Acta Botanica 1-2015 - za web.indd ACTA BOT. CROAT. 74 (1), 2015 1 Acta Bot. Croat. 74 (1), 1–17, 2015 CODEN: ABCRA 25 ISSN 0365-0588 eISSN 1847-8476 Morphometric variation and taxonomic identifi cation of thirteen wild rose populations from Tunisia ZOHRA BEN CHEIKH-AFFENE1*, FAOUZI HAOUALA2, FETHIA HARZALLAH-SKHIRI3 1 Department of Horticultural Science and Landscape, Higher Agronomic Institute, University of Sousse, 4042 Chott-Mariem, Sousse, Tunisia 2 Department of Agronomy and Plant Biotechnology, National Institute of Agronomy of Tunisia, University of Carthage, 43 avenue Charles Nicolle, 1082 Tunis Mahrajène, Tunisia 3 Laboratory of Genetic, Biodiversity and Valorisation of Bioresources (LR11SE14), High Institute of Biotechnology of Monastir, University of Monastir, rue Tahar Haddad, Monastir 5000, Tunisia Abstract – Thirteen populations of wild roses (Rosa L.) growing in northern and central Tunisia have been used for studies on the discrimination between accessions and popula- tions. Thirty-eight morphological characters related to the branches, prickles, leaves and corymbs were measured on the collected accessions to study the phenotypic diversity among and within species. Principal component and hierarchical cluster analyses (PCA and HCA) separated rose accessions into two distinctive groups and eight subgroups. A taxonomic interpretation of the morphological variability has shown that Tunisian rose populations belong to two sections (Synstylae and Caninae) of the genus Rosa. Moreover, they have been identifi ed as seven separated taxa: R. sempervirens L., R. sempervirens var. submoshata Rouy., R. sempervirens var. prostrata Lindl. belonging to Synstylae section and R. canina L., R. agrestis Savi., R. micrantha Smith. and R. dumetorum Thuill. belong- ing to Caninae section. PCA and HCA proved that morphological characters used in taxo- nomic identifi cation such as styles form, leaf and leafl ets length, number of fl owers by corymb, leafl et serration, presence of glands in leafl et, peduncle, receptacle and sepal have a high value of discrimination, and have been very successful in morphological identifi ca- tion. Keywords: Caninae section, morphometry, Synstylae section, taxonomic differentiation, wild roses. * Corresponding author, e-mail: bencheikhz@hotmail.fr Copyright® 2015 by Acta Botanica Croatica, the Faculty of Science, University of Zagreb. All rights reserved. BEN CHEIKH-AFFENE Z., HAOUALA F., HARZALLAH-SKHIRI F. 2 ACTA BOT. CROAT. 74 (1), 2015 Introduction Roses are native to diverse habitats within the northern temperate hemispheres (Europe, Asia, the Middle East, North Africa and North America) and have been introduced and naturalized through the world. They have been planted around human habitats for at least two millennia for the ornamental and medicinal values of their fl owers (fl owering land- scape, cut fl owers, perfume, oil and rose water) (HUMMER and JANICK 2009). The genus Rosa L. is characterized by an enormous phenotypic plasticity that has challenged bota- nists. Indeed, the taxonomic treatment of this highly diverse subgenus is complicated and it is caused by an enormous phenotypic, genotypic and ecological variability and plasticity due to some evolutionary process, such as hybridization, introgression, etc. (DE COCK 2008). Rose species exhibit a highly diverse ploidy level from diploid to octoploid, while decaploidy was observed in R. prealucens (JIAN et al. 2010) and pentaploidy (2n = 5x = 35) in species from section Caninae. Those species evolved a mode of reproduction unique in the plant kingdom a distinctive unbalanced meiosis termed ‘canina meiosis’(KLASTERSKA and NATARAJAN 1974, KOVARIK et al. 2008). The evidence for apomixis in wild Rosa L. is extremely limited and confi ned to the section Caninae (DICKINSON et al. 2003). CZAPIK (1994) mentioned that apomictic taxa are highly polymorphic with numerous microspecies leading to a diffi cult and controversial taxonomic treatment. The classifi cation system ad- opted by REHDER (1940), divided this genus in four subgenera, three of which are mono- typic; Hulthemia (Dumort) Focke, Platyrhodon (Hurst) Rehder and Hesperhodos Cocker- ell. Rosa, the fourth subgenus, contains about 95% of all species (about 200 species) and was subdivided into 10 sections: Pimpinellifoliae (DC.) Ser, Gallicanae (DC.) Ser, Cani- nae (DC.) Ser, Carolinae Crép, Cinnamomeae (DC.) Ser, Synstylae (DC.), Chinensis (DC.) Ser, Banksiae Lindl, Laevigatae Michx. and Bracteatae Thory., each of the four latter sec- tions have only one to three species (RITZ et al. 2005). Rehder’s system was widely accept- ed and is still used as a basis for modern treatises (BAILEY 1963, WISSEMAN 2003, NYBOM 2009). During recent years, wild roses have acquired great importance in research, not only in genetic but in other domains. In fact, rose hips, the fruits of the rose plant are in many Euro- pean countries used in food products (GAO et al. 2005) and for medicinal purposes (WAR- HOLM et al. 2003, REIN et al. 2004, BOSKABADY et al. 2006). Indeed, rose hips are rich in phyto-nutrients such as phenols, carotenoids (lycopene), vitamins (C, A, E and P) (EREN- TURK et al. 2005, OLSSON et al. 2005, BARROS et al. 2010) and carbohydrates (DEMIR and OZCAN 2001), as well as other bioactive compounds that have benefi cial medical effects. Recently, anti-infl ammatory properties (WINTHER et al. 1999), anti-ulcerogenic (GURBUZ et al. 2003), anti-oxidant (GAO et al. 2005, OZMEN et al. 2005) and anti-mutagenic activities have been demonstrated (KARAKAYA and KAVAS 1999). In Tunisia, native rose bushes used to grow in the ravines, in brush fi elds and hedges and in the forest from the North to the Tunisian Dorsal (Jebel Chaâmbi) (POTTIER-ALAPETITE 1979). But in the last ten years, climate changes and the programs of rural development and the expansion of the cities have threatened the biodiversity in Tunisia and caused the risk of the extinction of some species particularly Rosa accessions from northern and central hu- mid zones. Those populations were subjected to genetic erosion and the information about the species is limited. In this way it is urgent to prospect the Tunisian rose populations, to identify and essentially to conserve them for subsequently valorisation. MORPHOLOGICAL DIVERSITY OF TUNISIAN WILD ROSES ACTA BOT. CROAT. 