wat ar of Boal See fens 1 Fe t= Des 208 16119 Rapid in vitro Propagation of Medicinally nportant Rtinacandhus rnasutus (L) Kurz and Screening for its Antibacterial AC “vty “Shanthi H. Rojabegam,N. Yogamanth 18. Bhakyara, A. Chanthurw ane Arum Sas) 1G and Reser Deparment of Micon. 1J, College Ar Seen 2 Pasa du an» 62240 seorespondiigcithor: biogas ®gmaikcom Aeceved July 30, 2068 reviewed November 4, 2008; accepted November 10,2008 Abstract Calcul wc sinned ae tm expan Rhinacontas asin Lon Mush ge ed Stooge nleenta with 24-eNophntne asic ad 4D) nda seni MA) opr ni ail NAA) Benyamine BAP) and Kitna) Te get fy (enya) fen indoction wes bev in MS medio coetiag 1 mg” 2D +03 mi et ‘Spit Deopref avant ecard wie te pad and ode eens ee He srs edum uplemeaied wik BAP Ki, 18d NAA, Shot dilrestaton wa ex C6 set) a esl segments on MS rei coming 15 mg’ BAP+05 mg" TAA, Rhincrntas nous 8 ranma etal aii api ow importa han pape. Rode af mates ex vette diferent slvesiCRoseram) iacaste + Cicer soles sing SOA Pert “ino te two eee Ey oe +ilaroonena bowed iia ivi whe comers te cflotor stent erode Cals nto Shoo inton Anberlin nats Metal et Introduction The shrub Reinacaarhus nasatus (L) Kurt (Acanthacene) is a medicinal plant that is widely ibe in Sout Asia. The leaves and ses of is plane are often wed forthe treatment of hepatitis, isbeties, hypertension and sin disease (Wu tal, 1998).The trate repos that ens of vrs prs of Rinocamhas nasa La Rhine commis sewed forteament gaat ringworm and teres shin isases, as mle exe, pulmonary beri nl cance Kodo, 199A. Nagao 19) od Rojanapo ot af 1990), Although Havas amtrogunons. iterpens, sterol andoter comps have beet isolated from Rhinacanthns nasuits (Subramsisn and aga, 1981, Ho Dac and Ho Des. 1979 and Wa ta, 1988, the ony bilogeally ave contents reponed ae paphthoginenes (Ubi 1998. The ot cont Rinaanhin compound, hich possesses anne and antiparasitic propre. The plant i ich i potasium salts and sls0 contains ‘onymeshyl_anthroguinone. Rhinacanthin-A <0 Rican B wer alte rom he leaves ads ‘of Rhinacanthus aasatus L, by Dioassay~ suedci tae ect ult Bolg San, Ilse 2 Dn 08 9.119. ‘rctionation using cyst asa. The yi smed sho aplicion of plan tse ele ech for nesting high quality sonacones of Rhinacamhus asus L. witha cousane and increased level of ilgili ase compound Materials and Methods Apical ef mode an sen were cla fon cove year heathy ans, Explants waned with ‘using tp water f¢'1-2 min and sufve sod. sitally im 70% e1 ol followed by 0.1% (WHY) uerum chloride fra few seconds and thoroughly ised heer is i sind isi wee Explants were cultured on MS mediuiy plememed wih various soaenrations fms (2.4. > AA and NAA), loins (BAP an Kien, The lung was aud to $8 pi wo wating at [21°C oe 1Smin. Cultures wee mained wr cok hie recent ight (2.00 us) a 2522 with 6 oe pox Foc anibactetlatiy, the hey ans wee collected and shed in taper an she ded t oom temperate for 5 das, 25 the powder was fled in he thimble and exacted sucesively with ofc and tact for 48 bs ing» Soxhlet xo Al be extracts woe concentra using try ‘pra and preserved a SC in siti bls unt ora we us Bacal pathogens were: obiined fen Stay Medial College, Chenu, Inds, Amongst the four rmsreorganiss investigated ome Ic Gran positoe bce namely Stpocecis wareas ee se Gramacratie astra ancy Sano Pritas spand Escherichia col The antibacterial asay was pve by agar wll ifson met, A well was pepe nthe plas, wit the hp of or bre (038 m5, 109 an 130 poe st compounds were indeed fm she well. Me plates were inabsed ovenight it 37°C, Mico growt was drsincd by measuring he diameter of the zone of inibion, For ach ctrl train, onto as mite with pre soles ean instead oe exis. The els were hited hy mesaring the one of niin ‘Results and Discussion Calls induction was observed in MS media onan iret concen ad combats of 24, A. NAA, BAP and Kin Calis induction was ered i ef and tem ets within 10-15 ys ‘The pewcemage of ells induction je lea explana (68.00%) vas high io MS mam supplement wath 150mg! 24-0 +0 mgL? Kin wich was flowed ty 6500 cals induction in MS edu coming 1.50 ing! 2.40 +015 gL BAP ste xpi. he hes petecmtaye fells induction (61.008) was ered in MS meum containing | SOL! 2.40 4 (05 gL" BAP followed by 608 observed ia 150ml: "2:0 +05 mg Kin (Tal RTEewe al fail