PHYTOTHERAPY RESEARCH Phytother. Res. 19, 733–739 (2005) Published online in Wiley InterScience (www.interscience.wiley.com). DOI: 10.1002/ptr.1653 COMMON MULLEIN (VERBASCUM THAPSUS L.) 733 REVIEW ARTICLE Common Mullein (Verbascum thapsus L.): Recent Advances in Research Arzu Ucar Turker and Ekrem Gurel* Abant Izzet Baysal University, Faculty of Science and Arts, Department of Biology, 14280 Bolu, Turkey Common mullein (Verbascum thapsus L.) is a medicinal plant readily found in roadsides, meadows and pasture lands and has been used to treat pulmonary problems, inflammatory diseases, asthma, spasmodic coughs, diarrhoea and migraine headaches. Although it has been used medicinally since ancient times, the popularity of common mullein has been increasing commercially for the past few years. Today, the dried leaves and flowers, swallow capsules, alcohol extracts and the flower oil of this plant can easily be found in health stores in the United States. The use of common mullein extracts in folk medicine begun recently to be supported by an increasing number of research studies. This paper thoroughly reviewes all the scientific research related to Verbascum thapsus L. including plant tissue cultures and the biological properties of this plant. Copyright © 2005 John Wiley & Sons, Ltd. Keywords: Verbascum thapsus L.; mullein; medicinal herb; Scrophulariaceae. INTRODUCTION BOTANY The Scrophulariaceae family is an important family of plants comprising over 200 genera and about 2500 species. They occur mostly in temperate and subtropical regions, and many of them produce flowers of great beauty in either a garden setting or as roadside ‘weeds’. The family includes Mimulus, Penstemon, Digitalis, Veronica and Verbascum (Grieve, 1981). A number of the Scrophulariaceae are, or have been, valued for their curative properties and are widely employed both in domestic and regular medicine. At least 250 species of Verbascum are known. Of the species formerly used in medicine, the most important is Verbascum thapsus L. Common names include mullein, common mullein, great mullein, wooly mullein, candlewick plant, velvet plant, blanket leaf, white mans footsteps, Aaron’s rod, Jacob’s staff, hedge taper, high taper, old man’s flannel, lady’s foxglove and sı˝ır kuyru˝u (in Turkish) (Strange, 1977). The generic name of this plant, Verbascum, is believed to be a corruption of barbascum, from the Latin barba, meaning a beard, referring to the shaggy appearance of the genus, while thapsus, its specific name, may refer to the Greek island of that name, where the species originally thrived. The word ‘mullein’ comes from the Middle English moleyne and the Old French moleine, and originally from the Latin mollis, meaning ‘soft’ and referring to the leaves (Strange, 1977). Verbascum thapsus L. is a biennial, or rarely an annual plant, with a deep tap root. In its first year, it produces a low vegetative rosette up to 60 cm in diameter, which overwinters and is followed in the succeeding growing season by a stout flowering stem 0.3–2.0 m tall (Gross and Werner, 1978). The basal leaves are oblong-obovate to obovate-lanceolate and 10–40 cm long including the petiole. Cauline leaves are elliptic-lanceolate, decurrent, gradually reduced up the stem and densely woolly with branched hairs (Millspaugh, 1974). The leaf system is so arranged that the smaller leaves above drop the rain onto the larger ones below, which direct the water to the roots. This is a necessary arrangement since mullein grows mostly on dry soils. The stellately branched hairs, which cover the leaves so thickly, act as a protective coat, thus reducing moisture loss, and also providing a defence. They prevent attack by creeping insects and set up an intense irritation in the mucus membrane of any grazing animals that may attempt to browse upon them. The hairs are not confined to the leaves alone, but are also on every part of the stem, on the calyces and on the outside of the corollas, so that the whole plant appears whitish or grey (Gross and Werner, 1978; Whitson, 1991). According to Muzik (1970) these epidermal hairs hold away the droplets from the leaf surface so that they can protect the leaves from an aqueous solution of 2,4-D. The homely but valuable ‘mullein tea’, a remedy of the greatest antiquity for coughs and colds, must always be strained through fine muslin to remove any hairs that may be floating in the hot water, which was poured over the flowers or leaves. They can cause intolerable