Phytochem RevDOI 10.1007/s11101-013-9295-3 What makes Allium species effective against pathogenic microbes? Virginia Lanzotti • Giuliano Bonanomi Felice Scala • Received: 18 January 2013 / Accepted: 17 April 2013 Ó Springer Science+Business Media Dordrecht 2013 Abstract The antimicrobial activity of garlic (Allium sativum L.) has been known since ancient times. The first citation dates back to the Egyptian period of fifteenth century BC when garlic was reported to be used in folk medicine as a remedy for microbial infections. Scientific investigations on garlic started in 1858 with the work of Pasteur who first noted antibacterial properties of garlic extracts. From that date to the discovery of antibiotics, garlic has been used against amoebic dysentery and epidemic diseases such as typhus, cholera, diphtheria, and tuberculosis. But what makes garlic and Allium species effective against pathogenic microbes? The volatile allicin and other thiosulfinates, giving pungency to Allium plants, are well-studied antimicrobial agents. The thiosulfinates can decompose to form additional sulfur constituents, including diallyl, methyl allyl, and dipropyl mono-, di-, tri- e tetra-sulfides, and (E)- and (Z)-ajoene without losing antimicrobial activity. Besides these compounds, onion and garlic are characterized by polar compounds of steroidal and phenolic origin, often glycosilated, not pungent and more stable during cooking, showing also antimicrobial activity. Recently, there has been increasing scientific attention given to such compounds. Nitrogen organic compounds, like alkaloids and polypeptides, V. Lanzotti (&) G. Bonanomi F. Scala Dipartimento di Agraria, Universita` di Napoli ‘‘Federico II’’, Via Universita` 100, 80055 Portici, Napoli, Italy e-mail:
[email protected] have also been isolated from these plants and have shown antimicrobial activity. In this paper, the literature about the major volatile and non-volatile organic compounds of garlic and other Allium plants has been reviewed. Particular attention is given to the compounds possessing antimicrobial activity and to the correlation between the observed activity and the chemical structure of the tested compounds. Keywords Garlic Onion Thiosulfinates Saponins Phenolics Antimicrobial activity Introduction A great variety of chemical compounds with antimicrobial activity, traditionally known as secondary metabolites, are produced by higher plants. Thousands of diverse natural products, often involved in plant defense against pest and pathogens, have been identified including terpenoids, saponins, phenolics and phenylpropanoids, pterocarpans, stilbens, alkaloids, glucosinolates, tiosulfinates and indoles among many others (Dixon 2001). The phytochemical diversity of plant antimicrobial compounds has been previously reviewed by examining their involvement in constitutive (Wittstock and Gershenzon 2002) and inducible chemical defenses (Hammerschmidt 1999), mechanisms of plant resistance (Morrisey and Osbourn 123 Phytochem Rev 1999) and fitness cost (Heil 2002). The potential exploitation of such molecules for the control of plant pathogens (Dixon 2001) has also been evaluated. The genus Allium includes a great number of species (between 600 and 750) (Stearn 1992; Jiemei and Kamelin 2000), from which many antimicrobial compounds have been extracted and identified. The antimicrobial activity of extracts and oils from garlic (Allium sativum), the most studied species belonging to this genus, has been known since ancient times. The first citation of this plant is found in the Codex of Ebers (1550 BC), an Egyptian medical papyrus reporting several therapeutic formulas based on garlic as a useful remedy for a variety of diseases such as heart problems, headache, bites and worms (Block 1985). Cloves of garlic have been found in the tomb of Tutankhamen and in the sacred underground temple of the bulls of Saqqara. Egyptians thought garlic increased endurance and they assumed large quantities of it. Raw plants were routinely given to asthmatics and to those people suffering bronchial-pulmonary complaints. Later on, garlic was known by the Greeks and Romans, who used it like an important healing agent just as it is still used today by most of the people in the Mediterranean region (Lanzotti 2005). Besides garlic, other Allium species (e.g. onion, shallot, leek, chive, elephant garlic) are a rich source of phytonutrients, useful for the treatment or prevention of a number of diseases, including cancer, coronary heart disease, obesity, hypercholesterolemia, diabetes type Fig. 1 Schematic representation of major chemical classes of natural compounds from Allium species with antimicrobial activity. These natural compounds have been isolated from the different plant tissues (roots, bulbs, leaves and flowers) and seeds 123 2, hypertension, cataract and microbe infection (Lanzotti 2006, 2012). Scientific research on these plants started in the second half of the nineteenth century with the work of Louis Pasteur who in 1858 first noted antibacterial properties of garlic (Pasteur 1858). In 1932, Albert Schweitzer treated amoebic dysentery in Africa with garlic and, before the discovery of antibiotics, it was also used for several epidemic diseases (e.g. typhus, cholera, diphtheria, and tuberculosis). Recent research has shown that garlic extracts and oils are effective against a plethora of saprophytic, human- and plantpathogenic virus (e.g. Weber et al. 1992), bacteria (e.g. Naganawa et al. 1996), fungi (e.g. Yoshida et al. 1987) and protozoa (e.g. Millet et al. 2011). However, only in the middle of the last century, Cavallito and Bailey (1944) identified the most abundant and, among the most potent, antimicrobial compound from garlic, the thiosulfinate allicin (Fig. 1). Subsequent investigations attributed the antimicrobial properties of garlic to allicin and other organosulfur compounds. However, more recent studies on Allium species identify natural compounds with antimicrobial activity of different chemical classes, including saponins, flavonoids, phenols, alkaloids and also peptides and proteins (Fig. 1). In this work we examine the chemical nature and complexity of antimicrobial compounds discovered in Allium plants, giving particular attention to the correlation between their activity and chemical structure. Thiosulfinates (allicin) N-oxides & N-amides Saponins & Sapogenins (alliogenin) Flavonoids (quercetin) when tissue is damaged. Figure 2 shows allicin biosynthesis starting from L-cysteine that for further reaction with L-glutamic acid and S-2-propenyl carboxylic acid. Fig. from one to three carbon atoms.CO2 Glu CO2H double bond Isomerization O NH2 S Alliin ( 1a) O Alliinase S CO2H O Sulfenic acid Intermediate Fig. Fig. Fig. oxidation and isomerization gives rise to the close precursor alliin. These compounds originate from allicin (1b) and thiosulfinates (e. because of a sigmatropic rearrangement (Block 1985). In nature. did not have any pungency which. obtained by this procedure. after decarboxylation. These results demonstrate that the pungency of garlic and Allium plants is associated with the sulfur compounds which constitute up to 5 % of the dry weight of the plant. which are quite unstable because of their thioester bond. immediately is transformed into allicin by the enzyme alliinase. the precurson alliin (S-allylcysteine sulfoxide) (1a.g. ajoene and other organosulfur compounds with a hydroalcoholic solution. that has been isolated in the E and Z isomers (2 and 3. store antimicrobial compounds in the inactive forms of non-toxic precursors. All these compounds are characterized by small terminal alkyl chains.g. However. Fig.Phytochem Rev Allicin. however. 