74 (1), 2015 3 In this work we aim to study the variability within the genus Rosa accessions growing wild in the northern and the central part of Tunisia. To accomplish this we have analysed a plant material collection from 13 populations. The main objectives of this work were: 1) evaluation of morphometric variability and 2) to place the identifi ed groups in a hierarchi- cal classifi cation. Materials and methods Plant material Field collections of rose populations were carried out in April and May 2007–2010. The collection sites were selected according to the Flora of Tunisia (POTTIER-ALAPETITE 1979). From fi ve provinces, nine localities were chosen to cover the geographical range of wild roses. In the Northwest regions of Tunisia, nine populations were found to be naturally dis- tributed in seven localities. In each of the localities Tabarka, Aïn Drahem, Tebaba, Wechta- ta, Djeba and Kessra we found one type of population but in the Teboursouk locality we found three types. In the Northeast we found two populations in one locality (Sidi Median from Zaghouan). In the Center we found also two populations in the locality of Haffouz (Kairouan). The latitude longitude and altitude of each location were determined using the geographical positioning system (GPS) using a Magellan Explorist (Tab. 1). Morphological characters Studies were based on vegetative and fl oral characters. In each population fi ve acces- sions were sampled. Thirty samples of one year old branches of 100 cm of length (we re- spect all different positions and directions) were collected randomly in the spring (April– May). Thirty samples of leaves, infl orescences and fl owers in full bloom were taken to measure morphological characters (qualitative and quantitative characters). All voucher specimens are deposited in the High Institute of Agronomy of Chott-Meriem Botanic Labo- ratory (Herbarium ISACH) and each one was assigned a corresponding number and code RR110-RR123. Twenty one vegetative characters that are associated with the branches, prickles, leaves and leafl ets and gland density on those organs were defi ned. Seventeen fl oral characters that are associated with the corymb and with all other fl ower parts, and gland density on those parts were also defi ned (Fig. 1). Taxonomic differentiation Hierarchical classifi cation was based on Baily’s, Crépin’s and Maire’s classifi cations (BOULENGER 1924, 1932, 1937, CRÉPIN 1869, BAILEY 1963, MAIRE 1980, SILVESTRE and MONT- SERRAT 1998). In addition to the characters already established and to defi ne sections, we are also interested in styles (styles exerted beyond the mouth of receptacle, long or short, ex- erted styles free or connate into a column), in stipules (free, adnate only at the base or ad- nate more than one half of their length) and in the kind of infl orescence (solitary or corym- bose). BEN CHEIKH-AFFENE Z., HAOUALA F., HARZALLAH-SKHIRI F. 4 ACTA BOT. CROAT. 74 (1), 2015 Ta b. 1 . Pr ov in ce , l oc al ity , s pe ci es , c or re sp on di ng c od e an d ec o- ge og ra ph ic al p ar am et er s of s ite s ha rv es t. Pr ov in ce L oc al ity Sp ec ie s C od e N um be r of s am pl es B io cl im at ic st ag e L at itu de L on gi tu de A lti tu de (m ) Ja nd ou ba (N or th w es t) Ta ba rk a R . s em pe rv ir en s Ta b 10 H um id 36 °5 2’ 8 °4 3’ 9 1 A in D ra he m R . s em pe rv ir en s A in D 10 H um id 36 °4 0’ 8 °4 0’ 29 0 B ej a (N or th w es t) Te ba ba R . s em pe rv ir en s Te b 10 Su bh um id 36 °5 7’ 8 °5 7’ 11 4 W ec ht at a R . s em pe rv ir en s W ec h 10 Su bh um id 36 °5 4’ 9 °0 8’ 7 3 Te bo ur so uk R . c an in a Te bs 1 5 Su bh um id 36 °2 9’ 9 °1 8’ 22 8 R . a gr es tis Te bs 2 5 Su bh um id 36 °2 9 9 °1 8’ 22 8 R . s em pe rv ir en s Te bs 3 10 Su bh um id 36 °2 8’ 9 °1 6’ 29 0 D je ba R . s em pe rv ir en s D jb . 10 Su bh um id 36 °2 8’ 9 °0 5’ 29 8 Se lia na (T ra ns iti on z on e be tw ee n th e N or th w es t a nd th e N or th ea st ) K es sr a R . c an in a K es s 5 Se m i- ar id 35 °5 0’ 9 °2 2’ 94 2 Z ag ho ua n (N or th ea st ) Si di M ed ia n R . s em pe rv ir en s Z ag h1 10 Se m i- ar id 36 °2 0’ 10 °2 0’ 33 5 R . c an in a Z ag h2 5 Se m i- ar id 36 °2 0’ 10 °0 4’ 22 8 K ai ro ua n (C en te r) H af fo uz R . m ic ra nt ha H af z1 10 Se m i- ar id 35 °3 8’ 9 °3 9’ 26 4 R . a gr es tis H af z2 5 Se m i- ar id 35 °3 8’ 9 °3 9’ 26 4 MORPHOLOGICAL DIVERSITY OF TUNISIAN WILD ROSES ACTA BOT. CROAT. 74 (1), 2015 5 Statistical analysis The data were analysed using analysis of variance (ANOVA) and the signifi cance of the differences between means were determined at p < 0.05 using Duncan’s multiple range test. To evaluate the approximation or the removal of species all the data were subjected to prin- cipal components analysis (PCA) and hierarchical clusters analysis (HCA) (CRISCI and LO- PEZ ARMENGOL 1983, COTTIN 1988) using the SPSS 16 software (Statistical Package for the Social Sciences) and Excel 2007 software, allowing classifi cation of individuals into groups (MOLINA CANO 1977). Results Morphological diversity Mean values of morphological characters studied are reported in Tabs. 2, 3, 4. Data show large variability among accessions for almost characters. Among the examined characters diameter and length of prickles (Dia. Pr, L. Pr) were highly correlated with the length of prickle’s diagonal (L. Pr. dig) (75 and 88%, respectively) (data not reported). The length of leaf (L. Lf) was highly correlated with the width (W. Lf) and the area of the leaf (Lf. a) and with the length of rachis (L. ra) (84, 81 and 73%, respectively). The number of fl owers per corymb (Nb. Fl. co) was correlated with the length of corymb (L. co) (78%), and this last Fig. 1. Morphological and fl oral characters of wild roses (Rosa L.) evaluated in this study. BEN CHEIKH-AFFENE Z., HAOUALA F., HARZALLAH-SKHIRI F. 6 ACTA BOT. CROAT. 