Antibacterial activity of leaves and inter-nodal callus extracts of Mentha arvensis L
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摘要
<正>Objective:To determine the anti-bacterial efficacy of chloroform,ethanol,ethyl acetate and water extracts of inter-nodal and leaves derived calli extracts from Mentha arvensis(M.arvensis) against Salmonella typhi(S.typhi),Streptococcus pyogenes(S.pyogenes),Proteus vulgaris(P. vulgaris) and Bacillus subtilis(B.subtilis).Methods:The inter-nodal and leaves segments of M.arvensis were cut into 0.5-0.7 cm in length and cultured on Murashige and Skoog solid medium supplemented with 3%sucrose,gelled with 0.7%agar and different concentration of 2,4-Dichlorophenoxyacetie acid(2,4-D) either alone or in combinations.The preliminary phytochemical screening was performed by Brindha et al method.Antibacterial efficacy was performed by disc diffusion method and incubated for 24 h at 37℃.Results:Maximum percentage of callus formation(inter-nodal segments 84.3±0.78;leaves segments 93.8±1.27) was obtained on Murashige and Skoog's basal medium supplemented with 3%sucrose and 1.5 mg/L of 2,4-D.The ethanol extracts of leaves derived calli showed the maximum bio-efficacy than other solvents.The leaves and stem derived calli extracts on Proteus sp.showed that the plants can be used in the treatment of urinary tract infection associated with Proteus sp.Through the bacterial efficacy studies,it is confirmed that the in vitro raised calli tissue was more effective compared to in vivo tissue.Conclusions:The bio-efficacy study confirmed that the calli mediated tissues showed the maximum zone of inhibition.The present study paved a protocol to establish high potential cell lines by in vitro culture.
Objective:To determine the anti-bacterial efficacy of chloroform,ethanol,ethyl acetate and water extracts of inter-nodal and leaves derived calli extracts from Mentha arvensis(M.arvensis) against Salmonella typhi(S.typhi),Streptococcus pyogenes(S.pyogenes),Proteus vulgaris(P. vulgaris) and Bacillus subtilis(B.subtilis).Methods:The inter-nodal and leaves segments of M.arvensis were cut into 0.5-0.7 cm in length and cultured on Murashige and Skoog solid medium supplemented with 3%sucrose,gelled with 0.7%agar and different concentration of 2,4-Dichlorophenoxyacetie acid(2,4-D) either alone or in combinations.The preliminary phytochemical screening was performed by Brindha et al method.Antibacterial efficacy was performed by disc diffusion method and incubated for 24 h at 37℃.Results:Maximum percentage of callus formation(inter-nodal segments 84.3±0.78;leaves segments 93.8±1.27) was obtained on Murashige and Skoog's basal medium supplemented with 3%sucrose and 1.5 mg/L of 2,4-D.The ethanol extracts of leaves derived calli showed the maximum bio-efficacy than other solvents.The leaves and stem derived calli extracts on Proteus sp.showed that the plants can be used in the treatment of urinary tract infection associated with Proteus sp.Through the bacterial efficacy studies,it is confirmed that the in vitro raised calli tissue was more effective compared to in vivo tissue.Conclusions:The bio-efficacy study confirmed that the calli mediated tissues showed the maximum zone of inhibition.The present study paved a protocol to establish high potential cell lines by in vitro culture.
Objective:To determine the anti-bacterial efficacy of chloroform,ethanol,ethyl acetate and water extracts of inter-nodal and leaves derived calli extracts from Mentha arvensis(M.arvensis) against Salmonella typhi(S.typhi),Streptococcus pyogenes(S.pyogenes),Proteus vulgaris(P. vulgaris) and Bacillus subtilis(B.subtilis).Methods:The inter-nodal and leaves segments of M.arvensis were cut into 0.5-0.7 cm in length and cultured on Murashige and Skoog solid medium supplemented with 3%sucrose,gelled with 0.7%agar and different concentration of 2,4-Dichlorophenoxyacetie acid(2,4-D) either alone or in combinations.The preliminary phytochemical screening was performed by Brindha et al method.Antibacterial efficacy was performed by disc diffusion method and incubated for 24 h at 37℃.Results:Maximum percentage of callus formation(inter-nodal segments 84.3±0.78;leaves segments 93.8±1.27) was obtained on Murashige and Skoog's basal medium supplemented with 3%sucrose and 1.5 mg/L of 2,4-D.The ethanol extracts of leaves derived calli showed the maximum bio-efficacy than other solvents.The leaves and stem derived calli extracts on Proteus sp.showed that the plants can be used in the treatment of urinary tract infection associated with Proteus sp.Through the bacterial efficacy studies,it is confirmed that the in vitro raised calli tissue was more effective compared to in vivo tissue.Conclusions:The bio-efficacy study confirmed that the calli mediated tissues showed the maximum zone of inhibition.The present study paved a protocol to establish high potential cell lines by in vitro culture.