4). Fifty years later. oxidation and isomerization. 2) (Cavallito and Bailey. This may also happen for inner propenyl chains as in the case of ajoene. that is present in garlic bulbs at a concentration ranging from 5 to 14 mg g-1. Fig. many plants. 18–19. the enzyme was inactivated and the disulfide compounds were not found. The terminal alkyl chain could be also unsaturated thus giving the 1-propenyl (named allyl from Allium) or the 2-propenyl residue. This last group is present with Z and E configurations and thus the compound exists as a mixture of the Z and E isomers (e. The garlic powder. to avoid self-toxicity. 2). 16–23. respectively. and give rise to further reaction during the extraction and isolation procedures to form more stable sulfoxides and sulfides (2–15. 1944) is not present in garlic bulbs. This last by enzymatic H H2O condensation S S Allicin (1b) catalysis of alliinase affords the corresponding sulfenic acid intermediate whose autocondensation in water phase results in the formation of allicin 123 . Figure 3 reports the chemical structure of the antimicrobial sulfoxide and sulfide compounds (2– 15). gives rise to the close precursor alliin. This last by enzymatic catalysis of alliinase affords the corresponding sulfenic acid intermediate whose autocondensation in water phase results in the formation of allicin. 2 Allicin biosynthesis starting from L-cysteine that for further reaction with L-glutamic acid and S-2-propenyl carboxylic acid after decarboxylation. connected to sulphur atoms at different oxidation states. 3). This class of compounds was found first by Wertheim (1844) and later on by Semmler who identified the correct disulfide structure of the main component of garlic (1982a) and onion (1982b) distilled oil. and extracted with acetone. a sulfoxide and sulfide compound. 20–21 and 22–23. When the powder was treated NH2 HS NH L-Glu CO2H HS L-Cys Glu CO2H S CO2H S NH Glu NH Ox CO2H Glu CO2H L-Glu S-2-carboxylpropyl L-Cys L-Glu L-Cys O NH S . During pathogen attacks Allicin (diallyl thiosulfinate) (1b. 3) and of its analogue 10-devinylajoene also isolated in the two E and Z forms (14 and 15). 4). pulverized. it was demonstrated that these compounds are absent when the bulbs are frozen in dry ice. appeared after the addition of water. Fig. Escherichia coli. Allicin (1b. many fungi. 3 Sulfoxide and sulfide compounds from Allium and/or when tissues are damaged. enzymes that convert precursors into antimicrobial compounds are made available (Hallahan 2000). Salmonella typhimurium. Fig. is a potent antimicrobial compound.. antimicrobial compounds are locally produced and accumulated only when and where it is necessary for defense against pathogens. It is able to inhibit at low concentrations. 2). The bactericidal activity of allicin has been reported against at least 25 saprophytic and pathogenic bacteria such as Bacillus spp. pathogenic protozoa and also viruses (Table 1). (1999) reported a synergistic interaction between the antibiotic vancomycin and allicin in the inhibition of several antibiotic-resistant enterococci. and allicin in the inhibition of the yeast Saccharomyces cerevisiae (Ogita et al. largely used as a broadspectrum fungicide.. 4 Thiosulfinate compounds from Allium 123 study found that streptomycin and chloramphenicol synergize with allicin in inhibiting Mycobacterium tuberculosis (Gupta and Viswanathan 1955). Cutler and Wilson (2004) reported that allicin was effective against methicillin-resistant S. Allicin can act synergistically with synthetic fungicides. quite unstable and having a low water solubility. with an LD50 often within the range of 3–20 lg ml-1.Phytochem Rev Fig. Recently. Cryptococcus. Helicobacter pylori. In this way. aureus strains. Enterococcus spp. A similar synergistic interaction was reported between the cuprous ion. . although volatile. gram-positive and gram-negative bacteria. 2005). Epidermophyton and Microsporum. Trichophyton. An early Fig. Staphylococcus aureus and Vibrio cholera (Table 1). Antifungal activity of allicin was clearly demonstrated by Yamada and Azuma (1977) against several important genera of human pathogens including Candida. Jonkers et al. Culter and Wilson (2004) Bacillus paratyphosus Bacillus subtilis Bacillus typhosus Enterococcus durans Enterococcus faecalis Enterococcus faecium E. Jonkers et al. typhimurium Shigella flexneri Shigella dysenteriae S. aeruginosa S. cerevisiae Torulopsis glabrata Trichophyton ferrugineum Trichophyton mentagrophytes Trichophyton rubrum Antibacterial Acinetobacter baumanii Bacillus dysenteriae Bacillus enteriditis Bacillus morgani Cavallito and Bailey (1944). coli H. Gupta and Viswanathan (1955). Ankri and Mirelman (1999). Shadkchan et al. pylori Klebsiella pneumonia Mycobacterium tubercolosis Proteus mirabilis P. (2000). ursinum Aspergillus flavus Aspergillus fumigatus Aspergillus niger Aspergillus terreus Candida albicans Candida krusei Candida neoformans Candida tropicalis Cryptococcus sp. (2005).Phytochem Rev Table 1 List of organic sulfur compounds from the genus Allium with related antibacterial. (2004). antifungal. Feldberg et al. antiparasitic and antiviral activities Species name Antimicrobial compounds Activity References A. (1987). (1999). Bagiu et al. cholera 123 . Epidermophyton foccosum Microsporum gypseum S. Yoshida et al. sativum Allicin (1b) Antifungal Yamada and Azuma (1977). Ogita et al. (1988). (2012) A. O’Gara et al. aureus Streptococcus hemolyticus Streptococcus pyogenes Streptococcus viridans V. Ledezma et al. (1996). albicans Candida valida Colletotrichum sp.Phytochem Rev Table 1 continued Species name Antimicrobial compounds Activity References Antiparasitic Mirelman et al. Millet et al. (1998) . (2000) A. (1996). Naganawa et al. coli K. Fusarium lini F. (1990). 3) Antifungal Alternaria solani Alternaria tenuissima Alternaria triticina Yoshida et al. cereus B. (1999). Yoshida et al. (1987). (1987). niger C. Singh et al. (2011) Crithidia fasciculate Entamoebe histolytica Giardia lamblia Leishmania major Leptomonas colosoma Spironucleus vortens Antiviral Weber et al.and Z-Ajoene (2. Ledezma et al. luteus Mycobacterium phlei 123 Naganawa et al. subtilis E. (1992) Herpes simplex virus type 1 Herpes simplex virus type 2 Parainfluenza virus type 3 Rhinovirus type 2 Vaccinia virus Vesicular stomatitis virus A. pneumoniae Lactobacillus plantarum M. sativum E. cerevisiae Schizosaccharomyces pombe Tinea corporis Tinea cruris Tinea pedis Antibacterial B. San-Blas et al. oxysporum Fusarium semitectum Fusarium udum Hanseniaspora valbyensis Kluyveromyces lactis Paracoccidioides brasiliensis Pichia anomala S. Curvularia sp. (1998). Ankri and Mirelman (1999). Yoshida et al. (1989). Chen et al. cepa A. (2004) K. Perez et al. aeruginosa S. niger C. Millet et al. (1994). pneumoniae L. fumigatus A. fumigatus Tsao and Yin (2001). Yin and Cheng ((2003)). flavus A. aureus 123 . sativum Diallyl monosulphide (DAS) (4) Antifungal Tsao and Yin (2001) A. (2000). Tsao and Yin (2001). albicans C. Kim et al. vortens Trypanosoma cruzi Antiviral Weber et al. (1992) Herpes simplex virus type 1 Herpes simplex virus type 2 Parainfluenza virus type 3 Rhinovirus type 2 Vaccinia virus Vesicular stomatitis virus A. pneumoniae L. Yin and Cheng (2003) K. krusei Antibacterial Camplobacter jejuni Chen et al. (2011) S. typhimurium S. monocytogenes S. monocytogenes S. albicans Candida glabrata C. O’Gara et al. (1993). (1996). aureus Streptomyces griseus Xanthomonas maltophilia Antiparasitic Plasmodium berghei Urbina et al. cereus Camplobacter jejuni H. krusei Candida utilis Antibacterial B. sativum Diallyl disulphide (DADS) (5) Antifungal Allium odorum