itching in the mouth (Grieve, 1981). The inflorescence is a spike-like raceme 20–50 cm long and approximately 3 cm in diameter. It is usually very dense; rare axillary racemes may arise from the * Correspondence to: Dr E. Gurel, Abant Izzet Baysal University, Faculty of Science and Arts, Department of Biology, 14280 Bolu, Turkey. E-mail: [email protected] 734 A. U. TURKER AND E. GUREL upper leaves. The sessile flowers are usually one per axil with pedicels less than 2 mm and slightly irregular with rotate corollas (Millspaugh, 1974). Individual flowers of mullein are ephemeral, opening before dawn and closing before mid-afternoon of the same day. They are protogynous, the style maturing first and then bending downward once the anthers appear. The flowers are also autogamous, self-pollination occurring at the end of the day if cross-pollination has not occurred. The style returns to its original position and the corolla closes, pushing the still receptive stigma against the anthers. The calyx consists of five lanceolate or ovate sepals, 7–9 mm long with caudate tips. The corolla is 20–25 mm broad consisting of five yellow (rarely white) petals. Stamens are irregular and attached to the corolla, three upper filaments are shorter and densely white-villous, the lower two are longer and glabrous. Anthers are larger and coloured. The ovary is superior and two-celled. The fruit is an ovoid, stellatepubescent, capsule 3–6 mm long, longer than the calyx and splits into two valves at maturity. There are numerous brown seeds, 0.5–1.0 mm long which are six-sided and have angular lateral surfaces with rows of pits (Abrams, 1951; Davis, 1965–1985; Munz and Keck, 1973; Gross and Werner, 1978; Radford et al., 1968). Chromosome counts from plants collected in Ottawa and British Colombia gave 2n = 36 (Packer, 1964; Mulligan, 1961). Other counts from European material have given 2n = 34 and 2n = 36 (Darlington and Wylie, 1955; Love and Love, 1961) and n = 9, 11, 17 (Love and Love, 1961). DISTRIBUTION V. thapsus L. is native to Europe and Asia. It was probably introduced into North America several times as a medicinal herb. It was introduced in the mid-1700s to Virginia as a piscicide (fish poison) and spread rapidly (Semenza et al., 1978). In Turkey, common mullein is distributed in Black Sea region (Kastamonu, Ordu, Trabzon, Rize, Çoruh) and commonly found in riversides, forests, Corylus and Quercus scrub and volcanic tuff (Davis, 1965–1985). V. thapsus L. grows wild on stony ground in wasteland, woodland clearings and roadsides. It does well in shallow, well-drained, nitrogen-rich soils (Semenza et al., 1978). It occurs in areas where the mean annual precipitation is 50–150 cm and the growing season is at least 140 days (Gross and Werner, 1978). MEDICINAL OR HISTORICAL USES Historically, mullein has been used as a remedy for the respiratory tract, particularly in cases of irritating coughs with bronchial congestion (Hoffman, 1988). Mullein leaves and flowers have expectorant and demulcent properties which are used by herbalists to treat respiratory problems such as bronchitis, dry coughs, whooping cough, tuberculosis, asthma and hoarseness (Grieve, 1981; Mabey, 1988; Berk, 1996). The flowers are mildly diuretic and have a soothing and antiinflammatory effect on the urinary tract (Mabey, 1988). The leaves are also diuretic, helping to reduce inflammation of the urinary system and to counter the irritating effect of acid urine (Ambasta, 1986; Tyler, 1993; 1994). Some herbal texts extend the therapeutic use to pneumonia and asthma (Grieve, 1981). The leaves, roots and the flowers are also anodyne, antiseptic, antispasmodic, astringent, emollient, nervine, vulnerary, analgesic, antihistaminic, anticancer, antioxidant, antiviral, bacteristat, cardiodepressant, oestrogenic, fungicide, hypnotic and sedative (Lucas, 1969; Harris, 1972; Null and Null, 1972; Grieve, 1981). The demulcent and emollient properties come from the polysaccharide mucilage and gums that soothe the irritated tissue. The expectorant property is the result of saponins that stimulate fluid production. The antiinflammatory property is due to iridoid glycosides and flavonoids that decrease inflammation (Grieve, 1981). The mullein combines the expectorant action of its saponins with the soothing effect of its mucilage, making this a most useful herb for the treatment of hoarseness, tight coughs, bronchitis, asthma and whooping cough (Mabey, 1988). The dried leaves are sometimes