引文
[1]Tomoko N,Takashi A,Hiromu T,Yuka I,Hiroka M,MuneKazu I, et al.Antibacterial activity of extracts preparated from tropical and subtropical plants on methicillin-resistant Staphylococcus aureus. J Health Sci 2002;48:273-276.
[2]Ben Sassi AF,Barzallah-Skhiri,Aouni M.Investigation of some medicinal plants from Tunisia for antimicrobial activities.Pharm Biol 2007;15(5):421-428.
[3]Coruh I,Gomez AA,Ercisli S.Total phenolics,mineral elements, antioxidant and antibacterial activities of some edible wild plants in Turkey.Asian J Chem 2007;19(7):5755-5762.
[4]Veronika S,John P M,Maita L,Michael C,Adam C,Carol Y, et al.Dual effect of plant steroidal alkaloids on Saccharomyces cerevisiae.Antimicrobial Agents Chemother 2006;50(8):2732- 2740.
[5]Doughari JH.Antimicrobial activity of Tamarindus indica Linn. Trop J Pharm Res 2006;5(2):597-603.
[6]Maghrani MN,Zeggwah JM,Eddouks M.Antihypertensive effect of Lepidium Sativum in spontaeneously hypertensive rats.J Ethnopharam 2005;102(1&2):193-197.
[7]Balandrin MJ,Klocke JA.Medicinal,aromatic and industrial materials from plants.Berlin:Bajaj Springer-Verlag;1988,p. 1-36.
[8]Samy RP,Ignacimuthu S.Antibacterial activity of some folklore medicinal plants used by tribals in Western Ghats in India.J Ethnopharmacol 2000;69:63-71.
[9]Omonkhelin JO,Eric KIO,Osohon O.Antifungal and antibacterial Activities of the ethanolic and aqueous extract of Kigelia africana (Bignoniaceae) stem bark,Afr J Biatechnol 2007;6(14):1677- 1680.
[10]Ogbulie JN,Okoli IC,Anyanwu BN.Antibacterial activities and toxicological potentials of crude ethanolic extracts of Euphorbia hirta.Afri J Biatechnol 2007;6(13):1544-1548.
[11]Mahesh B,Satish S.Antimicrobial activity of some important medicinal plant against plant and human pathogens.World J Agric Sci 2008;4(S):839-843.
[12]Selvamaleeswaran P,Wesely EG,Johnson M,Velusamy S, Jeyakumar N.The effect of leaves extracts of Clitoria tematea Linn against the fish pathogens.Asian Pac J Trop Med 2010; 3(9):723-726.
[13]Franzios G,Mirotsou M,Hatziapostolou E,Kral J,Scouras ZG, Mavragani-Tsipidou P.Insecticidal and genotoxic activities of mint essential oils.J Agric Food Chem 1997;45:2690-2694.
[14]Kumar GS,Jayaveera KN,Ashok Kumar CK,Sanjay UP, Vrushabendra Swamy BM,Kishore Kumar DV.Antimicrobial effects of Indian medicinal plants against acne-inducing bacteria. Trop J Pharm Res 2007;6(2):717-723.
[15]Ayfer Call,Gokcen Yuvali Celik,Hikmet Katircioglu. Antimicrobial effect of natural dyes on some pathogenic bacteria. Afr J Biotechnol 2009;8(2):291-293.
[16]Yadegarinia D,Gachkar L,Rezaei MB,Taghizadeh M,Astaneh SA,Rasooli I.Biochemical activities of Iranian Mentha piperita L. and Myrtus communis L.essential oils.Phytochem 2006;67(12): 1249-1255.
