. Current address: Dept. of Horticulture, Clemson Univ., Clemson, SC 29634. We gratefully acknowledge the Fred C. Gloeckner Foundation for HPLC equipment and Yoder Brothers for providing rooted cuttings. We also thank Larry Kasperson for plant care, Della
Susan E. Trusty and William B. Miller
J.G.M. Cutting, D.K. Strydom, G. Jacobs, D.U. Bellstedt, K.J. Van Der Merwe, and E.W. Weiler
Abbreviations: ELISA, enzyme-linked immunosorbent assay; DNOC, dinitro-o-cresol; HPLC, high-performance liquid chromatography; RIA, radioimmunoassay. 1 Present address: Dept. of Horticultural Science, Univ. of Natal, P.O. Box 375, Pietermaritzburg
Arthur D. Wall, Marisa M. Wall, and Joe N. Corgan
1 Graduate research assistant. 2 Associate professor. 3 Professor emeritus and Jose Fernandez chair for crop production. We thank Cindy Waddell for support in the HPLC analysis, Jose Luis Mendoza, Helen Redden, Melodie Borden, and M.R. Doyle for
Richard Grazzini, David Hesk, Ellen Yerger, Diana Cox-Foster, June Medford, Richard Craig, and Ralph O. Mumma
Composition of anacardic acids (phenolic acids known to be associated with small pest resistance in Pelargonium ×hortorum) was examined in 13 diploid and 25 tetraploid cultivars by high-performance liquid chromatography (HPLC). The presence of an unusual desaturation (omega (ω)-5) in the alkyl tail of anacardic acids present only in glandular trichome exudate of pest-resistant diploid inbred lines had previously been associated with a sticky-trap pest-resistance phenomenon. In this study, we examine Pelargonium cultivars for variability in anacardic acid composition to assess the distribution of ω5 desaturation among commercial cultivars, to determine possible interactions between ω5 desaturation and other plant desaturation mechanisms, and to examine the possible impact of ploidy on ω5 desaturation. An unsaturation index (UI) is derived to compare exudates differing widely in composition yet which may provide a similarly effective sticky-trap pest-resistance mechanism based on exudate viscosity. ω-5 Anacardic acids were observed in the glandular trichome exudate of all 38 commercial cultivars examined. No diploid cultivar produced ω5- and ω9- anacardic acids, although the simultaneous production of ω5 and ω9- anacardic acids was observed in three tetraploid cultivars. Total ω5- anacardic acids comprised from 42.4% (tetraploid cultivar Perlenkette-syn. Snowhite, Weiss) to 86.8% (tetraploid cultivar Amanda). Commercial P. ×domesticum cultivars had no ω5 anacardic acids. UIs ranged from 60.9 (tetraploid cultivar Dixieland) to 103.4 (diploid cultivar Pinto White). In contrast, anacardic acids collected from a pest-susceptible inbred line contained no ω5- anacardic acids and had a UI of 38.7. No significant differences among ploidy levels were observed for UIs or for most specific anacardic acid components, with the exception of 24:1 ω5- anacardic acid, in which the mean diploid value (32.1%) was significantly higher than that of the mean tetraploid value (27.6%). We conclude that ω5- anacardic acid production occurs in all Pelargonium cultivars observed and that these cultivars are predicted to exhibit resistance to small arthropod pests. Significant genetic variability in specific anacardic acid composition appears to exist among Pelargonium cultivars, suggesting that breeding for pest resistance can be readily monitored by HPLC of anacardic acids.
Hendrik van Gorsel and Adel A. Kader
Internal breakdown (IB) is the limiting factor in the storage and postharvest handling of stone fruits. The symptoms of IB appear when fruits are kept for prolonged periods at temperatures below 10C and include leatheriness, mealiness, browning and bleeding of the flesh, and failure to ripen normally. We investigated the changes in phenolic compounds associated with IB of stone fruits. Twenty-eight phenolic compounds were separated by HPLC. Ten of these components were significantly affected by chilling temperatures. The concentration of six phenols changed in response to ripening after chilling temperatures, parallel to the appearance of IB symptoms. Most phenols showed a concentration gradient from the inside to the outside of the fruit, Comparison between peach cultivars showed characteristic differences in phenol metabolism during ripening. In both cultivars the most predominant phenol, chlorogenic acid, showed little change in concentration during storage. The structure of key phenolic compounds will be determined in order to elucidate the biochemical relationship between the phenols and the related enzymes. In this respect, a method was developed to detect phenylalanine ammonia-lyase (PAL) activity in peach fruit.
F.A. Tomás-Barberdán, J. Loaiza-Velarde, and M.E. Saltveit
Mechanical wounding and exposure to ethylene induces an increase in phenylpropanoid metabolism in lettuce and an increase in the concentration of several soluble phenolic compounds that are easily oxidized to brown substances by polyphenol oxidase. To study the early response of lettuce to wounding and ethylene, leaves of iceberg, butter leaf, and Romaine lettuces were either wounded or exposed to ethylene at 10 μL·L–1 in flows of humidified air at 5 or 10°C. Soluble phenolic compounds were extracted at intervals up to 72 hours and were analyzed by HPLC. After 72 hours, wounded leaves of all three lettuce types showed elevated levels of caffeoyl tartaric acid, Chlorogenic acid, dicaffeoyl tartanc acid, and 3,5-dicaffcoyl quinic acid at both temperatures. In contrast, there were no significant increases in soluble phenolic compounds in iceberg lettuce exposed to ethylene at 10°C. At 5°C for iceberg, and at both temperatures for the other two types, there was the same pattern for ethylene treated and wounded leaf tissue. The kinetics of wound and ethylene-induced phenolic metabolism are different and will be discussed in relation to phenolics produced and browning susceptibility.
