International Journal of Science and Research (IJSR) ISSN (Online): 2319-7064 Index Copernicus Value (2013): 6.14 | Impact Factor (2013): 4.438 Germination Variability in Different Populations of Cork Oak ( Quercus suber L.) in

ABDESSAMAD. Abdessalem1, 2, KSONTINI. Mustapha1, et SALHI HANNACHI Amel2

1Laboratory of Physiology and Ecology Forestry, National Institute for Research in Rural Engineering & Forestry INRGREF , Tunisia 2Laboratory of Molecular Genetics, Immunology and Biotechnology, Faculty of Sciences of Tunis, 2092 El Manar Tunis, Tunisia

Abstract: The cork oak (Quercus suber L.) is a Mediterranean species, having great ecological and socioeconomic importance. In Tunisia this species occupies considerable areas including Kroumirie Mogods. Germination is a phenomenon that leads to the inert seed to a plantlet can emit its first leaves. It has various modes of expression. In the present work we have undertaken a study of germination performed semi-controlled conditions (nursery) of INRGREF (Tunis) different populations of cork oak in Tunisia: El Feidja (), Beni M'ttir, Hammam Bourguiba, Dar Fatma, Oued Zena and Mejen Essef (), Ain Zana (Bouselm) Jebel Zouza () Keff El Rand (Haouria), Steam Jdidi (Hammamet) and Jebel Kroufa and Bellif () located Kroumirie Mogods and Cap Bon. Our results showed that the maximum germination rate is very high (98-100%), respectively, for the people of Jebel Kroufa, Oued Ezzen, Keff El rand, Blessed Mtir and Hammam Bourguiba, Jebel Zouza and Mejen Essef of lesser degree for the people of El Feidja (95.83%) and Ain Zena (90.27%) by cons it is lower for the population of Bellif, Dar Fatma (86.41%) and Hammam Jdidi (72.83%). The lag phases recorded are of the order of 42 days, respectively, for the population of Dar Fatma, Hammam Bourguiba, Jebel Kroufa, Steam Jdidi, Ain Zena and Mejen Essef, 49 days for the people of Jebel Zouza 56 days for public of Bellif, Blessed Mtir and Keff El rand, 63 days for the people of Oued Ezzen and about 70 days for the people of El Feidja. The difference in germination between populations can be attributed to ecological and climatic conditions of the original site and / or specific biological conditions of the case study.

Keywords: Germination, population, cork oak, variability, Tunisia

1. Introduction 2. Materials and Methods

Forests, although they cover only one tenth of the surface 2.1 Site Collection Seed of the planet, representing almost 90% of all biomass dominated by trees such as oaks and account for nearly The cork oak acorns L were collected from different half of the primary production biosphere [4]. The global locations in Kroumirie, Mogods and Cap Bon in Tunisia distribution of these forests obeys great ecological (72 individuals from each populations of Hammam gradients among current major explanatory factors, the Bourguiba [HB], Dar Fatma [DF], Oued Zeen [OZ], Aîn temperature and water data provide a couple of prime Zana [AZ]; Mejen Essef [ME]; Béni Mtir [BM]; Djebel importance, as well as genetic data that identify the Zouza [DZ] (Ain Draham) El Feidja [EF] (Gar Dimaou); variation between and within species population. The Bellif [B] (Nefza); Djebel Khroufa [DK] (Tabarka); Keff study of the germination of this species for assessing El Rand [KR] (El Haouaria) and Hammam Jdidi [HJ] populations and help conservation by providing better (Hammamet) (Tab 2 and Tab 3). The choice of the sites production quantitatively and qualitatively. was carried out according to the geographical distribution, the bioclimat and the relief in December 2011(Table.1). This is a first step in identifying the existence of the strongest sources and newer that seem best suited to the 2.2 Germination Experiments environment in Tunisia and help the conservation of the forest genetic heritage: the optimization of conduct In semi-controlled conditions in the nursery INRGREF cultivation of a variety of interest and selection of Ariana under semi arid bioclimate higher. The acorns are genotypes having outstanding characteristics. In this sown in polythene bags of 12 cm diameter and 20 cm deep context a comparative study of germination of twelve filled with a substrate (mixture containing 1/3 2/3 sand populations of cork oak El Feidja (GHARDIMAOU), and humus cork oak) and whose composition Beni M'ttir, Hammam Bourguiba, Dar Fatma, Oued Zena physicochemical is given by (Table.1). and Mejen Essef (Ain Draham), Ain Zana (Bouselm) Jebel Zouza (Nefza) Keff El Rand (Haouria), Steam Jdidi Seeds were considered to be germinated when the radical (Hammamet) and Jebel Kroufa and Bellif (Tabarka) the emergence. After seed sowing, the germinated seeds were purpose of identifying the within population of cork oak. counted and eliminated every week for 140 days.

