Fruit morphological and micro-morphological study of the Anchusa species (Boraginaceae) in Iran

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1 PHYTOLOGIA BALCANICA 19 (1): 47 52, Sofi, Fruit morphologil nd miro-morphologil study of the Anhus speies (Borginee) in Irn Ftemeh Nsrollhi 1, Mrym Keshvrzi 1 & Msoud Sheidi 2 1 Biology Dept., Fulty of Siene, Alzhr University, Vnk, Tehrn, Irn, e-mil: nesht112000@yhoo.om (orresponding uthor) 2 Fulty of Biologil Sienes, Shhid Beheshti University, Evin, Tehrn, Irn. Reeived: July 20, 2012 Aepted: Mrh 15, 2013 Astrt. Keywords: With its four speies, Anhus is one of the weedy gener of Borginee in Irn. Its speies re distriuted in different hitts. In this study, the fruit morphologil nd miro-morphologil fetures of the Anhus tx hve een onsidered, so s to evlute their dignosti vlue. Eight qulittive nd quntittive fruit fetures from 12 essions of A. itli, A. strigos, A. rvensis nd A. egypti hve een studied y stereomirosope nd SEM. Sttistil results hve shown tht this set of hrters provides n effiient identifition key. The most importnt ftors omprise suh hrters s nutlet length nd width, nutlet length-to-width rtio, nutlet shpe nd position (eliosome ondition) whih exerise mjor effet on the oserved vritions. This is the first study of the Anhus speies of Irn. Anhus, luster nlysis, Irn, nutlet miro-morphology Inrodution Anhus L. (Borginee) is lrge genus with out 170 nnul nd perennil speies in the temperte nd sutemperte zone of the Old World. The Blkn Peninsul is the min enter of diversifition of this genus (Selvi & Bigzzi 2003). The speies re distriuted in Europe, North nd South Afri nd West Asi (Akçin & l. 2010). The plnts of Anhus speies re overed with roust hirs nd hve ornmentl, mediinl nd edile uses in different prts of Europe nd Asi (Akçin & l. 2010). In Irn these speies re used s sustitution for the fmous mediinl Borgo plnt. Mny miro fetures of the merirps (Akçin & Ulu 2008; Bigzzi & Selvi 2000; nd Selvi & Bigzzi 2003) nd leves (Selvi & Bigzzi 2001) hve een studied in the Anhus tx ross the world nd their dignosti importne hve een proved. The morphologil nd ntomil studies of this genus re not so extensive (Bigzzi & Selvi 2000; Selvi & Bigzzi 2000) nd hve minly foused on the leves fetures (Nyuwme & Gill 1990; Selvi & Bigzzi 2001). Inter-speifi nd intr-speifi vritions of the speies hve een studied y different uthors (Selvi & Bigzzi 1998; Akçin & l. 2010), ut there hs een no suh study in Irn. In this study, Anhus essions re gthered from different prts of Irn to study the speies reltionship y mens of the fruit miro-morphologil fetures. A tetrploid speies, suh s Anhus itli Retz., hs vst distriution pttern in Irn nd shows some polymorphisms. The fruit surfe fetures re of txonomi importne in this fmily ut in mny txonomi revisions they hve een ignored or not mentioned s key fetures. Considering the ft tht these hrters re onstnt, the merirp of the Anhus tx in Irn hs een studied in the present study. Mteril nd methods In the present study, 12 popultions (5 to 10 individuls per eh ession) from four Anhus speies were olleted from different prts of Irn nd studied. Fresh mterils of the Anhus speies nd herrium speimen

