of Hered. 32(3):83-91. Hered. 54(2):55-60. Proc. Fla. State Hort. Soc. 92:298-300. 1979. LiPID MARKERS IN CHEMOTAXONOMY OF TROPICAL FRUITS: PRELIMINARY STUDIES WITH CARAMBOLA AND LOQUAT H. E. Nordby and N. T. Hall gotic seedlings. Because these lipid markers have not been U.S. Citrus and Subtropical Products Laboratory,,1 investigated to any extent in subtropical and tropical fruits P. O. Box 1909, other than Citrus, we undertook this preliminary investiga Winter Haven, Florida 33880 tion on the lipid composition of two fruits, the carambola (Averrhoa carambola L.) and the loquat (Eriobotrya Abstract. Long-chain hydrocarbons, desmethyl sterols and japonica Lindl.). A study was conducted (12) previously on fatty acids have previously been demonstrated to be suit the organic acids, total soluble solids, flavor and texture able markers for the chemotaxonomy of citrus. We have of 18 carambola cultivars. extended these studies to include two tropical fruits: caram- bola (Averrhoa carambola L), 5 cultivars, and loquat (Erio- Materials and Methods botrya japonica Lindl.), 2 cultivars. The profiles of these Samples. Fruits from 5 cultivars of carambola and 2 three lipids in the 7 cultivars were determined by gas- cultivars of loquat were obtained from the USDA Sub liquid chromatography. C21 to C31 long-chain hydrocarbons tropical Horticulture Research Unit (Miami, Florida), were present in both species but C22 to C29 monoenes were frozen and stored at 0°F until extracted (within 2 weeks). present only in carambola. Major sterols for both fruits were Lipid Extraction. Four fruits of each carambola cultivar /3-sitosterol, campesterol, and isofucosterol in order of their were thawed and then individually cut, separated from their prevalence. Both fruits contained primarily the four major seeds and weighed (total fruit minus seeds). Fruits from each plant fatty acids—palmitic, oleic, linoleic and linolenic; how cultivar of loquat were pooled, separated from their seeds, ever, the ratios of these and 3 other acids were quite differ divided into four replicates and weighed (40 to 60 g). The ent in the two fruits. lipid of each sample was extracted with Folch reagent as reported by Nordby and Nagy for citrus lipid (6), weighed Chemotaxonomy is used in the delimitation of closely and stored at —40°F in benzene/ethanol, 4:1. related species, cultivars, selections, etc., of plants, fungi Fatty Acid Analyses. An aliquot of each lipid sample and other organisms. The markers generally used in these was evaporated to dryness on a rotoevaporator and studies include flavanoids, coumarins, phenolics, terpenes, transesterified with MeOH-NaOH-BF3; the resultant methyl long-chain hydrocarbons, sterols and fatty acids. The last esters were purified by thin-layer chromatography (TLC) 3 types of compounds are generally classified as lipids since and then analyzed by gas liquid chromatography (GLC) these compounds are soluble in organic solvents. Other than on 3% SP-1000 (11) and on 10% Apolar 10C at 160°C. reports of the fatty acid composition of seeds (2), very little Desmethyl Sterol Analyses. An aliquot of each lipid work has been done on the chemotaxonomy of tropical and sample was evaporated to dryness as above, saponified with subtropical fruits. 