Mediterranean Pines (Pinus Halepensis Mill. and Brutia Ten.)

Chapter 5 1

Mill. and brutia Ten.) 3

[AU1] Maria Regina Chambel, Jose Climent, Christian Pichot, and Fulvio Ducci 4

5.1 Introduction 5

Pinus halepensis Mill. (Aleppo pine) and P. brutia Ten. (Brutia pine or Turkish red 6 pine) together cover more than 7 million hectare around the Mediterranean Basin 7 and play major ecological and economical roles in low- to mid-elevation Mediter- 8 ranean forests. Both species are well adapted to dry summer conditions and are able 9 to successfully colonize abandoned arable lands and burned areas. Apart from 10 the high ecological value of natural stands, these species can create highly resilient 11 forest covers in limiting dry conditions for both production and protection purposes. 12 Although large adaptive genetic diversity was demonstrated in both species, Aleppo 13 pine materials are globally more tolerant to water stress while Brutia pine is, in 14 general, less susceptible to frost damage. 15 Even when genetic improvement of these species is less developed compared to 16 other species of the genus in Europe, such as Scots pine or Maritime pine, provenance 17 testing has been carried out in most countries thanks to international collaborative 18

With contributions of: Ernesto Fusaro (CRA PLF, IT), Eduardo Notivol (CITA, ES), Francisco Auñón (INIA, ES), Paraskevi Alizoti (AUTH, GR), Şükran Gökdemir (CAFRD, TUR), Leonid Korol (Volcani C., IL), Mohamed Larbi Khouja (INRGREF, TUN), Hassan Sbay (FRC, MOR). M.R. Chambel • J. Climent (*) Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria (INIA), 28080 Madrid, Spain e-mail: [email protected] C. Pichot Institut National de la recherche agronomique (INRA), Avignon, France F. Ducci Consiglio per la ricerca e Sperimentazione in Agricoltura – Istituto Sperimentale per la Selvicoltura (CRA), 52100 Arezzo, Italy e-mail: [email protected]

L.E. Pâques (ed.), Forest Tree Breeding in Europe: Current State-of-the-Art and Perspectives, Managing Forest Ecosystems 25, DOI 10.1007/978-94-007-6146-9_5, © Springer Science+Business Media Dordrecht 2013 M.R. Chambel et al.

19 initiatives (FAO Silva Mediterranea, IUFRO and EU projects). Standard breeding 20 programmes based on seedling and clonal seed orchards have been developed in 21 Spain (among TreeBreedex partners) and particularly in Greece for Aleppo pine 22 and in Turkey for Brutia pine, outwith TreeBreedex. The target of these programmes 23 has been not only yield improvement, but especially adaptability and endurance 24 under limiting environmental conditions, particularly summer drought.

25 5.2 Taxonomy, Biology and Ecology of Aleppo Pine

26 5.2.1 Taxonomy and Species Distribution

27 Pinus halepensis Mill. and Pinus brutia Ten. are indeed two very close taxa, formerly 28 included in a separate section or subsection Halepenses (Price et al. 1998; López 29 et al. 2002). However, recent classifications of subgenus Pinus (Dyploxylon or 30 “hard” pines) tend to group these two species with P. heldreichii, P. pinaster, P. pinea, 31 P. canariensis and P. roxburghii within subsection Pinaster, also called the 32 Mediterranean pine group (Gernandt et al. 2005, 2008). 33 Both species have circum-Mediterranean ranges of distribution, sometimes 34 consisting of isolated populations, but occupy different geographical ranges and 35 bioclimatic niches along the Mediterranean basin (Panetsos 1981; Nahal 1986; 36 Vidakovic 1991; Fig. 5.1). Nevertheless, natural hybrids exist through unidirec- 37 tional pollen flow from P. halepensis as the pollen donor to P. brutia acting as the 38 female tree. This occurs in some Greek stands (Chalkidiki peninsula, where Pinus 39 halepensis Mill. meets the easternmost limit of its distribution in the Mediterranean 40 basin, Panetsos 1975; Panetsos et al. 1997) and in several Turkish stands. Distinction 41 between P. halepensis, P. brutia and their natural hybrids has long been discussed, 42 both in distant and sympatric populations (Panetsos 1975; Panetsos et al. 1997; 43 Tozkar et al. 2009). Controlled pollination experiments suggest that partial repro- 44 ductive barriers exist between the two species (Panetsos 1975). 45 P. halepensis Mill. occupies the southernmost area of the Mediterranean pines 46 (with the exception of P. canariensis) and it is widespread in the western part of the 47 Mediterranean Sea (ranging from 45° to 31°N), including North Africa (Morocco, 48 Algeria, Tunisia, Libya), the southern regions of France, Italy, eastern Spain, Greece 49 and Malta. It is also present in coastal areas of Croatia and Albania. There are some 50 natural and artificial populations in Turkey, Jordan, Israel, Lebanon and Syria (hence 51 the name Aleppo pine). 52 The natural range of Pinus brutia Ten. is the Eastern Mediterranean (ranging 53 from 44° to 35°N), from Greece to the eastern Crimea, Georgia, northern Iraq, west- 54 ern Syria, Lebanon and Cyprus. It is particularly abundant in Turkey, justifying its 55 English common name (Turkish red pine). In Italy, P. brutia is not indigenous, 56 although it is naturalized in some hilly areas of southern Calabria and Salento where 57 it was probably introduced by the Romans (who called it Calabria: Bruttium). 58 In the margins of its distribution area, three varieties (besides the type) and 59 one subspecies are recognized (Frankis 1999): P. brutia var. pityusa (Steven) Silba 5 Mediterranean Pines (Pinus halepensis Mill. and brutia Ten.)

