J" Phytol. Res. 23(2) : 239-245, 2010 SCAIYNING ELECTRON MICROSCPIC STTTDIE S OF M,INN,qYA KOENIGII L ()

VINEETA CHAUDIIARY Tnd ANITA RANA Department of Botany, School of Life Sciences, Khandari Campus, Dr, B.R. Ambedkar University, Agra - lndia. 2g2002 (u.p.), e-mail : [email protected]

SEM studies showed tl. lt n Murrrya koenigiil. the anther was dorsifixed, dithecous and bilobed. They dehisce longitudinaily, pistil was shortand slmcarpous. stigma *", ,ii.r.y, .rpL[+;;ilr. Pollen grains were spherical, mgno to tetracalporate with reticulited exine putt.-. rn. unicettutar hichomes were present on the inner and outer surface of calyx, .oroiiu, pedicel,.anther, ovarian surface and lamina of leaves. Stomata were prelent-on upp", *i lower surfaces ofpetals, ,ppr, *a lower surfaces of leaves.

Keywords : Anther; pollen Murrrya koenigiiL.; grains; sEM studies; stomata; Trichomes. boduction I for 4 hours at 4oC. I boyo koenigii L. commonly known as Kurrypatta, d) Ished in 75o/o ethyl alcohol and kept in the same for ifrrreem and Barsanga belongs to family Rutaceae is a 12 hours at room temperature. lrrllom€r short and shrubby trie of about 2 - 5 meters e) Passed through the graded series ofethyl alcohol for l{L. It is a native of India. In India 16 genera and 75 I hour at room temperature. +cis of family Rutaceae are recordedr 0 Placed in I 00% ethyl alcohol (prepared by the action ftegenus Munayacomprisesof spriciesof , ll ofheated copper sulphate in absolute alcohol) for or small trees 12 tr distributea in ttre tropics and sub- hours at 4oC. regions2. In India, only two species of Munrya g) Transferred t, 5cat to a mixture of l:l 100% ethyl alcohol il,zaaics and koenigii are reported3 . Murrrya koenigii and isoamyl acetate (3-methyl butyl acetate) and kept L fu common and found in Himachal pradesh , in the areas at room temperature for 30 minutes. tsB between 800-1450 meters above the sea level. [ h) Transferred to pure isoamyl acetate making 2 changes tndian species of Murrryahave t long been used at an interval of 30 minutes at room tempirature. inaigenous system ofmedicine as tonic, carminative, I tt r) Samples are placed in small basket made up of iron- [ -rtive to treat vomiting, diarrhoea, dysentery, wire fordrying. ftmatism, hysteria, body-aches and venereal [ j) Drying ofsamples is done with liquid carbon dioxide &+stto and as an important ingredient ofseveral [ anti_ in a HCP-2 Hitachi critical point dryer at l000lb per [ +r*q herbal formulation and drugsrlz.Since Murraya inch. L*r, is ofconsiderbale medicinal I importance, scanning k) Dry samples are placed in a dessiCator. fuon microscopic studies were undertaken [ for its D Samples were mounted on brass stubs with the help [ fuisarion. ofboth side adhesive tapes. I and Methods m) are coated with gold (20 mrq coating) in a *n re of anther and pistil was llpples [ -ril studied by SEM SCD 0.2 sputter coating unit (polaron E{uipment - Ltd., I f-ing Electron Microscopic) studies. The structure Walford, Englantl). I Jritomes was also studied. n) Observations and photo$aphs were taken in LEO lUmwing srepsareraken : I EM-SEM iitAll India Institute of Medical Sciences I f Soples are fixed in 3yo gtutaraldehyde in 0.1 ' (AIIMS),New-Delhi g-20 E ,f6ehate bufler at pH 6.8 for hours at room Results and l)iscussion Emperature. Aspirator was used for quick penefiation. [ , Antho and P dlen Morphologt : the SeM studies showed are ished inthe samebufferbythree I t lry]es changes. that in Murraya koenigii L. the anther was dorsifxed, I { k fixation is done in l7o osmic acid in same buffer dithecous and t bilobed. The anther dehisce longitudinally . 23e IT Il T E 240 Chaudhary & Rana polycarpellary. In Murroya exotica, the stigma was (Flgs. 4-5). Pistil was short and syncarpous. Stigma was and or trilobed with small papillae on its surface' Style sticky, capitate, papillate with a terminal style and ovary bilobed was solid and bilobed or trilobed. Ovary was bilobed or was bilocular and bicarPellary. lemonts, stigma was wet, capitate, Pollen grains (Fig. 1-3) were spherical, mono to trilobed in Citrus globose and yellowish. Style was long, yellowish and tetracolporate with reticulated exine pattern' Pollen grains Ovary was superior, polycilpeflary, in Rutaceae were 3-6 colporate,*suboblate-perprolate; iylindrical. multilocular and contains l0 locules and each longest axis l6p n Phlebalium; l7p in Lunasia amara; syncarpous, locule