Table 1 : Soil texture culture

clay (en%) 10% pH 7.5 coarse sand 48% fine silt 3% total limestone 2% total nitrogen 560 ppm coarse silt 5% P2O5 15 ppm organic matter 1.8% sandy 32% K2O 70 ppm

Volume 4 Issue 4, April 2015 www.ijsr.net Paper ID: SUB151595 Licensed Under Creative Commons Attribution CC BY 1497 International Journal of Science and Research (IJSR) ISSN (Online): 2319-7064 Index Copernicus Value (2013): 6.14 | Impact Factor (2013): 4.438 2.3 Methods of Germination Expression 3.4 Mean Germination Time (MTG)

The characteristics of seedling emergence were (FG), Mean Germination Time are recorded in the order of 1.14; number determined: final germination percentage of days 1.11; 1.07; 1.07; 1.05; 1.04; 1.03; 1.03; 1.00; 1.00; 0.89 to first germination (delay of germination), critical limits and 0.89 days respectively for populations Bellif; of germination and mean time to germinate (MTG). (i) Hammam Jdidi; Oued Ezzen; Mtir blessed; Jebel Zouza; Final germination percentage (FG) was calculated as the Keff El rand; Dar Fatma; Hammam Bourguiba; Jebel cumulative number of germinated seeds with normal Kroufa; El Feidja; Mejen Essef and Ain Zana (Fig.3). radicles. 3.5 Coefficient velocity GP = Σ푛, where 푛 is the number of seeds that had germinated at each counting. (ii) Mean Time of The coefficients of velocity are stored in the order of germination: MTG was estimated according to the 99.09%; 96.92%; 96.82%; 96.13%; 95.23%; 93.86%; formula: 90.62%; 88.62%; 88.47%; 88.17%; 83.05% and 78.81% respectively for the people of Jebel Kroufa; Hammam MTG = Σ (n. 푑푖) Bourguiba; Dar Fatma; Keff El rand; Jebel Zouza; Mtir blessed; Hammam Jdidi; Mejen Essef; Ain Zana; Bellif; where 푛 is the number of seeds germinated at day 푖,푑 the El Oued and Ezzen Feidja (Fig.4). incubation period in days, and 푁 the total number of germinated seeds in the treatment. 3.6. Energy germination

2.4 Statistical Analysis The germination energy are stored in the order of 98.77%; 98.77%; 98.61%; 97.53%; 97.22%; 96.30%; 93.06%; Statistical analysis was performed using two-way analysis 87.50%; 85.19%; 83.95%; 71.60% and 71.60% of variance (ANOVA and DUNCAN) to test the respectively for the population of Hammam Bourguiba; variability of germination characteristics for different Jebel Kroufa; Oued Ezzen; Jebel Zouza; Mejen Essef; populations. All statistical methods were performed using Mtir blessed; El Feidja; Ain Zana; Keff El rand; Dar SPSS, version 17. Fatma; Bellif and Hammam Jdidi. (Fig.5).