2 48 Nsrollhi, F. & l. Fruit morphologil study of Anhus from Irn were used (Tle 1). Vouher speimens were deposited in the Herrium of Alzhr University (AUH). Mesured nd evluted fetures were extrted from literture (Riedl 1967; Chmerlin 1979; Chter 1972) nd the uthors filed oservtions. A totl of eight qulittive nd quntittive fetures were dopted for fruit omprisons (Tle 2). Tle 1. Colletion dt for popultions used in this study. Speies Origin Vouher No Colletor A. itli vr.itli Tehrn, Vnk villge AUH Nsrollhi A. itli vr. itli Tehrn, Sdt Ad AUH Nsrollhi A. itli vr. itli Tehrn, Evin, Drke AUH Gnjivhed A. itli vr. kurdi Kurdistn, Snndj to Sghez AUH Assin A. itli vr. kurdi Kurdistn, Snndj, Mt Aidr AUH Nsrollhi A. rvensis susp. orientlis Alorz, SW Krj, Mrdd AUH Soudi A. rvensis susp. orientlis Tehrn, West of Tehrn AUH Ldi A. rvensis susp. orientlis Tehrn, Vnk AUH Khlili A. strigos Alorz, Mt Bghestn AUH Mosferi A. strigos Alorz, Rejii Shhr, Bghestn AUH Mosferi A. strigos Quzvin, Mohmdiyeh 15 km to Quzvin AUH Nsrollhi A. egypti Bushehr, Genveh Port AUH Nsrollhi Tle 2. Studied fruit fetures in the Anhus speies of Irn. Chrters Nutlet length Nutlet width Nutlet length-to-width rtio Angle of nutlet pex Nutlet shpe Nutlet position Nutlet olor Eliosome ondition Stte of hrter Eret (1), urved ovte (2), sueret ovte (3) Eret (1), urved (2) Green (1), ple-green to yellow (2), green to rown (3), rown (4) Exerted (1), not exerted (2) In order to detet signifint differenes in the studied hrters mong the vrious studied speies, n nlysis of vrine (ANOVA) ws performed. To revel the speies reltionships, we hve used luster nlysis nd prinipl omponent nlysis (PCA) (Ingrouille 1986). For multivrite nlysis, the men of the quntittive hrters ws used, while qulittive hrters were oded s inry/multi-stte hrters. Stndrdized vriles were used for multivrite sttistil nlysis. The verge txonomi distnes nd squred Euliden distnes were pplied s dissimilrity oeffiient in the luster nlysis of morphologil dt. In order to determine the most vrile miromorphologil hrters mong the studied speies, ftor nlysis sed on prinipl omponents nlysis ws performed y using SPSS ver. 19 (2010). Results To evlute the speies reltionships, the methods of numeril txonomy were used. Cluster nlysis sed on morphologil fetures hs reveled reltionships etween the essions. A dendrogrm plotted y the WARD method hs shown two min lusters (Fig. 1). The first luster omprises two sulusters ontining popultions of A. strigos Lill. nd oth vrieties of A. itli. The min seond luster omprises two susets: the first with popultions of A. rvensis (L.) Nordh. nd the seond with popultions of A. egypti. These two re relted t Level 8. Mture nutlets were studied y digitl stereo mirosope (Dino-Lite, AM413T) nd SEM without ny tretment. Coting with gold y mens of the physil vpor deposition method (PVD) ws restrited to 100 A (Hking & l. 2007). The SEM exmintion ws rried out on Philips XL30 mirosope. The mesurements were sed on 1020 redings for eh speimen (five smples for eh SEM oservtion were studied). For nutlet sulptures, the terminology of Buhner & Weer (2000) ws followed. Fig. 1. A WARD dendrogrm sed on the nutlet morphologil trits of four Anhus speies. Speies lel: It-it = A. itli vr. itli; str = A. strigos; it-kur = A. itli vr. kurdi; rv = A. rvensis susp. orientlis; eg = A. egypti.