Fig. 5.1 Range of distribution of Pinus halepensis (green) and P. brutia (blue) (adapted from Euforgen)

(syn: P. pityusa Steven), is widespread in Georgia and along the Russian coasts of 60 the Black Sea; P. brutia var. stankewiczii (Sukaczev) Frankis, is present in the 61 Crimea and the Ukraine; P. brutia var. pendulifolia Frankis. is widespread in coastal 62 regions of southern Turkey and P. brutia subsp. eldarica (Medw.) Nahal (syn: 63 P. eldarica Medw.) is typical of the Caucasus (mainly in Azerbaijan, but also in 64 Georgia, Iran and Turkey). It is considered a separate species (Pinus eldarica) by 65 some authors, adapted to drier and colder climates, with rainy summers. It was 66 probably introduced from Azerbaijan into the Mediterranean during the rule of 67 Alexander the Great. 68

5.2.2 Biology 69

Secondary needles of P. brutia are darker, longer (10–16 cm) and thicker (1.5 mm) 70 than those of Aleppo pine, and with a signiﬁcantly higher leaf mass per area (Pardos 71 et al. 2009). They have three lines of hypodermal cells and resin canals, while in 72 P. halepensis resin canals are marginal (Vidakovic 1991). Primary needles, very 73 similar in both species, are retained longer in Aleppo pine than in Brutia pine. 74 The more precocious formation of secondary needles and ﬁrst bud set in Brutia pine 75 is one of the traits distinguishing both species more clearly at early developmental 76 stages (Climent et al. 2011). 77 M.R. Chambel et al.

Fig. 5.2 Extremely different winter buds of Brutia pine (left) and Aleppo pine (right) taken at the same moment in 3-year-old plants at the nursery. Shoot elongation in Aleppo pine does not actually stop during mild winters (J. Climent)