smoked in an ordinary tobacco pipe to relieve the irritation of the respiratory mucus membrane, and will completely control the hacking cough of consumption. The leaves are employed with equal benefit when made into cigarettes, for asthma and spasmodic coughs. The flowers placed in a bottle and set in the sunshine are said to yield a fatty matter valuable as a cure for haemorrhoids. Fomentations and poultices of the leaves have been found serviceable in haemorrhoidal complaints. Mullein is said to be of much value in diarrhoea, from its combination of demulcent with astringent properties, this combination strengthening the bowels at the same time (Grieve, 1981; Mabey, 1988). In Europe, a sweetened infusion of the flowers strained in order to separate the rough hairs is used as a domestic remedy for mild catarrhs and colic. A conserve of the flowers has also been employed against ringworm, and a distilled water of the flowers was long reputed to be a cure for burns and erysipelas (Millspaugh, 1974; Grieve, 1981). A decoction of leaves was used as a hearth stimulant. A decoction of roots febrifuge is used to alleviate toothache and also to relieve cramps, convulsions and migraines. The juice of the plant and powder made from the dried roots is said to quickly remove rough warts when rubbed on them (Tyler, 1993; 1994). An oil produced by macerating mullein flowers in olive oil, stored in a corked bottle during prolonged exposure to the sun, or by keeping it near the fire for several days, is used as a local application in country districts in Germany for piles and other mucus membrane inflammations, and also for frost bite and bruises. Mullein oil is recommended for earache and discharge from the ear, and for any eczema of the external ear and its canal (Mabey, 1988; Yarnell, 1997). Mullein oil is a valuable destroyer of disease germs (Chopra et al., 1956; Milspaugh, 1974). The fresh flowers, steeped for 21 days in olive oil, are said to make an admirable bactericide. An alcohol tincture is prepared by homoeopathic chemists, from the fresh herb with spirits of wine, which has proved beneficial for migraines or sick headaches of long standing, with oppression of the ear (Bianchini and Corbetta, 1977; Lewis and Elvin-Lewis, 1977). The seeds of mullein are said to be toxic and should not be used in any of these preparations (Berk, 1996). The seeds when thrown into the water are said COMMON MULLEIN (VERBASCUM THAPSUS L.) to intoxicate fish, and are used by poachers for that purpose, being slightly narcotic. Major toxic elements affecting the circulatory, respiratory and central nervous systems of the fish include saponin, rotenone and glycoside. The common mullein causes fish to have difficulty in breathing (Wilhelm, 1974). Swedish settlers called it wild tobacco and tied the leaves around their feet and arms when they had the ague. Some prepared a tea from the leaves for dysentery. A decoction of the roots was injected into the wounds of cattle when afflicted with worms, which caused them to die and fall out. Also some American Indian tribes (Mohegans, Penobscots, Catawbas, Chochtaws, Creeks, Forest Potawatomis and Menominees) used common mullein as a medicinal herb (Moerman, 1986). The Mohegans smoked them to relieve asthma and sore throat, and the Penobscots smoked the dried and powdered leaves for asthma. The Catawbas boiled the root and sweetened it to make syrup for croup in children. The leaves were mashed and applied as a poultice for pain and swelling, sprains, bruises and wounds. The Chochtaws put the leaves on the head as a headache poultice. The Creeks boiled the roots with those of Button Willow for a drink used for coughs. The leaves were also boiled and the patient bathed in the infusion while it was hot. The Forest Potawatomis smoked the dried leaves for asthma, but it is not certain whether they learned the practice from the whites or vice versa. A smoke smudge was made of the leaves and the fumes inhaled for catarrh and to revive an unconscious patient. The Menominees smoked the root for pulmonary diseases. Whites smoked the leaves for asthma and bronchitis and that the flowers were believed to be diuretic and had been used for tuberculosis (Moerman, 1986; Vogel, 1990). The flowering stem was used dried by Greeks and Romans as a taper dipped in tallow for light. Mullein torches were said to repel witches. There is evidence that at one time it was a ‘magical plant’ of the ancients. Agrippa, a general and minister under