Development of In Vitro Protocol & Evaluation of Molecular Diversity in Nardostachys Jatamansi Using RAPD Fingerprinting Collected form Hills of Uttranchal

Abstract

i\ardoJtudl)'' j.J/<1m<~n•1 L)( as small, herbaceous, pcrcnmal speaes of farruh \'alcrianaceae. It IS dlstnhutcd randomly up 10 51KHJ mclcrs tn rcsmcted area of Himala1; and used m modern mcdactnc, pt:rfumcry and 3> ctheno·mcdlane. To come.r\'e thJs cnocall) endangered mnhcm.tl herb, an m t1ffrl propagauon and di,·ersuy csumauon by It \PO markers has been optim.t7C<.l G oo seed gemunaoon was achieved by the use of G.\. (IOUppm). The germtnalcd seeds and explancs u·cre stcrW7ed u~rh ",lh s•. >OWum hYpochloote for 12 mmutc. Shoot establishment wa' Jchaned u,rh B.\P (1.0 ppm) culturing on :\f$ mediA .md c.tllus mducrion ob,en cd when supplemented 1\llh :-=. \;\ (3.0ppm) and B \ P (() 25ppm) A modified and rcpmductblc protocol was de\'clopcd for DN.-\ tsolauoo from rhizome of Jatamanst by man1pulaung the concentrauon of 1'1riOus compo nents 111 the original CTAB method. Quallay DN1\ was obtained with CTi\13 (2.5 %), Nad (501!11), ~ mcrkeptocthanol (1°o) and PVP (1%). PCR condidons for Jntamansi during RAPD analysis were o purrutcd b)· 'umc mod1ficaoons an 1hc onginal protocol. Guod amplificauon was obtruned '' nh magncsaum chlonJc (2.5 m\1), dl'TPs (0. - rn.\1), I "4 ON.\ polyrnernse (2.5 U/j.tl} kccptng -12 crcb. The genetiC Surular:il}' based on ::-;c:a's sarrulantY mdex was c<lcul.ttcd . \lithe crght acce>stons sho11cd smulaol)· coefficient 03K to 0.52. The low level of gencuc s1rrul:ulry among rhc :1cccssaons of Jatarnansa could broadh· cxplatn a' a result of .tbiouc and biooc factors. Dendrogram consuucted b) these data usmg t;PG~l \ rhrn• clu~tc~'> were obtained. ,\,cc:ssaons collected from HKO and T' showed max unum samaLtruy due to high gcncuc flm,, where as accesstons collected from l\lS ,-wed ~tgnrficantl} when compared to other acccssaons, thJs lS due to non significant gene flow. 'l'hl· other rc.lsons of dlYcrsll} might be due ro reproducuvc asolauon, entomoplulous pollinauon, dtsper.-al of hair) seeds b) wmds. habitat ch.mgcs, and small populauon ,izc tn dafti:rentlocation of Hamala)3·