Anthocyanin and Potential Therapeutic Traits in Clitoria, Desmodium,orchorus, 2atharanthus and Hibiscus Species

J.B. Morris and M.L. WangUSDA, ARS, PGRCU1109 Experiment St., Griffin, GA 30223-1797USA

Keywords: medicinal plants, nutraceuticals, phytochemicals, health enhancingcompounds, germplasm, phytopharmaceutical

AbstractThe USDA, ARS, Plant Genetic Resources Conservation Unit curates several

important nutraceutical and medicinal plant species. Anthocyanins are responsiblefor flower, leaf and seed coat color in plants, and are antioxidants as well. However,little is known about anthocyanin content in C/ito na tern atea, Desniodiumadscendens, Corchorus olitorius, Catharanthus roseus, Hibiscus sabdariffa and Acersaccharum. This study was conducted to identify anthocyanin indexes and additionalhealth enhancing components in these species. An anthocyanin meter with an LEDdiode of 520 nm was used to measure the anthocyanin index of leaves and flowersfrom plants growing in the field during July, September and October 2006. Wefound anthocyanin indexes ranging from 4.1 to 52.4 for leaves and 0.8 to 26.8 forflowers in all five species. The highest leaf anthocyanin index average of 14.6 wasfound in D. adscendens (P1 316623) followed by sugar maple (A. saccharum) with aleaf anthocyanin average index of 13.0. Catharanthus roseus (P1 608581) also had thehighest flower anthocyanin index average of 17.3. These data were recorded at orprior to 50% maturity. In addition, potential health uses for these species arediscussed. Phytochemicals identified include but are not limited to an antimicrobialprotein from C. ternatea, anthocyanin rich extracts from H. sabdaniffa for potentialuse as a chemopreventive agent, and antitumor promoters in C. olitorius leaves.These anthocyanins and phytochemicals have potential value in the nutraceuticaland medical industries. These variable anthocyanin indexes among these species willassist breeders and other scientists with valuable germplasm for development ofanthocyanin enriched cultivars.

INTRODUCTIONSeveral genera of economic importance to the nutraceutical as well as the

medicinal plant markets are conserved and curated at the USDA, ARS, Plant GeneticResources Conservation Unit (PGRCU), Griffin, GA. Anthocyanins are responsible forflower, plant, seed coat color, and recently known to be an antioxidant as well as freeradical scavenging (Polya, 2003). Not only have several new anthocyanins been reportedin C. ternatea (Terahara et al., 1998) but an antimicrobial and insecticidal protein knownas finbtin has been discovered as well (Kelemu et al., 2004). The objectives of thisresearch were to compare anthocyanin indexs from these field grown species and toreport some additional phytochemicals found in C. ternatea, D. adscendens, C. olitorius,C. roseus and H. sabdar(ffa for potential use as a nutraceutical or medicinal plant.

MATERIALS AND METHODS-.Sed from each of eight legumes including five C. ternatea, three D. adscendens,

as well as one C. olitorius, one C. roseus and one H. sabdariffa accessions were plantedin Pro-Mix HP potting soil duritig mid to late-March inside a greenhouse. After 45-50 days, seedlings from all species were transplanted to field plots of a clayey, kaolinitic,thermic typic kanhapludults soil series at Griffin, GA, in a completely random design.Each plot consisted of 1 row at 6

in 3 in 3 in rows. Data were analyzedusing analysis of variance (P<0.001) (SAS Institute, Cary, NC). Mean separations were

Proc. IS on Med. and Nutraceutical Plants 381Ed.: A.K. YadavActa Hort. 756, ISHS 2007

Inconducted using Duncan's multiple range test (P<0.05).

An Opti Sciences CCM-200 chlorophyll content meter was converted to anexperimental hand held anthocyanin meter. The manufacturer replaced the 655 nm lightemitting diode (LED) of the CCM with a 520 nm LED in order to measure absorbancenear the wavelength at which free anthocyanin aglycones in beans, cyanidin andpelargonidin monoglucosides absorb (Macz-Pop et al., 2004).

Prior to and near 50% seed maturity, anthocyanin indexes were recorded fromeach of three leaves using this modified anthocyanin meter on four different dates(18 July, 25 September, and II or 24 October) from each accession of C. lernatea,D. adscendens, C. oliioriu.v, C. roseus, H. sabdari/ja and sugar maple growing in thefield. Anthocyanin index data recorded on II and 24 October were combined becausesome accessions produced unsuitable leaves due to senescence. Anthocyanin indexesfrom sugar maple (Acer saccharurn) leaves were included because the similaranthocyanin, cyanidin can be detected with the 520 nm LED (van den Berg and Perkins,2005). For the first time, additional species including C. olitorius, C. roseus andH. sabdariffa were analyzed for their anthocyanin indexes also using this 520 nm LED.

Leaf tissue from C. ternatea (P1 226265), D. adscendens (P1 237955 and 316623),Hibiscus sabdarifjà (P1 291126) and C. olitorius (P1 404028) were prepared for HPLCanalysis for the flavonoids myricetin, daidzein, quercetin, genistein and kaempferolfollowing a similar procedure by Wang and Morris (2007).