74 (1), 2015 character was correlated with the length of pedicel (L. ped) and of stylar column (L. sty) (86 and 75%, respectively). This last character (L. sty) was negatively correlated with the length of sepal (L. s), the number of lobes per sepal (Nb. lob. s) and number of lobulated sepals (Nb. s. lob) (87, 71 and 70%, respectively). The presence of glands in pedicel, recep- tacle and sepal (Gl. ped, Gl. rep, Gl. s) was positively correlated with the length of corymb (L. co) (74%) and the length of stylar column (L. sty) (95%) and negatively correlated with the diameter of prickles (Dia. Pr) (71%), number of lobulated sepals (Nb. s. lob), number of lobes per sepal (Nb. lob. s) and length of sepal (L. s.) (78, 91 and 72%, respectively). Principal component analysis and hierarchical cluster analysis The 38 morphological characters were used for the PCA and HCA. The HCA based on the Euclidean distances between groups indicated two groups of accessions: A and B, with six and seven accessions, respectively, identifi ed by their fl oral and vegetative characters with a dissimilarity > 20 (Figs. 2, 3). When the dissimilarity was higher than 16, the A group was divided into two sub groups, A1 (Hafz1, Hafz2 and Tebs2) and A2 (Zagh2, Tebs1 and Kess). With a dissimilarity > 5, subgroup A1 was further divided into two subgroups A11 (Tebs2, Hafz2) and A12 (Hafz1) and the second subgroup A2 was divided in two subgroups A21 (Tebs1, Kess) and A22 formed by one accession (Zagh2). The B group with dissimilarity > 8 was divided into 2 subgroups: B1, B2. When the dis- similarity was higher than two, the subgroup B1, was divided in three subgroups B11 (Tebs3, Tab. 2. Mean values of vegetative characters related to branches and prickles of each rose accessions growing wild in Tunisia. For the abbreviations of rose accessions (codes) see Tab. 1; Dia. Br – diameter of branches, Dt. Pr – density of prickles on a branch length of 10 cm, Dia. Pr – diameter of prickle’s base, L. Pr – length of prickle, L. Pr. dig – length of prickle’s diagonal. Values with the same letters within the same column are not different at p > 0.05. Accessions Dia. Br (cm) Dt. Pr Dia. Pr (mm) L. Pr (mm) L. Pr. dig (mm) L. Pr/ Dia. Pr Hafz2 0.6±0.1cd 6.2±6.8e 6.8±1.5d 4.8±0.6a 6.1±0.9ab 0.7±0.1a Tebs2 0.5±0.2b 15.6±4.4cd 7.5±1.4e 6.0±0.7bcd 7.9±0.9e 0.7±0ai0 Hafz1 0.7±0.1cd 25.1±6.5e 8.6±1.5g 6.8±0.9d 8.7±1.1f 0.8±0.1ab Kess 0.6±0.1c 11.9±5.3b 6.7±1.0d 6.2±0.9cd 7.0±1.0cd 0.9±0.1bc Tebs1 0.6±0.2c 13.1±3.7c 8.2±1.3ef 11.0±3.2f 10.8±2.2g 1.5±0.3g Zagh2 0.7±0.2d 36.5±13.9f 7.9±1.7ef 9.2±2.7e 10.3±2.2g 1.2±0.3ef Tebs3 0.4±0ai0 23.3±4.0e 4.9±0.7ab 6.3±1.8cd 6.5±1.1bcd 1.3±0.3f Djb 0.5±0.1a 17.3±3.4d 4.7±0.7ab 4.9±1.3a 5.9±0.8ab 1.0±0.3cde Teb 0.4±0.1a 16.5±5.3d 5.2±0.8bc 5.0±1.0a 6.2±1.0ab 0.9±0.1cd Tab 0.4±0.1a 18.3±3.7d 5.7±0.7c 6.2±1.4cd 7.2±0.6d 1.0±0.2de AinD 0.4±0.1a 24.2±5.6e 5.2±0.8bc 5.4±1.6abc 6.5±1.0bc 1.0±0.3cde Wech 0.6±0.1bc 16.8±5.2d 5.6±0.7c 6.5±1.9d 7.0±0.9cd 1.1±0.3ef Zagh1 0.4±0.1a 21.7±6.9e 4.5±0.6a 5.1±1.0ab 5.7±0.8a 1.1±0.2ef MORPHOLOGICAL DIVERSITY OF TUNISIAN WILD ROSES ACTA BOT. CROAT. 74 (1), 2015 7 Ta b. 3 . M ea n va lu es o f v eg et at iv e ch ar ac te rs re la te d to le af , l ea fl e t, gl an d an d br is tle o f e ac h ro se a cc es si on s gr ow in g w ild in T un is ia . F or th e ab br ev ia tio ns of r os e ac ce ss io ns ( co de s) s ee T ab . 1 ; N b. L ft – n um be r of le afl e ts , L . L f – le ng th o f le af , W . L f – w id th o f le af , L . r a – le ng th o f ra ch is , L . L ft – le ng th o f l ea fl e t, W . L ft – w id th o f l ea fl e t, L . p et – le ng th o f p et io le , L . s t – le ng th o f s tip ul e, L f. a – le af a re a, N b. P r. ra – n um be r o f p ri ck le s on th e ra ch is , L ft . s er – le afl e t s er ra tio n (n um be r of te et h in 1 c m o f le afl e t m ar gi n) , G l. uL ft – g la nd p re se nt ( gl an du la r; 5 ) or a bs en t ( eg la nd ul ar ; 0 ) on up pe r s id e le afl e t, B s. u L ft – w ith (p ub es ce nt ; 5 ) o r w ith ou t ( gl ab ro us ; 0 ) b ri st le o n up pe r s id e le afl e t. V al ue s w ith th e sa m e le tte rs w ith in th e sa m e co lu m n ar e no t d iff er en t a t p > 0 .0 5. A cc es - si on s N b. L ft L . L f (c m ) W . L f (c m ) L . r a (c m ) L . L ft (c m ) W . L ft (c m ) L . L ft / W . L ft L . p et (c m ) L . s t (c m ) L f. a (c m 2 ) N b. Pr . r a L ft . se r G l. uL ft B s. uL ft H af z2 6. 8± 0. 6e 8. 4± 0. 9c de 6. 7± 1. 0d 4. 6± 0. 5c de 3. 8± 0. 6d e 2. 2± 0. 4c d 1. 7± 0. 0d 1. 1± 0. 3a 1. 1± 0. 1b c 25 .0 ± 7. 5c d 4. 9± 2. 0d e 10 .2 ± 2. 2d 0a 0a Te bs 2 6. 0± 1. 0c d 8. 5± 1. 1c de 6. 0± 0. 8c 4. 9± 0. 6e 3. 6± 0. 5d 2. 5± 0. 4e f 1. 4± 0. 1c 1. 7± 0. 4c d 0. 9± 0. 1b 24 .9 ± 7. 1c d 6. 5± 2. 3f 11 .5 ± 2. 1e 0a 0a H af z1 7. 0± 0e 8. 0± 1. 1b cd 5. 9± 0. 8c 5. 0± 0. 8e 2. 1± 0. 4a 1. 9± 0. 2b 1. 0± 0. 2a 1. 3± 0. 2b 0. 8± 0. 1a 24 .7 ± 6. 0c d 5. 3± 1. 5e 9. 2± 1. 2c 5b 5b K es s 5. 2± 0. 6a 7. 4± 0. 7a b 5. 4± 0. 6c 4. 3± 0. 5a bc 3. 0± 0. 4c 2. 3± 0. 2d e 1. 3± 0. 1b 2. 2± 0. 3f 1. 7± 0. 32 f 18 .9 ± 4. 7b 1. 7± 1. 5a 6. 9± 1. 7b 0a 5b Te bs 1 5. 8± 1. 0c d 7. 8± 0. 8b c 3. 1± 0. 6a 4. 7± 0. 5d e 5. 6± 0. 5j 2. 0± 0. 6b c 2. 8± 0. 4g 1. 9± 0. 5e 1. 0± 0. 1b c 18 .0 ± 2. 9b 3. 6± 1. 3b c 7. 4± 1. 6b 0a 5b Z ag h2 6. 7± 0. 6e 6. 9± 1. 3a 4. 3± 1. 1b 4. 4± 0. 8b cd 2. 5± 0. 5b 1. 4± 0. 3a 1. 7± 0. 2d 1. 6± 0. 3c 1. 0± 0. 2b c 12 .9 ± 5. 5a 6. 5± 2. 7f 9. 3± 3. 2c 0a 0a Te bs 3 5. 6± 0. 9b c 8. 5± 1. 6c de 7. 1± 1. 5d ef 4. 4± 0. 9b cd 4. 1± 0. 7e f 2. 2± 0. 2b cd 1. 9± 0. 3e 1. 7± 0. 3c 1. 1± 0. 2c 24 .7 ± 8. 8c d 3. 8± 1. 5b c 7. 4± 1. 2b 0a 0a D jb 5. 3± 0. 7a b 8. 6± 1. 2d e 7. 5± 1. 2e f 4. 4± 0. 6b cd 4. 2± 0. 6f 2. 6± 0. 3f 1. 6± 0. 1d 1. 9± 0. 4d e 1. 1± 0. 1c 30 .7 ± 8. 0e 3. 7± 1. 2b c 7. 7± 0. 8b 0a 0a Te b 5. 2± 0. 7a b 8. 5± 0. 9c de 6. 9± 0. 8d e 4. 0± 0. 5a 4. 6± 1. 3g h 2. 1± 0. 3b cd 2. 1± 0. 3f 1. 5± 0. 3c 1. 0± 0. 1b 21 .6 ± 8. 6b c 3. 8± 1. 4b c 5. 8± 1. 0a 0a 0a Ta b 5. 2± 0. 6a b 9. 4± 0. 4f 8. 9± 1. 5g 4. 0± 0. 6a b 5. 2± 0. 8i 2. 5± 0. 4e f 2. 1± 0. 2f 1. 6± 0. 2c 1. 1± 0. 2c 28 .8 ± 10 .5 de 2. 9± 1. 2b 5. 8± 1. 1a 0a 0a A in D 5. 3± 0. 7a b 8. 8± 0. 9e f 7. 8± 0. 7f 4. 3± 0. 5a bc 4. 3± 0. 5f g 2. 5± 0. 3e f 1. 7± 0. 2d 1. 9± 0. 2d e 1. 4± 0. 2d 25 .4 ± 5. 4c d 3. 7± 1. 2b c 5. 8± 0. 8a 0a 0a W ec h 5. 2± 0. 6a b 10 .5 ± 1. 5g 9. 7± 1. 5h 4. 9± 0. 7e 5. 3± 0. 8i j 2. 5± 0. 4e f 2. 1± 0. 2f 1. 9± 0. 2e 1. 5± 0. 2d e 30 .3 ± 9. 8e 4. 4± 1. 5c d 5. 5± 1. 1a 0a 0a Z ag h1 6. 1± 1. 0d 10 .4 ± 1. 8g 9. 3± 1. 6g h 5. 4± 0. 8f 5. 