78 P. brutia varieties are not very distinct from the typical form. Var. pityusa has 79 shorter needles; var. stankewiczii has solitary cones and seeds with shorter wings 80 and var. pendulifolia has 20–29 cm long weeping needles. By contrast, the Eldar 81 pine (P. brutia subsp. eldarica) has a more slender growth habit, 5–6 cm long cones 82 with a short stalk and shorter and thicker needles (8.5–10 cm) (Vidakovic 1991). 83 Both species have the ability to produce multiple ﬂushes during the growing 84 period (polycyclic shoot growth), but this phenomenon is more extreme in Aleppo 85 pine (Isik et al. 2002; Pardos et al. 2003). Shoots of Aleppo pine can restart elonga- 86 tion very early in winter, and continue growing well into late autumn in mild climates 87 (Fig. 5.2). Multiple female ﬂowering in the same year is not rare, increasing the 88 amount of cones of the same yearly cohort (Climent et al. 2008; Pardos et al. 2003) 89 (Fig. 5.3a). 90 Cones of P. brutia are sessile or sub-sessile, while the cones of P. halepensis have 91 a 2–3 cm long petiole (Fig. 5.3b). Seeds of Brutia pine have a length of between 6 92 and 9.5 mm, with a 15–20 mm wing, while Aleppo pine seeds are 5–6 mm long with 93 a wing of 20 mm. Seeds of Aleppo pine are the lightest among Mediterranean pines: 94 between 45,000 and 65,000 seeds/kg, versus about 20,000 seeds/kg in Brutia pine. 95 Aleppo pine is one of the most reproductively precocious pine species, starting 96 to produce female cones as early as 3 years of age (Tapias et al. 2001, 2004). Besides, 97 high female fecundity and partial serotiny or xeriscence ensure the precocious 98 maintenance of an important aerial seed bank (up to one million seeds per ha; 99 Ne’eman et al. 2004; Tapias et al. 2001) (Fig. 5.3c). Brutia pine is less precocious 100 and serotinous, but it is still able to keep a variable aerial seed bank. Both species 101 start their reproductive life as females, while male strobili appear in lower or 102 secondary branches when the crown attains a certain complexity level (Ne’eman 103 et al. 2004; Shmida et al. 2000). 5 Mediterranean Pines (Pinus halepensis Mill. and brutia Ten.)

Fig. 5.3 Reproductive features of Aleppo pine: (a) multiple female ﬂowering cycles of the same vegetative period, (b) mature cones, (c) serotinous cone in a young tree showing the characteristic thin light grey bark and (d) male strobili (L. Santos del Blanco)

Bark aspect and thickness is another major feature distinguishing Pinus halepensis 104 and P. brutia. While Aleppo pine bark is extremely thin; silver grey at early ages and 105 later ash-grey, Pinus brutia shows a thicker reddish bark from young ages. 106

5.2.3 Ecology 107

Pinus halepensis is a thermophilic, drought-tolerant species; in fact it is probably 108 one of the pines most tolerant to high temperatures and drought (Magini 1955; 109 Scarascia-Mugnozza 1986). 110 In the northern part of the natural distribution range it grows in coastal areas 111 (from sea level up to 600 m), while in the south it can climb up to 1,400 m (the 112 absolute maximum is 2,600 m in the Atlantic mountains of Morocco; Fady et al. 113 2003). The optimum climatic conditions for this species are 350–700 mm annual 114 rainfall (semi-arid and sub-humid climate) and between −2 and 10 °C absolute 115 minimum temperatures. P. brutia is less thermophilic (minimum temperature 116 between −5 and 10 °C), but more water-demanding (it needs humid/sub-humid 117 climates, with annual rainfall ranging from 400 to 1,300 mm). The altitude limits of 118 this species are between 0 and 600 m, but in the southern part of its natural range it 119 can reach 1,200–1,400 m (1,650 m on the Taurus Mountains in Turkey). 120 Pinus halepensis is one of the European tree species able to survive in shallower 121 soils, particularly on limestone bedrocks, provided it has a favourable temperature 122 range. In Spain, France, Italy, Balkans and Greece 90 % of Aleppo pine stands 123 grow on calcareous soils, chalk and limestone shallow soils; however, it can grow 124 well on moderately acid soils, although on these soils it is frequently outcompeted 125 by other species, such as stone pine or maritime pine. 126 M.R. Chambel et al.