A. pylori Naganawa et al. (2004) A. niger C. flavus A. (1999). Tsao and Yin (2001). Kim et al. aureus A. (1999). typhimurium S. glabrata C.Phytochem Rev Table 1 continued Species name Antimicrobial compounds Activity References P. histolytica Lang and Zhang (1981). Liu et al. Tsao and Yin (2001). sativum Diallyl tetrasulphide (DATTS) (7) Antifungal Allium odorum A. (2004) S. lamblia Trichomonas vaginalis Trypanosoma brucei Trypanosoma congolese Trypanosoma equiperdum Trypanosoma evansi A. Dimethyl trisulfide (9). utilis Antibacterial H. cepa A. aureus A. Diproyl trisulfide (11). (2009) Allium odorum A. Kim et al. aureus Kim et al. (2004) A. fumigatus A. Diethyl tetrasulfide. (1994) G. flavus A. flavus A. krusei C. pylori Tsao and Yin (2001). (2004) . glabrata C. fumigatus Tsao and Yin (2001). sativum A. pylori Shen et al. niger C. O’Gara et al. niger C. Kim et al. glabrata C. utilis Antibacterial S. (1996). (2000). cepa 123 Dipropyl disulfide (8). sativum Diallyl trisulphide (DATS) (6) Antifungal Yoshida et al. O’Gara et al. albicans C. albicans C. (2004) S. Lun et al. aureus Antiparasitic E. Dipropyl tetrasulfide (13) Antifungal C. Kim et al. utilis Penicillium expansum Antibacterial Cryptococcus neoformans H. krusei C. cepa A. Diethyl trisulfide (10). (2004). (1987). (2000). Dimethyl tetrasulfide (12).Phytochem Rev Table 1 continued Species name Antimicrobial compounds Activity References A. Kim et al. Tsao and Yin (2001). E)-1-propenyl ester (18. (1992) Herpes simplex virus type 1 Herpes simplex virus type 2 Parainfluenza virus type 3 Rhinovirus type 2 Vaccinia virus Vesicular stomatitis virus A. maltophilia A. luteus P. maltophilia 123 . 21). (1998). pombe Antibacterial B.Phytochem Rev Table 1 continued Species name Antimicrobial compounds Activity References A. sativum E-10 devinylajoene (14). sativum Methyl allyl thiosulfinate (17) Antiviral Weber et al. cereus B. coli K. Z-10 devinylajoene (15) Antifungal Yoshida et al. sativum Allyl methyl thiosulfinate (16) Antiviral Weber et al. Yoshida et al. pneumoniae M. aureus X. 2-propenesulfino thioic acid Smethyl ester (20. phlei P. aeruginosa S. pneumoniae M. (1999b) S. cerevisiae S. sativum S-(Z. valida H. anomala S. luteus M. lactis P. subtilis E. cerevisiae S. subtilis E. 19). 23) Antifungal Yoshida et al. (1999a) C. methanesulfino thioic acid S-(Z. E)-1-propenyl ester (22. (1992) Herpes simplex virus type 1 Herpes simplex virus type 2 Parainfluenza virus type 3 Rhinovirus type 2 Vaccinia virus Vesicular stomatitis virus A. aeruginosa S. cereus B. valbyensis K. coli K. pombe Antibacterial B. aureus X. 2000) and protozoa (Perez et al. likely as a result of the interaction between the amino acid and the disulfide bonds of the compound (Naganawa et al.e. The majority of available studies focus on common. the few available studies indicate that ajoene has a more potent antiviral activity compared to allicin and other Allium derived organosulfur compounds (Weber et al. Surprisingly. 3) and seems to be related to the number of disulfide bounds. DATTS [ DATS [ DADS [ DAS (Tsao and Yin 2001). porrum and Allium nigrum. the addition of both cysteine and glutathione reduced the inhibitory effects of allicin because these two compounds react with allicin disulfide bond. we found that only 9 out of 62 studies concerning Allium saponins reported significant antifungal activity (Table 2). In particular. . Millet et al. This can be also obtained by hydrolysis of the furostanol aglycon with the releasing of the sugar unit(s) at C-26. Allium leucanthum. including Botrytis cinerea. they are C27 sterols in which cholesterol has undergone modifications to produce a spiroketal. Table 2). studies on rare or wild species are also available (e. onion (Allium cepa). 2007). 1998. The antimicrobial activity of allicin has been associated with the inhibition of sulfhydryl-dependent enzymes (Wills 1956) such as alcohol dehydrogenase. 1984). Saponins from Allium species has many biological properties including antispasmodic. only a few studies have investigated their antimicrobial properties. From the data in the literature it seems that the spirostanol skeleton is typical of garlic taxa. It is formed when allicin is dissolved in several solvents including edible oils (Block et al. Fig. Allium cyrilii. 1996). Antimicrobial activity has been reported for several allyl sulfide compounds (Table 1 and 4–7. minutoside B (29. (2007) reported three saponins. Ajoene takes its name from the Spanish word ajo which means garlic. In this review. Structurally. the addition of cysteine greatly reduces the inhibitory effect of ajoene. Barile et al. 1987. The different responses of microbial and mammalian cells to allicin is likely related to the action mechanisms of this compound. as well as within the genus Allium. while the furostanol skeleton is typical for onion taxa. and then producing a ketone function at C-22. named minutosides A-C tested against several fungal plant pathogens. 1996). fungi (Yoshida et al. leek 123 (Allium porrum). Both E. Among these. that is the spirostanol aglycon. 7) was the major saponin plant tissue (83. Moreover. Fusarium oxysporum and Rhizoctonia solani. (Table 1). Hydroxylation on the skeletal core has been commonly found at position 2. Allium minutiflorum. chinese onion (Allium chinense) and chives garlic (Allium tuberosum).and Z-ajoene exhibited broad-spectrum antimicrobial activity against grampositive and gram-negative bacteria (Yoshida et al. and RNA polymerase. Naganawa et al. 1992). cultivated species such as garlic (A. In particular.5 mg kg-1) and also showed the highest activity in inhibiting spore germination and hyphal growth of the tested fungi. i. sativum). Recent studies further reported that several saponins from the common garlic var. 5 and 6 as shown in Fig.Phytochem Rev Allicin has a low toxicity towards mammalian cell cultures with negative effects generally observed at concentrations higher than 60 lg ml-1 (Ankri and Mirelman 1999). Vincken et al. In many cases. while the other skeletons are both present at almost the same level (Lanzotti 2012). anti-inflammatory and antitumour activity (Sparg et al. thioredoxin reductase. the spiroketal function is derived from the cholesterol side-chain by a series of oxygenation reactions affording hydroxylation at C-16 and at one of the terminal methyls. 5) (Lanzotti 2012). As reported for allicin. cytotoxic. haemolytic. Saponins Saponins are widespread in the plant kingdom (Sparg et al. furostanol and cholestanol aglycon (Fig. 2011. However. an unstable intermediate that undergoes glycosilation reaction at C-26 thus obtaining the spirostanol aglycon or further reaction giving rise to the corresponding spiroketal. Figure 6 reports the more common sapogenins (24–28) found in these plants that constitute the spirostanol aglycon of the corresponding saponins. Fig. 3. 1994. 