Caesar Augustus, claimed that the scent from the leaves had an overpowering effect on demons. Mullein was thought to be an ingredient in brews and love potions, and mentioned in incantations used by witches during the Middle Ages. The women of Rome also infused the flowers and mixed the resulting liquid with lye, using it as a wash to turn their hair golden yellow (Strange, 1977). BIOLOGICAL STUDIES Williams and Kemp (1976) showed that seedlings of V. thapsus L. collected from a range of cold to warm temperature habitats (based on altitude and latitude) exhibit similar rates of photosynthesis within a temperature range 20°–35 °C. Only at the highest temperature tested, 40 °C, the seedlings from the warmest habitat (low altitude and latitude) exhibit higher photosynthetic rates than those from the coolest habitat. They concluded that the ability of an individual plant to photosynthesize over a broad range of temperature has contributed to mullein’s success across a diversity of habitats (Gross and Werner, 1978). Williams et al. (1975) reported a CO2 compensation point of 58 vpm CO2 for V. thapsus L. and on this basis 735 concluded that the species had a C3 photosynthetic pathway. Wuenscher (1970) shaved the dense trichomes of the leaves of V. thapsus L. and found that the unshaved half of the leaf was consistently warmer than the shaved half. The hairs must, then, affect the leaf energy exchange, since two halves of the same leaf, differing only in the presence of hairs, reached different equilibrium temperatures. Convection and the latent heat loss are reduced by the hairs although the hairs have little effect on radiation absorption. The transpiration rate is reduced in hairy leaves. All of these effects are explained by an increase in boundary layer thickness caused by the hairs. The boundary layer forms above the surface of the hair coating rather than directly over the leaf surface. This thickens the boundary layer by the distance to which the hairs extend from the leaf surface. Increased transpiration resistance has obvious ecological significance for V. thapsus L., which grows on dry, exposed sites. The dense hair coating is an efficient water-conserving mechanism. Parkhurst (1976), evaluating the work of Wuenscher (1970), showed by calculation that the main effect of the trichomes was to increase stomatal resistance with only a slight increase in boundary layer resistance. Lortie and Aarssen (1997) demonstrated that clipping the shoot apex of V. thapsus L. resulted in significantly more branches, and the branching intensity could not be increased by greater resource levels in mullein when the apical meristem was intact. Branching was stimulated by the addition of nutrients only when the shoot apex was damaged. These results indicated that nutritional status does not solely determine the degree of branching expressed in mullein. Glier and Caruso (1973) demonstrated that decreasing temperatures were observed to induce starch degradation in the roots of mullein. Virtually no starch remains in the roots when the acclimation period of decreasing temperature was followed by an extended period of exposure to 4 °C. The breakdown of starch which was presumed to occur in late autumn under field conditions may provide cryoprotective chemicals for the over-wintering rosette. Such chemicals might also serve as an energy source for the bolting process, which occurs during the following spring or early summer. Later, they showed that the starch content in roots of V. thapsus L. was reduced when rosettes were exposed to low temperatures because of the increased activities of the starch degradative enzymes (Glier and Caruso, 1974). They also (Glier and Caruso, 1977b) reported that two nonspecific enzymes, namely, acid and alkaline phosphatase increased their activities in Verbascum roots exposed to decreasing temperatures; alkaline phosphatase was of more importance than acid phosphatase in over-wintering rosettes. It was known that mullein had a cold-requirement for bolting and that applied gibberellin substituted for this requirement (Caruso and Glier, 1970). Glier and Caruso (1977a) also described the influence of gibberellin on the activities of starch degradative enzymes and phosphatase. There was a sharp decrease in starch in the roots of Verbascum as a result of treating rosettes with GA3 and all three starch degradative enzymes revealed increases in their activities. Also, application of GA3 resulted in an increase in the activity of alkaline phosphatase in roots and rosettes showed an early response to applied gibberellin. 