3 Phytol. Bln. 19(1) Sofi In order to determine the most vrile hrters mong the studied speies, ftor nlysis sed on PCA ws performed, reveling tht the first two ftors omprise over 81 % of the totl vrition. In the first ftor with out 61 % of the totl vrition fetures, suh hrters s nutlet length nd width, length-to-width rtio, nutlet shpe, eretness nd olor hve shown more orreltion. In the seond ftor with out 20 % of the totl vrition, the eliosome ondition showed more orreltion (Tle 3). Tle 3. Results of the ftor nlysis sed on the fruit morphologil hrteristis of the Anhus popultions in Irn. Chrters 1 st ftor 2 nd ftor Nutlet length 0/96 Nutlet position 0/89 Nutlet olor 0/88 Nutlet length-to-width rtio 0/82 Nutlet shpe 0/77 Nutlet width 0/73 Eliosome ondition 0/90 Angle of nutlet pex A PCA grph sed on two min ftors (Fig. 2) revels the reltions etween studied speies nd is in onordne with the luster nlysis. (kurdi nd itli) hd spine-like ornmenttions. A. itli vr. kurdi hs shown more pointed spines. In A. strigos, the merirp surfe ws smooth nd flt, with identil spines. In A. egypti, the merirp surfe ws tuerulte with wrinkled sustrte, while in A. rvensis the surfe ws merely tuerulte. Fig. 2. PCA ordintion of the Anhus speies sed on the fruit morphologil hrteristis. Speies lel: it it = A. itli vr. itli; str = A. strigos; it kur = A. itli vr. kurdi; rv = A. rvensis susp. orientlis; eg = A. egypti. Tle 4. Dignosti fetures of SEM mirogrph of the nutlet. Speies Dignosti fetures A. itli vr. itli A. itli A. strigos A. rvensis vr. kurdi susp. orientlis spine like spine like spine like tuerulte with wrinkled sustrte A. egypti merely tuerulte In order to find further dignosti fetures of the nutlet, SEM mirogrph of the nutlet ws studied. All smples were wrinkled, oth vrieties of A. itli Fig. 3. SEM mirogrphs of the merirps of A. itli vr. itli. ) Lterl view. ) Ornmenttion with 250. ) Ornmenttion

4 50 Nsrollhi, F. & l. Fruit morphologil study of Anhus from Irn Fig. 4. SEM mirogrphs of the merirps of A. itli vr. kurdi. ) Lterl view. ) Ornmenttion with 250. ) Ornmenttion Fig. 5. SEM mirogrphs of the merirps of A. strigos. ) Lterl view. ) Ornmenttion with 250. ) Ornmenttion

5 Phytol. Bln. 19(1) Sofi Fig. 6. SEM mirogrphs of the merirps of A. rvensis. ) Lterl view. ) Ornmenttion with 250. ) Ornmenttion Fig. 7. SEM mirogrphs of the merirps of A. egypti. ) Lterl view. ) Ornmenttion with 250. ) Ornmenttion

6 52 Nsrollhi, F. & l. Fruit morphologil study of Anhus from Irn Disussion Merirp miro-morphologil evlution of the Anhus speies hs shown the dignosti vlue of these hrters. Merirp eretness is one of the dignosti fetures. Pkrvn & l. (2009) nd Akçin & Binzet (2011) hve pointed out the dignosti vlue of suh fetures in None medius nd Onosm L. respetively. The merirp hs two shpes: eret nd urved. In A. strigos nd oth vrieties of A. itli, the merirp is eret nd ylindril. A. egypti hs suurved merirp. The merirp is horizontl only in A. rvensis. So it is lerly distinguished. Riedle (1967) nd Khtmsz (2002) used suh fetures in their identifition key. These uthors onsidered three sugener in the Anhus, nmely Buglossoides (Reihen.) Guşule, Anhus nd Lyopsis L. The uthors sw the fruit shpe s very