BIODIVERSITY OF ECTOMYCORRHIZAS IN CONIFERS FROM HIMALAYAN MOIST TEMPERATE FORESTS OF PAKISTAN A THESIS SUBMITIf,D TO THE UNTVERSITY OF THE PUNJAB IN FULFILMENT FOR THtr DICREE OF DOCTOR OF PIIILOSOPIIY IN EOTANY BY ABDUL REHMAN KHAN NIAZI DEPARTMEN'I OF BOTANY UNIVERSITY OF THE PUNJAB, LAHORf, MAY.2008 frfrfrfrftTfrfr Prof.Dr.S.H,lqbaf "n/r"t,* ^. S-fe"*. 5.f &F"c6 &,e*rnd-, 6utlr^o lq,e o'{et"l,tltg t&cErr an6lJr'vt'J CONTENTS CONTENTS Poge t{o. ACKNOWLEDGEMENTS I ST'MMARY 0l INTRODUCI'ION 03 AIMS AND OBJECTIVES l3 MATER,IAIS AND METIIOD li RESULTS l9 DISCUSSION REFERENCES 239 ANNEXURB )249 ACKNOWLEDGEMENTS I ACKNOWLEI'CEMENTS I sircdely tiiols b nry Fpecd ta.hds; Dr.Abdul Nair Khali4 my Pb-D sup€dho, for suggsting ihe lopic ed for his moEl luppon at every occ6io!, Prot Dr- S'Il. lqb.l tu Fovidirg me th. clr$@ ro r.conpoly bim d fidd rips, $courugmdt dd praycit. Ttdls e .lso duc to Prot Dr. Zhuli.g Y6t, (Chha), Prof. Di. Reidst Agc (c'dncry), D.. Yau.ki Munk.mi (hpan), Dr. Ahtd&r Bq:@ (Arg6tim) , Dr. Xavid Pdlade (Sp!in) dd Dr. Stavomn Adamcik (Slovakia) for dci help ir ildlifotior/verifi€tio of tbe rnu6tt1)(m e *!ll s E brrycdhia! dd ploviding nany valuabl€ research parqs. I .xt6d Dy SrNtitd€ b ny lovirg pldts and fiDily ftmbqs csp€ialy my oldd bbthq Prct Shd Khm Niui, for my fndcid suppdr dd prar€s fd I rmain lhdrftl ro ny tAb Glo* Ms. Nljeul-Satar AAh.n, fq tq l€lp in &e ficld, lab, ed Soing rltoud ln. th6is. ABDUL REHMAN XHAN NIAZI SUMMARY SUMMARY This work d€als wifi the invesrigarioi of biodiv€cily of ecbnycofthizb in cof,ifm fon fie ttimdryan noisl ropedre fo6r5 of patista!, Dunng $is supey, lh€ sel@led sn6 of th6e foBl3 w@ visned duri.g rh€ niny se@m (Jun€ 2005. province O.t, 200?). The* @llecdng eir.s w@ North W6tch Fmntior {NrVFp); Khans9uFAFbia, Kuag.li, Khain8ati, Narhagati, Kaehd Valler Shtu, and Nonh€m Pakisrani Fai.y nqdo*s, Duing ofs invetigation, 40 rax. betonging lo 16 eromycorrhizt ger@ of dik!ryomwota were coll€t€d, gbwing in Gociation with ,.t6r.6 fu,rbo||, ahus dadhtu, Pi.u snithiu"a Md Pinlr M/rt i/,u. Anonts urs m&rcm'€eB, onc spei6 ot Anarito (A. lld@o tes nom. pov.) dd threq spci€s or R6eta (R. ahntutlianz nom. proy., R. hiualcbit non. prc!. R pat pbv.l ^uu.a non. s@n nw to si@c. Four ,l,ra,i,z spcicByiz. ,t lat@o,birts .on. prct.. n. Iav?$. /. p&is1ahi.4 ond )1. rubs.ets wqe idenliUed by DNA sequencing ot their ITS rcgioN patisle for h. fi6l rine in Bolet6 harrcwsii, D. rctietd4, A. tub.Uri, Det@lea lla@nnulata, R^srta brdip* nd R saklontu The r@red jmlud. r.ra 7 srdi6 of Andaita (A. cinfunofu*cens, A. llaw.onoid.s M. prcy, A. ltdrips,.t. pakbtani@, L pakj, A. rubae$, /. wgnda), 5 ot Botets (8. Vd6 &nmvsii, B. catopB, R. .trtk, B. tuietur&, B. tubel6) one sp€ci6 .e\ ot co"thaleltf (C. cibatiB), Ctaturittl.tt)hus (C ttrneta). ctito.lbe lc..f. inlqdibttilor,,k), cottiwi8 (c. Ai,dule,a). [email protected] (D ce^tun (c. lawntutnta), bipt.rr, emph6 (G qo.cotus). thtwu( \rt oispaJ, Htnntn (H. rcpadun, two sr4is of LadarnB (L. delici66, t,. hutsudalz), I of Lyoplr,tun (r,. potphrr. de6ts), I of us (p. ptphr&porus), 2 or Raktuiu (R. aurca, R stictu),9 ol RBsutu (R. dtnudiuna non, prc\., R ahcthtdi'ta, R. toitEs. R hiMtqsis rcn. pr6!., R. pansrarr.a nom. pmv., R inloria, R. vcleno$hy4 Rusula pat<t & R_ pat2), I of Sarco.l@ (5. dttorin lis),2 ot Stiltus (li. td,e dB, S. sihi.,.6,) dd onc spei6 ofl.ro torn (r.. tutd,tirD). As Fgards th€ biodiv.Eny oferonycornizat norphorr?6, lh€ bsidiocarps 6*iated wirh conifw wde ll&ed dietly |o lhe nor?noryps. rony (40) nolphot,P6 ofrhe m&Dny.€t6 wft re.d ad idorified by lhis mdhod. Th* cnifi ed octonrorhiat norpholDes w.rc€h&acrnzed nrcbmorpnobs,ca y. AnonS th6e, norphottTes ofallrtt tix /tmariro fiei6, Aolat4 hoiowsii, D. Ldlotlus, B rcticulutus, B. tuhcls, Cortirurns hhnncu! Cktvtit.lelpltu! truret*, Det@l@ lawnul,to, G6tnn tripla, Co,1ph6 IMc66, E Jwllo tispa, E, rcpd.ld, La.la ^ hatsudd*e, Ltoplltllun d..6tes, Potphtrcfut ptpbhsto6 Rusula ahna.lidna rcn. pb!., R. anethtstina, R. hihalckris nn, ptor.. R. tlatisani@ nm. prc\. R. wlendsryi, R. pakl, R. p*J, Sarcodu dmynrl s, Tricholond ourantiun ds dc6*ibed lisl rime, Thc norphotrls ot aromy@nhiu of Botets edvtis, Ca"tha,.tt6 cibtiu, Rwsula b,.ripa & R, tanlonia rrc d6cnbed s new rc€rds 16r Pakisr.n. Amonssr thc oniacious nosl lrc$, a gMtc. divenily oa €clonrteirhizs qs obeded sirh ,1r,6 pt./De i-e. 22 noAh'ny96. Pin6 @Iichiufu, pipu shithiana 4nd Crdtut tlbtlzt tt vc I3,04,dd0l norphot}?$ Es!€clivcty. These anif@6 tre spei6 d.eqr C. .leo.bm rc rd *tonrlorhiz.l h6rs non This i! lhe fi'sl sriols altenpt in pakisld lo isolatc, chatuteri& (moryholoeically & oatoDicalty) &d idmlify €tomrerrnial norphor'!6 and ddelopinB an invdlory of elomttoEnizal mNhrcohttund asscialcd rilh thc conir€n of Him.l.ya hoisl t@pe6te ro@rs ofpakist&. Th€ Publish€d wo* out of lhn invBlieation hs ben app.nd€d s Annqure INTRODUCTION INTRODUCTION Pakislar lis bctqen 24o md l7'Nonn lalitldes ad 6l'and 75o lsl longitude. lt t locil€d in Egion whft $rce nountain d.gcs, Himalata, Hindllosh md Kdatorun net The l6sr Hitulaya od Hitululush 10g6 exr.nd dep into ln€ coutry &d fom h!s. complex of nounlains &d plardu. Aboul 6010 rc. oa Patiste is €ov€r€n by nounlainous cohplcx (Pakistan onlin€ cncyclopcdia, 2OO4). ln lhis, moutainous @mdq Hindlys e rhc major sutr of focrs in patjsta.. TIE for€sts of Pakisld r€flet grest physiogr.phic, clinatic ed €daphic @nttuts n Ue @untry Tn@ @ $elve typ6 ot foBts bscd o! rhe climrre, allltud€ a.d composilion of forqt speies. Th6e