127 The relationship between Pinus halepensis and fire has been the subject of 128 intensive research (Daskalakou and Thanos 1996; Ne’eman et al. 1992, 2004; 129 Pausas et al. 2004; Tapias et al. 2001; Thanos et al. 1996) following widespread 130 concern after the highly destructive fires that occurred in some European 131 Mediterranean countries over recent decades. Briefly, Aleppo pine is capable of 132 having a very intense recruitment after stand-replacing fires, with seedling densities 133 as high as 50,000–200,000 seedlings/ha (Ne’eman et al. 1992); but, on the other 134 hand, it is very flammable and its capability of withstanding low-intensity fires is 135 very low compared to, for example, Pinus pinea (Pausas et al. 2008; Rigolot 2004). 136 In fact, Aleppo pine can be considered a model of an evader strategy in the face of 137 fire, characterized by a low investment in adult endurance combined with the previ- 138 ously mentioned precocious high fertility (Keeley and Zedler 1998; Tapias et al. 139 2004). This extreme behaviour is less marked in Pinus brutia, but this species is also 140 capable of recruiting successfully after destructive crown fires; for example, densi- 141 ties of 20,000 seedlings/ha 5 years after a stand-replacing fire have been reported in 142 Greece (Spanos et al. 2000). 143 Aleppo pine is one of the first species used for afforestation in dry conditions. 144 The most emblematic programme is probably the “Green Belt” programme in South 145 Algeria where 100,000 ha were planted with Aleppo pine over 20 years. However, 146 the extensive use of Aleppo pine for afforestation in highly degraded soils in the 147 western Mediterranean has been the object of strong criticism by ecologists and 148 environmentalists for various reasons; the species has been blamed for increasing 149 the risk of fire, reducing biodiversity and diminishing water resources (Bellot et al. 150 2004; Maestre et al. 2003). This negative view starts with the lack of recognition of 151 the Aleppo pine as a true native species out of very narrow limits, a gross scientific 152 error that still lingers within some collectives (see a discussion in Gil et al. 1996). 153 Aleppo pine can rapidly colonize burned areas covered by other species, less 154 adapted to frequent fires, like Pinus pinaster or Pinus nigra. This high colonizing 155 capacity, based on the already mentioned reproductive traits, makes this species one 156 of the major invasive alien trees in South Africa where, together with P. pinaster and 157 P. radiata, it constitutes a conservation problem in large areas of the Cape peninsula 158 covered with fynbos, the highly diverse native shrubland (Richardson 1998; 159 Richardson et al. 1990). In south-western Australia, Aleppo pine has been declared 160 as an invader weed (www.environment.sa.gov.au) to be eradicated but, at the same 161 time, it has been promoted for low-input forestry farming.

162 5.3 Aleppo and Brutia Pines in Treebreedex Countries

163 5.3.1 Native and Cultivated Area

164 Considering the ﬁgures for natural and cultivated areas of Aleppo pine within 165 TreeBreedex countries (Table 5.1), it could be considered as a minor species with 166 the exception of Spain, where it represents as much as 45 % of the total forest 5 Mediterranean Pines (Pinus halepensis Mill. and brutia Ten.)

Table 5.1 Native and cultivated area covered by Aleppo pine in Treebreedex countries t1.1

Native Cultivated Total range Total area of conifers Total forest t1.2 Country range (ha) range (ha) (ha) (ha) area (ha) t1.3 Spain 2,345,663 649,211 2,994,874 6,374,650 (+ 3,498,643 27,872,829 t1.4 mixed conifer and t1.5 broadleaf stands) t1.6 France 250,000 ~0 250,000 4,800,000 16,100,000 t1.7 Italy 226,101a 20,000 143,300 1,388,000 8,860,701 t1.8 aArea occupied by Mediterranean pine forests: P. halepensis, P. pinaster and P. pinea t1.9

area. In all three countries, Aleppo pine remains essentially a wild species, with 167 a low percentage of cultivated forests (22 % in Spain, 14 % in France and 8.8 % 168 in Italy). 169

5.3.2 Products and Major Uses 170

Intrinsic wood quality of Aleppo and Brutia pines is very similar to that of other 171 [AU2] Mediterranean pines, for example P. pinaster (Giordano 1976; Thibault et al. 1992). 172 However, the wood of both species is of variable interest in different countries 173 depending on the availability of other pine wood of better quality. In general, the 174 wood provided by P. brutia has a higher quality due to the better stem form and 175 branching, but this depends largely on site quality. Mean productivity is approximately 176 1–3 m3 ha−1 year−1 for Aleppo pine, and 2–3 m3 ha−1 year−1 for Brutia pine. According 177 to the French Forest Inventory Service (Anonymous 2010), mean productivity in 178 France averages 2.3 m3 ha−1 year−1 but it can reach 5 m3 ha−1 year−1 in favourable 179 conditions. It must be stressed that this productivity was strongly increased (+40 %) 180 over the last 25 years. Maximum yield can reach 12–15 m3 ha−1 year−1 in high- 181 quality stands for both species. 182 Some non-wood products are also obtained from these species. Seeds are used 183 for making pastry (particularly in Tunisia). Resin was extracted in some countries 184 during the twentieth century, but this use is currently practically abandoned, except 185 in Greece, were it is still used for wine production (retsina). Bark was used for 186 tannin extraction, but nowadays this use is limited to North Africa. A curious, yet 187 economically relevant, non-wood production of these pines is the honey produced 188 in Greece and Turkey from the honeydew released by the sap-sucking insect 189 Marchalina hellenica (Bacandritsos et al. 2004; Santas 1983). 190 In Spain, Pinus halepensis is widely used for land protection and afforestation of 191 degraded mountain areas. Extensive planting of this species took place from the 192 early twentieth century for land reclamation and soil protection both within and 193 outside its natural range. In particular, moderate success was achieved in the refor- 194 estation of gypsum-limestone marls at areas of extreme Continental climate in the 195 M.R. Chambel et al.