Joe-Ann McCoy, Mark Widrlechner, and Jeff Carstens
Echinacea is becoming a well-established, high-value crop, both as an ornamental and a dietary supplement. A comprehensive collection of Echinacea germplasm is conserved by the USDA-ARS North Central Regional Plant Introduction Station (NCRPIS) in Ames, Iowa, and is available via seed distribution for research and educational purposes (ars-grin.gov/npgs). Representing all nine species collected throughout their respective North American geographic ranges, the Echinacea collection includes 179 accessions. Extensive morphological characterization data associated with this collection have been compiled and are available to researchers on the Germplasm Resources Information Network (GRIN) database to aid in selection criteria. The collection has been used extensively for various research projects, ranging from ornamental breeding studies to HPLC analyses of metabolites of interest to the phytopharmaceutical industry. This poster will summarize the Echinacea collection conserved at the NCRPIS, including a list of available accessions by species, illustrations of seed, and control-pollinated cage propagation methods; and facilities utilized for seed cleaning, testing, and storage. In addition, instructions on how to use the GRIN database to view evaluation data and acquire germplasm will be provided.
Malkeet S. Padda and David H. Picha
Phenolic compounds and antioxidant activity were quantified in the principal sweetpotato cultivars marketed in the European Union. Total phenolic content, individual phenolic acids, and antioxidant activity in each cultivar were determined using Folin-Denis reagent, reversed-phase HPLC, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) methods, respectively. Significant differences in phenolic composition and antioxidant activity were found between cultivars. A Jamaican-grown, white-fleshed cultivar had the highest total phenolic content [4.11 mg·g-1 chlorogenic acid (dry tissue weight)], while the highest antioxidant activity [3.60 mg·g-1 Trolox (dry tissue weight)] was observed in the orange-fleshed California-grown cultivar Diane. Chlorogenic acid and 3,5-dicaffeoylquinic acid were the predominant phenolic acids, while caffeic acid was the least abundant in most cultivars. The highest content of chlorogenic acid (0.42 mg·g-1 dry tissue weight); 3,5-dicaffeoylquinic acid (0.43 mg·g-1 dry tissue weight); and 3,4-dicaffeoylquinic acid (0.25 mg·g-1 dry tissue weight) was present in the white-fleshed Jamaican cultivar. The orange-fleshed cultivars Diane and Beauregard had the highest content of caffeic acid (0.13 mg·g-1 dry tissue weight) and 4,5-dicaffeoylquinic acid (0.32 mg·g-1 dry tissue weight), respectively.
Carol Auer, Jerry D. Cohen, and Todd Cooke
The uptake and metabolism of exogenous tritium-labelled benzyl adenine was studied during the shoot induction period of petunia leaf explants in tissue culture. Transfer experiments with Petunia `MD1' leaf explants (1 cm2) on MS media with 2.2 uM BA show that 27% and 100% of the leaf explants are committed to shoot induction on days 6 and 10, respectively. To study BA uptake and metabolism, leaf explants were placed on media containing tritium-labled BA for 1, 3, 6 and 10 days. BA was taken up from the media on days 1-6. BA metabolizes were analyzed using HPLC, a UV absorbancc detector and enzymatic techniques. Metabolizes produced include: BA, BAdo, BA 7G, BA 9G, BAdoMP, BAdoDP, BAdoTP and 3 unidentified compounds. BA and BAdo were detected on days 1 and 3 but not during day 6-10, the time of shoot induction. The pool of ribotide metabolites decreased from days 1 to 10, from 26.5% of all metabolites to 1.6%. Glucosylated compounds, BA 7G and BA 9G, increased continuously from 24.9% to 69.8% between days 1 and 10. An unidentified compound C increased from 13% on day 3 to 24.8% on day 10. In separate experiments, BA uptake and metabolism were compared in two Petunia hybrida lines, St40 and TLV1, with different shoot organogenic responses in tissue culture. These data show interesting patterns of BA metabolism in relationship to shoot induction and organogenesis.
Howard F. Harrison, Joseph K. Peterson, and Maurice E. Snook
Bioasssay-guided investigation of constituents possibly contributing to the allelopathic potential of sweetpotato led to the isolation of a nonpolar seed germination inhibitor in sweetpotato (Ipomoea batatas L.) roots. Mass spectral data supported by HPLC s pectroscopic analyses and data obtained from hydrolysis products revealed the presence of three monogalactosyl-diglycerides (MGDGs) (galactosyl-di-linoleneoyl glyceride, galactosyl-linoleneoyl-linoleoyl glyceride, and galactosyl-di-linoleoyl glyceride) in storage roots. The compounds inhibited proso millet germination, and at 100 ppm inhibition was about 90%. MGDG with fully saturated fatty acids (galactosyl-distearoyl glyceride) was not inhibitory in the bioassay. An efficient method for quantitation of individual MGDGs was developed, and the contents of each compound in the storage root tissues of 12 genetically diverse cultivars and breeding lines were determined. On a dry weight basis, total MGDG contents ranged between 107 and 452 μg/g in the periderm, 298 and 807 μg/g in the cortex, and 296 and 755 μg/g in the stele. Also, large differences in the ratios of the three compounds between clones and between tissues within a clone were noted. The differences between clones indicate that manipulating total content and ratios of MGDGs through plant breeding is feasible.