[17]Adilson Sartoratto,Ana Lucia M Machado,Camila Delarmelina, Glyn Mara Figueira,Mart a Cristina T Duarte,Vera Lucia G Rehder.Composition and antimicrobial activity of essential oils from aromatic plants used in Brazil.Braz J Microbiol 2004;35: 275-280.
[18]Bassam abu-shanab,Ghaleb Adwan,Naser Jarrar,Awni Abu-Hijleh, Kamel Adwan.Antibacterial activity of four plant extracts used in Palestine in folkloric medicine against methicillin-resistant Staphylococcus aureus.Turk J Biol 2006;30:195-198.
[19]Belsem Marzouk,Mouna Ben Hadj Fredj,Imed Chraief,Maha Mastouri,Kamel Boukef,Zohra Maramk.Chemical composition and antimicrobial activity of essential oils from Tunisian Mentha pulegium L.J Food Agric Environ 2008;6(1):78-82.
[20]Jazani NH,Ghasemnejad-Berenji H,Sadagpoor S.Antibacterial effects of Iranian Mentha pulegium essential oil on isolates of Klebsiella sp.Pak J Biol Sci 2009;12(2):183-185.
[21]Ministry of Health and Family Welfare.Anonymous Pharmacopiea of India(The Indian Pharmacopiea),3rd ed.New Delhi:Ministry of Health and Family Welfare;1996.
[22]Budavari S,O'Neil MJ,Smith A,Heckelman PE.The merck index: an encyclopedia of chemicals,drug,and biologicals.11th ed. Rahway:Merck & Co;1989.
[23]Gupta R.Agrotechnology of medicinal plants.In:Wijesekera ROB.The medicinal plant industry.Boca Raton:CRC press;1991, p.43-57.
[24]Dorman HJ,Kosar M,Kahlos K,Holm Y,Hiltunen R.Antioxidant prosperities and composition of aqueous extracts from Mentha species,hybrids,varieties and cultivars.JAgric Food Chem 2003; 51:4563-4569.
[25]Oudhia P.Traditional and medicinal knowledge about pudina (Mentha Sp.family:Labiatae) in Chhattisgarh,India.[Online]. Available from:http://botanical.com.
[26]Zakaria Z,Aziz R,Yoga Latha Lachimanan,Sreenivasan S, Rathinam X.Antioxidant activity of Coleus hlumei,Orthosiphon stamineus,Ocimum basilicum and Mentha arvensis from Lamiaceae family.Int J Nat Sci 2008;2(1):93-95.
[27]Clark GS.Menthol-an aroma chemical profile.Perfum Flavor 1998;23:33-46.
[28]Alvi MN,Ahmad S,Rehman K.Preparation of menthol crystals from mint(Mentha arvensis).Int JAgric Biol 2001;3(4):527-528.
[29]Aflatuni A.The use of plant origin substances against Galerucella sagitaria.Proceeding of the Nordic Association of Agricultural Scientists 22nd Congress "Nordic Agriculture in Global Perspective",July 1-4 2003,Turku,Finland[Online].Available at:www.njf.dk/njf/reports/njfreports.htm.
[30]Asai I,Yoshihira K,Omoto T,Sakui N,Shimomura K.Growth and monoterpene production in shoot culture and regenerates of Menthe arvensis.Plant Tissue Cult 1994;11(3):218-225.
[31]Shasany AK,Khanujia SPS,Dhawan S,Yadav U,Sharma S, Kumar S.High regenerative nature of Mentha arvensis internodes. JBiosci 1998;25(5):641-646.
[32]Raja DH,Arockiasamy DI.In vitro propagation of Mentha viridis L from nodal and shoot tip explants.Plant tissue cult Biotechnol 2008;18(1):1-6.
[33]Chishti N,Shawl AS,Kaloo ZA,Bhat MA,Sultan P.Clonal propagation of Mentha arvensis L through nodal explant.Pah J Biol Sci 2006;9(8):1416-1419.
[34]Chakraborty A,Chattopadhyay S.Stimulation of menthol production in Mentha piperita cell culture.In Vitro Cell Dev Biol Plant 2008;44:518-524.
[35]Dhawan S,Shasany AK,Naqvi A,Kumar S,Khanuja SPS. Menthol tolerant clones of Mentha arvensis:approach of in vitro selection of menthol rich genotypes.Plant Cell Tissue Organ Cult 2003;75:87-94.
