New Boranssr 12 (2-4) : 117-127 : 1985 FLORAL NECTARIES IN TWO SPECIES OF TURNERA L. (TURNERACEAE) BIR BAHADUR, N. RAMA SWAY, ARTHI CHATURVEDI Aw 5. M. FAROOQUI Deparinentof Botany, Kakaiya University, Warangal, India (Received for Publication : 3 December 1984) ABSTRACT Floral nectariesin_hetero-dstylous Tarra subulata J.E. Smith and homostylous 7. wmiftia Lehave been studied, Floral nectaries are five and located at the base of ovary, alternating with the petals and ate integtal part of the basal portion ofeich filameat. The basal swollen region of adaxial side ofegch filsieat form a shallow depression. The nectar pouch and this extends laterally ‘until fares with the lora tbe forming the nectar pocket, The nectar pocket is more conspicuous in T. inifoi in comparison to thote of pin and thrum eyed flowers of T.subulte- Nectar pouch characterised by anomocytic stomata. in both the species and are epstomatic with the exception pin form of T.tubuata, The yellow conical ovarian nectary at the base of the filament is provided ‘with several anomoyetie stomata. Nectar i gecreted through thece and pouch stomata and is stored in the nectar pocket and last for more than & hrs. Nectar in oth the species comprises of licose, Feactose, sucrore and mannitol, but lest sucrose ws in the pin form of T. slate, 0 acid composition has ano bee studied in Tabula, 5 = Key words : Floral nectaries—Hetero-dstylous--Homostylous- Nectar pocket—Pin eyed— “Theum eyed —Anomocytie stomata —Eplstomatic— Amico seid composition, INTRODUCTION Urban (1883) first reported floral nectaries in Mathurina pendwiflora (Turmera eae). During this century Brown (1938) investigated 3000 species of angiosperms for their nectaries but there is no mention of the nectaries in Tumera L., Capuron (1963) reported nectaries in Pirigueta ansingyae, Recently, Elias ¢t al, (1975) have studied the structure of foliar and floral nectaries as well as nectar composition of Tumera ulnifelia L, but the floral nectaries have not been thoroughly studied by them. The amino acid composition of the nectar from foliar and floral nectaries of T. umifolia has been studied by Baker et al. (1978). The morphology and embryology of T. ulmifolia has been studied in the past by several workers but ‘one commented on the floral nectaries (Rao, 1949; Johnson, 1958; Vijayaraghavan & Kaur, 1966 ; Rajagopal ef al 1973). Barrett (1978) studied the distyly and ‘the pollination biology of T. ulmiflia. Various aspects of morphology, heterostyly and incompatibility in Twrnera subulata J. E. Smith including unilateral interspecific hybridization have been studied (Rama Swamy & Bahadur, 1981, 1983, 1984a & bj Bahadur et al. 1981; Bahadur & Rama Swamy, 1982). Recently Bahadur & Rama Swamy (1984) have briefly commented on the nectaries while studying the pollination biology of 7. subulata, This paper is an extension. of the same and describes the nectaries of T. subulata and T. wlnifolia in detail and also reports related aspects of nectar secretion and composition.118 ‘New Borawisr 12 (2-4) +1985 MATERIAL AND METHODS For the present studies, the flowers of Turnera subulata and T. ulmifolia were collected from plants growing in the Botanical Gardens, Kakatiya University, Warangal. Epidermal peelings from both the surfaces of freshly collected stamen and nectar pouch (shallow depression of basal region of filament) were stained in 1% aniline blue in lactophenol and mounted in 70% glycerin for observations. Hand cut transverse section of the lower portion of the flower was mounted in 1% slycerin for the study of nectar pocket, nectariferous stomata, secretory tissue and trichomes around the ovary. Nectar was collected with 1 co plastipak microsyringe (USA) from freshly opened flowers at 8.30 hrs and also at 12.00 hrs. Nectar was analysed to study the various carbohydrates and amino acids present by the paper chromatography following the method of Purvis et al. (1966). OBSERVATIONS AND