0± 1. 0h i 2. 5± 0. 6e f 2. 0± 0. 3e f 1. 9± 0. 3d e 1. 6± 0. 2e f 42 .5 ± 17 .1 f 4. 8± 1. 4d e 7. 3± 1. 2b 0a 0a BEN CHEIKH-AFFENE Z., HAOUALA F., HARZALLAH-SKHIRI F. 8 ACTA BOT. CROAT. 74 (1), 2015 Ta b. 4 . M ea n va lu es o f fl o ra l c ha ra ct er s of e ac h ro se a cc es si on s gr ow in g w ild in T un is ia . F or th e ab br ev ia tio ns o f ro se a cc es si on s (c od es ) se e Ta b. 1 ; N b. Fl co – n um be r o f fl o w er s pe r c or ym b, L . c o – le ng th o f c or ym b, L . p ed – le ng th o f p ed ic el , N b. L ft . B lt – nu m be r o f l ea fl e t i n fl o w er in g br an ch le ts , L . r ep – le ng th o f re ce pt ac le , W . r ep – w id th o f re ce pt ac le ( m ed iu m ), L . s ty – le ng th o f st yl ar c ol um n, W . s ty – w id th o f st yl ar c ol um n, D ia . r ep . o – di am et er o f r ec ep ta cl e or ifi ce , N b. s . l ob – n um be r o f s ep al lo bu la te d, N b. lo b. s – n um be r o f l ob es p er s ep al , L . s – le ng th o f s ep al , W . s – w id th of s ep al , L . p – le ng th o f p et al , W . p – w id th o f p et al , G l. pe d – gl an du la r ( 5) o r e gl an du la r ( 0) p ed ic el , G l. re p – gl an du la r ( 5) o r e gl an du la r ( 0) re - ce pt ac le , G l. S – gl an du la r ( 5) o r e gl an du la r ( 0) s ep al s. V al ue s w ith th e sa m e le tte rs w ith in th e sa m e co lu m n ar e no t d iff er en t a t p > 0 .0 5. A cc es si on N b. Fl co L . c o (c m ) L . p ed (c m ) N b. L ft . B lt L . r ep (c m ) W . r ep (c m ) L . s ty (m m ) W . s ty (m m ) D ia . r ep . o (m m ) N b. s. lo b N b. lo b. s L . s (c m ) W . s (c m ) L . p (c m ) W . p (c m ) G l. pe d G l. re p G l. s H af z2 2. 8± 1. 1a 3. 7± 0. 8a 1. 5± 0. 7a 5. 1± 0. 5a 0. 9± 0. 1e 0. 5± 0. 0d e 2. 6± 0. 2a 1. 9± 0. 0a 4. 6± 0. 7c 2. 5± 0d 7. 1± 0. 9f 2. 6± 0. 6g 0. 5± 0. 4c 2. 1± 0. 2e f 2. 4± 0. 2e 0a 0a 0a Te bs 2 2. 8± 1. 1a 3. 7± 0. 7a 1. 5± 0. 7a 5. 1± 0. 4a 0. 9± 0. 1e 0. 5± 0d e 2. 3± 0. 2a 2. 7± 0a 5. 1± 0. 7e 2. 5± 0d 7. 0± 0. 9f 2. 6± 0. 6g 0. 6± 0. 4c 2. 1± 0. 2e f 2. 4± 0. 1e 0a 0a 0a H af z1 2. 9± 1. 1a 3. 6± 0. 9a 1. 2± 0. 5a 7. 0± 0c 1. 1± 0. 1g 0. 4± 0c 1. 9± 0. 2a 2. 6± 0a 4. 2± 1. 2b d 2. 5± 0d 5. 0± 0. 6d 1. 9± 0. 3e 0. 6± 0. 3d 2. 0± 0. 2d e 2, 0± 0. 2d 0a 0a 0a K es 2. 9± 1. 5a 4. 0± 1. 2a 1. 3± 0. 8a 6. 6± 0. 7b 1. 0± 0f 0. 5± 0d e 2. 3± 0. 3b 2. 6± 0. 3e 5. 1± 0. 4e 2. 4± 0d 6. 1± 1. 2e 1. 7± 0. 1d 0. 5± 0a b 1. 7± 0. 2b 2. 0± 0. 2c d 0a 0a 0a Te bs 1 2. 9± 1. 4a 4. 0± 1. 2a 1. 3± 0. 8a 6. 6± 0. 7b 1. 0± 0f 0. 5± 0d e 2. 3± 0. 2b 2. 6± 0. 2e 5. 1± 0. 4e 2. 4± 0. 1d 6. 1± 1. 1e 1. 7± 0. 1d 0. 5± 0a b 1. 7± 0. 1b 2. 0± 0. 2c d 0a 0a 0a Z ag h2 3. 0± 1. 2a 4. 8± 1. 0a 1. 4± 0. 6a 5. 0± 0a 1. 1± 0h 0. 5± 0e 2. 6± 0. 4c 4. 0± 0. 3f 5. 2± 0. 3e 2. 5± 0d 5. 2± 2. 3d 2. 1± 0. 2f 0. 4± 0a 1. 9± 0. 2c d 1. 9± 0. 2b c 0a 0a 0a Te bs 3 4. 7± 1. 7b c 8. 3± 2. 3c 2. 7± 1. 4b 5. 0± 0a 0. 8± 0d 0. 4± 0a 5. 6± 0. 5d 1. 8± 0. 2c 3. 9± 0. 3a 0± 0a 0± 0a 1. 3± 0. 2c 0. 5± 0a b 2. 0± 0. 1c d 2. 3± 0. 1d e 5b 5b 5b D jb 4. 1± 1. 7a b 8. 0± 1. 7c 3. 2± 1. 2c 5. 0± 0a 0. 7± 0b c 0. 4± 0a 5. 6± 0. 7d e 1. 6± 0. 2b 3. 7± 0. 4a 0. 2± 0. 4b 0. 2± 0. 4a 1. 1± 0. 1b 0. 5± 0a b 1. 7± 0. 2b 1. 8± 0. 2b 5b 5b 5b Te b 8. 4± 5. 5d 12 .2 ± 3. 1e 4. 6± 1. 3d 5. 0± 0a 0. 7± 0. 1a b 0. 4± 0b c 6. 1± 0. 7f g 1. 6± 0. 2b 4. 0± 0. 5a b 1. 1± 1. 0c 0. 8± 0. 8b 0. 9± 0. 1a 0. 4± 0. 1a 1. 9± 0. 2c d 2. 0± 0. 2b cd 5b 5b 5b Ta b 6. 0± 3. 2c 9. 5± 2. 8d 3. 7± 1. 2c 5. 0± 0a 0. 7± 0c 0. 4± 0b c 6. 0± 0. 8f 1. 6± 0. 1b 3. 8± 0. 3a 0± 0a 0± 0a 1. 3± 0. 1c 0. 6± 0b 2. 1± 0. 3f 2. 3± 0. 2e 5b 5b 5b A in D 8. 1± 4. 3d 12 .3 ± 2. 9e 3. 4± 0. 7c 5. 0± 0a 1. 0± 0f 0. 5± 0d 6. 3± 0. 6g 2. 1± 0. 2d 4. 3± 0. 3c 2. 5± 0. 0d 2. 2± 0. 5c 1. 5± 0. 1c 0. 5± 0a b 2. 4± 0. 2g 2. 7± 0. 2f 5b 5b 5b W ec h 7. 9± 3. 5d 10 .5 ± 2. 5d 3. 5± 0. 8c 5. 0± 0a 0. 7± 0a 0. 5± 0a c 5. 9± 0. 7e f 1. 7± 0. 3b c 4. 2± 0. 3b 0. 4± 0. 8b 0. 3± 0. 6a b 0. 9± 0. 1a 0. 5± 0a b 1. 6± 0. 21 a 1. 6± 0. 1a 5b 5b 5b Z ag h1 3. 9± 2. 3a b 6. 4± 2. 3b 2. 2± 1. 0b 5. 0± 0a 0. 7± 0a 0. 4± 0b 5. 6± 0. 6d e 1. 6± 0. 2b 4. 3± 0. 4b d 0± 0a 0± 0a 1. 1± 0. 1b 0. 5± 0a 1. 8± 0. 1b c 1. 9± 0. 1b c 5b 5b 5b MORPHOLOGICAL DIVERSITY OF TUNISIAN WILD ROSES ACTA BOT. CROAT. 74 (1), 2015 9 Fig. 2. Repartition of the vegetative and fl oral characters on the two principal axes of the principal components analysis. Identifi cation of the groups and subgroups corresponding to the 13 Tunisian rose accessions studied. For the abbreviations of rose accessions (■) and the char- acters (�) see Tabs. 1, 2, 3, 4. Fig. 3. Dendogram of the 13 Tunisian rose accessions studied, obtained by cluster analysis based on the Euclidean distances relating to the fl oral and vegetative characters. For the abbreviations of rose accessions see Tab. 1. BEN CHEIKH-AFFENE Z., HAOUALA F., HARZALLAH-SKHIRI F. 10 ACTA BOT. CROAT. 74 (1), 2015 Djb), B12 (Tab, Teb), and B13 (AinD). The subgroup B2 was divided in 2 subgroups B21 (Wech) and B22 (Zagh1). The PCA horizontal axis explained 34.42% of the total variance and the vertical axis 11.58%. The groups A and B clearly stand out, forming separate groups in PCA (Fig. 2) and a deep dichotomy in HCA (Fig. 3). The horizontal axis (PC1) was positively correlated with groups A1 and A2 and nega- tively correlated with group B. The variables most highly correlated were diameter of prickles (Dia. Pr), the presence of glands and of bristles on the upper side of leafl et (Gl. uLft, Bs. uLft), length of receptacle (L. rep), number of lobulated sepals and of lobes per sepal (Nb. s. lob, Nb. lob. s), the length and the width of sepals (L. s, W. s) (On-line Supple- ment Tab. 1). This axis 1 was moderately correlated with Dia. Br, L. Pr. dig, Lft. ser, Nb. Lft. Blt, W. rep, W. sty. The width of leaf (W. Lf), the length of leafl et (L. Lft), the lengths of corymb, peduncle and stylar column (L. co, L. ped, L. sty) and the presence of glands in pedicel, receptacle and sepal (Gl. ped, Gl. rep, Gl. s) were highly negatively correlated with PC1. The vertical axis (PC2) was positively correlated with subgroup A2 and negatively with A1. It was positively correlated with the length of prickle and the ratio of length and diam- eter of prickle (L. Pr, L. Pr/Dia. Pr) (On-line Supplement Tab. 1, Fig. 2) and negatively with