196 Duero basin (northern Castillian plateau). Annual fellings are about 206,250 m3 197 (2007 data). Timber of this species is not appreciated because the density is too 198 high, and there are numerous knots and resin pockets, even when other mechanical 199 properties (MOE) are adequate. There is some use for pallets and chipping for par- 200 ticleboards; it is also used for boat making at a local scale. Aleppo pine is frequently 201 planted in rain-fed suburban parks and road lines. 202 In France this species is mainly used for soil preservation in erodible calcareous 203 slopes and for recreation forestry. However, there are very few plantations and the 204 increase of Aleppo pine forests is mostly due to natural regeneration in abandoned 205 agricultural lands. Although it is sometimes used as fuel or in the pallet industry, its 206 wood is mostly used for paper production (200,000 tm year−1). The species is also 207 used as an ornamental tree and for road line plantations. 208 In Italy, afforestation with Aleppo pine is carried out in coastal areas and in the 209 adjacent hills. This species was widely planted between 1930 and 1970 in 210 Mediterranean areas for soil protection and wind breaking near the coasts and for 211 resin extraction, staining products and wood manufacture. Indeed, the wood of this 212 species was used for boat construction; nowadays it is mainly used for dune protec- 213 tion, for ornamental proposes, for road line plantations and as raw material for the 214 pulp and paper industry. Resin production, tannin extraction for leather and ﬁshing 215 nets were also important traditional uses. However, the recreational use of this 216 species is nowadays the dominant use along the coasts.

217 5.3.3 Plant Needs for Reforestation

218 There is a marked lack of statistics within TreeBreedex countries concerning the 219 actual number of plants consumed for afforestation with Aleppo pine, due to the 220 highly dispersed information between countries and regions. The EU Common 221 Agricultural Policy has increased the use of this species for afforestation of private 222 former arable lands. For example, in Spain between 2003 and 2006 a total of 223 29,113 ha were planted with Aleppo pine. This gives us about 28 million plants 224 (considering an average use of 800 plants/ha and replanting rates of 20 %, Fig. 5.4). 225 In France and Italy plant production is presently very low due to the lack of affores- 226 tation with this species and the limited use for ornamental purposes. 227 In Italy, during the forest fires of 2007, about 10,000 ha of natural and afforested 228 areas planted with Pinus halepensis and P. brutia were destroyed in the Abruzzo 229 and Puglia regions. Reforestation programmes to recover these areas have started 230 at least in the Puglia region, due to the touristic and economic importance of forests 231 in this area. 232 To complete reforestation of these and other burned areas as in Greece (Greece 233 is certainly the country the most affected by the great fires of recent years), the global 234 potential demand for seedlings could be to the tune of several million in the next 235 years. Intensive exchange of reproductive materials occurs between Italian and 236 Greek nurseries, but there are no statistics on these figures. 5 Mediterranean Pines (Pinus halepensis Mill. and brutia Ten.)

Fig. 5.4 Aleppo pine plant stock at TRAGSA nurseries, Spain (J. Climent)

5.4 The Current Situation as a Starting Point for Breeding 237

Within the TreeBreedex consortium, there are no active breeding programmes 238 of Aleppo or Brutia pine at the same level of those of major species (Norway spruce, 239 Sitka spruce, Scots pine, etc.). Provenance research can be considered as part of 240 low-intensity breeding programmes in France, Italy and Spain aimed mainly at 241 assuring self-maintenance of low-production and protective forests. 242