3. Results

3.1 Germination rate

The maximum germination rate are recorded in the order of 100%; 100%; 100%; 100%; 100%; 98.77%; 98.61%; 95.83%; 90.28; 86.42%; 86.42% and 72.84% respectively for the population of Beni Mtir, Hammam Bourguiba, Jebel Kroufa, Oued Ezzen, Keff El rand, Jebel Zouza, Mejen Essef El Feidja, Ain Zana Bellif, Dar Fatma and Hammam after Jdidi 140 days (Fig.1)

3.2 Speed germination

The lag phases are recorded in the order of 42, 42, 42, 42, 42, 42, 49, 56, 56, 56, 63 and 70 days respectively for the people of Dar Fatma, Hammam Bourguiba, Jebel Kroufa, Steam Jdidi, Zena and Mejen Essef Ain, Jebel Zouza, Bellif, Blessed Mtir and Keff rand El Oued and El Ezzen Feidja

3.3 Germination capacity

The germination capacity are recorded in the order of 100%; 100%; 100%; 100%; 100%; 98.77%; 98.61%; 95.83%; 90.28; 86.42%; 86.42% and 72.84% respectively for the population of Beni Mtir, Hammam Bourguiba, Jebel Kroufa, Oued Ezzen, Keff El rand, Jebel Zouza, Mejen Essef El Feidja, Ain Zana Bellif, Dar Fatma and Hammam after Jdidi 140 days (Fig.2).

Volume 4 Issue 4, April 2015 www.ijsr.net Paper ID: SUB151595 Licensed Under Creative Commons Attribution CC BY 1498 International Journal of Science and Research (IJSR) ISSN (Online): 2319-7064 Index Copernicus Value (2013): 6.14 | Impact Factor (2013): 4.438 Table 2: Characteristics geographical, bioclimatic and relief of different populations of oak cork (Quercus suber L) in Tunisia

Geographical Bioclimat (bioclimatic Altitude Characteristics Site Abbreviation Area Relief distribution stage) (m) Geographical

32 43 84 Western North Wet superior at moderate 780 El Feidja EF Ghardimaou Montagne 22 E (Kroumirie) winter m 40 40 26 7 N 32 47 48 Western North Wet superior at moderate 800 36 E Beni M'ttir BM Ain Drahem Montagne (Kroumirie) winter m 40 65 89 8 N

32 46 76 Hammam Western North Wet superior at moderate 570 HB Ain Drahem Montagne 08 Bourguiba (Kroumirie) winter m 40 71 96 0 N 32 47 84 Western North Wet superior at moderate 909 09 E Dar Fatma DF Ain Drahem Montagne (Kroumirie) winter m 40 73 59 6 N 32 48 22 Western North Wet superior at moderate 675 49 Oued Zena OZ Ain Drahem Montagne (Kroumirie) winter m 40 75 63 0 N 32 48 17 Western North Wet superior at moderate 800 Mejen Essef ME Ain Drahem Montagne 17 E (Kroumirie) winter m 40 69 20 4

32 48 77 Western North Wet superior at moderate 851 Aîn Zana AZ Bouselm Montagne 03 E (Kroumirie) winter m 40 64 69 4 N 32 49 96 Western North Wet superior at moderate 542 49 E Djebel Zouza DjZ Nefza Montagne (Kroumirie) winter m 40 77 41 2 N 32 65 77 Sub-wet superior at hot 642 63 E Keff El Rand KR Haouaria North Est (Cap Bon) Montagne winter m 40 77 15 8 N

32 62 08 North Est Semi-arid lower than hot 174 Hammam Jdidi HJ Hammamet Montagne 79 E (Cap Bon) winter m 40 35 84 9 N

32 49 41 Western North Chaine 160 Djebel Khroufa DjK Tabarka Wet inferior at hot winter 25 E (Mogods) tellienne m 40 90 49 5 N

32 50 79 Western North Chaine Bellif B Tabarka Wet inferior at hot winter 88 m 87 E (Mogods) tellienne 40 98 60 9 N