2004. The isolated saponins are based on spirostanol. This intermediate is transformed into the hemiacetal. In this review we found that saponins have been extracted and identified from at least 38 different Allium species (Table 2). The cholesterol aglycon has been only found in A. 6. thus preventing cell damage. Glutathione occurrence in mammalian cells may explain their limited sensitivity to allicin compared to microbial cells which contain a very small amount of this compound (Davis 2005). Ledezma et al.g. 2004). oxysporum Mucor sp. sativum One furostanol saponin Antifungal Matsura et al.sp. leucanthum Five spirostanol saponins Antifungal Mskhiladze et al. (2007) A. (1996b) A. porrum Four spirostanol saponins Antifungal Carotenuto et al. ursinum Two spirostanol saponins Antifungal Sobolewska et al. (2012) A. solani Sclerotium cepivorum Trichoderma atroviride T. (1988) C. (1999) F. cinerea T. minutiflorum Two furostanol and three spirostanol saponins Antifungal Barile et al. lycopersici Fusarium solani P. alternata Alternaria porri B. (2008b) A. solani T. sativum Ten furostanol and two spirostanol saponins Antifungal Lanzotti et al. cepa Three furostanol saponins Mortierella ramanniana Cytotoxic Antifungal Lanzotti et al. cinerea F. (2006) C. niger B. glabrata Candida parapsilosis Microsporum canis Trichophyton mentagrophytes Cytotoxic Allium vineale Nine spirostanol saponins Antifungal Chen and Snyder (1989) P. ultimum R. expansum Molluscicidal Biomphalaria pfeifferei Allium fistulosum Nine furostanol saponins Antihypoxic Lai et al.Phytochem Rev Table 2 List of saponins from the genus Allium and related biological activities Species name Compounds Activity References Allium ampeloprasum Six spirostanol saponins Antifungal Sata et al. cinerea F. harzianum A. harzianum A. albicans A. (1998) A. (2012a) B. cepa Eleven furustanol saponins Antispasmodic Corea et al. Phomopsis sp. (2005) 123 . (2010) Allium macrostemon Three furostanol saponins Antiplatelet Ou et al. oxysporum F. sativum One furostanol and three spirostanol saponins Antiplatelet Peng et al. harzianum A. oxysporum f. culmorum A. R. (2012b) Alternaria alternata A. (1993) 123 . (1994) A. Ada˜o et al. (2002) A. (2012) A. (1999c) Mskhiladze et al. (2011a). (2005) Allium hirtifolium Five furostanol and four spirostanol saponins Antispasmodic A. tropeana Eight furostanol saponins Not assessed Dini et al. (1976) Allium albiflorus One saponin Not assessed Ismailov and Aliev (1976) Allium albopilosum Two spirostanol and four cholestanol saponins Not assessed Mimaki et al. giganteum One spirostanol saponin Not assessed Kawashima et al. (2005) A. (2011b) A. (1999a) Allium karataviense One furostanol. (2006) Allium aflatunense Five spirostanol saponins Cytotoxic Mimaki et al. cepa var.Phytochem Rev Table 2 continued Species name Compounds Activity References Barile et al. macrostemon Two furostanol saponins Cytotoxic. (2008a) A. chinense Two furostanol saponins Not assessed Peng et al. (1995) A. (1996a) Allium cyrillii Two spirostanol saponins Not assessed Tolkacheva et al. antitumour Chen et al. nigrum Eight spirostanol and two cholestanol saponins Cytotoxic Jabrane et al. (2006) A. (1992) A. (1991) A. leucanthum Seven spirostanol saponins Cytotoxic A. macrostemon Nine furostanol saponins Cytotoxic. (1991) Allium albanum Five saponins Not assessed Ismaiiov et al. (1976) Allium schubertii One furostanol and three spirostanol saponins Not assessed Kawashima et al. gastroprotective Ada˜o et al. anti-inflammatory. (2011) A. (1970) Allium jesdianum One spirostanol saponins Not assessed Mimaki et al. (2000) Allium macleanii Five spirostanol and one cholestanol saponins Antitumor Inoue et al. antitumour Chen et al. karataviense Four spirostanol saponins Not assessed Vollerner et al. five spirostanol saponins and one sapogenin Cytotoxic Mimaki et al. (2007) A. (2000) A. (2004) Allium erubescens One spirostanol saponins Not assessed Chincharadze et al. (1995) A. (1993) Allium rotundum One furostanol and one spirostanol saponin Not assessed Maisashvili et al. fistulosum Five spirostanol saponins Not assessed Do et al. (2012) Allium elburzense Eight furostanol and five spirostanol saponins Not assessed Barile et al. (2007) Allium atroviolaceum One sapogenin and one spirostanol saponin Not assessed Zolfaghari et al. chinense Two spirostanol saponins and one sapogenin Antitumor. chinense Six spirostanol saponins ATPase inhibition Allium giganteum One spirostanol and two furostanol saponins cAMP phosphodiesterase inhibition Mimaki et al. (1997) Allium senescens Two spirostanol saponins Antitumor Inoue et al. aflatunense One spirostanol saponin Not assessed Kawashima et al. ampeloprasum One spirostanol saponin Haemolytic. (2009) A. karataviense Three spirostanol saponins Not assessed Gorovits et al. (2012) Allium rubellum Four saponins Not assessed Ismaiiov et al. (1978) Allium narcissiflorum Two furostanol saponins Not assessed Krokhmalyuk and Kintya (1976) Allium nutans Two furostanol and two spirostanol Not assessed Akhov et al. (1995) Kuroda et al. (1993) A. (1999) Allium ostrowskianum One cholestanol saponin Not assessed Mimaki et al. ascalonicum Four furostanol saponins Not assessed Kang et al. (1979) A. fistulosum Seven sapogenins Not assessed Lai et al. (1999b) A. giganteum One sapogenin Not assessed Khristulas et al. porrum Six spiristanol and two cholestanol saponins Cytotoxic Fattorusso et al. porrum Four sapogenins Antitumor Carotenuto et al. tuberosum One spirostanol saponin Cytotoxic Sang et al. macrostemon Eight furostanol saponins Cellular Ca2? activity Chen et al. (1973) A. Cytotoxic Baba et al. (1996) Allium triquetrum Ten furostanol saponins Not assessed Corea et al. This contrasts with saponins from other plant species Fig. 2012b) have antifungal activity versus several plant pathogens. tuberosum Three furostanol saponins Not assessed Sang et al. (1979) Allium waldsteinii Three sapogenins and five saponins Not assessed Gugunishvili et al. their role in plant resistance against pathogens still has to be elucidated. 2012a). Allium species can produce a great diversity of saponins that are stored in plant cells and have in vitro antifungal activity.Phytochem Rev Table 2 continued Species name Compounds Activity References Allium sphaerosephalon One furostanol saponin Not assessed Mimaki et al. (1999a) A. Sobolewska et al. 5 Aglycon biosynthesis in Allium saponins 123 . three furostanol. and onion (Lanzotti et al. However. (2003) A. 7 the chemical structures of voghieroside A (30) and ceposide B (31) are reported both based on furostanol aglycon. Indeed. (2006) demonstrated that two steroidal saponins from Allium ursinum inhibited in vitro two Candida species. tuberosum One spirostanol saponins Not assessed Zou et al. (2003) A. and one cholestanol saponins Not assessed Sang et al. (2001) Allium turcomanicum Five sapogenins and one saponin Not assessed Pirtskhalava et al. tuberosum Two spirostanol saponins Not assessed Sang et al. In Fig. (2006) Note that only in few cases (9 out of 62) antifungal activity was assessed Voghiera (Lanzotti et al. (1999b) A. tuberosum Four spirostanol. Phytochem Rev Fig. Bacillus mycoides and P. avenae is able to infect oat because it produces the enzyme avenacinase that detoxifies avenacins. cepa 123 Hernandez et al. A. syringae. campestris. 2000).Voghiera. Clavibacter michiganensis pv. and A. Streptomyces turgidiscabies. sativum var. Agrobacterium tumefaciens. future studies should assess if these compounds accumulate in plant tissues at concentrations which can inhibit pathogens in vivo. In contrast with organosulfur compounds. MartinFig. The different response of fungi. fluorescens) and an oomycete (Pythium ultimum). oomycetes and bacteria to saponins is likely related to . Bowyer et al. (1995) reported that the fungus Gaeumannomyces graminis var. seven bacteria (Xanhomonas campestris pv. and identify the molecular mechanisms involved in their antimicrobial activity. Pseudomonas syringae pv. Allium saponins have weak or negligible antibacterial activity. To clarify the biological significance of Allium saponins in plant defense. michiganensis. Barile et al. The five saponins inhibited the growth of all fungi but were ineffective against Pythium and the tested gram-negative and grampositive bacteria. the antifungal oat saponins. minutiflorum. 