the width of sepal (W. s) and the presence of glands in the upper side of leafl et (Gl. uLft). All accessions from group A were correlated with those characters: Dia. Br, Dia. Pr, L. Pr. dig, W. rep, L. rep, W. s, L. s, W. sty, Gl. uLft, Bs. uLft, Lft. ser, Nb. lob. s, Nb. s. lob, Nb. Lft. blt, (Figs. 2, 3) and were characterized by large branches (Dia. Br = 0.5–0.7 cm), dotted with a large prickle having a long diagonal (Dia. Pr = 6.7–8.6 mm; L. Pr. Dig = 6.1– 10.8 mm), leaves with a high number (Nb. Lft = 5.2–7.0) of short leafl ets (L. Lft = 2.1–5.6 cm) with high serration (6.9–11.5). Leaves on fl owering branchlets were formed by a high number of leafl ets (Nb. Lft. Blt = 5 to 7), short corymbs (L. co = 3.6–4.8 cm) with few fl owers (Nb. Fl. co = 2.8–3.0). A short pedicel (L. ped = 1.2–1.5 cm) bearing a fl ower also having a short column stylar (L. sty = 1.8–2.6 mm), but a long receptacle (L. rep = 0.9–1.1 cm), long sepal (1.7–2.6 cm) with many appendix (Nb. slob = 2.4–2.5; Nb. Lobs = 5.0– 7.1). Receptacle, pedicle and sepals are all eglandular. The subgroupA1, was represented by Hafz1, Hafz2 and Tebs2, all characterized by the highest values for the characters related to the serration of leafl et and sepals and the high values for length and wide of sepals (Lft. ser, Nb. Lob. s, Nb. s. Lob, L. s, W. s). This sub- group was divided into two subgroups: Subgroup A11: Hafz2 and Tebs2. Those accessions have eglandular and glabrous leafl ets with short petiole, rachis with numerous prickles, short corymb (L. co = 3.7 cm) with the lowest number of fl owers (Nb. Fl. co = 2.8), the fl ower has a long petal, the longest sepal longer than petal (L. s = 2.6 cm, L. p = 2.1 cm). Hafz2is characterized by highly serrated sepal (Nb. lob. S = 7.1), the narrowest and the shortest scattered and shorter prickles (Dt. Pr = 6.2, L. Pr = 4.8mm), the lowest number of fl owers by corymbs (Nb. Fl. co = 2.8), while Tebs2 is characterized by rachis with the highest number of prickles (Nb. Pr. ra = 6.5), and highly serrated leafl et (Lft. ser = 11.5). Subgroup A12: Hafz1 include accessions with a robust bush exhaling an apple scent. It differs in having its highest number of leafl ets in bloomy branchlets (Nb. Lft. Blt = 7.0), in the largest thorny branches (Dia. Br = 0.7 cm, Dt. Pr = 25.1), with the shortest leafl et (L. Lft MORPHOLOGICAL DIVERSITY OF TUNISIAN WILD ROSES ACTA BOT. CROAT. 74 (1), 2015 11 = 2.1 cm), glandular and pubescent (Gl. uLft = 5, Bs. uLft = 5), fl owers in the shortest cor- ymb (L. co = 3.6 cm), dotted with the shortest pedicel (L. ped = 1.2 cm), the longest recep- tacle (L. rep = 1.1 cm), a high number of lobulated sepals (Nb. s. lob = 2.5) and of lobes per sepal (Nb. lob. s = 5.0). The subgroup A2 reported in the positive section for PC1 and PC2 (Fig. 2), was corre- lated with diameter of branches and of prickle’s base (Dia. Br, Dia. Pr), lengths of prickle and of their diagonals (L. Pr, L. Pr. dig), number of leafl ets in fl owering branchlets (Nb. Lft. Blt), length and width of receptacle (L. rep, W. rep), width of styles (W. sty), and number of lobulated sepals (Nb. slob). This subgroup is divided into two subgroups: Subgroup A21: Kess and Tebs1. Those accessions were mainly characterized by an eg- landular (Gl. uLft = 0) and pubescent leafl et (Bs. uLft = 5) and a long receptacle (L. rep = 1.0 cm). Those accessions were characterized by bristly leafl ets with low serration (Lft. ser = 6.9 and 7.4, respectively) with a rachis with few prickles. Kess accession was character- ized by the longest petiole (L. pet = 2.2 cm) and rachis with the lowest number of prickles (Nb. Pr. Ra = 1.7). Tebs1 accession is characterized by the longest prickle and the longest prickle diagonal (L. Pr = 11.0 mm; L. Pr. dig = 10.8 mm), the narrowest leaf (W. Lf = 3.1 cm), the longest leafl et (L. Lft = 5.6 cm), and the highest ratios for leafl et and prickle (L. Lft/W. Lft = 2.8; L. Pr/Dia. Pr = 1.5). Subgroup A22: Zagh2 is an upright shrub, characterized by the largest thorny branches (Dia. Br = 0.7 cm, Dt. Pr = 36.5), rachis with numerous prickles (Nb. Pr. ra = 6.5), leafl ets eglandular (Gl. uLft = 0), glabrous (Bs. uLft = 0), and doubly serrated (Lft. ser = 9.3), fl ower with the longest (L. rep = 1.2 cm) and the broadest (W. rep = 0.5 cm) receptacle, the largest receptacle orifi ce (Dia. rep. o = 5.2 mm) and the widest style (W. sty = 4.0 mm). The group B was represented by Zagh1, Wech, AinD, Tab, Teb, Djb and Tebs3, which were correlated with the length and diameter of prickle ratio (L. Pr/Dia. Pr), the length of stipule (L. st), the number of fl owers per corymb (Nb. Flco), the length and the width of leaf (W. Lf, L. Lf), the length of leafl et, pedicel and stylar column (L. Lft, L. ped, L. sty) (Figs. 2, 3). All accessions from group B were characterized mainly by a long stylar column (L. sty. = 5.6 to 6.3 mm. They were also characterized by slender branches (Dia. Br = 0.4– 0.6 cm), long (8.5–10.5 cm) and width (9.7–6.9 cm) leaves, mainly with lanceolate (L. Lft = 4.1–5.3 cm and W. Lft = 2.1–2.6 cm), eglandular and glabrous leafl ets (Gl. uLft = 0, Br. ulft = 0), the lowest number of leafl ets by leaves in fl owering branchlets (Nb. Lft. Blt = 5), long corymbs (L. co = 6.4–12.3 cm) with many small fl owers (Nb. Fl. co = 3.9–8.4), long and glandular pedicel (L. ped = 2.2–4.6 cm). Receptacle (L. rep = 0.7–1.0 cm) and sepals (L. s = 0.9–1.5 cm) were glandular too but short. Group B was divided into 2 subgroups: B1, B2. The subgroup B1was divided in three subgroups: B11, B12 and B13. Subgroup B11 is represented by Tebs3 and Djb accessions which were characterized mainly by a medium number of fl owers (Nb. Fl. co = 4.1–4.7), and short leafl et (L. Lft = 4.1–4.2 cm) with high leafl et serration (Lft. ser = 7.4–7.7). Djb accession presents a medi- um density for prickles, the broadest leafl et (W. Lft = 2.6 cm), a low number of lobultated sepals and of lobes per sepal (Nb. s. lob = Nb. lob. s = 0.2), fl ower with the shortest recep- tacle (L. rep = 0.7 cm), having the lowest diameter for the receptacle orifi ce (Dia. rep. o = 3.7 mm). Tebs3 accession has a fl ower with a sepal with entire edges (Nb. lob. s = 0, Nb. s. lob = 0) and the narrowest branches (Dia. Br = 0.4 cm) but with high density of prickles (Dt. Pr = 23.3). BEN CHEIKH-AFFENE Z., HAOUALA F., HARZALLAH-SKHIRI F. 12 ACTA BOT. CROAT. 