5.4.1 Provenance Research Within TreeBreedex Countries 243

5.4.1.1 FAO International Network 244

Between 1977 and 1982, FAO/Silva Mediterranea, within the project FAO/SCM/ 245 CRFM/4 bis, promoted a large network of provenance trials with species of the 246 halepensis group. This programme allowed the collection of seeds from 57 prove- 247 nances (33 of Aleppo pine, 17 of Brutia pine and seven of Eldarica pine) covering 248 the natural range of the species (Fig. 5.5) and supported the establishment of several 249 ﬁeld trials across the Mediterranean and in other areas with a Mediterranean climate 250 like Australia (Spencer 1985) or New Mexico (Fisher et al. 1986). Twelve of these 251 trials are located in TreeBreedex countries (10 in Italy and 2 in the south-east of 252 France; Tables 5.2 and 5.3; Fig. 5.6). 253 M.R. Chambel et al.

Fig. 5.5 Location of the provenances sampled for the FAO/Silva Mediterranea trial network (dots) and location of the trials in TreeBreedex countries (ﬂags). Red dots represent provenances of Pinus halepensis and orange dots represent P. brutia. Dark dots represent provenances not included in TBX countries trials

254 Considering the age of these trials, the results can now be regarded as deﬁnitive. 255 In general, growth traits displayed high plasticity and signiﬁcant species × site 256 interaction across trial-sites; however, provenance rankings tended to remain rather 257 stable (i.e. there is quantitative but not qualitative g × e interaction). On the other 258 hand, traits such as stem form and frost resistance showed lower levels of interaction 259 both between species and between provenances within species. 260 In Italy, in the trial sites situated at higher altitudes, a long and hard frost 261 occurred in 1985 which had devastating consequences for most of the Aleppo 262 pine provenances, with the French provenance A24 – Gemenos being the least 263 affected (Bariteau 1992; Eccher et al. 1987; Ducci and Guidi 1998). In sites 264 where temperatures reached −24 °C for as long as 2 weeks, only Pinus brutia 265 provenances managed to survive. Therefore, Pinus halepensis provenances 266 should be planted only in truly Mediterranean conditions, while P. brutia should 267 preferably be used in interior and more continental sites. In these conditions, the 268 best performing and most stable provenances were those form the Chalkidiki 269 (A4 and A5) peninsula (North-eastern Greece), A27-Vico del Gargano and 270 A28-Patemisco (Italy), A6-Shaharia and A7-Elkosh (Israel). Furthermore, these 271 provenances, together with the French provenance Gemenos, were also characterized 272 by good stem quality. 5 Mediterranean Pines (Pinus halepensis Mill. and brutia Ten.) 15.5 15.0 Mean annual temp. (°C) 12.0 15.5 16.7 17.7 12.7 16.0 14.7 16.6 14.8 11.8 569 786 976 685 625 502 828 834 714 544 655 Mean annual (mm) rainfall 1,020 1978 1976 Planting year 1979 1981 1982 1982 1977 1975 1975 1976 1976 1976 60 40 50 200 446 Altitude (m) 700 175 165 140 300 225 575 ¢ ¢ ¢ ¢ ¢ ¢ ¢ ¢ ¢ ¢ ¢ ¢ 5° 15 5° 40 8° 57 9° 33 13° 52 14° 50 11° 56 12° 21 12° 09 17° 14 11° 07 11° 48 Long. (E) ¢ ¢ ¢ ¢ ¢ ¢ ¢ ¢ ¢ ¢ ¢ ¢ 43° 28 43° 13 Lat. (N) 41° 13 41° 49 38° 58 39° 10 43° 27 41° 54 41° 53 40° 28 42° 21 44° 07 d’Azur d’Azur Provence Côte Provence Provence Côte Provence Region Abruzzo Molise Sardinia Sardinia Tuscany Lazio Lazio Apulia Tuscany Emilia-Romagna Rhône Rhône Bouches-Du- Province Pescara Campobasso Cagliari Cagliari Arezzo Rome Rome Taranto Grosseto Forlì Bouches-Du- Provenance trials installed within the framework of FAO/Silva Mediterranea between 1977 and 1982 in TBX countries Mediterranea between 1977 and 1982 in of FAO/Silva trials installed within the framework Provenance