DISCUSSION Flowers in both the Tumera species ate epiphyllous, large, showy and penta- ‘merous. They are hcterostylous bearing thrum and pin eyed flowers in T. subulata and homostylous in T. ulniflia. The calyx is united below forming a floral tube which is cylindrical but broadened at the apex. ‘The campanulate corolla is bicoloured, the limb being whitish yellow while the violet basal portion is marked by streaks, that extends in. subulata whereas in T. ulmifoia the corolla is sule Phureous yellow. ‘The five petals are obovate and short clawed and adnate to the calyx tube, The five stamons are inserted near the base of the floral tube and are rectangular in transverse section; the base of each lament becoming broadened and this is adnate laterally to the floral tube near the base forming a nectar pocket where the secreted nectar is stored (Figs. 1-5, 18-15, 21-23). ‘The nectar pouch is at the base of each filament in both the Tunera species studied (Figs. 4,5, 14, 15, 22, 23). The pouch is comparatively smaller jn bud condition i.c., one day prior to flowering and is prominent and slightly longer in T. wffolia than in both the pin and thram lower forms of T. subulata, ‘The epidermal cells of nectar pouch are cuboidal and cpistomatic in both the species with the exception of pin flower form of T. swbulatz, The stomata occur irregularly and 3-5 stomata have been observed in the middle region and the rest are scattered along the sides ofthe nectar pouch. These stomata are anomocytic and remain wide open throughout the day. The guard eells are devoid of chloro- plasts. Higher stomatal index has been observed and high stomatal frequency on the adaxial side of pin form of T. subulata and larger stomata was noted in T. uimifolia (Table 1). Bach nectar pouch is parenchymatous and secretory in nature, stomatiferous, rich in starch grains and with granular contents, The granular contents and stomata are absent on the abaxial sutface with the exception of pin flower form of T subulata. ‘The secretory cells are cuticularised, the cuticle breaks down releasing the nectar into the nectar pocket confirming the observations of Elias «fal, (1975). These secretory cells of the pouch facing the ovary are vasculated and this continues farther up to the anther lobe, Just above the nectar pouch the cells are etra-to hexagonal without starch grains and the cells are somewhat longer on both the surfaces of the filament compared to the cells of‘Tamu 1. Stomatal features ofnectary pouch, ovarian nectary and associated insect vsitorsin two species of Tarra Species Flower Nectary pouch Ovarian Awociated itcet siwems) Stomatal—-Stoma Pore ise stomatal Tadex feequeney) sae nam Sie mm in tm inam Tarnea ubulte 65 en 4 36.0%99.2 16.088 26.64x26.27 —_Apiserana india, Bombus (Pin eyed) Adasial & p. 14.93%5.2—Componotas >.» Abssial 4a 2 280x280 136x586 Catiohraerthrcsthao, (Thrum eyed) Adaxial 6.6 178 1 BLAX92.8 16.0% 10.8 25.56 x28,0 Pieis brassicae, Papillis & fonulet, 18.0x6.0 Opedaes sp. and Three sp. Abatiat G = = uy Tomere viola (Homostylous) ‘Adaxiat 50 47 2 892x376 160x144 $2.98 11.07 Apis corane indica, & Bombus sp. Companatr ap. 1287x846 Celliphoreertinecphale, Abarial i = = = Piers brascee an Tire ep. Adasial=nectar pouch (or shallow portion ofthe base of the filament) ViraNny, 40 sumIVioay TWHOTY SIV 15 wavaVE mE, 6120 ‘New Boranisr 12 (2-4) : 1985 Figs 1-12. Tina sbulte. Pig. 1. Theuma eyed lower after the removal of calyx and eorolla he potitioa of nectar pockets. Fig. 2. T.S, off poste to cay lobes, Fig.3. Developing capsule showing position of nectaries Adaial side of the swol ‘the filament showing nectary pouch 6. A portion of the epidermis from ‘A portion of the epidermis fom allavial surface, stoma enlarged from the surface of the nectar pouch respectively. Fig. 10. Nectarferous anomocytic 12, Amultteriate glandular and amall vesicular glandular hairs respectively (Gocorolla; ffilement; La-long anther; np=nectar pocket; p=nectar pouch).Bim Bawapur er at.: Florat Necraries oF TURNERA 121 axial etary pouch he ofthe neetary pouch and stoma fom the same (Ls=boag styles peace nectar pouch (Figs. 6—9, 16—20, 24—27). Abnormal stomata with one guard cell rarely occur on the adaxial side of nectar pouch of T. ulmifolia (Fig. 28). The five pale yellow conical nectaries are situated little above the base of the ovary in both T. subulata and T: ulmifolia (Figs. 1-3) and are located opposite to the five calyx lobes, alternating with the petals. Each floral nectary is a conical122 Naw Botanasr 12 (24) +1905 Fiss.21.28. Tirta alaiflia, Fig. 21. Homostylous flower afr the removal ofcalyx and corallasiowing the position of mectar pockets. Figs. 22-23. Adaxial side of the swllen bate of the filyneat showing nectary pouch in bud esndition and in the flower respectively. Fig. 24. A portion of the enidermis from the flamcat, Fig.25-27. A portion ofthe abavial, adax'al side of the nestary poach and stoma enlarge from the aime. Fig.28, Abnocimal stoma of adaxial surface. (peneetary pouch). structure with a depression in the centre. The basal region of swollen part of the filament partially overlaps with the outer rim of the nectary, so that the nectary becomes visible only whien the filament and petals are filly removed. The lateral Side ofeach filament is flattened and extended laterally into a wing until it fuses with the wall of the floral tube. This adnation of the extended portion of the filament (but not the middle portion of the filament), to the floral tube forms the nectar pocket approximately 35 mm deep in T. ulmifolia and 1-3 mm deep in T. subulata. The nectar pocket is conspicuous in T. ulmifolia whereas it is not much pronounced in both the lower forms of T. subulata. Nectar pockets alternate with the petals which are readily accessible to- honey bees and other pollinators studied earlier by Bahadur & Rama Swamy (1984). Irrespective of their size difference, the quantity of nectar produced by the flowers of the Turera species is negligibly small and hence quantitative differences were not studied. Each ovarian nectary is provided with several anomocytic stomata surrounded i glBin Banapur er AL: Frorat Nucrariss oF TURNERA 123 by 4-6 epidermal cells (Fig. 10). These stomata remain open throughout the day and the guard cells are non-chlorophyllous. Larger stomata have been noted in T. ulnifolia as in nectar pouch (Table 1). Stomata are surrounded by parenchy- matous cells of secretory nature, small in size, cuboidal, filled with numerous dark staining bodies containing loosely arranged numerous small vacuoles, ‘The topography of the nectar pouch and distribution of nectar stomata of the ovarian nectary suggest that the nectar oozes out through these stomata immedia- tely after the flowers open between 6.30 to 8,00 hrs and accumulates in the nectar pocket and becomes available to a wide vatiety of visiting insects (Table 1). More nectar secretion was observed at 10.30 to 13.00 hrs and a high frequency of insect visitors has been observed during that time (Bahadur & Rama Swamy, 1984). The nectar secretion lasts upto 15.30 hrs in both the flower forms of T. subulata and T. ulmifolia, Nectar in both the Tummera species studied is some what acidic (pH=6.5), colourless, sweet and turbid. Nectar production in flowers of both the Turnera species is scanty and even with the use of a microsyringe it was not possible to quantify. However, careful observations showed that the flowers of T ulmifolia produce comparatively more nectar than those of T- subulala since the nectar ouch in the former is bigger than the latter (see Figs. 4, 5, 14, 15, 22, 28). The analysis of carbohydrates and amino acids was done from the morning (8.30 hrs) as well as in the noon (12 hrs) collected nectar. ‘The presence of four sugars viz., glucose, fructose, sucrose and mannitol has been recorded. Slightly more sucrose was observed in the nectar of T. ulmifolia and also in thrum form of T. subulata (Table 2). But xylulose was recorded only in. pin form of. subulata. However, equal concentration of glucose, fructose and mannitol has been observed in both the speeies of Tumera, It may be pointed out that Elias etal. (1975) have reported the occurrence of more sucrose in T. ulnifaia. They have analysed the floral nectar of T. ulmifolia in the morning (10.00 hrs) and in the afternoon (15.00 hrs) and noted more suctose in the afternoon than in the morning collected nectar. Further, they observed decrease in concentration of glucose and fructose jin the nectar collected after noon. Traces of melizitose were also. recorded in T. ulmifotia by them. Nectar collected from freshly opened flowers, spotted on chromatographic paper and treated with 0.2% Ninhydrin (in acetone) gave intense violet coloration (Baker and Baker, 1973) not only in T- subulata (both pin and thrum) but also in T. ulmifolia indicating the presence of e-amino acids in both the species. In Table 3, the composition of various amino acids present in pin and thrum forms of T. subulata are summarised. A perusal of the data shows that the nectar of thrum is characterised by 7 amino acids, while the pin has only 5 amino acidss Phenylalanine, Tryptophan and Isoleucine are absent in pin, while amino-n- captoic acid is absent in thrum. In both the pin and thrum forms a-amino bbutysic acid was detected in higher concentration than other amino acids. No difference in the composition of carbohydrates and amino acids was detected the nectars collected during morning and afternoon in both the species the amino acid composition in the nectars of floral and extra floral nectaries of T. ulmifolia and noted 7 amino acids in extrafloralit New Boranisr 12 (2-4) + 1985 Tanta 2, Carbohyate composition of the iétarin Tare specet Sk. Gatbobyate Tamera bale Tora te a Pin eyed Thum eyed (omenyiow)— 1. Ghucwe Ge ie ++ 2, acie = = 7 2. Malte = - “ ieee + + + Ramen = = a 6 Soxbito - - o es ; +4 ++ 8. Manso . + + 8. Stach ~ s - 10. Dentrn - 2 = 1M, Melee” F = . 12. Xylelse + - - 13. Unknown & = = =Abent 4 Present $4 = Comparatively more Data of Elia a. (1975) ‘Taaix 3. Amino acid composition ofacctar in distylous Turner subulte Alanine Leucine Tsoseucine Phenylalanine Tryptophan x-amino butyic acid Amnino-n-caproie acidBm Bauapur er at,: FLorat Neorantes or TUnnena 125 nectar but 21 amino acids and 2 non protein in the floral nectar. The composition ‘of amino acids in floral nectar as shown by them may be exprested as fellows : Proline and Serine >Alanine, Arginine, Isoleucine, Leucine and Lysine > Aspar- gine, Aspartic, Cysteine, Glutamic, Glutamine, Glycine, Phenyl alanine, Threo- nine, Methionine, Tryptophan, Tyresine, Valine, cc-amino butyric acid -+ one unknown and two non protein amino acids. Further, they did not detect any quantitative difference in amino acids in vatious geographical races of T. ulmifolin, ‘The observed differences in the composition of nectar, both sugars and amino acids particularly in distylous 7. subulata perhaps isan indication of the dflerences in the food of foraging insects, since nectar composition enables to determine the taste and naturally the visitor type and this has been discussed elsewhere, Clusters of multseriate glandular hairs oceur alternating with the five nectar Pockets (Figs. 1, 12). The cells of the distal part of the glandular hairs are compact and are densely cytoplasmic but the stalk is sparsely cytoplasmic, The small vesicular hairs are also densely cytoplasmic but are provided with a terminal veticle around the glandular region, ‘These vesicular glandular haire occur alternating with the conical nectaries in Tumera, ‘These hairs probably are secre- tary and are comparable to the nectary glands on the corolla of Lonicea japonica studied by Fahn & Rachmilevitz (1970). Whether these glandular baits of Turnera secrete nectar is presently not known. According to Brown (1938) in Gesearia (Flacourtiaceae), the stamens alternate with glands in circles, the latter are interior to the stamens whose lower part secretes nectar internally and forms a cup around the base of the ovary. This situation is partly comparable to the one observed in Tumera presently studied. From the foregoing account it is evident that the nectar is seereted through the nectar stomata of ovarian nectar glands and nectar pouch in bath the species of Tumera. A wide variety of insect visitors feed on the nectar of T- subulate vie, Apis cerana indica, Camponots sp. (Formicidae), Bonbus sp. Skipper and Thrips sp Bahadur & Rama Swamy, 1984). Barrett (1978) also reported Apis melijore and Trigna species on the flowers of T. uimifolia var. elegans while studying the pollina- tion biology of the species Similar findings of the nectar secretion through the nectariferous stomata have been earlier reported in various Rubiaceae (Bahadur et al, 1971), Vince mse, V. naj and Gitrs sinensis cv. calenia (Rachmilevite & Fahn, 1973), some Compositae (Gopinathan & Varatharajan, 1982) and in four species of Kalanchoe (Bahadur tal, 1984), Elias e al. (1975) have studied the floral nectaries of T. ulmifolia but neither describe the structural details nor mention the presence of stomata, According to them the nectar is secreted by the rupture of the cuticle of the secretory cells, present below the filament, In the light of the observations presented, itis thus clear that nectar comes out through the nectariferous stomata and accumulates in the nectar pocket. ‘This nectar enables the insect visitors to feed on it and in the process, the visitors bring about pollen flow in distylous T: subwlata (see Rama Swamy & Bahadur, 1964b) and alto pollinates the homostylous T. ulnifola. Although no information exits on the nectaries_of Turneraceae, the following information on the closely related ‘Theales and Guttiferales is relevant. "According to Brown (1988) in some Guttferales the nectar glands seem to be modified groupe126 New Boranisr 12 (2-4) : 1985 of stamens, while in some Theales, the more basal parts of the stamens secrete nectar. In Eurya japonica nectariferous tissue adheres to the basal portion of the ovary while in Theales there isan aggregation of nectariferous tissue as a ring below the stamens on the inner side. Thus, the nectariferous basal part of filae rents is comparable to the nectary pouch in Tunera presently studied. Fahn (1953, 1979) classified the nectaries in the flowering plants into 9 types. Under type 2, ie. staminal nectaries, he distinguished 3 sub-types viz. (i) on filament (ji) transformed anthers and (ii) appendages/appendix. of connectives, Elias et al. (1975) classified the floral nectaries of T. ulmifolia under the broad category of staminal nectaries, On the basis of present observations, the staminal nectaties of Twmnera are considered as a special nature and therefore are designated as “modified filament pouch nectaries”, which respresents the base of filament. Apart from these, five yellowish conical concave ovarian nectaties which are also stomatiferous and have been discused earlier. ‘The five filament pouch nectaries are superimposed on the five conical concave ovarian nectaries and are held together. ‘The floral tube falls off on the following day exposing the five concave nectaties which gradually become prominent after 3 to 5 days of growth. ‘These are classified as ovarian nectaries which lic immediately below the filaments; but to our knowledge are not described in the literature, This can be accommo- dated under the 5th category of Fahn (1979) i... ovarial nectary but the glandular nature of these does not fit into any of the categories; hence we propose new category i.e, ‘Nectary around the ovary immediately below the floral tube’. ACKNOWLEDGEMENTS We thank the Head, Department of Botany. Kakatiya University, Warangal for the facilities. 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