74 (1), 2015 Subgroup B12 is represented by Tab and Teb accessions, characterized by prickles with a medium density (16.5–18.3) and a prickle base of medium diameter (Dia. Pr = 5.2–5.7 mm), the shortest rachis (L. ra = 4.0 cm) and short petiole (L.pet = 1.5–1.6 cm). Its fl owers having the shortest and the narrowest receptacle (L. rep = 0.7 cm, W. rep = 0.4 cm). Teb is characterized by the longest pedicel (L. ped = 4.6 cm), the highest number of fl owers by corymb (Nb. Flco = 8.4), the shortest (L. s = 0.9 cm) and narrowest (W. s = 0.4 cm), low number of lobulated sepals (Nb. s. lob = 1.1) and low lobes per sepal (Nb. lob. s = 0.8). In contrast, Tab has a fl ower with long pedicel (L. ped = 3.7 cm) and sepals with entire edges (Nb. s. lob = 0, Nb. lob. s = 0). Subgroup B13 is represented by AinD accession, and it was characterized by thorny branches with a high number of prickles (Dt. Pr = 24.2), the longest corymbs (L. co = 12.3 cm), the broadest receptacle (W. rep = 0.5 cm), the longest stylar column (L. sty = 6.3 mm), the highest number of lobulated sepals (Nb. s. lob = 2.5), the longest and widest petals (L. p = 2.4 cm, W. p = 2.7 cm). The subgroup B2 was divided in two subgroups each of them with one accession (B21; Wech and B22; Zagh1) characterized by long (L. Lf = 10.5; 10.4 cm, respectively) and broad leaves (W. Lf = 9.3; 9.7 cm, respectively), ratio of length and diagonal of prickles tends to- wards 1 (L. Pr/Dia. Pr = 1.1, which means the length and the diagonal of prickles are equal) and the shortest receptacle (L. rep = 0.7 cm). Wech accession has the longest (L. Lf = 10.5 cm) and the broadest (W. Lf = 9.7 cm) leaf, the broadest receptacle (W. rep = 0.5 cm), the lowest number of leafl ets (Nb. Lft = 5.2), a low leafl et serration (Lft. ser = 5.5), the shortest sepal (L. s = 0.9 cm) with low number of lobulated sepals (Nb. s. lob = 0.4), and lobes per sepal (Nb. lobs = 0.3), the shortest and narrowest petal (L. p = 1.6 cm, W. p = 1.6 cm). Zagh1 accession was characterized by the lowest diameter (Dia. Pr = 4.5 mm), and length of diagonal prickles (L. Pr. dig = 5.7 mm). The highest length of rachis (L. ra = 5.4 cm) and the highest leaf area (Lf. A = 42.5 cm2). A medium length for pedicel (L. ped = 2.2 cm) and corymb (L. co = 6.4 cm), with few fl owers (Nb. Fl. co = 3.9), sepal with entire edges (Nb. s. lob = Nb. lob. s = 0). Discussion Accessions from group A were characterized by a large branches dotted with long prick- les, leaves, in general, with seven short leafl ets, the margins of which were highly serrated, leaves in fl owering branchlets with fi ve to seven leafl ets, short corymb with few fl owers, pedicel short too, bearing fl owers having short styles but long receptacles, sepals also long with many appendices. The receptacle, the pedicel and the sepals of those accessions were eglandular. All this characteristics described species of section Caninae, according to the taxonomy of roses (BAKER 1871, BOULENGER 1937, BAILEY 1963, MAIRE 1980). In this group, Hafz2 and Tebs2 accessions are characterized by big fl owers, leafl et pre- sumed with few glands in the under surface but eglandular in the upper surface, a glabrous pedicel. According to BAKER (1871) and MAIRE (1980) those accessions taxonomically should belong to Rosa agrestis Savi. Hafz2 and Tebs2, The slight variability within the two taxa can be related to the effect of the environment, indeed accession of Tebs2 was collect- ed from a river bank but Hafz2 was collected from an arid region. The Hafz1 accession is a robust bush that gives off a smell like apples. Leaves with seven short leafl ets, glandular and pubescent in both sides, fl owers clustered in short cor- MORPHOLOGICAL DIVERSITY OF TUNISIAN WILD ROSES ACTA BOT. CROAT. 74 (1), 2015 13 ymb with short pedicel. Flowers with the broadest sepal are dotted with many appendices. This accession presents the same characteristics of R. villosa L. but differs by a glabrous pedicel and receptacle (BAILEY 1963). MAIRE (1980) describes R. micrantha Smith. with glandular leafl ets on both sides but did not give any information about the coloration and the scent of glands of leafl ets. Nevertheless, this accession may be a hybrid, specifi c to the Tunisian region, of R. canina L. and R. villosa. Accessions Tebs and Kess were characterized by an eglandular leafl ets on both sides with glabrous pedicel, receptacle and sepal. Those characteristics correspond to R. canina taxa (MAIRE 1980), in addition those accessions present a pubescent leafl et on both sides and bristles are oppressed in the upper side of leafl ets which characterize R. canina ssp. dumetorum Thuill. (syn R. dumetorum Thuill.), (syn R. corymbifera Borkh.) (LE FLOC’H et al. 2010). The slight variability within the two species can be related to the effect of the environment, indeed accession of Tebs1 was collected from a river bank. Zagh2 accession is an upright shrub, characterized by the largest thorny branches, gla- brous leafl ets, and doubly serrated edge. Zagh2 accession should belong to Rosa canina ssp. vulgaris Gams (syn R. caninasensu strict) because of its glabrous leafl ets (MAIRE 1980). Accessions from group B are characterized by the longest stylar column, this character was a systematic determinant character only for Synstylae section, indeed the styles connate into a slender exerted column (CRÉPIN 1869, BOULENGER 1932, BAILEY 1963, MAIRE 1980, SILVESTRE and MONTSERRAT 1998). They were also characterized by slender branches, leaves, generally, with fi ve lanceolate and glabrous leafl ets, leaves of fl owering branchlets with fi ve leafl ets also. They have a long corymb with many small fl owers. The pedicel, re- ceptacle and sepals were glandular. Those characters characterized Rosa sempervirens L. (SILVESTRE and MONTSERRAT 1998). In this group fi ve subgroups were defi ned with HCA and we defi ned two varieties. Tebs3 and Djb accessions were characterized mainly by a medium number of fl owers and short leafl et with high leafl et serration. In addition, the stylar column was densely pu- bescent throughout the column