contained 3 ovules. Similar observations have also l00p in Ravenia rullioides. Singhss in Pyrostegia venusta and According to Sharmar3 rn Aegle marmelos' the been reported by grandiflora. The stigma inpresently SEM studies showed that the anther was basifixed, Shakyao in Campsis studied was bilobed or trilobed and was of wet type' dithecous and bilobed. The anther dehisce longitudinally' The structural and physiological feature of pollen The pollen grains were tetracolporate, spherical with 60 between familiesat' p* fo diu*et"t i Mrrroyo exotica, the SEM studies capturing surface vary considerably Chauhan et al.zs and Ranao reported that ievealed that intlrer was dorsifixed, dithecous and dehisce However, fruitless of Cresentia longitudinally. The pollen grains were spheroidal, stigmatic surface ofcompletely pinnata, respectively growing at Agra ricolporate with finely reticulate exinera and m Citrus cuj ete and Kigelia medium sized and compactly arranged papillae with lemon, the SEM shrdies showed that the anthers were has swollen tips. According to them, this may be the cause of basifixedrs. Anther dehiscence was of longitudinal type' normal .in vivo pollen germination and The pollen grains were numerous, tetracolparate and inhibitio; of fruitlessness m C' cuiete and.Kigelia pinnata around 45 pm rn size. ultimately in the morpholory Mohfu is first to initiate the studies in pollen growing atAgra. Similmly, differences papillae in Tecoma sfars plants showing morphology of Bignoniaceae. Pollen morphology of of stigmatic sterility have been reported by Singh Bigroniaceae has received considerable attention in the seasonally transient ,""*t pu"t -a several palynologists have made sigrificant and Chauhanar. contribution to the knowledge of pollen morpholory of Trichomes trichomes , Sh4 studies showed the presence ofunicellular several members of Bignoniac eaetl'z7 . 'on parts and extra floral parts of Murrtya Tricolpate pollen grains have also been reported variors floral i.e. calyx, coroll4 pedicel, anther, ovarian by Mehra and kukarni2s tn Millingtonia hortensis and koenigii L. (Figs- I I I 8). iabebuio rosea, Singh2e in Withania somnifera, surface and lamina of leaves ' to Rathorera m Munaya uorica, SEM tricolporate rn Haplophragmo adenophyllumlo, Bombac According ptesence of unicellular trichomes on ceibi, q-S colporate in Crataeveq religiosd2. Elliptical studies showed the parts and extra floral parts of Murroya to sub circular pollen with spinous reticulate exine was various floral qeotica i.e. calyx, corollq anther lobe, stigma, style, reported by Tomar and Chauhanst fui Sqlvadora persica' L. upper and lower surfaces of leaves' Similar observations are made by Vrjayaraghavan and ovarian surface and ln Aegle marmelos, glandular and non glandular trichomes Sudeslrr in Ps ophocarpus tetragonolobtts, Tabermontana on various floral partst3. SEM observations divarticatds and n Cuctmis' species36. are observed the presence of unicellular and Lim and Tinggie3T have reported that pollen in present study showed peltate trichomes on the outer and inner surface of calyx, grains of Sever ini a btniiblia @utaceae) are tetracolporate and lower ovarian surface. It was also noted that LO biceUea. Prakash3s has reported that pollen grains of iorolla surface of corolla showed the presence of sunken prostrata (Rutaceae) were tricolpate and bicelled' inner also showed the presence of Pistil trichomes. Anther surface and peltate trichomes. SEM studies showed that the pistil was short, syncarpous' unicellular Bahadur# have reported the presence offloral The stigmawas short, bright, sticky, capitate and papillate various floral parts and which are used in with aierminal style. Style was elongate, cylindrical, thick nectaries on phylogenetic consideration. and articulate. Ovary was hypogynous, bilocular, taxonomic and is characterized,by the presence bicarpellry and was seated on the disk, two celled with Bignoniaceae of both floral and extra floral trichomesaltr. Four types of one or rarely two stigma (Figs. 6-10). of non- glandular trichomes have According to Sharrnals i*Aegle marmelos, the glandular and two types reported in 5 species of Bignoniaceaea7. Recently pistil was short, syncarpous. Stigma was sticky, capitate, been the presence ofnuptial and extra nuptial nectaries in 15 papillate with a terminal style and ovary was multilocular J. Phytol. Res. 23(2) : 239-245, 2010 241