Sens Innocent t= D2 111 \ Tat WS320 07 2 er | Ms 2403 wie |e | MS+240 16 ois lg se24D 15 2 |e Ms+24D20 oa Ms +2425 6 |Dg MGsianer i tig MS“AAtS Blo S148 10 so joa MSHIAALS pe Jog MBSTIAA2@ 4 (pa | Ms-Iaa2s |b MS + NAAay nly | MS NAADS j> fs jug MS+ Na Lo jo fe [is MS NAA LS 3 la (tg MS + Naaz0 jp |z os st NARS 2 ie foo Mss24D0s+uaros [3p |i |p Mss24Dt0rparos [sz (te [bg Me-iapisemaras |e lie |e use2enz0snaras |S |S bg, Mssapascoapas le [5 og Mec2eoso+mpos | |e lt MSw24p0s+Ka0s [asa Lg Msr24pinsKeos la | [po MS+24D 15 +Kinas lat [gee MS+t4pz0eKnas |e (58 Ibu MB+24p25 Kens ay | [DG MS:24D30+Ke03 [39 lat fog UG LIGHT GREEN, bo = nave GREEN, ‘The overall percentage of calls induction was | oud to higter in le expiants tha ten expan, ‘The colar ofthe calls was mos light green dak 7 Simic fxourable rest ofthe combine effec. of auxin (NAA) and cylin (Kin) were reported in ‘fgohara nace Mba Faisal ast Moanin is, 2008), Narn 9p Sires an Langton, 1990, pin ese Saas a Crisps. 588) aus Yee lana (Beate a, 198), Fe erent erenee 6 "oe nein Wy pat inc moan (Ka omg i Adc Any “Table Let of ifeem auxin and cyst on cals indoction nn texans in MS msm ‘The apical bas and noel segments wer cate (08 MS medium supplemented with 4 ‘oncentiom and combination of BAP. Kin, JAA an NAA, Shoot formation was observed i il homme bination, Homer, the bet reponse was bina when the explants were cultured on MS mediam ‘comaining (50-mgL" BAP +0 mg TAA. In hese ‘ombnations, 2 to 4 shots were developed irom exh upland his mamber was found to be highs (8), reatsea oa a alge Sees Taste Fa Ot Do 3, 1419, ren the cute is mintaind in sme etiam tn te merle nodal expan roduc mare soos eben compare with piel bad Tale 2 Pe). 5. ress ere report for many medina plots ie, hinacomtins nasuus (ota ta, 2072) Pure -rplfolia Jeyakumar and Jayablon, 2092) Motus ines (Bhsshioson, 1982) Mera nigra Yel t (90 sod Piper sp (Bhat a, 19835). Pate Shoat ination From the naga expos of hinacanhnsnosisu Lon MS meson ‘Table 2 Eifot of fees ctokinn und auxin sho into frm nodal explants i MS mem ‘ues repeat = staid evan 10 eis er tenne in tee eaiedexeineas ‘The sty of he anita aivity of the two soles amass ead tht th on with eorform nett acetate howe more igi ci aga taro pathogen acter, nthe present Soy the exact ohfshowstenly era inert sath actvty when compared with cloofonn and oy asetateexrats. Among the four human phages tested, Sanonela ht was hgh Sensi revealing 209 iton of 26min 100 and 150, followed by Suplylacirce ares reveling» zone of inition of am in 15 Table 3 al Figcilia staan seccncmmaaitent ech mal opal cess tes eet, Ot B28. 114319. Tod vie Point Medina Rach nat) Kart ie Soin is tcl Ai ‘Table 3, Anvibucterial activity of lea extracts of hinacanthus rasutus “Giratorm avast | Hiiglaceiateand eroform Concent ‘am tract) = FF ee 7 Ed 16 7 Ww By is 1010 ‘oof feo fa 1501 iw Ba Tis 2 B 16 __ 136 4 ‘Spores ra Puen, ST: Sleep BC: hee ig ‘4; Syocosus aues B ‘yp: Esch cl Ait prove tbe main tase for wean of uci infeston, However th high penetic variability of bacteria enables them 10 apy eva the acon of mlbitiesistanee. Thus, there hasbeen conning seach for new and more potent antibiotics (Het 2001), Therefore, in the present investigation kinacadias manus Lean important medicinal pi vas evaluated for its amtibucteril potential against human pathogenic bates. The presen! investigation cel eval he antibacterial ii ofthis pla amo Anibal activity of leaf entets of Rhimacons manus Preudomoness Seiwa ‘tha dus pant could be exploited inthe gest management of diseases caused by these bacteria ia ‘ran system, “To concine, the present study poits tx apd net forage sale mas propagation of Rinacahas asus L hugh various explants an the protocol overt |e eficen and could be used as 2 means of propagation nc mpi of kinacarrus asus [L.— a medicinal lant wih potential for commercial exploitation, uecss cers eewth if ila Seen, fetes Plt 2 De 208.1119 References ‘Bem SF. Patan B Talbot 1 8 Akerman W (A988), Foto fecting vio propa uf Tes lance, Plant Celt, iene ond Ofc Cale, 14111. ‘Bha SR Chel K PS nd Mai SK 9% Plt egenoation from varios explants of cui Pier spose, Plan Col Rep, 14:395-402. ‘ise P (200), lan mee, 6112. Ho Dac A and Ho Dae K (1979). 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