RESULTS AND DISCUSSION

AnthocyaninsLeaf anthocyanin indexes ranged from 5.5 to 13.9 on 18 July 2006 (Table 1). Thespecies C. olitorius leaves produced the highest but not significantly average anthocyanin

index at 12.4 followed by H. sandarffa leaves (11.6), sugar maple (11.0) and C. ternateaP1 209591 (10.0) (Fig. I). During 25 September 2006, sugar maple leaves produced asignificantly higher anthocyanin index (22.2) than C. roseus. Interestingly as the seasonprogressed on II October 2006, P1 3 16623 of D. adscendens produced a significantlyhigher leaf anthocyanin index (19.6) than all other species. During 24 October 2006, bothP1 316623 and P1 237955 of D. adscendens produced significantly higher leafanthocyanin indexes (17.4 and 16.9, respectively) than all other species. The threeD. adscendens accessions increased by an average of 60% during October in leafanthocyanin index. Cot-chorus olitorius produced greater than 100% more leafanthocyanin indexes in July and September than during October. Anthocyanin indexeswere generally stable during July and September with the exception of sugar maple whichdramatically increased in leaf anthocyanin index by 100% in September.

For all months combined, anthocyanin flower and leaf indexes ranged from 1.0 to17.3. The accession P1 608581 (C. roseus) produced the significantly highest flower (darksalmon) anthocyanin index (17.3) followed by D. adscendens (P1 316623) leaves (14.6),dark blue flower of C. ternatea P1 283232 (13.5), sugar maple leaves (13.0),D. adscendens P1 237955 (12.7) and H. sabdar(ffa P1 291126 (11.1). The white flower ofC. ternatea P1 209591 produced the significantly lowest anthocyanin index of 1.0 whilethe blue flower produced by C. ternatea (P1 226265) and leaves from C. roseus yieldedsignificantly lower anthocyanin indexes of 3.1 and 5.9, respectively, as well (Fig. 2).

Corchorus olitorius produced the most quercetin (578 ng/pl) followed byH. sabdar(ffa (280.8 ng/jil) and C. ternatea (161.6 ng/pi). Clitorai ternatea leavesproduced the most kaempferol (658.6 ng/pl) followed by H. sabdarffa (183.4 ng/.tl).Negligible amounts of quercetin and kaempferol were found in both D. adscendensaccessions (Fig. 3). Interestingly, quercetin and kaemferol are isoflavones with similarbiochemical pathways as the anthocyanin, cyanidin and pelargonidin.

Potential Nutraceutjcals and MedicinalsThe African traditional medicinal plant, D. adscendens exerted analgesic effects in

mice (N'gouemo et al.. 1996) and is used for asthma in Ghana, Africa plus ovarianinflammation in Mato Grosso, Brazil (Barreto, 2002). A soluble dietary fiber fromC. olilorius suppressed blood gluscose elevation in rats and humans (lnnami et al., 2005).Corchorus olitorius has been found to inhibit the malaria parasite (Plasmodiumflulciparuin) above 96% (Sathiyamoorthy et al., 1999).

The common ornamental plant. periwinkle (Catharanthus roseus) has been shownto be effective against leukemia, skin cancer, lymph cancer, breast cancer, and Hodgkin'sdisease (Kintzios et al., 2004). Hibiscus sabdari/ja has anticancer properties (Hou et al.,2005) as well as inhibition of LDL oxidation plus potential service as a chemopreventiveagent (Chang et al., 2006). See Table 2 for additional potential nutraceuticals andmedicinals from these species.

CONCLUSIONSEven though some significant differences for leaf and flower anthocyanin indexes

occur among these species tested, several species decrease in anthocyanin index whileothers increase as the season progresses from 18 July till 24 October. Anthocyaninindexes remained stable for the majority of the species, however sugar maple increaseddramatically on 25 September. During plant senescence on 24 October, anthocyaninindexes for both C. olitorius and sugar maple dropped while H..andari/jà andD. adscendens increased. Anthocyanins provide chemical defenses against pests and mayhelp protect plants during excessively cold days. Anthocyanins also increase in plants asthe season progresses and can be observed in some species. For example, D. adscendensand H. sabdari/j?j produced green leaves containing large areas of purple leaf color withprogressively darker purple leaf areas as the season extended into October.

Dark flower colors in C. roseus and C. ternatea produced the greatest anthocyaninindexes overall during the entire season while green with purple tinges in leaves ofD. adscendens and H. sabdarifftu produced high anthocyanin indexes as well.Anthocyanin indexes were very low from those species producing light colors such aswhite or blue flowers and green leaves. Further testing should be considered forenvironmental effects on anthocyanin indexes. However, in this first report foranthocyanin indexes from these species, we suggest that several accessions from withinthese species need testing to verify potential use in breeding programs with eventualcultivar development for enriched anthocyanin indexes. This hand held anthocyanin metercan be successfully used to estimate anthocyamin indexes from these species tested.

Nutraceutical and phytopharmaceutical species do exist in the USDA, ARS,National Plant Germplasm System at the Plant Genetic Resources Conservation Unit,Griffin, GA. Not only do some of these species provide current nutraceutical uses butmany others have potential to be used in this type of market as well as for medicinal uses.

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