736 A. U. TURKER AND E. GUREL Gross (1981) showed that the rosette sizes of V. thapsus L. gives a reliable estimate of an individual’s fate the following year. A minimum size must be reached before a plant is capable of flowering and the probability of flowering increases steadily with rosette size. For mullein, all rosettes with a diameter greater than 41 cm flower the subsequent year and rosettes less than 9 cm in diameter do not flower. Conversely, the probability of death decreases with increasing rosette size. According to Reinartz (1980; 1984a), plant size has a large impact on plant fitness; within V. thapsus L. populations large plants develop faster, flower earlier and produce more seed. Compared with other monocarpic species, V. thapsus L. has an extremely low reproductive effort (11%–23% of biomass) and seed output (4%–8%). The reproductive efforts and seed output of other monocarpic species average 40% and 25%, respectively (Reinartz, 1984b). Differences between cohorts of mullein in plant size and the year of flowering were primarily environmentally induced; however, the variation between cohorts in a number of other characters appeared to have a genetic basis (Reinartz, 1984c). The seeds of V. thapsus L. are contained in a capsule with two cells. Salisbury (1942) reported that the mean number of capsules per explant was 226 ± 42 (SD; n = 37), with an average of 596 ± 30 (SD; n = 16) seeds per capsule. This gives an approximate average of 10 000– 180 000 seeds per individual plant, each seed averaging 0.067 mg (Gross, 1980; Gross and Werner, 1982). The seeds possess no specialized morphological adaptations for dispersal by wind or animals. The capsules split open along the longitudinal axis when mature, and movement of the stalk by wind or a large animal is required to release the seeds from the parent. Seeds are dispersed as far as 11 m, although 93% of them fall within 5 m and 75% of them fall within 1 m of the parent plant. Darlington and Steinbauer (1961) recorded that the seeds of mullein remain viable for up to 35 years but Kivilaan and Bandurski (1981) noticed that seeds may remain viable for over 100 years and in Denmark viable seeds of V. thapsus were collected from soil samples archaeologically dated from 1300 A.D. (Ødum, 1965). Mullein seeds may germinate under a wide variety of environmental conditions. Germination is completely inhibited below 10 °C and at constant temperatures above 40 °C (Semenza et al., 1978). Chilling during the winter lowers the temperature requirement for germination; thus, seeds brought to the surface in the autumn by soil disturbance are able to germinate early the next spring (Baskin and Baskin, 1981). Semenza et al. (1978) found that only 35% of seeds germinated in the dark, compared with 93% germination in the light. This light sensitivity varies seasonally, but on the whole, only those seeds, which lie at, or near the soil surface (0.5 cm or less) will be able to germinate. If seed burial occurs due to subsequent disturbance or heavy rains rapidly sifting the seeds below the soil surface, germination may be reduced or prevented through light deprivation (Gross, 1980). There are some natural enemies (insects and pathogens) of common mullein such as the curculionid weevil (Gymnaetron tetrum Fab.) which is specific to V. thapsus L. and was introduced to North America from Europe (Burcham, 1937). The larvae mature in the capsules (Sleeper, 1954) and destroy up to 50% of the seeds. Weevils were responsible for the predation of 57% of mullein capsules (Pottmeyer, 1985). The other insect, the mullein moth (Cucullia verbasci) feeds and develops on mullein species. There are some pathogens which cause disease in common mullein such as Erysiphe cichoracearum (powdery mildew) and Phymatotrichum omnivorum (root rot) (Gross and Werner, 1978). Although extracts from mullein can inhibit the growth of wheat seedlings, it is not a serious agricultural weed, since it can be controlled by cultivation. In overgrazed or poor pastures, the presence of common mullein represents a further degradation of the pasture because grazing animals avoid eating mullein. This species is not allelopathic, allergenic or poisonous to humans (Gross and Werner, 1978). Filippini et al. (1990) examined the commercial powdered samples of V. thapsus L. flowers and powdered samples of V. thapsus L. cultivated in the