7 Examples of saponins isolated from A. 6 Sapogenins isolated from Allium such as oat (Avena sativa) and tomato (Salanum lycopersicum) for which the role of avenacin and atomatine in plant defense has been clearly demonstrated (Morrisey and Osbourn 1999. (2007) tested five saponins on eight fungi (Table 2). However. The limited toxicity of saponins to pathogens such as the oomycetes Pythium and Phytophthora may depend on the lack of sterols in their membranes (Arneson and Durbin 1968. although the mechanisms underlying the different sensitivities are still unclear. saponins. 2004) discovered ascalin and allicepin. A recent study reported that the antifungal activity of some saponins from A. for oxygenation at C-5. respectively. while negative effects have been found for a furostane aglycone. 2006.Phytochem Rev differences in their membrane structure. Hedera (Cioaca et al. For instance. most studies have addressed only the antifungal activity of single molecules. Because the infusion of garlic roots is used in traditional medicine as antielminthic. Barile et al. (2009) discovered from A.g. Data obtained from antifungal tests carried out with analogous saponins (Barile et al. Medicago (Avato et al. O’Donnell et al. 2012a. 2000) can be inhibitory against bacteria. cinerea and the biocontrol agent Trichoderma harzianum can be synergically enhanced when they are supplied in combination (Lanzotti et al. and increasing number of sugars at C-3. Barile et al. further studies are needed to clarify the mechanism of action of saponins and to determine how these compounds are detoxified by pathogenic fungi resulting in the degradation of their chemical structure with the loss of activity. N-feruloytyrosine (32) and N-feruloyltyramine (33) (Fig. Despite the fact that in Allium species organosulfur compounds. In particular. Avato et al. Several proteins and peptides with antimicrobial activity have been found. Lanzotti et al. Fig. Wang and Ng (2002. and a spirostane aglycone. it has been found in garlic and leek roots and bulbs the presence of high amounts of two N-feruloyl amides. where the pharmacophoric elements for this activity are highlighted. Both compounds were active against the pathogen Fusarium culmorum with ED50 of 20 and 22 lg/ml. 9) (Fattorusso et al. comparative studies between fungi and bacteria usually indicate that fungi are more sensitive than bacteria (e. respectively. 1999). thus damaging the membrane with consequent cell collapse (Morrisey and Osbourn 1999). 1995). 1978). Table 3). alkaloids and antimicrobial proteins occur together within plant tissues. the possible antimicrobial activity of these two compounds was evaluated. 2012b). more recently. the 10 kD AceAMP1 protein from A. positive effects have been found for hydroxyls at positions 3 and 6. 2007). ascalonicum and A. b) allowed the definition of some structure– activity relationships that are summarized in Fig. 8 Key pharmacophoric elements for the antifungal activity of Allium saponins 123 . Recently. to saponins. It has been hypothesized that saponins exhibit their antimicrobial activity by forming complexes with the sterols that are present in the membrane of microorganisms. 2007). Some reports indicate that saponins from non-Allium plant species such as Acacia (Mandala et al. cepa against the plant pathogen B. However. 2007. other antimicrobial compounds that belong to different chemical classes have been discovered (Fig. cepa. 8. In addition. cepa inhibited twelve fungal and two bacterial species (Phillippe et al. 1. stipitatum Other organic compounds with antimicrobial activity Although the antimicrobial activity of Allium species has been ascribed to organosulfur compounds and. 2006) and Yucca (Wang et al. phenols. 2005). two peptides with antifungal activity from A. (2010) B. oxysporum Mycosphaerella arachidicola Physalospora piricola A. lycopersici Nectria haematococca Phoma betae Pyrenophora tritici-repentis Pyricularia oryzae Verticillium dahliae Antibacterial Bacillus megaterium Sarcina lutea Allium fistolosum Fistulosin (octadecyl 3-hydroxyindole) Antifungal F.sp. cepa Allicepin (peptide) Antifungal B. oxysporum F. Tassin et al. pisi F. ascalonicum Ascalin (peptide) Antifungal Wang and Ng (2002) B. monocytogenes M.Phytochem Rev Table 3 List of other organic compounds with antimicrobial activity extracted from the genus Allium Species name Compounds Activity References A. aeruginosa S. aureus A. cinerea A. cereus E. (2010) M. luteus S. aureus A. cepa Kaempferol (Flavon) (39) Antibacterial Santas et al. coli L. culmorum F. (1995). solani Fusarium moniliforme Verticillium dahliae Penicillium roqueforti Aspergillus oryzae Magnaporthe grisea Rhizopus oryzae 123 Phay et al. (1998) B.sp. cepa Quercetin (Flavon) (41) Antibacterial Santas et al. luteus P. cepa Protein (10 kD—AceAMPl) Antifungal Alternaria brassicola Ascochyta pisi Phillippe et al. cinerea Colletotrichum lindemuthianum F. (1999) . oxysporum f. cinerea Wang and Ng (2004) F. oxysporum f. flavonoids. e. quercetin (41) and kaempferol (39). Listeria monocytogenes. sativum N-amides (32–33) A. (1999) F. culmorum Pyridine-N-oxide alkaloids (34–36) Antibacterial O’Donnell et al. oxysporum M. that make garlic and Allium species effective against pathogenic microbes. the major flavonoids from A. although other mechanisms could not be excluded. tuberosum Chitinase-like protein (36 kDa) Lam et al. cinerea Coprinus comatus F. Although preparations containing these compounds have been used for centuries to treat human diseases. E.Phytochem Rev Table 3 continued Species name Compounds Activity References A. it seems that the activity of quercetin and the flavonoids with B-ring hydroxylation. aureus M. garlic has been used as an antimicrobial remedy in folk medicine. Conclusions From the ancient Egyptian period to the discovery of antibiotics. saponins. arachidicola Physalospora piricola A. 9) with potent antibacterial activity on fast-growing species of Mycobacterium. (2000) Antifungal B. and many groups have isolated and identified the structures of flavonoids possessing antifungal. Pseudomonas aeruginosa and S. nitrogen compounds and peptides. porrum Allium stipitatum Antifungal Fattorusso et al. methicillin-resistant S. Concerning the mechanism of action. sativum Allivin (36 kDa protein) Wang and Ng (2001) Antifungal B. (2009) S. Flavonoids are ubiquitous in photosynthesising cells and are commonly found in fruits and vegetables. Isolation and identification of other antimicrobial compounds should be pursued to meet the urgent need for new classes of drugs. However. 1993). lindemuthianum Trichoderma viride Trochoderma hamatum A. porrum Chitinase protein (34 kDa) Antifungal Vergawen et al. and a multidrug-resistant (MDR) variants of S. Scientific investigations have demonstrated that besides the well-studied sulphur compounds there are other classes of compounds. arachidicola R. only recently this class of natural products is becoming the subject of research about infectious diseases. phlei M. the development of more effective antimicrobial agents or a group of agents can also be achieved through 123 . Micrococcus luteus.g. coli. antiviral and antibacterial activity. In particular. could be partially attributed to the inhibition of DNA gyrase (Ohemeng et al. The common flavonoids found in garlic and onion are reported in Fig. showed antibacterial activity to Bacillus cereus. aureus (Table 3). smegmatis Mycobacterium fortuitum A. aureus. aureus. cepa. Fig. (1998) Cercospora musae C. solani three pyridine-N-oxide alkaloids (34–36. Despite the great number of antimicrobial compounds isolated. 10 (37–42) with their content in fresh bulbs. Allium species may still be an important source of such molecules. cinerea M. Phytochem Lett 4:306–310 Akhov LS. Future studies can address this issue by testing combinations of antimicrobial compounds (e. For instance. characterization of the interaction between the antimicrobial compounds and their target sites could reduce resistance mechanisms thus allowing the 123 design of second generation inhibitors that are more active and less toxic. da Silva BP. Microbes Infect 1:125–129 . da Silva BP. Fitoterapia 82:1175–1180 Ada˜o CR.d.g. Further studies are also required to completely understand the mechanism of action of the known compounds. Pizza C. it has been shown that these compounds may synergistically increase their activity when they work together. Existing structure–activity data suggest that it might be possible.d. n.d. Quercetin (41) OH OH OH OH H OH 1497 47 Myricetin (42) OH OH OH OH OH OH n. n. porrum. Parente JP (2011a) A new steroidal saponin with antiinflammatory and antiulcerogenic properties from the bulbs of Allium ampeloprasum var.d. Musienko MM.a. porrum with antiinflammatory and gastroprotective effects.d. References Ada˜o CR. Oleszek W (1999) Structure of steroidal saponins from underground parts of Allium nutans L. 693 ____________________________________________________________________________________ m. In addition. to prepare potent antibacterial saponins by obtaining compounds with a spirostanol skeleton and oxygenation at C-3 and C-6. 9 Aromatic N-amides and N-oxides from Allium Fig. Piacente S. 10 Flavonoids in onion and garlic and their contents Compound Substituents Onion content Garlic content 3 3’ 4’ 5 5’ 7 mg/Kg mg/Kg _________________________________________________ ___________________________________ Apigenin (37) H H OH OH H OH n. Parente JP (2011b) A new steroidal saponin from Allium ampeloprasum var. 217 Isorhamnetin (38) OH H OH OH H OMe m.d.Phytochem Rev Fig. Mirelman D (1999) Antimicrobial properties of allicin from garlic. This is an important step to be able to use these antimicrobial compounds to the maximum of their potential. Luteolin (40) H OH OH OH H OH 391 n. organosulfur compounds and saponins) that co-occur in plant tissues. minor amounts. for example. Kaempferol (39) OH H OH OH H OH 832 n. J Agric Food Chem 47:3193–3196 Ankri S. not detected structural modifications of the already known compounds.a. Maisashvili MR (2006) Steroidal saponins from Allium waldsteinii. Sajjadi SE. Shibata S. senescens. Abubakirov NK (1973) Steroid saponins and sapogenins from Allium. Nishino A. Clarke BR.: structure–activity relationship. Wang N. Lanzotti V (2004) Saponins of Allium elburzense. Sajjadi SE. Dini A (2005) Furostanol saponins in Allium cepa L. Durbin RD (1968) The sensitivity of fungi to atomatine. J Asian Nat Prod Res 8:21–28 Chen H. Science 267:371–374 Carotenuto A. Pharmazie 33:609–610 Corea G. Kishi N. Jain MK. Ohmura M. Ahmad S. I. Vlaicu B. physical properties and antibacterial action. Chem Nat Compd 42:499–500 Gupta KC. new antiproliferative sapogenins from Allium porrum. Zolfaghari B. tropeana seeds. Kelginbaev AN. Okuyama T (2000) Saponins isolated from Allium chinense G. Var. Isolation. Lanzotti V. Snyder JK (1989) Diosgenin-bearing. Lanzotti V (2003) Saponins and flavonoids of Allium triquetrum. Gorovits MB. Bucci R. Sci Am 252:114–119 Block E. Tenore GC. Wang G. Trimarco E. J Agric Food Chem 53:935–940 Cutler RR. Mycoses 48:95–100 Dini I. var. Carnuccio R. Cucu V (1978) The saponins of Hedera helix with antibacterial activity. Antibiot Chemother 5:24–27 Hallahan DL (2000) Monoterpenoid biosynthesis in glandular trichomes of Labiate plants. Son KH (1992) Steroidal saponins from the subterranean part of Allium fistulosum. Eristavi LI. Mimaki Y. Int J Mol Sci 13:1426–1436 Barile E. Phytochemistry 68:596–603 Block E (1985) The chemistry of garlic and onions. Fattorusso E. Scala F. Zolfaghari B. Gorovits TT. Phytopathology 58:536–537 Avato P. Tropea. J Am Chem Soc 66:1950–1951 Chen S. Lanzotti V. Fattorusso E. J Nat Prod 66:1405–1411 Corea G. Plant Biosyst 133:199–203 Fattorusso E. Kotik AN. and their inhibitory activity on tumour 123 . Tava A. Cruz MR (1984) (E. Streptomycin and allicin. Sajjadi SE. 15. Sundstrom DC. Chung JG. Zolfaghari B (2005) Structure-activity relationships for saponins from Allium irtifolium and Allium elburzense and their antispasmodic activity. Izzo A. Apitz-Castro R. Rosato A. Lin JG (1999) Effects of the garlic compounds diallyl sulphide and diallyl disulphide on arylamine N-acetyltransferase activity in Klebsiella pneumoniae. 3-seco-porrigenin. Daniels MJ. Tetrahedron 53:3401–3406 Carotenuto A. Curr Opin Plant Biol 5:1–6 Inoue T. Eruboside-B from Allium erubescens. Wang Z. Lanzotti V. Viswanathan R (1955) Combined action of streptomycin and chloramphenicol with plant antibiotics against tubercle bacilli. Crecely RW. Karatavigenin. Gvazava LN. Jung KY. Yao X (2007) New furostanol saponins from the bulbs of Allium macrostemon Bunge and their cytotoxic activity. Chem Nat Compd 6:747–749 Gugunishvili DN. Wang NL. Butnariu M (2012) Chemical composition and in vitro antifungal activity screening of the Allium ursinum L. IV. Bonanomi G. Khristulas FS. Nadler M. Brit J Biomed Sci 61:71–74 Davis SR (2005) An overview of the antifungal properties of allicin and its breakdown products—the possibility of a safe and effective antifungal prophylactic. Peng J. Ho HC. I. Pharmazie 62:544–548 Chen HF. J Org Chem 54:3679–3689 Chen GW. Capasso R. Jurzysta M (2006) Antimicrobial activity of saponins from Medicago sp. Luo Q. Wang X. II. Lunness P. Don and antitumor-promoting activities of isoliquiritigenin and laxogenin from the same drug. J Am Chem Soc 106:8295–8296 Bowyer P. Lanzotti V (2007) Saponins from Allium minutiflorum with antifungal activity. Magno S (1999) Spirostanol saponins of Allium porrum L. Wang NL. Abubakirov N (1979) Steroidal saponins and sapogenins of Allium. stable. J Nat Prod 67:2037–2042 Barile E. Lanzotti V. Eristavi LI. Food Chem 93:205–214 Dixon RA (2001) Natural products and plant disease resistance. Yao XS (2009) Two new steroidal saponins from Allium macrostemon Bunge and their cytotoxity on different cancer cell lines. J Nat Prod 55:168–173 Fattorusso E. Phytother Res 20:454–457 Baba M. Lanzotti V. the antibacterial principle of Allium sativum. Antignani V. Bailey JH (1944) Allicin. Capasso R. Bialy Z. Fattorusso E. aqueous extract of allicin against methicillin-resistant Staphylococcus aureus. Osbourn AE (1995) Host range of a plant pathogenic fungus determined by a saponin detoxifying enzyme. J Agric Food Chem 48:3455–3462 Feldberg RS. Molecules 14:2246–2253 Chincharadze DG. Sashida Y. Izzo AA (2005) Antispasmodic saponins from bulbs of red onion. Nature 411:843–847 Do JC. Ianaro A (2000) Cytotoxic saponins from bulbs of Allium porrum L. Thompson NH (1988) In vitro mechanism of inhibition of bacterial cell growth by allicin. D’Acquisto F (1997) 12-Keto-porrigenin and the unique 2.Phytochem Rev Arneson PA. J Appl Toxicol 19:75–81 Chen HF. Planta Med 71:1010–1018 Barile E. Biol Pharm Bull 23:660–662 Bagiu RV. Antimicrob Agents Chemother 32:1763–1768 Gorovits MB. Streptomycin and chloramphenicol with cepharanthine. Wilson P (2004) Antibacterial activity of a new. Z)-Ajoene: a potent antithrombotic agent from garlic. Lanzotti V. Allium cepa L. Adv Bot Res 31:121–151 Hammerschmidt R (1999) Phytoalexins: what have we learned after 60 years? Annu Rev Phytopathol 37:285–306 Heil M (2002) Ecological costs of induced resistance. Nishino H (1995) Steroidal glycosides from Allium macleanii and A. Magno S. Fattorusso E. Chang SC. Sun HL. Vitali C. Chem of Nat Comp 15:442–446 Cioaca C. Satomi Y. Wang GH. Taglialatela-Scafati O (1999) Antifungal N-feruloyl amides from roots of Allium species. Neuwirth Z. and their bioactivity on [Ca2?] increase induced by KCl. Margineanu C. Okada Y. Taglialatela-Scafati O. Yang M. molluscicidal saponins from Allium vineale: an NMR approach for the structural assignment of oligosaccharide units. Di Rosa M. (Liliaceae). Phytochemistry 51:1077–1082 Cavallito CJ. Yang BF. a new sapogenin from Allium karataviense. Yao XS (2006) Novel furostanol saponins from the bulbs of Allium macrostemon B. Nikaido T (1995) Steroidal saponins from Allium chinense and their inhibitory activities on cyclic AMP phosphodiesterase and Na? K? ATPase. Bonanomi G. Voghiera. Chem Pharmaceut Bull 42:710–714 Mimaki Y. Mimaki Y. Harzallah-Skhiri F. Osbourn A (2000) Effects of targeted replacement of the tomatinase gene on the interaction of Septoria lycopersici with tomato plants. Ushiroguchi T. Kameyama A. J Chromat A 1112:3–22 Lanzotti V (2012) Bioactive polar natural compounds from garlic and onion. Sinha Babub SP. Saunders RA. Antimicrob Agents Chemother 43:3045 Kang LP. Wu Z. Fukasawa T. In: Zhengyi W. Chem Pharm Bull 36:3659–3663 Millet COM. Harada N (1996) A new furostanol saponin with six sugars from the bulbs of Allium sphaerosephalon. Phytochemistry 40:521–525 Ismailov AI. Science Press. Sashida Y (1999b) Steroidal glycosides from the bulbs of Allium jesdianum. J Nat Prod 73:1053–1057 Lai W. Barile E. Ohmoto T (1994) New steroidal saponins from the bulbs of Allium giganteum exhibiting potent inhibition of cAMP phosphodiesterase activity. Chen WS (2012) New steroidal sapogenins from the acid hydrolysis product of the whole glycoside mixture of Welsh onion seeds. Chen W (2010) Antiischemia steroidal saponins from the seeds of Allium fistulosum. Sun LN. Melto R. Chin Chem Lett 23:193–196 Lam YW. Zhang L. Luchanskaya VN. Sashida Y (1999c) Steroidal saponins from the bulbs of Allium karataviense. Phytochemistry 32:1267–1272 Khristulas FS. J Nat Prod 62:194–197 Mimaki Y. Yang YB. Menzinger M. bulbs of Allium sativum L. Sashida Y (1993) Steroidal saponins from the bulbs of Allium schubertii. Zhao Y. Mirjolet JF. Cable J (2011) Effect of garlic and Alliumderived products on the growth and metabolism of Spironucleus vortens. Kuroda M. Ng TB (2000) A robust cysteine-deficient chitinase-like antifungal protein from inner shoots of the edible chive Allium tuberosum. J Am Acad Dermatol 43:829–832 Liu P.). Kyung KH (2004) Antimicrobial activity of alk (en) yl sulfides found in essential oils of garlic and onion. Phytochemistry 34:799–805 Mimaki Y.. Structural elucidation by modern NMR techniques. ostrowskianum. Chem Nat Comp 12:46–47 Kuroda M. Mol Plant-Microbe Interact 13:1301–1311 Matsuura H. Evans G. Sun L. Biochem Biophys Res Commun 279:74–80 Lang YJ. Lanzuise S. Mandal NC (2005) Antimicrobial activity of saponins from Acacia auriculiformis. De Sousa L. Hayashi N. Mimaki Y. Li H. Hugouvieux V. Stobberingh E (1999) Effect of garlic on vancomycin resistant enterococci. Farmatsiia 25:17–20 Ismailov AI. Scala F (2012a) Antifungal saponins from bulbs of garlic. Zhang KY (1981) Studies on the effective components of garlic (Allium sativum L. Williams D. var. Mimaki Y. Chen HB. Kyung SH. Fitoterapia 76:462–465 Martin-Hernandez AM. Fuwa T (1988) A furostanol glycoside from garlic. Exp Parasitol 127:490–499 Mimaki Y. Entamoeba histolytica and Giardia lamblia) in vitro. The structure of glycosides A and B. Chai Y. Li X. Phytochem Rev 11:179–196 123 Lanzotti V. Kuchukhidze DK. Magn Reson Chem 45:725–733 Kawashima K. Apitz Castro R (1999) Ajoene in the topical short-term treatment of Tinea cruris and Tinea corporis in humans. Allium cepa L. Fukasawa T. Kameyama A. Phytochemistry 40:1071–1076 Lai W. Jorquera A. Long C. Arzneimittelforschung 49:544–547 Ledezma E. Cheng Y. Gvazava LN (2012) Steroidal glycosides of gitogenin from Allium rotundum. Liu H. Abubakirov NK (1970) New steroidal sapogenin from Allium giganteum. Phytochemistry 30:3063–3067 Kawashima K. Marin P. Lacaille-Dubois MA (2011) Spirostane and cholestane glycosides from the bulbs of Allium nigrum L. Deng B (2009) Diallyl trisulfide (DATS) effectively induced apoptosis of postharvest disease Penicillium expansum of citrus. Nat Med 53:88–93 Mimaki Y. Romano A. albanum. Randomized comparative study with terbinafine. Sashida Y. Louis Jonkers D. Chem Nat Compd 48:86–90 Mandala P. Deyuan H (eds) 1994–2009: Flora of China 24. Romero H. Kikoladze VS. Apitz-Castro R (2000) Efficacy of ajoene in the treatment of Tinea pedis: a double-blind and comparative study with terbinafine. Guo J. Missouri Botanical Garden. Kawashima K. Gorovits MB.Phytochem Rev promoter-induced phospholipid metabolism of hela cells. Chem Lett 25:431–432 Mimaki Y. Burri C. Beijing. Sashida Y. Phytochem Rev 4:95–110 Lanzotti V (2006) The analysis of onion and garlic. Dufresne M. Chin Herb Med 4:4–6 Lanzotti V (2005) Bioactive saponins from Allium and Aster plants. Phytochemistry 74:133–139 Ledezma E. Ferrara G. Li T. Wang HX. Khim Prir Soed 4:550–551 Jabrane A. Aliev AM (1976) Determination of the steroid saponins in the onion (Allium albiflorus) that grows in Azerbaijan. Matsumoto K. aflatunense. Raven PH. Lin H. Bonanomi G. Food Sci Biotechnol 13:235–239 Krokhmalyuk VV. Antignani V. Itakura Y. Marcano K. Ann Microbiol 59:675–679 Lun ZR. Huh JE. Glycosides of Allium narcissiflorum glycosides. Food Chem 125:447–455 Jiemei X. Satou T. Liu ZJ. Sluimer J. Phytochemistry 78:126–134 Lanzotti V. Tagiev SA. Kintya PK (1976) Steroid Saponins 10. Kaminsky R (1994) Antiparasitic activity of diallyl trisulfide (Dasuansu) on human and animal pathogenic protozoa (Trypanosoma sp. Wu ZJ. Sashida Y. Allium sativum L. Nikaido T. Wang B. Kanmoto T. Williams C. Kuroda M. Kuroda M. Lloyd D. Ma BP (2007) New furostanol saponins from Allium ascalonicum L. Pulgar M. Ben Jannet H. De Sousa L. Miyamoto T. Sashida Y (1993) Steroidal glycosides from Allium albopilosum and A. Chem Pharmaceut Bull 47:738–743 . Lopez JC. Padilla M. Khim Prir Soedin 6:489–490 Kim JW. Rasulov EM (1976) Steroidal saponins and sapogenins from Allium rubellum and A. Kuroda M. Ann Soc Belg Med Trop 74:51–59 Maisashvili MR. Kamelin RV (2000) Allium Linnaeus: 165–202. Sashida Y (1999a) Steroidal saponins from the bulbs of Allium aflatunense. Scala F (2012b) Antifungal saponins from bulbs of white onion. Tan DW. St. Sashida Y (1991) Steroidal saponins from Allium giganteum and A. Duchamp O. Zhao Y. Jorquera A. Janeczko Z. Ma LR. Kikuchi T (1996a) Furostanol glycosides from bulbs of Allium chinense. Yao Xue Xue Bao 31:607–612 Perez HA. a sulfur-containing compound derived from garlic. Fujino T. Neidle S. Elias R. Eggermont K. Chen Z. Podolak I. Almajano MP. Fu KP. Fukushi Y. in vitro. Broekaert WF (1995) A potent antimicrobial protein from onion seeds showing sequence homology to plant lipid transfer proteins. Chen ZL (2002) Tuberoside M. Chem Nat Compd 15:446–452 San-Blas G. Apitz-Castro R (1989) Inhibition of growth of the dimorphic fungus Paracoccidioides brasiliensis by ajoene. Int J Food Sci Technol 45:403–409 Sata N. Piras R (1993) Inhibition of phosphatidylcholine biosynthesis and cell proliferation in 123 . Microbiol Mol Biol R 63:708–724 Mskhiladze L. Osbourn AE (1999) Fungal resistance to plant antibiotics as a mechanism of pathogenesis. Biochemistry 37:3623–3637 Tolkacheva NV. Lao A. Wilchek M. Am J Med Sci 344:261–267 Pasteur L (1858) Memoire sur la fermentation appele´e lactique. Toxicology 215:205–213 Ohemeng KA. Wang H. Lao AN. Chincharadze D. Zhong Y. Light ME. Takamura S. Wang H. and their diallyl constituents against Helicobacter pylori. Monheit D. Zimhony O. J Med Microb 50:646–649 Urbina JA. Favel A (2008a) In vitro antifungal and antileishmanial activities of steroidal saponins from Allium leucanthum C Koch-a Caucasian endemic species. Visbal G. Guerbette F. the active principle of garlic extract (Allium sativum). Georgian Med News 154:39–43 Mskhiladze L. Zhang X. Ishikawa K. Lawson LD (1996) Enhanced diallyl trisulfide has in vitro synergy with amphotericin B against Cryptococcus neoformans. Marin˜o L. Matsuura H. Matsunaga S.) extracts. Nishikawa H. Chen Z (1999b) Two new spirostanol saponins from Allium tuberosum. and its relation to the role of alkyl hydroperoxide reductase 1 as a cell surface defense in Saccharomyces cerevisiae. Chen HF. Kobayashi H (1996b) Two new steroidal saponins from Allium sativum and their inhibitory effects on blood coagulability. Legault J. Yokota A. Saito T (1998) New antifungal and cytotoxic steroidal saponins from the bulbs of an elephant garlic mutant. Arch Pharm 230:443 Shadkchan Y. Proost P. Carbo´ R (2010) Antimicrobial and antioxidant activity of crude onion (Allium cepa. Can J Bot 68:1354–1356 Sobolewska D. Plant Physiol 109:445–455 Pirtskhalava GV. Bioorg Med Chem Lett 3:225–230 Ou WC. Planta Med 62:415–418 Singh UP. Tezuka Y. J Nat Prod 62:1028–1029 Sang SM. Fujita KI. Appl Environ Microbiol 62:4238–4242 O’Donnell G. Gorovits TT. Marchan E. Chem Nat Compd 48:272–275 Tsao SM. Hirooka K. J Asian Nat Prod Res 4:67–70 Sang S. a new cytotoxic spirostanol saponin from the seeds of Allium tuberosum. Acta Pol Pharm 63:219–223 Sparg SG. Bhakta S. Gibbons S (2009) Bioactive pyridine-N-oxide disulfides from Allium stipitatum. Suzuki A (1996) Inhibition of microbial growth by ajoene. Lenaerts A. Ptak M. Poeschl R. an allyl sulfur compound from garlic. Fusetani N. Narui T. Kisiel W. Turoside C from Allium turcomanicum. San-Blas F. Hendriks M. Evangelopoulos D. Pichette A (2008b) Cytotoxic steroidal saponins from the flowers of Allium leucanthum. Appl Environ Microbiol 66:2269–2273 Ogita A. Cammue BPA. the reactive molecule of garlic. Kutchukhidze J. Wallace JM. Suzuki H. J Ethnopharmacol 94:219–243 Stearn WT (1992) How many species of Allium are known? Curtis’s Bot Mag 9:180–182 Tassin S. Hill DJ. Abubakirov NK (1979) Steroid saponins and sapogenins of Allium. Aguirre T. Sodano P (1998) Solution structure of AceAMP1. Apitz R (1994) In vivo activity of ajoene against rodent malaria. Antimicrob Agents Chemother 38:337–339 Phay N. Pandey VN. Liu BR. and in a murine model of disseminated aspergillosis. Barrett JF (1993) DNA gyrase inhibitory and antibacterial activity of some flavones. Wagner KG. Yin MC (2001) In-vitro antimicrobial activity of four diallyl sulphides occurring naturally in garlic and Chinese leek oils. J Antimicrob Chemother 53:832–836 Shen J. Varon S (1987) Inhibition of growth of Entamoeba histolytica by allicin. Marion D. J Infect Dis 156:243–244 Morrisey JP. Molecules 13:2925–2934 Naganawa R. Trojanowska D (2006) Steroidal glycosides from the underground parts of Allium ursinum L. Kader JC.Phytochem Rev Mirelman D. Volvelle F. Shashkov AS. Piras MM. Lao A. Liu SM. Phytochemistry 52:271–274 Phillippe B. Biosci Biotechnol Biochem 62:1904–1911 Semmler FW (1982a) Oil of garlic. Boshoff HI. Broekaert WF. Mshvildadze V. Chen H. Kuchukhidze J. Maslin DJ (2000) Activities of garlic oil. Shemesh E. Singh K (1990) Antifungal activity of ajoene. XVI. Miron T. Cai Y. Yamamoto Y. Delmas F. Antimicrob Agents Chemother 33:1641–1644 Sang S. Tsuji M. Van Damme J. Goderis IJ. Schwender CF. Thevissen K. a potent antimicrobial protein extracted from onion seeds. Fukuda H. garlic powder. Taniguchi M. Acland DP. Mirelman D. Food Chem 83:499–506 Santas J. Rabinkov A. Apitz-Castro R. J Nat Prod 72:360–365 O’Gara EA. a constituent of garlic (Allium sativum). Elias R. Tomita F (1999) An antifungal compound from roots of welsh onion. Zou ML. Qiao YQ. Gorovits MB. Chirva VY (2012) Steroidal glycosides from Allium cyrillii bulbs. Lavoie S. De La Rosa M. Gil F. and their cytostatic and antimicrobial activity. Malkinson JP. Chen KJ (2012) Inhibition of platelet activation and aggregation by furostanol saponins isolated from the bulbs of Allium macrostemon Bunge. van Staden J (2004) Biological activities and distribution of plant saponins. Okuyama T. Ann Chim Phys Ser 52:404–418 Peng JP. in inhibiting Aspergillus spp. Phytochemistry 52:1611–1615 Sang S. Gunaratnam M. Mao S. Tsutsui N. Lao A. Osborn RU. Arch Pharm 230:434 Semmler FW (1982b) Onion oil. Chen Z (1999a) Furostanol saponins from Allium tuberosum. Davis LE. Higashiyama T. Iwata N. L. Moreira JB. Gil F. Tanaka T (2005) Synergistic fungicidal activity of Cu2? and allicin. Lazardi K. Yao XS. Osherov N (2004) Efficacy of allicin. Phytochemistry 41:283–285 Peng JP. Galanty A. Ho C-H (2003) New steroid saponins from the seeds of Allium tuberosum L. Abubakirov NK (1978) Steroid saponins and sapogenins of Allium XIV. Cheeke PR (2000) Effect of steroidal saponin from Yucca schidigera extract on ruminal microbes. Biosci Biotechnol Biochem 63:591–594 Zolfaghari B. Phytochemistry 68:275–297 Vollerner YS. Meat Sci 63:23–28 Yoshida S. Biochem J 63:514 Wittstock U. J Appl Microbiol 88:887–896 Weber ND. Ng TB (2004) Isolation of Allicepin. Suzuki A (1998) Antimicrobial activity of a compound isolated from an oil-macerated garlic extract. Gorovits MB. Fukuda H. Phytochemistry 57:1219–1222 . Ishikawa K. J Pept Sci 10:173–177 Wang Y. Katsuzaki H. Tindall & Cassel. Ushiroguchi T. Gorovits TT. and its antimicrobial effect. Chem Nat Compd 14:630–635 Wang HX. a new anti-fungal peptide with human immunodeficiency virus type 1 reverse transcriptase-inhibiting activity from shallot bulbs. Yu DQ. Biochem Pharmacol 45:2381–2387 Vergawen R. Yanke LJ. Ng TB (2002) Ascalin. Cong PZ (2001) A steroidal saponin from the seeds of Allium tuberosum. Hayashi N. Ohta R. Planta Med 58:417–423 Wertheim T (1844) Protozoology. Biosci Biotechnol Biochem 63:588–590 Yoshida H. Curr Opin Plant Biol 5:1–8 Yamada Y. Ohta R. p 251 123 Wills ED (1956) Enzyme inhibition by allicin. an antiplatelet compound isolated from garlic. a novel antifungal peptide from onion (Allium cepa) bulbs. Heng L. The structure of karatavoiside A. a novel antifungal protein from bulbs of the round-cloved garlic. Ng TB (2001) Purification of allivin. Gershenzon J (2002) Constitutive plant toxins and their role in defense against herbivores and pathogens. classification and occurrence in the plant kingdom. Biosci Biotechnol Biochem 62:1014–1017 Yoshida H. Lanzotti V (2006) The sapogenin atroviolacegenin and its diglycoside atroviolaceoside from Allium atroviolaceum. de Groot A.Phytochem Rev Trypanosoma cruzi by ajoene. Iwata N. Physiol Plant 164:175–182 Vincken JP. Suzuki A (1999a) An organosulfur compound isolated from oil-macerated garlic extract. J Nat Prod 69:191–195 Zou ZM. Capasso R. Cheng WS (2003) Antioxidant and antimicrobial effects of four garlic-derived organosulfur compounds in ground beef. Life Sci 70:357–365 Wang HX. Appl Environ Microbiol 53:615–617 Yoshida H. Lawson LD. Katsuzaki H. Izzo AA. Van Leuven F. Sajjadi SE. Gruppen H (2007) Saponins. the active principle of garlic. Baillie`re. Naganawa R. Murray BK. Fukuda H. North JA. Ishikawa K. McAllister TA. Andersen DO. Van Laere A (1998) Purification and characterization of strongly chitin-binding chitinases from salicylic acid-treated leek (Allium porrum). Matsuura H. Katsuzaki H. Azuma K (1977) Evaluation of the in vitro antifungal activity of allicin. Fujino T. Suzuki A (1999b) Antimicrobial activity of the thiosulfinates isolated from oil-macerated garlic extract. Kasuga S. Fukuda H. Hughes BG (1992) In vitro virucidal effects of Allium sativum (garlic) extract and compounds. Fujino T. Antimicrob Agents Chemother 11:743–749 Yin MC. Barile E. Ishikawa K. Peptides 23:1025–1029 Wang HX. Nakagawa S (1987) Antifungal activity of ajoene derived from garlic.