[36]Md Tariqul Islam,Semuel Lenufna,Philibert Dembele D,Joachim Keller ER.In vitro conservation of four mint(Mentha spp.) accessions.Plant tissue cult 2003;13(1):37-46.
[37]Haripriya D,Selvan N,Jeyakumar N,Periasamy R,Johnson M, Irudayaraj V.The effect of extracts of Selaginella involvens and Selaginella inaequalifolia leaves on poultry pathogens.Asian Pac J Trop Med 2010;3(9):678-681.
[38]Arun P,Purushotham KG,Johnsy Jayarani J,Kumari Vasantha. In vitro antibacterial activity of Oldenlandia umbellata an Indian medicinal plant J Pharm Sci Technol 2010;2(4):198-201.
[39]Salama HMH,Marraiki N.Antimicrobial activity and phytochemical analysis of Polygonum aviculare L.(Polygonaceae) naturally growing in Egypt.Aust J Basic Appl Sci 2009;3(3): 2008-2015.
[40]Mangesh Khond JD,Bhosale TA,Mandal TK,Padhi MM,Dabur R.Screening of some selected medicinal plants extracts for invitro antimicrobial activity.Middle-East J Sci Res 2009;4(4): 271-278.
[41]Ana(jrid,Branka Vinterhalter,Katarina Savikin-Fodulovic, Marina Sokovic,Vinterhalter D.Chemical analysis and antimicrobial activity of methanol extracts of Celandine (Chelidonium majus L.) plants growing in nature and cultured in vitro.Arch Biol Sci Belgrade 2008;60(1):7-8.
[42]Jain SC,Jain R,Vlietinck AJ.In vivo and in vitro antimicrobial efficacy of Mimosa hamata.Indian J biotech 2004;3:271-273.
[43]Maria del Rayo Camacho-Corona,Juan Manuel de Jesus FavelaHernandez, Omar Gonzalez-Santiago,Elvira Garza-Gonzalez, Gloria Maria Molina-Salinas,Salvador Said-Femandez,et al. Evaluation of some plant-derived secondary metabolites against sensitive and multidrug-resistant Mycobacterium tuberculosis.J Mex Chem Soc 2009;53(2):71-75.
[44]Murashige T,Skoog F.A revised medium for rapid growth and bioassays with tobacco tissue culture.Physiol Plant 1962;15: 473.
[45]Pathak P,Saraswathy,Vora A,Savai J.In vitro antimicrobial activity and phytochemical analysis of the leaves of Annona murkata.Int J Pharm Res Dev 2010;2(5):1-6.
[46JNCCLS.Performance standards antimicrobial disc susceptibility tests.Approved standard fifth edition.NCCLS Document M2-A5. Villanova:Nccls;1993.
[47]Shariff N,Sudharshana MS,Umesha S,Hariprasad P. Antimicrobial activity of Rauvolfia tetraphylla and Physalis minima leaf and callus extracts.Afr J Biotechnol 2006;5:946- 950.
[48]Manickam VS,Elangomadhavan R,Antonisamy R.Regeneration of Indian ginseng plantlets from stem callus.Plant Cell Tissue Org Cult 2000;14:55-58.
[49]Rout GR,Saxena C,Samantaray S,Das P.Rapid plant regeneration from callus culture of Plumbago zeylanica.Plant Cell Tissue Org Cidt 1999;56:47-51.
[50]Johnson M,Babu A.Somoclonal variation studies on Passiflora mollussima(H.B.K.) Bailey using phytochemical methods.Nat Prod An Indian J 2010;6(1):5-10.
[51]Johnson M.Somoclonal variation studies on Phyllanthus amarus Schum and Thonn.Iran J Biotechnol 2007;5(4):240-245.
[52]Johnson M,Maridass M,Irudayaraj V.Preliminary phytochemical and anti-bacterial studies on Passiflora edulis.Ethnobotanical Leaflets 2008;12:425-432.
[53]Castello M,Phatak A,Chandra N,Sharon M.Antimicrobial activity of crude extracts from plant parts and corresponding calli of Bixa orellana L.Ind J Exp Biol 2002;40:1378-1381.
[54]Afolayan AJ,Meyer JJ.The antimicrobial activity of 3,5, 7-trihydroxyflavone isolated from the shoots of Helichrysum aureonitens.J Ethnopharm 1997;57(3):177-181.
[55]Singh K,Sudharshana MS.Antimicrobial activity of Baliospermum axillare plant and callus extract.Asian J Microbiol Biotecnol Environ Sci 2003;5:571-574.
[56]Madigan MT,Martinko JM,Parker J.Brock biology of microorganisms.9th ed.New Jersey:Prentice-Hall,Inc;2000,p. 783-784.
[2]Ben Sassi AF,Barzallah-Skhiri,Aouni M.Investigation of some medicinal plants from Tunisia for antimicrobial activities.Pharm Biol 2007;15(5):421-428.
[3]Coruh I,Gomez AA,Ercisli S.Total phenolics,mineral elements, antioxidant and antibacterial activities of some edible wild plants in Turkey.Asian J Chem 2007;19(7):5755-5762.
[4]Veronika S,John P M,Maita L,Michael C,Adam C,Carol Y, et al.Dual effect of plant steroidal alkaloids on Saccharomyces cerevisiae.Antimicrobial Agents Chemother 2006;50(8):2732- 2740.
[5]Doughari JH.Antimicrobial activity of Tamarindus indica Linn. Trop J Pharm Res 2006;5(2):597-603.
[6]Maghrani MN,Zeggwah JM,Eddouks M.Antihypertensive effect of Lepidium Sativum in spontaeneously hypertensive rats.J Ethnopharam 2005;102(1&2):193-197.
[7]Balandrin MJ,Klocke JA.Medicinal,aromatic and industrial materials from plants.Berlin:Bajaj Springer-Verlag;1988,p. 1-36.
[8]Samy RP,Ignacimuthu S.Antibacterial activity of some folklore medicinal plants used by tribals in Western Ghats in India.J Ethnopharmacol 2000;69:63-71.
[9]Omonkhelin JO,Eric KIO,Osohon O.Antifungal and antibacterial Activities of the ethanolic and aqueous extract of Kigelia africana (Bignoniaceae) stem bark,Afr J Biatechnol 2007;6(14):1677- 1680.
[10]Ogbulie JN,Okoli IC,Anyanwu BN.Antibacterial activities and toxicological potentials of crude ethanolic extracts of Euphorbia hirta.Afri J Biatechnol 2007;6(13):1544-1548.
[11]Mahesh B,Satish S.Antimicrobial activity of some important medicinal plant against plant and human pathogens.World J Agric Sci 2008;4(S):839-843.
[12]Selvamaleeswaran P,Wesely EG,Johnson M,Velusamy S, Jeyakumar N.The effect of leaves extracts of Clitoria tematea Linn against the fish pathogens.Asian Pac J Trop Med 2010; 3(9):723-726.
[13]Franzios G,Mirotsou M,Hatziapostolou E,Kral J,Scouras ZG, Mavragani-Tsipidou P.Insecticidal and genotoxic activities of mint essential oils.J Agric Food Chem 1997;45:2690-2694.
[14]Kumar GS,Jayaveera KN,Ashok Kumar CK,Sanjay UP, Vrushabendra Swamy BM,Kishore Kumar DV.Antimicrobial effects of Indian medicinal plants against acne-inducing bacteria. Trop J Pharm Res 2007;6(2):717-723.
[15]Ayfer Call,Gokcen Yuvali Celik,Hikmet Katircioglu. Antimicrobial effect of natural dyes on some pathogenic bacteria. Afr J Biotechnol 2009;8(2):291-293.
[16]Yadegarinia D,Gachkar L,Rezaei MB,Taghizadeh M,Astaneh SA,Rasooli I.Biochemical activities of Iranian Mentha piperita L. and Myrtus communis L.essential oils.Phytochem 2006;67(12): 1249-1255.
[17]Adilson Sartoratto,Ana Lucia M Machado,Camila Delarmelina, Glyn Mara Figueira,Mart a Cristina T Duarte,Vera Lucia G Rehder.Composition and antimicrobial activity of essential oils from aromatic plants used in Brazil.Braz J Microbiol 2004;35: 275-280.
[18]Bassam abu-shanab,Ghaleb Adwan,Naser Jarrar,Awni Abu-Hijleh, Kamel Adwan.Antibacterial activity of four plant extracts used in Palestine in folkloric medicine against methicillin-resistant Staphylococcus aureus.Turk J Biol 2006;30:195-198.