r:s l. Showing spherical mbno to textracolporate pollen grains (586 X); Fig.2. Showing magnified view ofmonocolporate r- ;n grain (200 x); Fig.3. Showing magnified view of bicblporate pollen grain (200X); Fig.4. Anther showing r':rudinal dehiscence (a00 X); Fig.S. Anther showing magnified view of longitudinal dehiscence (170X); Fig.6. i'',:'A-ing style and papillate stigma (112X); Fig.7. Magnified view of stigma (180X); Fig.8. Magnified view of styte !.. \). 242 Chaudhary & Rana

(170x); Pedicel and showing Fig.9. Magnified view of ovary (200X); Fig.ro. Magnified view of filament Fig.ll. pedicel and sepal shorring the presence of the"presen-c" of unicellular trithomes (115x); rig.iz. Magrrified view of presence of unicellular trichomes (180x); unicellular trichomes (lz6X); Fig.13. Magnified view of sepal showing the (170X); showing the Magnified view of peafil showing the presence of unicellular trichomes Fig.15. Fig.l4. (85x). prJr"n oimicellulartrichomes (210x); rig.r6. Larninaof leafshowingthe presence ofunicellulartrichomes " J. Phytol. Res. 23(2) : 239-245, 2010 243

ovary t"t'' showing the presence of unicellular tri^choles (200x); ",t{*;;;#" ffifl the presence of mffi's,f;ffr,'Jl'Ji;11i"?:yll,:::H::,",r,r"":g;;;;a;;;;;;;ffi""(,ffi;rig.2o.Magnined orstomata showingrh" upp"."*?u"" orp;i;iii70x;ft:;i:;ffi":ffi;#ffi1.r1',,i1#fi:';f:flf:1 g;yi"il-'." (250x); Fig.23. Magnined view ffi a [i3lf,];T5'?3;yj::]::i::^1t"',"11:***,qshowing the upper surface of leaf (r69x); Fiis.2a.Lo*., surface;il#J#;#?;?;ffifl'J:?Jff#"rstomata or 244 Chaudhary & Rana Murraya koenigii (Curry leaf): a review' J' Med' species of l2 genera of Bignoniaceae has been reported Aromat. Plant Sci.2l 1139-1144. by Galetto4. The nuptial nectary is supplied by phloem Bhattacharjee S K 2000, Handbook of Medicinal branches. The extra-nuptial nectaries (devoid of vascular Plants, Pointer Publishers, Jaipur 302003 (India) tissue) are found on the outer and inner calyx surface' According to Gupta3o in Haplophragma 9. la ZO 01,Healing plants of P eninsular India, adenoplryllum,trichomes were present on the innersurface ffia CABI publishing, Walling ford, UK and New York, of corolta and on the ovarian surface. Unicellular trichomes p. were present on the outer surface of corolla, ovarian 917 frividi Pc20o7, Medicinal plants: Ethnobotanical surface and style. t0. Approach, Agrobios Publication (India) 287, 158p'- Stomata Grover J K, Yadav S and Vati V 2002, Medicinal SEM studies showed the presence of stomata on vqrious ll. plants of India with antidiabetic potential' J' parls of Murraya koenigiiL. i'e. upper and lower surfaces of p"tult, upper and lower ffi:*- of leaves Ethnopharmacol. 8 l-100. MK, Girish Kumar Ravindran JB (Figs. l9-24) t2. Vinuthan Y Jaya and Narayan K 2004, Effect ofextrapts Similar observationihave also been reported by Prakash of M. Koenigfi leaves on the levels of blood glucose Kumarrs n Citrus lemonL.According to him, numerous and plasma insulin in alloxan-induced diabetes rats' stomata were present on the style. Rathorela also reported Indian J. Physiol. P hqrmacol. 348'3 52. numerons stomata on leaves of Murraya exoiicaL' 13. Sharma L 2008, Reproductive biologt of Aegle Mehta4e found the stomata disnibuted all over marmelos L. (Rutaceae). M. Phil. Disser., Dr' B'R' the corolla surface n Adhatoda vasics Nees' According Ambedkar UniversitY, Agra. to her the mature stomata were typically caryophyllous 14. Rathore M 2008, Reproductive biologt of Murraya witlr two to four subsidiary cells; Yadavso n Colliondre exotica L. (Rutaceae). M. Phil. Disser., Dr' B'R' haemotocephala Hassk observed stomata on the outer Ambedkar UniversitY, Agra. surface of corolla; Singht' observed the stomata on the U 2008, Reproductive biologt of Citrus lemon ovarian surface of Clerodendraz species' Presence of I 5. Kumar L. (Rutaceae). M. Phil. Disser., Dr. B.R. Ambedkar stomata on the ovarian surface was also reported by Singh' Agra. inTecomaria caPensis- UniversitY, Mohl H I 835, Sur la Structure et lesformes des gains References 16. pollen. Ann. Des. Sciences Natureles. Seconde l. Shukla P and Misra S 1997, An introduction to de Vol. 2304-306. tconomy ofAngiosperms.Vikas Pub' House Pr4' Ltd' Serie. T. III morphology and plant NewDelhi. PP.466470. 17. Erdtman G 1952, "Pollen Angiosperms. J. Osmqnia University 2. Swingle WT and PC Reece l967,The botany of taxonomy". (Science FacultY). l0 166-174. Citrus andits wild relatives of the orange subfamily 1955, Contribucae a systematic das (family Rutaceae, sub family Aurantinoideae) In : 18. Gomex J C Brasileiras. I rg Do Serv. Foresffal 9 btrus tndustry(eds ReutherW. et al)Yol' f Pp 190- Bignoniaceae 430. 261-296. L 1962, Endemism and Isolation in the three 3. Nair K N and Nayar lvtP-1997, Rutacdde' In : Flora 19. Cranwell kings Iseland New Zealand, with notes on pollen and of India (eds Hajra PK et al.) Botanical Survey of tlpe of the endemics. Rec. Inst. Mu* 5 2l' Indiq Calcutta Vol. 4 259-408.' spore kievigii) 232. 4. Joseph S a4dPeterKv 1985, Curryleaf(M 1962, Pollen, d Asiac Tropical InsL perurnial nutritious, leafy vegetable' Econ' Bot' 39 20. Guinet P H Francis Pondicheri. Trave Sec. Scient. Tech. Torna 68-73. 5. XIOG PG and Wang NG 1991, Can V. original not seen cited by: Buurman J. 1977 ' Polla 19 447'519. ethnopharmacology contribute to the development et spore 1968, Pollen morpholory in Bigrroniacec ofanti- fertility drugs? J. Ethnopharmacologt32 (l' 21. Mitra K to ta

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