6% KOH and extracted into hexane (5). The desmethyl We have extensively studied the chemotaxonomy of the sterols were separated from the monomethyl sterols, di genus Citrus with these lipid markers (3, 7-11), and observed methyl sterols, hydrocarbons and other nonsaponifiables by trends in the lipid compositions of the various cultivars. silica gel TLC with chloroform as the developing solvent Each citrus species had intrinsic profiles, and hybrids (5). Underivatized sterols were analyzed by GLC on a 1% generally had profiles characteristic of both parents. Pro SP-1000 column at 220°C (5). files of nucellar seedlings were different from those of zy- Hydrocarbon Analyses. The hydrocarbons eluted from the TLC platings of the nonsaponifiables, were analyzed by iSouthern Region, U. S. Department of Agriculture, Science and programmed GLC in which monounsaturates (monoenes) Education Administration. were resolved from saturates (alkanes) (10). For verification Mention of a trademark or proprietary product is for identification of the monoene fractions, the total hydrocarbon fraction only and does not constitute a guarantee or warranty of the product by the U. S. Department of Agriculture and does not imply its ap was subjected to silver nitrate TLC (10), the isolated proval to the exclusion of others which may also be suitable. monoenes were hydrogenated (4), and the two saturated 298 Proc. Fla. State Hort. Soc. 92: 1979. fractions (original alkanes and the hydrogenated alkenes) Table 4. Relative percentages of alkanes in loquat fruit. were analyzed by GLC (10). Values in Tables 1-4 are the means of the four replicate Cultivar analyses for fatty acids, desmethyl sterols and hydrocarbons, Alkane M-18553 20 respectively. Table 1. Relative percentages of major fatty acids in carambola and 21 0.5 1.8 loquat fruits. 22 0.4 0.5 23 0.7 0.6 24 0.3 0.4 Fatty acidsz 25 0.7 0.6 Cultivar 14 16 16:1 18 18:1 18:2 18:3 26 0.1 0.4 27 12.1 9.8 28 2.0 2.0 Carambola 29 76.9 77.8 37 4.1 17.2 2.1 2.1 51.3 9.0y 14.2 30 1.3 0.9 44 6.1 16.7 2.1 1.8 51.8 9.2y 12.3 31 5.1 5.2 42 4.9 15.7 2.4 2.0 54.7 8.0y 12.3 Tean Ma 2.3 15.5 2.1 1.9 56.0 8.5y 13.7 17 7.1 15.4 2.1 1.6 50.9 10.4y 12.5 species are difficult to separate into edible (flesh) and non- edible portions, we used entire fruit minus its seeds in Loquat these taxonomic studies. No attempt was made to determine M-18553 0.9 22.5 0.8 7.0 20.0 36.9 11.9 20 0.9 24.4 0.5 8.7 13.7 38.5 13.3 the location of specific lipids. Generally, hydrocarbons are derived from the wax coating of fruits, sterols from the structural membrane and fatty acids from storage tissue ^Number of carbons in chain: number of double bonds. yMixture of linoleate and another 18:2 acid of unknown structure. as a component of triglycerides. Programmed GLG analyses revealed fatty acids from Table 2. Relative percentages of desmethyl sterols in carambola and C12 to C26 in both fruits; however, only the seven C14 to loquat fruits. C18:3 fatty acids listed in Table 1 were present at relative percentages greater than one. There were only minor Sterol differences between the fatty acid profiles of the five caram- Cultivar Cholesterol Campestero] I /?-Sitosterol 1 sofucosterol bolas, the differences being greatest for G14 and oleic acid (C18:1). Tean Ma and cultivar 42 had from 3.8 to 5.1 per centage points more oleic acid than the other three culti Carambola vars, whereas Tean Ma and cultivar 17 had the lowest and 37 0.5 18.3 75.8 5.4 44 0.5 14.8 80.1 4.6 highest values for C14 of the five carambolas. In Citrus, fatty 42 0.5 21.6 73.1 4.8 acid profiles of specific lipids in the fruit were more useful Tean Ma 0.5 16.2 79.0 4.3 for taxonomic studies than were profiles of other lipids (7).. 17 0.4 13.1 80.4 6.1 The same may be true for carambola, but further studies Loquat are required to determine whether the oleic acid triglyceride M-18553 1.5 3.1 90.4 5.0 levels in Tean Ma and cultivar 42 were significantly different 20 1.0 1.9 93.5 3.6 than those in the other three cultivars. In all five carambola cultivars the values for C18:2 in clude two fatty acids that could not be resolved on the Table 3.
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