[19]Belsem Marzouk,Mouna Ben Hadj Fredj,Imed Chraief,Maha Mastouri,Kamel Boukef,Zohra Maramk.Chemical composition and antimicrobial activity of essential oils from Tunisian Mentha pulegium L.J Food Agric Environ 2008;6(1):78-82.
[20]Jazani NH,Ghasemnejad-Berenji H,Sadagpoor S.Antibacterial effects of Iranian Mentha pulegium essential oil on isolates of Klebsiella sp.Pak J Biol Sci 2009;12(2):183-185.
[21]Ministry of Health and Family Welfare.Anonymous Pharmacopiea of India(The Indian Pharmacopiea),3rd ed.New Delhi:Ministry of Health and Family Welfare;1996.
[22]Budavari S,O'Neil MJ,Smith A,Heckelman PE.The merck index: an encyclopedia of chemicals,drug,and biologicals.11th ed. Rahway:Merck & Co;1989.
[23]Gupta R.Agrotechnology of medicinal plants.In:Wijesekera ROB.The medicinal plant industry.Boca Raton:CRC press;1991, p.43-57.
[24]Dorman HJ,Kosar M,Kahlos K,Holm Y,Hiltunen R.Antioxidant prosperities and composition of aqueous extracts from Mentha species,hybrids,varieties and cultivars.JAgric Food Chem 2003; 51:4563-4569.
[25]Oudhia P.Traditional and medicinal knowledge about pudina (Mentha Sp.family:Labiatae) in Chhattisgarh,India.[Online]. Available from:http://botanical.com.
[26]Zakaria Z,Aziz R,Yoga Latha Lachimanan,Sreenivasan S, Rathinam X.Antioxidant activity of Coleus hlumei,Orthosiphon stamineus,Ocimum basilicum and Mentha arvensis from Lamiaceae family.Int J Nat Sci 2008;2(1):93-95.
[27]Clark GS.Menthol-an aroma chemical profile.Perfum Flavor 1998;23:33-46.
[28]Alvi MN,Ahmad S,Rehman K.Preparation of menthol crystals from mint(Mentha arvensis).Int JAgric Biol 2001;3(4):527-528.
[29]Aflatuni A.The use of plant origin substances against Galerucella sagitaria.Proceeding of the Nordic Association of Agricultural Scientists 22nd Congress "Nordic Agriculture in Global Perspective",July 1-4 2003,Turku,Finland[Online].Available at:www.njf.dk/njf/reports/njfreports.htm.
[30]Asai I,Yoshihira K,Omoto T,Sakui N,Shimomura K.Growth and monoterpene production in shoot culture and regenerates of Menthe arvensis.Plant Tissue Cult 1994;11(3):218-225.
[31]Shasany AK,Khanujia SPS,Dhawan S,Yadav U,Sharma S, Kumar S.High regenerative nature of Mentha arvensis internodes. JBiosci 1998;25(5):641-646.
[32]Raja DH,Arockiasamy DI.In vitro propagation of Mentha viridis L from nodal and shoot tip explants.Plant tissue cult Biotechnol 2008;18(1):1-6.
[33]Chishti N,Shawl AS,Kaloo ZA,Bhat MA,Sultan P.Clonal propagation of Mentha arvensis L through nodal explant.Pah J Biol Sci 2006;9(8):1416-1419.
[34]Chakraborty A,Chattopadhyay S.Stimulation of menthol production in Mentha piperita cell culture.In Vitro Cell Dev Biol Plant 2008;44:518-524.
[35]Dhawan S,Shasany AK,Naqvi A,Kumar S,Khanuja SPS. Menthol tolerant clones of Mentha arvensis:approach of in vitro selection of menthol rich genotypes.Plant Cell Tissue Organ Cult 2003;75:87-94.
[36]Md Tariqul Islam,Semuel Lenufna,Philibert Dembele D,Joachim Keller ER.In vitro conservation of four mint(Mentha spp.) accessions.Plant tissue cult 2003;13(1):37-46.
[37]Haripriya D,Selvan N,Jeyakumar N,Periasamy R,Johnson M, Irudayaraj V.The effect of extracts of Selaginella involvens and Selaginella inaequalifolia leaves on poultry pathogens.Asian Pac J Trop Med 2010;3(9):678-681.