length and reached the length of the internal stamens, the calyx and the fl oral pedicel were glandular; all those characters should indicate R. semper- virens var. genuina Rouy. (syn R. sempervirens). Teb and Tab accessions are characterized by a medium density of prickles, prickle base with medium diameter, the shortest rachis and a short petiole, and by fl owers that have the shortest and the narrowest receptacle. According to MAIRE (1980) those accessions present the characteristics of the variety genuina but they differ by the globular form of the recep- tacle. Thus, we identifi ed them as the form f. scandens (MAIRE 1980). So Teb and Tab ac- cessions should be R. sempervirens var. genuina f. scandens (Mill) Batt. The accessions Tebs3, Djb, Teb and Tab were collected from the North East of Tunisia (longitude: 36°28’–36°57’; latitude: 9°05’–8°57’). Indeed, their descriptions suggest R. sempervirens and the slight morphological differences can be related to the effect of envi- ronment and we can consider those accessions as R. sempervirens ecotypes. AinD accession was distinguished by its biggest fl owers with the largest style and lobu- lated sepals. This accession presents the characteristics of the R. sempervirens var. sub- moshata Rouy. (SILVESTRE and MONTSERRAT 1998). Wech and Zagh1 accessions presented a prostrate habit, which characterize R. semper- virens var. prostrate (DC) Desv. (DE CANDOLE 1815, GORE 1832, BOULENGER 1932). BEN CHEIKH-AFFENE Z., HAOUALA F., HARZALLAH-SKHIRI F. 14 ACTA BOT. CROAT. 74 (1), 2015 As a result of this study, we can conclude that among the vegetative and fl oral charac- ters studied, those showing the highest discriminating value were the sizes of prickle, leaf and leafl et, leaf area, lengths of style, pedicel and receptacle, sizes of sepal, petal, and re- ceptacle, pedicel and sepal gland density. Moreover, those discriminant characters are used in the taxonomic identifi cation of the genus Rosa. The study revealed considerable pheno- typic diversity in the Tunisian wild rose population. Accessions from the 13 populations studied belong to two different sections. The fi rst one is section Caninae, into which the ac- cessions from Teboursouk1, 2, Kessra, Haffouz1, 2 and Zaghouan2 were grouped, being related by some cases of synonymy but differing in some morphological traits such as the presence or absence of bristles and glands in leafl et. Thus, this morphological criterion sub- divided the six Caninae accessions into four species such as R. canina (Zagh2), R. agrestis (Hafz2, Tebs2), R. micrantha (Hafz1) and R. dumetorum (Tebs1, Kess). The second section is Syntylae, which is represented by accessions from Tebaba, Aïn Drahem, Wechtata, Tabar- ka, Djeba, Teboursouk3 and Zaghouan1, identifi ed as R. sempervirens. Indeed, the morpho- logic analyses prove the presence of four ecotypes of R. sempervirens such as: ecotypeTebs 3, ecotype Djb, ecotype Teb and ecotype Tab and two varieties; R. sempervirens var. sub- moshata (AinD) and R. sempervirens var. prostrata (Zagh1, Wech). Phenotypic variation for quantitative characters is most likely non-genetic in origin. The most reliable estimates of genetic structure should therefore come from qualitative characters, especially when en- vironmental effects can be minimized. We are now in the process of developing suitable molecular markers through DNA fi ngerprinting with microsatellite DNA hybridization SSR. These markers will eventually provide an additional data set on those accessions. Determination key A. Exerted styles, connate into a column usually long as stamens (L. sty = 5.6–6.3 mm)….......................................................................... Section: Synstylae Leaves usually with 5 leafl e ts, lanceolate, long and slender branches..... Rosa sempervirens 1. Climbing shrub 1.1. Stylar column densely pubescent throw the column length and reach the length of the internal stamens, receptacle, calyx and fl oral pedicel were glandular.......................................................................... R. sempervirens (Tebs3, Djb, Teb and Tab) 1.2. The same characters than as for var. genuina but sepals were lobulated with many appendix……..................................................... R. sempervirens var. submoshata (AinD) 2. A prostrate habit................................................................................... R. sempervirens var. prostrate (Zagh1 and Wech) B. Style reaching only the mouth of the receptacle and stigmas forming a sessile head over it (L. sty = 1.8–2.6 mm), leaves of fl owering branches usually with 7 leafl ets (5.0–7.0), short pedicel with long and smooth receptacle (L. rep = 0.9–1.1 cm), glabrous sepals with many appendices...................................................................................... Section Caninae 1. Leafl et pubescent and glandular on both sides, copiously pinnate...... R. micrantha (Hafz1) 2. Leafl et glandular and pubescent in under side and longer than 2 cm..... R. agrestis (Hafz2 and Tebr2) 3. Leafl et eglandular................................................................................. R. canina (Zagh2) 4. Leafl et pubescent in both sides with bristles oppressed in the upper side of leafl et........................................................................................ R. dumetorum (Tebr1 and Kess) MORPHOLOGICAL DIVERSITY OF TUNISIAN WILD ROSES ACTA BOT. CROAT. 74 (1), 2015 15 References BAILEY, L. H., 1963: The standard encyclopaedia of horticulture. Macmillan Co., New York. BAKER, J. G., 1871: Monograph of British rose. The Journal of the Linnean Society. Vol XI, 197–243. BARROS, L., CARVALHO, A. M., FERREIRA, I., 2010: Exotic fruits as a source of important phy- tochemicals: Improving the traditional use of Rosa canina fruits in Portugal. Food Re- search International 44, 2233–2236 BOSKABADY, M. H., KIANI, S., RAKHSHANDAH, H., 2006: Relaxant effects of Rosa damascene on guinea pig tracheal chains and its possible mechanism(s). Journal of Ethnopharma- cology 106, 377–382. BOULENGER, G. A., 1924: Les Rose de l’Europe de l’herbier Crépin Vol 1. Bulletin du jardin botanique de l’état à Bruxelles, Vol. 10, Fasc. 1 (1924–1925), 1–192. Published by Na- tional Botanic Garden of Belgium 2010. BOULENGER, G. A., 1932: Les Rose de l’Europe de l’herbier Crépin Vol 1. Bulletin du jardin botanique de l’état à Bruxelles, Vol. 12, Fasc. 2/3, 193–542. Published by National Bo- tanic Garden of Belgium 2010. BOULENGER, G. A., 1937: Introduction à l’étude du genre Rosa: Les caractères mor- phologiques passés en revue au point de vue de leur valeur pour la systématique. Bul- letin du jardin botanique de l’état à Bruxelles, vol. 14, Fasc. 3, 241–273. Published by National Botanic Garden of Belgium 2012. COTTIN, R., 1988: La taxonomie numérique, application aux agrumes. Fruits 43, 721–733. CREPIN, F., 1869: Primitiae Monographiae Rosarum- Matériaux pour servir à l’histoire des roses. Bulletin de la Société Royale de Botanique de Belgique/ Bulletin van de Konin- klijke belgische vereninging, T. 8, No. 2, 226–235. Royal Botanical Society of Belgium 2010. CRISCI, J. V., LOPEZ ARMENGOL, M. F., 1983: Introduccion a la teoria y practica de la tax- onomía numerica. Monografi a nro. 26, Serie de Biologia, Programa de Monografi as Cientifi cas, OEA (ed.), Secreteria General de la Organizacion de los Estados America- nos- PRDCT, Washington, D.C. 128. CZAPIK, R., 1994: How to detect apomixis in angiospermae. Polish Botanical. Studies 8, 13–21. DE CANDOLE, M., 1815: Flore Française, V6. Paris. DE COCK, K., Genetic diversity of wild roses (Rosa spp.) in Europe, with an in-depth mor- phological study of Flemish populations. PhD Thesis, Faculty of Bioscience Engineer- ing, Ghent University. DEMIR, F., OZCAN, M., 2001: Chemical and technological properties of rose (Rosa canina L.) fruits grown wild in Turkey. Journal of Food Engineering 47, 333–336. DICKINSON, T. A., LO, E., TALENT, N., 2007: Polyploidy, reproductive biology, and Rosaceae: understanding evolution and making classifi cations. Plant Systematics and Evolution 266, 59–78. ERENTURK, S., GULABOGLU, M. S., GULTEKIN, S., 2005: The effect of cutting and drying me- dium on the vitamin C content of rose hip during drying. Journal of Food Engineering 68, 513–518. BEN CHEIKH-AFFENE Z., HAOUALA F., HARZALLAH-SKHIRI F. 16 ACTA BOT. CROAT. 74 (1), 2015 GAO, X., UGGLA, M., RUMPUNEN, K., 2005: Antioxidant of dried and boiled rose hips. In: NYBOM, H., RUMPUNEN, K. (eds.), Proceeding of the fi rst international rose hip confer- ence, 239–249. Acta Horticulturae 690. ISHS. GORE, C. F., 1838: The roses Fancier’s manual. Henry Colburn, Pulisher, Great Marlbor- ough Street London. GURBUZ, I., OSTUN, O., YESILADA, E., SEZIK, E., KUTSAL, O., 2003: Anti-ulcerogenic activity of some plants used as folk remedy in Turkey. Journal of Ethnopharmacology 88, 93– 97. JIAN, H., ZHANG, H., TANG, K., LI, S., WANG, Q., ZHANG, T.,QIU, X., YAN, H., 2010: Deca- ploidy in Rosa praelucens Byhouwer (Rosaceae) endemic to Zhongdian Plateau, Yun- nan, China. Caryologia 62, 162–167. HUMMER, K. E., JANICK, J., 2009: Rosaceae: taxonomy, economic importance, genomics. In: FOLTA, K. M., GARDINER, S. E. (eds.), Plant genetics and genomics 6: Genetics and ge- nomics of Rosaceae, 339–351. Springer, USA. KARAKAYA, S., KAVAS, A., 1999: Antimutagenic activities of some foods. Journal of the Sci- ence of Food and Agriculture 79, 237–242. KLASTERSKA, I., NATARAJAN, AT., 1974: Cytological studies of the genus Rosa with special reference to the section Caninae. Heriditas 76, 97–108. KOVARIK, A., WERLEMARK, G., LEITCH, A. R., SOUCKOVA-SKALICKA, K., LIM, Y. K., KHAITOVA, L., KOUKALOVA, B., NYBOM, H., 2008: The asymmetric meiosis in pentaploid dog roses (Rosa sect. Caninae) is associated with a skewed distribution of rRNA gene families in the gametes. Heridity 101, 359–367. LE FLOC’H, E., BOULOS, L., VELA, E., 2010: Catalogue synonymique commenté de la fl ore de la Tunisie. Publications Scientifi ques Tunisiennes. MAIRE, R., 1980: Flore de l’Afrique du Nord, vol. XV Dicotyledonae: Rosales, Platanaceae. Lechevalier S. A. R. L. Paris. MOLINA CANO, J. L., 1977: Introduccion a la taxonomia numerica. Monografi as de la Es- cuela Tecnica superior de Ingenieros Agronomos de Madrid 80. NYBOM, H., 2009: Introduction to Rosa. In: FOLTA, K. M., GARDINER, S. E. (eds.), Plant ge- netics and genomics 6: Genetics and genomics of Rosaceae, 339–351. Springer, USA. OLSSON, M. E., ANDERSSON, S., WERLEMARK, G., UGGLA, M., GUSTAVSON, K. E., 2005: Carot- enoids and phenolics in rose hips. In: NYBOM, H., RUMPUNEN, K. (eds.), Proceeding of the fi rst international rose hip conference, 249–252. Acta horticulurae 690. ISHS. OZMEN, I., ERCISLI, S., HIZARCI, Y., ORHAN, E., 2005: Investigation of antioxidant enzyme activities and lipid peroxidation of Rosa canina and R. dumalis fruits. In: NYBOM, H., RUMPUNEN, K. (eds.), Proceeding of the fi rst international rose hip conference, 245–248. Acta horticulturae 690. ISHS. POTTIER-ALAPETITE, G., 1979: Flore de la Tunisie Angiospermes-Dicotylédones, Apétales Dialypétales. Publications Scientifi ques tunisiennes. REHDER, A., 1940: Manuel of cultivated trees and shrubs. Mac Millan. New York. REIN, E., KHARAZMI, A., WINTHER. K., 2004: An herbal remedy, Hyben Vital (stand. powder of subspecies of Rosa canina fruits), reduces pain and improves general wellbeing in patients with osteoarthritis, a double-blind, placebo-controlled, randomized trial. Phyto- medicine 11, 383–391. MORPHOLOGICAL DIVERSITY OF TUNISIAN WILD ROSES ACTA BOT. CROAT. 74 (1), 2015 17 RITZ, C. M., SCHMUTHS, H., WISSEMANN, V., 2005: Evolution by reticulation: European dog roses originated by multiple hybridisation across the genus Rosa. Journal of Heredity 96, 4–14. SILVESTRE, S., MONTSERRAT, P., 1998: Rosa. In: MUNOZ, F., NAVARRO, C. (eds.), Flora Iberica, vol VI, 143–195. Real Jardin Botanico. CSIC Madrid. WARHOLM, O., SKAAR, S., HEDMAN, E., 2003: The effects of standardized herbal remedy made from subtype of Rosa canina in patients with osteoarthritis: A double-blind, ran- domized, placebo-controlled clinical trial. Current Therapeutic Research 64, 21–31. WINTHER, K., REIN, E., KHARAZMI, A., 1999: Anti-infl ammatory properties of rose-hip. In- fl ammopharmacology 7, 63–68. WISSEMANN, V., 2003: Classifi cation/conventional taxonomy (Wild roses). In: ROBERTS, A. V., DEBENER, T., GUDIN, S. (eds.), Encyclopaedia of rose science, 111–117. Elsevier, Ox- ford.