Botanical Garden of the University of Padua by using SEM. Contamination of a commercial powdered sample of V. thapsus L. flowers with plant material not belonging to a Verbascum species was observed. Vegetative reproduction does not occur in V. thapsus L. (Gross and Werner, 1978; Gross, 1980). Caruso (1971) showed that excised internodal segments of flowering specimens of V. thapsus L. with vascular tissues grew and formed numerous buds on a simple nutrient medium which lacked added growth regulator. Pith explants without vascular tissues became brown in a matter of 2 to 3 weeks with no visible sign of growth. Endogenous growth regulators supplied by vascular tissues were believed to be major factors in bud formation in excised internodal segments of this species. An in vitro culture protocol for common mullein was established by Turker et al. (2001). Explants (leaf discs, petioles and roots) were cultured on Murashige and Skoog minimal organics (MSMO) medium with benzyladenine (BA) or kinetin (KIN). The best shoot proliferation was obtained from leaf discs and petiole explants with 3 mg/L BA. Leaf discs were cultured on MSMO medium with 3 mg/L BA in combination with naphthalene acetic acid (NAA) or 2,4dichlorophenoxyacetic acid (2,4-D). More shoot development was obtained with 3 mg/L BA and 1 mg/L NAA. Shoots were transferred to rooting media containing different levels of NAA or 2,4-D. Most shoots developed roots on medium with 1 mg/L NAA. Plants were transferred to vermiculite and subsequently to potting media and maintained in a greenhouse. ACTIVE INGREDIENTS OF V. THAPSUS L. The constituents of V. thapsus L. include polysaccharides; iridoid glycosides including harpagoside; harpagide and aucubin (especially in the leaf); flavonoids, including 3 methylguercitin, hesperedin and verbascoside; saponins and volatile oils (Pascual Teresa et al., 1978a; 1978b; 1980; Hattori and Hatanaka, 1958; Khuroo et al., 1988; Mehrotra et al., 1989; Warashina et al., 1991; 1992). COMMON MULLEIN (VERBASCUM THAPSUS L.) Pascual Teresa et al. (1978a) isolated veratric acid and a-spinasterol from the benzene extract of capsules of V. thapsus L. From the hydrolysed ethanol extract, the triterpene A, saikogenin A, benzyl alcohol and methylfurfural were isolated. They also showed that the seed oil from mullein (benzene extract) had the following components: fatty acids: palmitic, steriac, oleic, linoleic, linolenic, arachic and behenic. Unsaponifiable matter: b -sitosterol and ergosta-7-en-3-b -ol (Pascual Teresa et al., 1978b). Phenylethanoid and lignan glycosides (Warashina et al., 1992), sterones, iridoid glycosides and sesquiterpene acid (Khuroo et al., 1988; Warashina et al., 1991) and verbacoside, a new luteolin glycoside (Mehrotra et al., 1989) were obtained from whole plants of V. thapsus L. Bourquelot and Bridel (1910) discovered the verbascose, an oligosaccharide in the root of the mullein. Hattori and Hatanaka (1958) demonstrated the oligosaccharides in V. thapsus L. and distribution of mono- and oligosacharides in various organs of the plant in various stages was studied. Pascual Teresa et al. (1980) isolated four saponins from the capsules of V. thapsus L., thapsuine A, thapsuine B, hydroxythapsuine A and hydroxythapsuine B with a chromatographic separation on silica gel (TLC). Crushed capsules (5 kg) were extracted with benzene and ethanol and subjected to chromatography on silica gel and sephadex. They obtained 300 mg thapsuine A and 460 mg thapsuine B. Bom et al. (1998) purified a-galactosidase from the roots of V. thapsus L. using hybrid affinity chromatography with good recovery. An extraction and analytical protocol for saponins was established for Verbascum thapsus L. (Turker et al., 2003). Four different kinds of plant sample were analysed; (i) field-grown, (ii) in vitro cultured, (iii) commercially obtained leaves and (iv) field-grown capsules. A cleanup procedure with octadecyl (C18) solid phase extraction columns was used prior to HPLC (high pressure liquid chromatography) analysis. Ilwensisaponin A was used as an external standard and digitoxin as an internal standard. C18 reverse phase column and gradient elutions (acetonitrile with 0.1% orthophosphoric acid and water with 0.1% orthophosphoric acid) were used for HPLC analysis. Commercially obtained leaves had a higher concentration of saponin (0.215 mg/g tissue) than other leaves (0.081–0.198 mg/g tissue) or the capsule samples (0.016 mg/g tissue). SCREENING STUDIES McCutcheon