[38]Arun P,Purushotham KG,Johnsy Jayarani J,Kumari Vasantha. In vitro antibacterial activity of Oldenlandia umbellata an Indian medicinal plant J Pharm Sci Technol 2010;2(4):198-201.
[39]Salama HMH,Marraiki N.Antimicrobial activity and phytochemical analysis of Polygonum aviculare L.(Polygonaceae) naturally growing in Egypt.Aust J Basic Appl Sci 2009;3(3): 2008-2015.
[40]Mangesh Khond JD,Bhosale TA,Mandal TK,Padhi MM,Dabur R.Screening of some selected medicinal plants extracts for invitro antimicrobial activity.Middle-East J Sci Res 2009;4(4): 271-278.
[41]Ana(jrid,Branka Vinterhalter,Katarina Savikin-Fodulovic, Marina Sokovic,Vinterhalter D.Chemical analysis and antimicrobial activity of methanol extracts of Celandine (Chelidonium majus L.) plants growing in nature and cultured in vitro.Arch Biol Sci Belgrade 2008;60(1):7-8.
[42]Jain SC,Jain R,Vlietinck AJ.In vivo and in vitro antimicrobial efficacy of Mimosa hamata.Indian J biotech 2004;3:271-273.
[43]Maria del Rayo Camacho-Corona,Juan Manuel de Jesus FavelaHernandez, Omar Gonzalez-Santiago,Elvira Garza-Gonzalez, Gloria Maria Molina-Salinas,Salvador Said-Femandez,et al. Evaluation of some plant-derived secondary metabolites against sensitive and multidrug-resistant Mycobacterium tuberculosis.J Mex Chem Soc 2009;53(2):71-75.
[44]Murashige T,Skoog F.A revised medium for rapid growth and bioassays with tobacco tissue culture.Physiol Plant 1962;15: 473.
[45]Pathak P,Saraswathy,Vora A,Savai J.In vitro antimicrobial activity and phytochemical analysis of the leaves of Annona murkata.Int J Pharm Res Dev 2010;2(5):1-6.
[46JNCCLS.Performance standards antimicrobial disc susceptibility tests.Approved standard fifth edition.NCCLS Document M2-A5. Villanova:Nccls;1993.
[47]Shariff N,Sudharshana MS,Umesha S,Hariprasad P. Antimicrobial activity of Rauvolfia tetraphylla and Physalis minima leaf and callus extracts.Afr J Biotechnol 2006;5:946- 950.
[48]Manickam VS,Elangomadhavan R,Antonisamy R.Regeneration of Indian ginseng plantlets from stem callus.Plant Cell Tissue Org Cult 2000;14:55-58.
[49]Rout GR,Saxena C,Samantaray S,Das P.Rapid plant regeneration from callus culture of Plumbago zeylanica.Plant Cell Tissue Org Cidt 1999;56:47-51.
[50]Johnson M,Babu A.Somoclonal variation studies on Passiflora mollussima(H.B.K.) Bailey using phytochemical methods.Nat Prod An Indian J 2010;6(1):5-10.
[51]Johnson M.Somoclonal variation studies on Phyllanthus amarus Schum and Thonn.Iran J Biotechnol 2007;5(4):240-245.
[52]Johnson M,Maridass M,Irudayaraj V.Preliminary phytochemical and anti-bacterial studies on Passiflora edulis.Ethnobotanical Leaflets 2008;12:425-432.
[53]Castello M,Phatak A,Chandra N,Sharon M.Antimicrobial activity of crude extracts from plant parts and corresponding calli of Bixa orellana L.Ind J Exp Biol 2002;40:1378-1381.
[54]Afolayan AJ,Meyer JJ.The antimicrobial activity of 3,5, 7-trihydroxyflavone isolated from the shoots of Helichrysum aureonitens.J Ethnopharm 1997;57(3):177-181.
[55]Singh K,Sudharshana MS.Antimicrobial activity of Baliospermum axillare plant and callus extract.Asian J Microbiol Biotecnol Environ Sci 2003;5:571-574.
[56]Madigan MT,Martinko JM,Parker J.Brock biology of microorganisms.9th ed.New Jersey:Prentice-Hall,Inc;2000,p. 783-784.