4. Discussion Ezzen population; 56 days for the people of Bellif, Blessed Mtir and Keff El rand; 49 days for the people of Germination is often seen as a phenomenon that leads Jebel Zouza and 42 days for the people of Dar Fatma, from inert seed to a seedling capable of transmitting its Hammam Bourguiba, Jebel Kroufa, Steam Jdidi, Ain first leaves. This presents various modes of expression. Mejen Essef Zena and then a second acceleration of germination phase (ranges from 56-126 days in the Germination rates observed over a period of 140 days population of Bellif; 42-112 days with the population of were manifested in three phases: first a latency period of Dar Fatma, 49-91 days for the people of Jebel Zouza; 56- 70 days for the people of El Feidja; 63 days for the Oued 91 days in the population of Beni Mtir; 42-105 days with Volume 4 Issue 4, April 2015 www.ijsr.net Paper ID: SUB151595 Licensed Under Creative Commons Attribution CC BY 1499 International Journal of Science and Research (IJSR) ISSN (Online): 2319-7064 Index Copernicus Value (2013): 6.14 | Impact Factor (2013): 4.438 the population of Hammam Bourguiba, 70 to 105 days for rand, Jebel Zouza, Mejen Essef El Feidja, Ain Zana Bellif, the people of El Feidja; 42-98 days for the people of Jebel Dar Fatma and Hammam after Jdidi 140 days. In the Kroufa; 63-84 days for the people of Oued Ezzen; 56-84 nursery, the twelve populations of cork oak have high and days for the people of El Keff rand, and 42 through 119 varied maximum rate of germination obtained after one to days among the population of Hammam Jdidi; 42-98 days two months of storage (42 days to 70 days). These results for the people of Ain Zana and 42-112 days in the are in agreement with that found by [7] on the species of population of Mejen Essef) and a third phase oak. characterized by a plateau indicating the maximum rate of germination (from 126 days in the population of Bellif; Germination appears to be also dependent on the size 112 days in the population of Dar Fatma; 91 days among factor acorns; our results showed the highest rate (96%) the people of Jebel Zouza; 91 days in the population of for the species of oak afares length of about 4.09 cm in Beni Mtir; 105 days in the population of Hammam diameter on the order of 1.72 cm. For zeen oak species, it Bourguiba; 105 days in the population of El Feidja; 98 has the germination rate of 94.44%, it has at the same time days among the people of Jebel Kroufa; 84 days in the a length of 3.80 cm and 1.62 cm diameter. The population population of Oued Ezzen; 84 days in the population of El of cork oak (82.40%) has tassels length of about 2.93 cm Keff rand; 119 days in the population of Hammam Jdidi; and a diameter of about 1.56 cm. These results are 98 days among the people of Ain Zana and 112 days in consistent with those found by [3] attributed the size of the population of Mejen Essef). acorns or their weight to a positive influence on the germination and early growth stages (days); during this The last phase is relatively large for all populations of period of life, the seedling uses the reserves of acorns and cork oak with a more or less for the people of El Feidja more important are his best reserves will start germination (70j) compared to that of the population of Wadi Ezzen and growth. The difference in the size of acorns can be (63j) and populations Bellif, Blessed Mtir and Keff El attributed to environmental and climatic conditions of the rand (56j), the population of Jebel Zouza (49J) and original site and the specific conditions of the study populations of Dar Fatma, Hammam Bourguiba, Jebel species. Kroufa, Steam Jdidi, Ain Zena and Mejen Essef (42j). This can be explained by the time required for the seed to 5. Conclusion put in place mechanisms that allow it to adjust its osmotic potential with respect to the medium [8]; as well as its root As a result of the present study, we can conclude that development. This scope is a limiting factor of traits related to germination of Quercus suber.L constitute regeneration by planting cork oak especially when it is a useful approach to analyze the diversity in this important unable to provide a balance between the nutritional needs species. This variability related to the germination rate, of other wildlife [3] environment. speed germination, germination capacity, means germination Time (MTG), coefficient velocity, energy Note also that there is a differential dormancy within the germination present the most discriminating value. A case same species from the same population or the same seed of homonymy was detected in cultivar originating from lot of the same population of cork oak. Or even among Ain Drahem. seeds from the same mother tree. The dormancy intensity varies with latitude, population and harvest year. This has This study although revealed considerable variability in the effect of dormancy stagger the germination over a germination among oak cork accessions, accurate oak period of time or longer [6]. cork cultivars identification however, needs a large number of morphological markers and the exploration of [5] also described the germination difficulty by the other eco physiological, biochemical and molecular existence of embryo dormancy due to the mechanical markers such as sequencing at end to analyze genotypic action exerted by the pericarp to the output of the radical diversity. and that depends on the shaft which is a producer of characteristics physiological acorns mature and freshly Acknowledgments harvested. The slow germination is also recorded for [2] for the holm oak [1] and olive [9]. This work was supported by grants of the National Institute of Research in Agricultural engineering, Water Preney et al (1997) show that the germination rate and Drills (Institute Tunis ), and the Faculty of decreases with increasing shelf life because they reduce Sciences of Tunis (University Tunis El Manar). the vitality of acorns. This paper is dedicated to Professor Mokhtar Trifi, Our results showed that the maximum germination rate Director of the molecular Laboratory of genetics, are recorded in the order of 100%; 100%; 100%; 100%; immunology and biotechnology, and co-author of this 100%; 98.77%; 98.61%; 95.83%; 90.28; 86.42%; 86.42% work, who passed away in September 2010. May God and 72.84% respectively for the population of Beni Mtir, bless his soul and dedicated. Hammam Bourguiba, Jebel Kroufa, Oued Ezzen, Keff El