effiient in the seprtion of the Anhus sugener. Selvi & Bigzzi (2000,) showed tht A. itli nd A. strigos elong to sugenus Anhus nd mnifest lose reltions. They oth hve retngulr nd eret merirps. By the wy, these two speies re more distned from A. egypti nd A. rvensis. A. egypti elongs to Buglossoides nd hs n ovte, semieret merirp. A. rvensis elongs to sugenus Lyopsis nd hs n olique nd ovte merirp. Thus these sugener were distingushed y these fetures. These findings re inongruent with the results of Akçin & Ulu (2008) out the Anhus speies. By mens of SEM they hve found tht shpe nd surfe ornmenttions re txonomilly importnt for these tx. The otined lustering pttern (Fig. 1) shows lose reltionship etween A. itli vr. itli, A. itli vr. kurdi nd A. strigos, with spiny wrinkled sustrte on the merirp surfe, while A. rvensis nd A. egypti re tuerulte. These results support the mro-morphologil similrities. Aording to dt otined in the present study, it is evident tht the merirp hrters re useful for speies identifition. They showed signifint differenes mong the studied Anhus speies. Merirp morphology ould help in lrifying the systemtis of the Anhus speies. Referenes Akçin, Ö.E. & Binzet, R Miromorphologil studies on nutlets of Onosm L. (Borginee) speies from Turkey. Pkistn J. Bot., 43(2): Akçin, T.A. & Ulu, S Miromorphologil hrters of fruits of some Anhus L. (Borginee) speies from Turkey. Intl. J. Nt. Engin. Si., 2(1): Akçin, T., Ulu, S. & Akçin, A Morphologil, ntomil nd numeril studies on some Anhus L.(Borginee) tx from Turkey. Pkistn J. Bot., 42(4): Bigzzi, M. & Selvi, F Anhus smothri (Borginee), new speies from the islnd of Smothrki, Greee. Nordi J. Bot., 20(2): Chmerlin, D.F Anhus L. In: Dvis, P.H. (Ed.), Flor of Turkey nd the Est Aegen Islnds. Vol. 6. Edingurgh Univ. Press, Edingurgh. Chter, A.O Anhus L. In: Tutin, T.G., Heywood, V.H., Burges, N.A., Moore, D.M., Vlentine, D.H., Wlters, S.M. & We, D.A. (eds), Flor Europe. 3. Cmridge Univ. Press., Cmridge. Hking, S.A., Zurw, M. & Hrvey, E.J A physil vpor deposition method for ontrolled evlution of iologil response to iomteril hemistry nd topogrphy. J. Biomed. Mter Res., A, 82: IBM Corp. Relesed IBM SPSS Sttistis for Windows, Version Armonk, NY: IBM Corp. Ingrouille M. J The onstrution of luster wes in numeril txonomi investigtions. Txon, 35: Khtmsz, M Anhus. In: Assdi, M. & l. (eds), Flor of Irn (in Persin), 39, pp Reserh Institute of Forests nd Rngelnds, Tehrn. Nyuwme, H.G.K. & Gill, L.S Epiderml morphology nd ontogeny of stomt in some tropil Borginee. Feddes Repert., 101: Pkrvn, M., Fltouri, A.N. & Tvssoli, A Morphologil nd miromorphologil studies of None (Borginee: trie Borginee) in Irn. Irn. J. Bot., 15(1): Riedl, H Anhus L. In: Rehinger, K. H. (ed.), Flor Irni. Vol. 48, pp Akd. Druk-u. Verlgsnstlt, Grz. Selvi, F. & Bigzzi, M Anhus L. nd llied gener (Borginee) in Itly. Pl. Biosyst., 132(2): Selvi, F. & Bigzzi, M Removl of Anhus medoni from Anhus (Borginee). Evidene from phonetis nd kryotypi nlysis. Txon, 49(4): Selvi, F. & Bigzzi, M Lef surfe nd ntomy in Borginee trie Borginee with respet to eology nd txonomy. Flor, 196: Selvi, F. & Bigzzi, M Revision of genus Anhus (Borginee-Borginee) in Greee. Bot. J. Linn. So., 142: