Nikolić, D. et al.  The influence of orographical and bioclimatic factors … BIOLOGICA NYSSANA 6 (1)  September 2015: 1-9 Nikolić, D. et al.  The influence of orographical and bioclimatic factors … 1 Original Article Received: 15 July 2015 Revised: 18 August 2015 Accepted: 01 September 2015 The influence of orographical and bioclimatic factors on morphological variability of analyzed characters of Jovibarba heuffelii (Schott) A. Löve & D. Löve (Crassulaceae) Danijela Nikolić1*, Jasmina Šinžar-Sekulić2, Vladimir Ranđelović1, Dmitar Lakušić2 1University of Niš, Faculty of Sciences and Mathematics, Department of Biology and Ecology, Višegradska 33, 18000 Niš, Serbia 2University of Belgrade, Faculty of Biology, Institute of Botany and Botanical Garden „Jevremovac”, Takovska 43, 11000 Belgrade, Serbia * E-mail: danid@pmf.ni.ac.rs Abstract: Nikolić, D., Šinžar-Sekulić, J., Ranđelović, V., Lakušić, D.: The influence of orographical and bioclimatic factors on morphological variability of analyzed characters of Jovibarba heuffelii (Schott) A. Löve & D. Löve (Crassulaceae). Biologica Nyssana, 6 (1), September 2015: 1-9. The goal of this paper was to determine the extent of morphological variability in species J. heuffelii caused by orographic and bioclimatic factors. Samples were collected from 14 populations of species J. heuffelii, from the territories of Serbia, Macedonia, Bulgaria and Romania. For this purpose cluster analysis (UPGMA) on bioclimatic parameters and regression analysis were performed. The cluster analysis of bioclimatic factors has shown that the study area was influenced by semiarid temperate-continental or subcontinental climate, continental mountain climate and humid mountain climate. Among the orographic factors, the greatest influence on morphological characters of species J. heuffelii was determined for altitude, exposition and slope of the terrain. The mean temperature of the wettest quartile (BIO8) and temperature seasonality (BIO4) have shown the greatest influence on the morphological characters of J. heuffelii. Key words: Jovibarba heuffelii, morphological variability, environmental factors Apstrakt: Nikolić, D., Šinžar-Sekulić, J., Ranđelović, V., Lakušić, D.: Uticaj orografskih i bioklimatskih faktora na morfološku varijabilnost analiziranih karaktera Jovibarba heuffelii (Schott) A. Löve & D. Löve (Crassulaceae). Biologica Nyssana, 6 (1), Septembar 2015: 1-9. Cilj ovog rada je bio da se utvrdi u kolikoj meri je velika morfološka varijabilnost kod vrste J. heuffelii uslovljena uticajem orografskih i bioklimatskih faktora. Prikupljeni su uzorci 14 populacija vrste J. heuffelii sa područja Srbije, Makedonije, Bugarske i Rumunije. Urađena je klaster analiza (UPGMA) sa bioklimatskim parametrima i regresiona analiza. Klaster analiza bioklimatskih faktora je pokazala da je istraživano područje pod uticajem tri tipa klime: semiaridne umereno kontinentalne ili subkontinentalne klime, kontinentalne planinske klime i humidne planinske klime. Od orografskih faktora pored nadmorske visine, ekspozicija i nagib terena imaju najveći uticaj na morfološke karaktere vrste J. heuffelii. Srednja temperatura najvlažnijeg kvartala (BIO8) i temperaturna sezonalnost (BIO4) najviše utiču na morfološke karaktere J. heuffelii. Key words: Jovibarba heuffelii, morfološka varijabilnost, ekološki faktori 6 (1) • September 2015: 1-9 BIOLOGICA NYSSANA 6 (1)  September 2015: 1-9 Nikolić, D. et al.  The influence of orographical and bioclimatic factors … 2 Introduction J. heuffelii (Crassulaceae) is a succulent xerophyte primarily inhabiting rocky habitats at various substrates (limestone, serpentinite, silicate) in a range of altitudes from the coastline to 2500 m above sea level (B a r c a & N i c u l a e , 2005; L a k u š i ć et al., 2005; D i m i t r i j e v i ć et al., 2011). Distribution of this species in Balkan Peninsula and the Southern Carpathians indicates that this is a European endemic species (M e u s e l , 1965; J a l a s et al., 1999). According to the climatic division of southeastern Europe (H o r v a t et al., 1974) and division into main climate and biome types by W a l t e r and L e i t h (1964), there are two main climate types within the range of J. heuffelii complex: temperate-continental climate (type VI) and mountain climate (type X). The temperate- continental climate is represented by subtype VI 3 - semiarid temperate-continental climate (Moesian- Carpathian variant). Within the mountain climate there are two subtypes: X 2 - humid mountain climate of alpine type (Illyrian variant) and X 3 – continental mountain climate (Moesian-Carpathian variant). This species is characterized by a very high morphological variability, as evidenced by recent studies on variability of morphological characteristics of vegetative and reproductive organs (D i m i t r i j e v i ć et al., 2011) and analysis of morphological characteristics of nectaries (N i k o l i ć et al., 2015a). The impact of orographic and bioclimatic habitat factors on morphological variability was analyzed in paper by N i k o l i ć et al. (2015b), while this paper pertains only to populations from identical mountain habitats, while gorge and canyon populations were omitted from the analysis. Regression analysis and cluster analysis (UPGMA) based on matrix of bioclimatic parameters were performed in order to determine the extent of impact of orographic and bioclimatic factors on variability of morphological characteristics and differentiation of all analyzed populations. Table 1. The list of population of J. heuffelii used in this study. Vouchers are deposited in the Herbarium of the Institute of Botany, Faculty of Biology, University of Belgrade (BEOU) Population Coordinate Substrate Altitude Voucher Individuals 1. RO-Domogled 44°52'41.70"N 22°25'54.11"E limestone 1300 BEOU- 16510 20 2. SR-Gradac 44°15'17.98"N 19°53'23.00"E limestone 490 BEOU-16458 21 3. SR-Suvaja 44°10'54.98"N 19°52'11.08"E limestone 417 BEOU-16457 20 4. SR-Studenica 43°29'20.56"N 20°32'7.74"E serpentinite 486 BEOU-16461 20 5. SR- Nebeske stolice 43°15'34.14"N 20°49'33.59"E serpentinite 1907 BEOU-16468 22 6. SR-Treska 43°15'36.31"N 20°47'6.40"E serpentinite 1628 BEOU-16462 20 7. SR-Basarski kamik 43° 9'37.29"N 22°42'9.57"E limestone 1350 BEOU- 16460 25 8. SR-Radan 42°55'4.99"N 21°33'25.74"E silicate 802 BEOU-16456 20 9. SR-Pljačkovica 42°34'47.20"N, 21°53'31.09"E silicate 674 BEOU-16465 21 10. SR- Besna Kobila 42°31'45.08"N 22°13'51.10"E silicate 1900 BEOU- 16463 30 11. SR-Stara planina 43°23'23.41"N 22°38'1.98"E silicate 1840 BEOU-16459 20 12. BU-Trojanski prolaz 42°46'1.62"N 24°37'2.30"E silicate 1400 BEOU-16509 20 13. MA-Treskavec 41°24'14.73"N 21°32'14.44"E silicate 1250 BEOU- 16511 20 14. MA- Mavrovo 41°38'14.33"N 20°42'29.90"E limestone 1300 BEOU- 16512 20 BIOLOGICA NYSSANA 6 (1)  September 2015: 1-9 Nikolić, D. et al.  The influence of orographical and bioclimatic factors … 3 Table 2. Morphological characters investigated using the populations of J. heuffelii described in Table 1. MORPHOLOGICAL CHARACTERS ABBREVIATIONS 1 Diameter of rosette (mm) Ros_D 2 Number of leaves in rosette LeRos_N 3 Length of biggest leaf (mm) LeRos_L max 4 Width of biggest leaf (mm) LeRos_W max 5 Distance of widest part of leaf from the top of the leaf (mm) Apex_D_Ros 6 Length of spike (mm) LeRos_Sp_L 7 Length of cilia (mm) LeRos_Ci_L 8 Width of cartilaginous leaf edge (mm) LeRos_Ed_W 9 Height of stem to lowest flower branch (mm) Ste_H 10 Number of leaves at stem LeSte_N 11 Length of middle leaf on stem (mm) MidLeSte_L 12 Width of middle leaf on stem (mm) MidLeSte_W 13 Distance of widest part of leaf from the top of the leaf (mm) Apex_D_Ste 14 Number of floral branches FloBra_N 15 Number of flowers at stage of ripening fruit Flo_N 16 Length of longest branch in fruit (mm) FloBra_L 17 Length of sepal (mm) Sep_L 18 Width of sepal (mm) Sep_W 19 Length of petal (mm) Pet_L 20 Width of petal (mm) Pet_W 21 Length of longest filament (mm) Fil_L max 22 Height of ovary (mm) Ova_H 23 Height of stylus (mm) Sty_H 24 Height of central tooth on petal (mm) CenToo_H 25 Height of lateral tooth on petal (mm) LatToo_H 26 Height of fruit (mm) Fru_H 27 Width of fruit (mm) Fru_W 28 Length of rostrum (mm) Rost_L 29 Total seed length (mm) See_L 30 Total seed width (mm) See_W 31 Width of central longitudinal fold (costa) (mm) Cos_W 32 Width of nectary (mm) Nect_W 33 Height of nectary (mm) Nect_H 34 The angle between carpels and nectaries (degree) Nect-Ang Material and methods Plant material Fourteen populations (299 individuals) of J. heuffelii were collected for morphological analysis. The samples were taken from Serbia, Macedonia Bulgaria and Romania (during two growing seasons 2010, 2011). The voucher specimens are deposited in the Herbarium of the Institute of Botany and Botanical Garden “Jevremovac", Faculty of Biology, University of Belgrade - BEOU (Tab. 1). Morphometric analyses Morphometric analyses were performed on dissected plant organs (leaves, stems, flowers, fruits and seeds). Leaves and inflorescences were stored in a glycerol: 96% ethanol solution (50:50). For measuring of flowers, the microscope slides were used, where all flower parts (sepals, petals, stamens, carpels) were separated and individually placed on the microscope slide. Slides were first scanned (ScanExpress A3 USB, Mustek) and then measured by using the LeicaQWin image analyzing program (Leica image software). For purposes of measuring BIOLOGICA NYSSANA 6 (1)  September 2015: 1-9 Nikolić, D. et al.  The influence of orographical and bioclimatic factors … 4 the smallest details on leaves, flowers and nectaries, these organs were photographed with the LEICA DM 1000 microscope while seeds were photographed with the LEICA MZ16 A stereomicroscope. Thirty four morphological characters were used in these analyses (Tab. 2). Statistical analysis Cluster analysis (UPGMA) was performed in order to evaluate the bioclimatic differentiation between the habitats of the 14 investigated populations. Each location was characterized using 19 bioclimatic parameters, extracted from the WorldClim set of global climate layers (H i j m a n s et al., 2005). The extraction of bioclimatic parameters was done with DIVA-GIS 7.5 software (H i j m a n s et al., 2012). Regression analysis (linear regression) was performed in order to estimate the correlations between the variation of morphological characters of J. heuffelii and basic orographic, geological, and bioclimatic habitat characteristics, as well as the geographic position of each population. The geographic positions were recorded using a hand- held Global Positioning System (GPS Garmin eTrex Vista® C). Orographic characteristics including altitude, slope and aspect were calculated from the Shuttle Radar Topography Mission (Reuter et al., 2007) at an approximate 90 m pixel resolution using ARCGIS 10 Spatial Analyst. Prior to the regression analysis, habitat characteristics were tested for multicollinearity. Bioclimatic predictors that have shown significant correlations with other predictors were not used in regression analysis of morphological characters. All statistical analyses were performed using the package Statistica 5.1 (Statsoft, 1996). Results and discussion Cluster analysis of bioclimatic data Cluster analysis based on bioclimatic factors showed presence of 3 clusters (Fig. 1). The first cluster (C1) included localities with semiarid, temperate-continental climate or subcontinental climates. These habitats appear in gorges of rivers Gradac, Suvaja and Studenica as well as at Domogled in Romania. This cluster also included populations from Bulgaria (BU-Trojanski prolaz - central part of Stara planina) and the population from Mt. Stara planina in the E part of Serbia. The second cluster (C2) contained localities with continental mountain climates (SR-Besna Kobila, SR-Radan, SR-Basarski kamik, SR-Pljačkovica and MA-Treskavec). The third cluster (C3) includes localities with humid mountain climate (SR-Treska, SR-Nebeske stolice and MA-Mavrovo). Fig. 1. Results of cluster analysis for populations of J. heuffelii based on habitat climatic characteristics (C1, C2, C3, climate type, for details see Tab. 2.) BIOLOGICA NYSSANA 6 (1)  September 2015: 1-9 Nikolić, D. et al.  The influence of orographical and bioclimatic factors … 5 T a b le 3 . M in im a l, a v e ra g e a n d m a x im a l v a lu e o f 1 9 b io c li m a ti c p a ra m e te rs o f lo c a li ti e s st u d ie d . C 1 , C 2 a n d C 3 r e p re se n t c li m a te t y p e s c o rr e sp o n d t o t h e g ro u p in g s o b ta in e d f ro m t h e c lu st e r a n a ly si s o f b io c li m a ti c p a ra m e te rs ( se e F ig . 1 ). T h e b io c li m a ti c v a ri a b le s th a t w e re u se d a s p re d ic to rs i n r e g re ss io n a n a ly si s a re m a rk e d w it h a st e ri sk ( * ). B io c li m a ti c p a r a m e te r s C 1 C 2 C 3 se m i -a ri d m o d e ra te ly c o n ti n e n ta l c li m a te c o n ti n e n ta l m o u n ta in c li m a te h u m id m o u n ta in c li m a te M in A v g M a x M in A v g M a x M in A v g M a x A n n u a l m e a n t e m p e ra tu re ( 1 ) 3 .3 7 7 .7 1 1 1 .3 3 3 .3 0 7 .7 4 9 .7 2 2 .8 7 4 .2 3 4 .9 8 M e a n m o n th ly t e m p e ra tu re r a n g e ( 2 ) * 7 .9 8 9 .1 1 0 .1 8 .7 3 9 .1 1 0 .3 6 7 .4 8 8 .1 6 9 .0 4 Is o th e rm a li ty ( 2 /7 ) (* 1 0 0 ) (3 ) * 2 9 .4 9 3 1 .7 4 3 3 .6 7 3 1 .6 4 3 3 .3 0 3 4 .6 4 2 9 .3 4 3 0 .8 8 3 3 .4 9 T e m p e ra tu re s e a so n a li ty ( S T D * 1 0 0 ) (4 ) * 6 7 6 .6 0 7 2 4 .8 4 7 5 5 .9 2 6 6 1 .0 8 7 2 1 .5 2 7 5 0 .3 9 6 4 5 .8 4 6 6 8 .4 9 6 9 1 .6 4 M a x . te m p e ra tu re o f th e w a rm e st m o n th ( 5 ) 1 7 .2 2 2 .6 5 2 6 .8 1 7 .6 2 3 .4 8 2 4 .1 1 6 .4 1 8 .3 1 9 .6 M in . te m p e ra tu re o f th e c o ld e st m o n th ( 6 ) -3 .2 -5 .9 7 -8 .9 -4 .4 -5 .9 3 -9 -7 .4 -8 .1 -9 .1 A n n u a l te m p e ra tu re r a n g e ( 5 – 6 ) (7 ) * 2 6 .1 2 8 .6 2 3 0 .2 2 6 .6 2 9 .4 3 0 .4 2 5 .5 2 6 .4 2 7 M e a n t e m p e ra tu re o f th e w e tt e st q u a rt e r (8 ) 9 .6 1 4 .6 1 1 8 .4 5 3 .9 3 1 0 .2 9 1 3 .5 5 -1 .3 6 .3 6 1 1 .3 2 M e a n t e m p e ra tu re o f th e d ri e st q u a rt e r (9 ) * -0 .9 2 3 .4 5 9 .5 8 1 .8 1 1 .2 7 1 6 .5 2 -2 .7 5 6 .0 6 1 2 .6 3 M e a n t e m p e ra tu re o f th e w a rm e st q u a rt e r (1 0 ) 1 1 .4 8 1 6 .3 6 2 0 .1 7 1 1 .2 5 1 6 .3 5 1 8 .5 8 1 0 .8 5 1 2 .2 6 1 3 .1 2 M e a n t e m p e ra tu re o f th e c o ld e st q u a rt e r (1 1 ) -4 .9 5 -1 .4 2 2 .0 1 -3 .3 -1 .3 4 0 .1 7 -5 .3 8 -4 .0 2 -2 .9 A n n u a l p re c ip it a ti o n ( 1 2 ) * 7 7 8 8 1 5 .1 7 8 4 9 6 0 6 6 6 5 .7 5 7 4 2 9 5 0 1 0 0 1 .6 7 1 0 4 5 P re c ip it a ti o n o f th e w e tt e st m o n th ( 1 3 ) 8 7 1 0 0 .3 3 1 2 1 6 8 7 6 .7 5 8 4 1 0 4 1 1 3 .6 7 1 2 7 P re c ip it a ti o n o f th e d ri e st m o n th ( 1 4 ) * 4 2 5 0 5 5 4 0 4 2 .7 5 4 6 5 6 6 2 .6 7 6 9 P re c ip it a ti o n s e a so n a li ty ( C V ) (1 5 ) * 1 7 .5 7 2 3 .1 3 3 4 .4 0 1 6 .6 7 1 9 .1 0 2 1 .2 9 1 5 .1 3 1 8 .9 2 6 .2 4 P re c ip it a ti o n o f th e w e tt e st q u a rt e r (1 6 ) 2 5 5 2 6 9 .1 3 3 1 0 1 7 2 2 0 0 .7 5 2 3 1 2 8 5 3 1 2 .3 3 3 4 8 P re c ip it a ti o n o f th e d ri e st q u a rt e r (1 7 ) 1 4 7 1 6 2 .1 7 1 7 5 1 2 7 1 3 6 .2 5 1 4 7 1 7 8 2 0 2 2 2 2 P re c ip it a ti o n o f th e w a rm e st q u a rt e r (1 8 ) * 2 2 0 2 4 5 .1 7 2 9 1 1 3 6 1 6 1 .7 5 2 0 4 1 7 9 2 3 5 2 7 2 P re c ip it a ti o n o f th e c o ld e st q u a rt e r (1 9 ) * 1 5 8 1 7 7 .5 1 8 7 1 4 7 1 6 0 1 9 4 2 1 8 2 5 5 3 1 0 BIOLOGICA NYSSANA 6 (1)  September 2015: 1-9 Nikolić, D. et al.  The influence of orographical and bioclimatic factors … 6 T a b le 4 . S u m a ry s ta ti st ic s o f re g re ss io n ( R 2 ) fo r in d e p e n d e n t b io c li m a ti c , g e o g ra p h ic a l a n d g e o lo g ic a l v a ri a b le s. A c r o n y m A L T A S P S L O B IO 2 B IO 3 B IO 4 B IO 7 B IO 8 B IO 9 B IO 1 2 B IO 1 3 B IO 1 4 B IO 1 5 B IO 1 8 B IO 1 9 R o s_ D 0 .0 7 * * 0 .0 2 0 .0 1 0 .0 2 0 .0 7 * * 0 .0 5 0 .0 0 0 .2 6 0 .1 1 0 .0 1 0 .0 0 0 .0 5 0 .0 1 0 .1 6 0 .0 2 L e R o s_ N 0 .2 6 * 0 .1 3 * 0 .0 2 0 .2 5 * 0 .1 3 * 0 .1 9 * 0 .2 8 * 0 .0 8 * * 0 .0 1 0 .0 2 0 .0 0 0 .0 4 0 .0 2 0 .0 1 0 .0 2 L e R o s_ L m a x 0 .0 9 * 0 .0 2 0 .0 1 0 .0 0 0 .0 5 0 .1 3 * 0 .0 2 0 .2 2 * 0 .0 1 * 0 .0 0 0 .0 3 0 .0 0 0 .0 9 * 0 .1 6 * 0 .0 9 * L e R o s_ W m a x 0 .1 2 * 0 .0 2 0 .0 0 0 .0 0 0 .0 4 0 .1 6 * 0 .0 4 0 .2 0 * 0 .1 9 * 0 .0 1 0 .0 5 0 .0 2 0 .0 3 0 .1 4 * 0 .0 2 A p e x _ D 1 0 .0 6 0 .0 2 0 .0 0 0 .0 0 0 .0 0 0 .0 7 0 .0 2 0 .1 0 * 0 .0 4 0 .0 0 0 .0 0 0 .0 0 0 .0 2 0 .0 4 0 .0 4 L e R o s_ S p _ L 0 .0 0 0 .0 0 0 .0 3 0 .0 0 0 .0 0 0 .0 0 0 .0 0 0 .0 0 0 .0 1 0 .0 2 0 .0 2 0 .0 1 0 .0 0 0 .0 1 0 .0 2 L e R o s_ C i_ L 0 .0 0 0 .0 1 0 .0 7 0 .0 0 0 .0 1 0 .0 4 0 .0 1 0 .0 0 0 .0 1 0 .0 0 0 .0 2 0 .0 2 0 .1 0 * 0 .0 0 0 .0 2 L e R o s_ E d _ W 0 .0 3 0 .0 4 0 .0 4 0 .0 0 0 .0 0 0 .0 1 0 .0 1 0 .0 3 0 .0 1 0 .0 0 0 .0 3 0 .0 7 * * 0 .1 4 * 0 .0 1 0 .0 1 S te _ H 0 .1 7 * 0 .0 4 0 .0 1 0 .0 0 0 .0 4 0 .1 7 * 0 .0 5 0 .3 0 * 0 .1 2 * 0 .0 0 0 .0 0 0 .0 4 0 .0 1 0 .0 8 * * 0 .0 5 L e S te _ N 0 .0 0 0 .0 1 0 .0 0 0 .0 6 0 .1 2 * 0 .0 0 0 .0 1 0 .0 5 0 .0 7 * * 0 .0 3 0 .0 0 0 .1 6 * 0 .1 3 * 0 .0 3 0 .0 0 M id L e S te _ L 0 .0 0 0 .0 1 0 .0 0 0 .0 3 0 .0 5 0 .0 0 0 .0 1 0 .0 4 0 .0 3 0 .0 0 0 .0 2 0 .0 0 0 .0 4 0 .1 0 * 0 .0 2 M id L e S te _ W 0 .0 0 0 .0 0 0 .0 1 0 .0 9 * 0 .1 9 * 0 .0 2 0 .0 2 0 .1 0 * 0 .2 0 * 0 .0 6 0 .0 7 * * 0 .0 9 * 0 .0 0 0 .2 2 * 0 .0 0 A p e x _ D 2 0 .0 5 0 .0 0 0 .0 1 0 .0 0 0 .0 5 0 .0 7 * * 0 .0 1 0 .2 3 * 0 .0 8 * * 0 .0 1 0 .0 2 0 .0 1 0 .0 4 0 .0 7 * * 0 .1 0 * S e p _ L 0 .0 3 0 .0 0 0 .0 9 * 0 .0 0 0 .0 0 0 .0 1 0 .0 0 0 .0 8 * * 0 .0 2 0 .0 6 0 .1 0 * 0 .0 0 0 .0 6 0 .0 0 0 .0 6 S e p _ W 0 .0 2 0 .0 0 0 .0 7 0 .0 0 0 .0 1 0 .0 1 0 .0 0 0 .0 7 * * 0 .0 0 0 .0 1 0 .0 0 0 .0 0 0 .0 0 0 .0 1 0 .0 3 P e t_ L 0 .2 2 * 0 .0 6 0 .0 4 0 .0 2 0 .0 1 0 .1 9 * 0 .0 9 * 0 .3 1 * 0 .0 5 0 .0 1 0 .0 1 0 .0 2 0 .0 3 0 .0 3 0 .0 6 P e t_ W 0 .0 1 0 .0 1 0 .0 4 0 .0 3 0 .0 8 * * 0 .0 1 0 .0 0 0 .0 4 0 .0 4 0 .0 1 0 .0 0 0 .1 1 * 0 .0 9 * 0 .0 1 0 .0 0 F il _ L m a x 0 .2 3 * 0 .1 1 * 0 .0 0 0 .0 2 0 .0 0 0 .1 7 * 0 .0 8 * * 0 .3 4 * 0 .0 8 * * 0 .0 0 0 .0 1 0 .0 5 0 .0 5 0 .0 6 0 .0 3 O v a _ H 0 .2 3 * 0 .1 3 * 0 .0 4 0 .0 1 0 .0 1 0 .1 5 * 0 .0 7 * * 0 .3 1 * 0 .0 4 0 .0 0 0 .0 0 0 .0 3 0 .0 2 0 .0 4 0 .0 3 S ty _ H 0 .2 9 * 0 .1 5 * 0 .0 0 0 .0 4 0 .0 0 0 .2 9 * 0 .1 6 * 0 .3 6 * 0 .1 0 * 0 .0 0 0 .0 0 0 .0 2 0 .0 1 0 .0 9 * 0 .0 7 * * C e n T o o _ H 0 .0 2 0 .0 0 0 .0 5 0 .0 1 0 .0 0 0 .0 0 0 .0 1 0 .0 2 0 .0 1 0 .0 2 0 .0 9 * 0 .0 0 0 .1 2 * 0 .0 3 0 .0 1 L a tT o o _ H 0 .0 1 0 .0 4 0 .0 1 0 .0 0 0 .0 0 0 .0 1 0 .0 0 0 .0 1 0 .0 1 0 .0 1 0 .0 9 * 0 .0 0 0 .1 7 * 0 .0 8 * * 0 .0 0 F ru _ H 0 .3 1 * 0 .1 4 * 0 .0 2 0 .0 3 0 .0 0 0 .2 2 * 0 .1 2 * 0 .4 2 * 0 .0 6 0 .0 0 0 .0 2 0 .0 5 0 .0 7 0 .0 4 0 .0 5 F ru _ W 0 .3 1 * 0 .1 4 * 0 .0 4 0 .0 3 0 .0 0 0 .2 1 * 0 .1 2 * 0 .4 0 * 0 .0 5 0 .0 0 0 .0 2 0 .0 4 0 .0 7 0 .0 3 0 .0 5 R o st _ L 0 .2 5 * 0 .3 7 * 0 .0 8 * * 0 .0 4 0 .0 0 0 .2 6 * 0 .1 3 * 0 .1 9 * 0 .0 0 0 .0 0 0 .0 7 * * 0 .0 0 0 .2 1 * 0 .1 1 * 0 .0 6 F lo B ra _ N 0 .0 3 0 .0 2 0 .0 0 0 .0 1 0 .0 5 0 .0 3 0 .0 0 0 .1 1 * 0 .0 7 * * 0 .0 1 0 .0 1 0 .0 3 0 .0 0 0 .0 9 * 0 .0 0 F lo _ N 0 .0 9 0 .0 5 0 .0 0 0 .0 1 0 .0 0 0 .0 6 0 .0 3 0 .1 0 * 0 .0 4 0 .0 0 0 .0 1 0 .0 0 0 .0 1 0 .0 4 0 .0 1 F il _ L m a x 0 .1 1 0 .0 0 0 .0 0 0 .0 0 0 .0 6 0 .1 3 * 0 .0 2 0 .3 8 * 0 .2 0 * 0 .0 2 0 .0 7 0 .0 2 0 .1 1 * 0 .0 6 0 .1 0 * S e e _ L 0 .0 7 * * 0 .0 8 * * 0 .0 2 0 .0 0 0 .0 0 0 .0 2 0 .0 1 0 .0 7 * * 0 .0 6 0 .0 2 0 .0 6 0 .0 0 0 .0 6 0 .0 9 * 0 .0 0 S e e _ W 0 .1 3 * 0 .0 3 0 .0 0 0 .0 5 0 .0 2 0 .0 5 0 .0 6 0 .0 8 * * 0 .0 4 0 .0 0 0 .0 0 0 .0 0 0 .0 0 0 .0 0 0 .0 0 C o s_ W 0 .0 2 0 .0 0 0 .0 2 0 .0 1 0 .0 0 0 .0 1 0 .0 1 0 .0 7 0 .0 1 0 .0 1 0 .1 7 * 0 .0 3 0 .4 1 * 0 .0 1 0 .0 0 N e c t_ W 0 .0 8 * * 0 .1 0 * 0 .0 3 0 .1 7 * 0 .1 3 * 0 .0 5 0 .1 4 * 0 .0 0 0 .0 6 0 .0 6 0 .1 1 * 0 .0 2 0 .0 4 0 .1 2 * 0 .0 2 N e c t_ H 0 .0 4 0 .0 4 0 .0 0 0 .0 0 0 .0 1 0 .0 7 * * 0 .0 3 0 .0 4 0 .0 5 0 .0 3 0 .0 3 0 .0 3 0 .0 0 0 .0 4 0 .0 0 N e c t- A n g 0 .0 4 0 .0 1 0 .0 3 0 .0 8 * * 0 .0 8 * * 0 .0 1 0 .0 5 0 .0 0 0 .0 1 0 .0 5 0 .0 4 0 .0 4 0 .0 0 0 .0 5 0 .0 1 A b b re v ia ti o n s: A L T -a lt it u d e , A S P - a sp e c t, S L O -s lo p e , B IO - b io c li m a ti c c h a ra c te ri st ic s (s e e T a b le 3 ) * p < 0 .0 5 , * * p < 0 .0 1 BIOLOGICA NYSSANA 6 (1)  September 2015: 1-9 Nikolić, D. et al.  The influence of orographical and bioclimatic factors … 7 The analysis of the three clusters regarding the values of bioclimatic parameters (Tab. 3) shows that type C1 is characterized by the highest values of BIO1 bioclimatic parameter, the mean annual temperature within the range 3.37-11.33 °C. C2 type has shown variation in this character in range of 3.30-9.72 °C, while C3 climate type has shown the lowest values of this parameters, 2.87 - 4.98 °C (Tab. 3). A similar trend was noticed in bioclimatic parameters BIO5, BIO6, BIO8, BIO10 and BIO11, while in the bioclimatic parameter BIO9 (mean temperature of the driest quarter) the highest values were recorded for the second type of climate C2 (1.8-16.52 °C), followed by the third type C3 (–2.75 -12.63 °C) while the lowest values were recorded in the first climate type C1 (–0.92-9.58 °C). The analysis of these three clusters according to bioclimatic factors (Tab. 3) indicated that the greatest differences between clusters were present in the amount of precipitation, as measured by the factors Annual precipitation (BIO12), Precipitation of the driest month (BIO14) and Precipitation of the wettest quarter (BIO16). The largest amount of precipitation was present in the third cluster, the second had the lowest amount of precipitation, while the first cluster was intermediate. Considering the temperature, the highest values were recorded in canyons and river gorges (C1) and the lowest values in the third cluster (C3). There is an important question: to which extent do bioclimatic factors in analyzed habitats cause the differentiation in populations? The answer to this question may be provided by cluster analysis including both the bioclimatic characters and the morphological characteristics of individuals from the analyzed populations. If the position of populations in the cluster analysis of bioclimatic factors matches the position obtained from morphometric characters, it may be concluded that such grouping is a result of impact by various bioclimatic factors and vice versa (K u z m a n o v i ć et al., 2011). As cluster analysis of bioclimatic factors have shown that the study area is strongly differentiated into three main clusters: one with semiarid temperate continental or subcontinental climate, another with continental mountain climate and the third cluster with humid mountain climate, it might be expected that the cluster analysis of morphological characters would also produce three main clusters. However, the analysis has shown that within the cluster analysis of morphological characters all populations were divided into four clusters (N i k o l i ć et al., 2015b), while the distribution of individual clades (populations) was not matched to the distribution of populations obtained in the cluster analysis of bioclimatic parameters. This type of distribution indicates that bioclimatic factors are not the single reason for such morphological differentiation of populations. Results of the cluster analysis performed solely on mountain populations were similar. The effect of microclimatic differences caused by orography on morphological differentiation was avoided in this analysis, as all mountain populations inhabit open grassland habitats from classes Festuco-Brometea and Elyno-Seslerietea. Although both cluster analyses, of morphological characters and bioclimatic parameters, have shown separation into two groups, the placement of individual populations into these groups is different, indicating that bioclimatic parameters are not crucial in differentiation of populations (N i k o l i ć et al., 2015b). Regression analysis (Linear regression) Regression analysis showed that orographic factors (altitude, aspect and slope) influenced the morphological characters of the species J. heuffelii. The geological substrate was not analyzed, as this factor showed a high level of co-linearity with other factors (Tab. 4). Altitude had the greatest influence on the following morphological characters: length of rostrum, height of fruit, height of stylus, width of fruit, number of leaves in rosette, length of the longest branch in floral maturation stage, length of the longest filament. Exposition aspect is also one of the key abiotic factors, with the greatest impact on morphological characters of reproductive organs (ovary stylus, fruit and nectary). The morphological differentiation of the analyzed populations is also highly influenced by the bioclimatic factors precipitation and temperature. The greatest correlation with the analyzed characters was shown by Mean temperature of driest quarter (BIO8) and Temperature seasonality (BIO4). These two bioclimatic factors have the highest influence on the same characters that are highly influenced by altitude. There is a somewhat smaller but still significant impact of Precipitation seasonality (BIO15), with the greatest impact on following characters: length of petal, height of ovary, height of the stem to the lowest flower branch, width of the nectary, width of the middle leaf on the stem, width of the largest leaf, length of the largest leaf, width of the central longitudinal fold (costa). There is a somewhat smaller but still significant impact of Precipitation of warmest quarter (BIO18) and Mean Temperature of driest quarter (BIO9). BIOLOGICA NYSSANA 6 (1)  September 2015: 1-9 Nikolić, D. et al.  The influence of orographical and bioclimatic factors … 8 The geographic variability in plant morphology is the result of phenotypic changes expressed as an answer to local ecological conditions, genetic variability and evolution among the populations, as well as the biogeographic history of species (E l l i s o n et al., 2004). Some of the morphological characteristics have genetic origins, for example leaf shape, but they may also be strongly influenced by local conditions in which the plants develop (T h o m p s o n , 1991; S c h l i c h t i n g & P i g l i u c c i , 1998). Altitude is a factor with significant impact on variability of morphological characteristics of analyzed populations, as shown by regression analysis. Change in altitude causes changes in other factors, including temperature, air humidity, precipitation and partial pressure of gasses in the atmosphere, causing the adaptive response in plants and therefore also changes in morphology and physiology (K ö r n e r , 1999). Altitude may have a strong impact on both leaf morphology and physiology in individuals from various populations within the same species (H o v e n d e n & V a n d e r S c h o o r , 2004). With increase of altitude the length and width of leaves generally decrease (K ö r n e r et al., 1986), while thickness of leaves increases (K ö r n e r et al., 1989; R o d e r i c k et al., 2000). Regression analysis has supported the hypothesis that altitude has the greatest impact on variability of morphological characters in J. heuffelii populations. Conclusion The regression analysis has shown that morphological characteristics are under stronger influence by seasonal dynamics in temperature and precipitation than by the total amount of precipitation or mean annual temperature, which are the most commonly analyzed climatic factors for any geographic area. In addition to altitude, the orographic factors with the strongest impact on morphological characters of species J. heuffelii are exposition and slope of terrain. The bioclimatic parameters may also influence the variability in morphological characters. The greatest impact was recorded in mean temperature of wettest quartile (BIO8) and temperature seasonality (BIO4). They are mostly influencing the following characters: height and width of fruit, height of stylus, length of rostrum, number of leaves in the rosette, length of the longest flowering branch and length of longest filament of the stamen. The seasonal character of precipitation (BIO15) was shown to influence size of leaves, height of stem and reproductive characters: length of petal, height of ovary and width of nectary. These results suggest that temperature conditions and precipitation may contribute to plant phenotype in various habitat types. This trend was also recorded in the populations from canyons and river gorges with C1 type of climate. 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