et al. (1992) used the disc diffusion method to show the antibiotic activity of common mullein leaves and demonstrated that a methanol extract of leaves had antibacterial activity against Escherichia coli [inhibition zone (iz); 8.0–10.0 mm], Mycobacter phlei (iz; 8.0–10.0 mm) and Staphylococcus aureus methicillinresistant (iz; 10.1–15.0 mm). The methanol extract of common mullein leaves was found to partially inhibit the cytopathic effects of bovine herpesvirus type 1, two double-stranded DNA viruses, which causes respiratory, genital, conjunctival or encephalitic infections which become latent in the trigeminal ganglion (McCutcheon et al., 1995). Moreover, this extract had antifungal activity against Micro- 737 sporum cookerii (iz; 8.0–10.0 mm) and M. gypseum (iz; 8.0–10.0 mm) (McCutcheon et al., 1994). Pardo et al. (1998) isolated iridoid glycosides lateroside 1, harpagoside 2, ajugol 3 and aucubin 4 from an ethanol extract of the roots of mullein that exhibits antigermination activity on the seeds of barley (Hordeum vulgare). Bioassays indicated that at 3 mm concentration, compounds 1, 2 and 4 showed moderate inhibition of seed germination. Aucubin 4 was the most active against root elongation and ajugol 3 showed no activity in the bioassays on barley seed germination and growth. Biological activities of common mullein extracts and commercial mullein products were assessed using selected bench-top bioassays (Turker and Camper, 2002). Bioassays included antibacterial, antitumour, and two toxicity assays (i.e. brine shrimp and radish seed bioassay). Extracts were prepared in water, ethanol and methanol and their biological activities assessed. For extractions, three different sources of leaves (fieldgrown, in vitro cultured and commercially obtained) and capsules (from field-grown plants) were used. Commercial products of common mullein (tea bags, swallow capsules, an alcohol extract and flower oil) and purified saponins were also assessed. The disc diffusion assay (Kirby-Bauer method) and six different bacterial strains (Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae which are gram-negative bacteria, and Streptococcus pyogenes, Staphylococcus aureus and Staphylococcus epidermidis which are grampositive bacteria) were used for antimicrobial activity assessment. In general, aqueous extracts were the most effective and showed antibacterial activity against K. pneumoniae and S. aureus. Methanol extracts from leaf materials showed little antibacterial activity only against K. pneumoniae. Generally, commercial products and purified saponins showed little or no antibacterial activity against these bacteria (except for the flower oil sample showing inhibition to K. pneumoniae, E. coli, P. aeruginosa and S. aureus). Agrobacterium tumefaciensinduced tumours in potato tissue were inhibited by all common mullein extracts, commercial products and purified saponins. Methanol extracts of in vitro-grown leaves and commercially obtained leaves inhibited tumour formation better than the other methanol, ethanol and water extracts. The results of brine shrimp bioassay showed that extracts of mullein were toxic at higher doses (around 1000 mg/L). Among aqueous extractions, decoctions had more toxicity (LC50 < 1000 mg/L) than the infusion extracts. In a radish seed bioassay, at high doses (10 000 mg/L), all extracts affected the root length. However, at 1000 mg/L, the extracts did not affect the root length as much. Seed germination was also inhibited by mullein extracts at 7500 mg/L. Low doses (1000 mg/L) affected germination, but not as much as higher doses. Ethanol extracts inhibited seed germination more than other types of extracts. CONCLUSION Verbascum thapsus L. (common mullein) is a herb with a long history of use in folk medicine. The commercial popularity of this plant has been increasing for the past 738 A. U. TURKER AND E. GUREL few years with the growing interest in herbs and preferring the ‘greener’ lifestyle. Today in health food stores, one can easily find dried leaves and flowers, swallow capsules, alcohol extracts and flower oil of mullein in USA. The screening studies (bioassays results) mentioned here confirm the popular utilization of mullein extracts in folk medicine as a remedy for respiratory disorders, tumour formation, urinary tract infection and certain skin diseases. With these results, V. thapsus L. has some scientific justification as a medicinal herb. 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