range frcn Tbpicat wcr everSrctr 1o sub-'.lpinc &d alpii. aoBrs (Chupion dr dr, | 968)_ Moilt leftp€Dle FoEst a@ found along lhc moist Hinalayo Mounldns b.lw@ ltle dry ldpqat€ ed sub alpinc enq (t200-3,000 m). It6e e fo!.d jn Kshmi.j MufteHaea hilts leis, Swcr, t wq Dn, UpFr @ch€s of Kumm Agcnc, moisl pans of uppcr Swar, Uppd Di., Citgil, ed Ballistan dislricls.Io noisl Hinalat"n nouraiB pripirarion is ddiled fiw the Soulh-W6tcE nonmn, july whrch falls duing tiE honths oa to Sephber dd due io wcslcrly disturbdccs il placc de lat6 duing th€ winr.r srd rpring s€ME, Rainf. tug6 betwd 6jO, 750 mm or some rimes to aboul l5O0 mn. h fi6e trcas, tarSc anounr of P@'t'r.ton talG place ir rhc fom ot mw aid is a f&ror oa @mid@btc imponan e in d€temining rlE ryp.s ot forcsls, panicllarly wh€E summer,anrall is low. GFdul D.lling of snow in @ly summs potongs to thc seen doring which adequ.r€ noislw is avaitrbte. The m€d mdimun r€np€r.luE in rhc honths of Dachbd to F€b@y is 48,2oF, 24.3oF ed 29.5"F r6p4l,vely wtule man n'nrmun rdpcmtur€ lh6e nonths e 27.6qF,24.3oF ed 2t.5qF. 'n h tnc suomer nonlhs ofJunc, July dd AuCul mtu naxihun lcmpqarurc is 8S.4"F, 87.8.F and 36.loF and 63.4oF 6 d@ ninimm &mpmtuE. Ma dnual Fidta b 59,1", hr8l'c of which lakcs placc in lhc nonsoon pcriod of July rnd Ausud. Avcfuec acrial hunidity is 57%. Inponr fcarurs of noisr rcnrmrc rorqrs ,rc ltrc conilctus rt cs ol Abiet pin&ow R.yte l60lk)a C.dr^ deadqa (Roxb, cx Llmbcd) A. Dot (roo/ori Pnw wtuchiu"u A. piuu B_ ),.ks, l|)yr)., sn,nhiau 8ois. ( l5%) T66 vallihiana Zte. (2%). In rhe* aoq$, ^nd a iuhber oa Oats d€ quih connon tikc SktM inc^d Roxb, O .!ita!a!a Lindt., sd O smi.aryilolia Sn. Bcsidc ib6c olher bmadl€aved t@s rrc in largc nunber, likc ,4.et acuunutu wall. €x. D. Don. Anon8lhe shobs v&ious spei6 of]?6a L. snd,{!16 L. s wctl htligolaru hebepdalt Berth. ex Bak t, yihunu\t cotinlloli"h D. Don,, md Urrkd ^s di,id L, @ lrrical. A l.rge nubq of m.diciml pl&1s @ur in ln€ an€ of i Gr incltding zi,,1h6 lulgoris lan., Befietk lt.iu Royre, tkinnia luuteota (DC.) yiolo Sieb & Ztcc. .x. wal .. !pp. ald Manthd pipuild L. \Cl\Mpion et dt., t96P)_ Pakier4 h6 mqgd foct l$uEs. nE colntry duc lo iis sbarp clinaric vanaliotu rnd arid oonditids lack6 @sonabte t@ @vcr. Thcre is hardly 5.7% ot thc roral led aE of&e country @vcrtd wnh b6tr out ofiq rh€ prcduclilc aock are lss thm 2%, dd fie rare of depletion @nrinu6 to bc nigh. Conn€rcid logeing .nd ovcr cxpro ltion oa loresls by a sbwins pdput.lion for fuct, fo<ldo buiklnrts matrials, ..sin md charcoal hs ftsutled in.ripplins lh€ nc.ger foresl @ur€es ot thc country. Thc effftls of defo$tation o. biodivcBitt d€ crnical sincc non forcsl {osystens arc dBloyed. thc dhapp@ance of t@s dd shdbs n€.s lnar the a6ocrated 0or.