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Figure 1: Germination rate based on time acorns from different populations of cork oak (Quercus suber. L) in Tunisia (semi- controlled conditions)

Figure 2: Germination capacity acorns from different populations of cork oak (Quercus suber. L) in Tunisia (semi-controlled conditions)

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Figure 3: Germination mean time (GMT) acorns from different populations of cork oak (Quercus suber. L) in Tunisia (semi- controlled conditions)

Figure 4: Velocity coefficient (CV) acorns from different populations of cork oak (Quercus suber. L) in Tunisia (semi- controlled conditions)

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Figure 5: Germination energy (EG) acorns from different populations of cork oak (Quercus suber. L) in Tunisia (semi- controlled conditions)

Références robur L)et de chêne sessile (Quercus petraea Liebl) à l’office National des forêts.ONF Bultletin technique [1] Aissa D., 1983, Etude sur la germination des N°33 office national des forêts service graines et semences de chêne vert (Quercus ilex) I.Influence de plants Montraincon supt.39300. l’arbre producteur et de la taille des semences, Rev. [9] YSARD A., 1987, Etude expérimentale de la Cyto.Biol. Véget.Bot.6. Alaoui-Sossé et al., 1994). germination de deux chênes méditerranéens : le chêne [2] Chaussat R., Le Deunff Y., 1975, La germination pubescent (Quercus pubescens Willd.) et le chêne des semences, Gauthier. Villars, Paris. kermès (Quercus coccifera L), Thèse Doct.3e cycle [3] Hasnaoui B., 1992 Chênaies du Nord de la Tunisie. en Développement et amélioration des Végétaux, Ecologie et Régénération. Thèse de Doctorat, Univ. Université Aix-Marseille III De provence-Aix-Marseille I. [4] Ksontini M., 1996 Etude écophysiologique des réponses à la contrainte hydrique du chêne liège (Quercus suber) dans le Nord–Tunisie: comparaison avec le chêne Kermès (Q. coccifera) et le chêne zeen (Q.faginea) Thèse université Paris XII Val De Marne, 157p. [5] Merouani H., Camen B., Maria H A. et Joâo S.P., 2000, Comportement physiologique des glands de chêne liège (Quercus suber L.) durant leur conservation et variabilité interindividuel producteurs. Ann. For. Sci.58 (2001) 143.153. [6] Mokrani A., 2002 Etude de la germination des semences des espèces : Casuarina glauca, Eucalyptus gomphocephala, Cupressus sempervirens et Atriplex halimus, les plus utilisées dans le reboisement en Tunisie.. D.E.A. Physiologie Vég. Fac. Sci. Tunis, 79p. [7] Ncibi R., 2005 Sénescence et rajeunissement des subéraies de Tabarka. Aîn Draham avec approche écologiques et biotechnologiques. Thèse Doc. Phaseolus vulgaris cultivars in response to induces drought stress. Environ Exp Bot; 43 : 227-337. [8] Preney S.,Bonvicini M.P et Conche J., 1997 La récolte des glands de chêne pédonculé (Quercus

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