. Collemezzanotte was destroyed by fire in 2003 destroyed . Collemezzanotte was A . Castel di Guido . Fornari . Ceyreste . Collemezzanote . Pixinamanna . Castiadas . Vitrolles . . Lentiscete Arezzo . . Ovile . Montebello . Castiglioncello Test Test L Table 5.2 Table Site A B C D E F G H I J K t2.1 t2.2 t2.3 t2.4 t2.5 t2.6 t2.7 t2.8 t2.9 t2.10 t2.11 t2.12 t2.13 t2.14 t2.15 t2.16 t2.17 t2.18 t3.1 Table 5.3 List of provenances used in the FAO/Silva Mediterranea international network of field trials t3.2 Code Provenance name Country Longitude Latitude t3.3 Pinus halepensis t3.4 A 1 Albania Albania 19°25¢ E 40°37¢ t3.5 A 2 Elea Greece 21°32¢ E 37°46¢ t3.6 A 3 Euboea Greece 23°18¢ E 38°58¢ t3.7 A 4 Chalkidiki 1 Greece 23°21¢ E 40°11¢ t3.8 A 5 Chalkidiki 2 Greece 23°44¢ E 40°03¢ t3.9 A 6 Shaharia Israel 34°50¢ E 31°36¢ t3.10 A 7 Elkosh Israel 35°18¢ E 33°01¢ t3.11 A 8 Sakiet Sidi Y. Tunisia 08°25¢ E 36°15¢ t3.12 A 9 Qoum Djeddur Tunisia 08°57¢ E 35°38¢ t3.13 A10 Djebel Selloum Tunisia 08°40¢ E 35°05¢ t3.14 A12 Zaouia Ifrane Morocco 05°23¢ W 33°15¢ t3.15 A13 J. Afra Selm.te Morocco 07°55¢ W 30°44¢ t3.16 A14 Quardane B. Morocco 05°08¢ W 35°03¢ t3.17 A15 Tamga Zaonia Morocco 06°07¢ W 32°02¢ t3.18 A17 Guadameldina Spain 02°15¢ W 37°02¢ t3.19 A24 Gemenos France 05°40¢ E 43°25¢ t3.20 A25 Imperia Italy 08°03¢ E 43°54¢ t3.21 A26 Otricoli Italy 12°38¢ E 42°24¢ t3.22 A27 Vico del Garg. Italy 16°00¢ E 41°54¢ t3.23 A28 Patemisco Italy 17°20¢ E 40°39¢ t3.24 A29 Aures Beni M. Algeria 06°50¢ E 35°10¢ t3.25 A30 Senalba Algeria – – t3.26 A31 Telagh Algeria – – t3.27 A32 Ourasensis Algeria 05°04¢ E 35°05¢ t3.28 A33 Lebanon Lebanon – – t3.29 Pinus brutia t3.30 B 1 Chania Greece 23°57¢ E 35°17¢ t3.31 B 2 Kavala Greece 24°42¢ E 40°48¢ t3.32 B 3 Lassithiou Greece 25°32¢ E 35°06¢ t3.33 B 5 Cyprus Cyprus 33°17¢ E 35°08¢ t3.34 B 6 Marmaris Turkey 28°18¢ E 37°00¢ t3.35 B 7 Isparta Turkey 29°32¢ E 38°04¢ t3.36 B 8 Duzlerçani Turkey 30°25¢ E 37°03¢ t3.37 B 9 Pamuçak Turkey 30°41¢ E 37°40¢ t3.38 B10 Bozburun Turkey 30°45¢ E 37°21¢ t3.39 B11 Bakara Turkey 32°43¢ E 36°09¢ t3.40 B12 Silifke Turkey 33°43¢ E 36°13¢ t3.41 B13 Gamgolu Turkey 35°20¢ E 41°50¢ t3.42 B14 Baspinar Turkey 35°15¢ E 37°48¢ t3.43 B15 Kisildag Turkey 35°58¢ E 36°21¢ t3.44 B16 Zawita Iraq 44°20¢ E 36°35¢ t3.45 B17 – Lebanon – – t3.46 B25 Unknown t3.47 B27 Unknown t3.48 B29 Unknown t3.49 Pinus brutia subsp. eldarica t3.50 E 1 Karaj Iran 51°00¢ E 35°56¢ t3.51 The longitude is given as the first coordinate, given its importance in the range distribution 5 Mediterranean Pines (Pinus halepensis Mill. and brutia Ten.)

Fig. 5.6 Field trial belonging to the FAO/Silva Mediterranea network at Ovile, Rome (E. Fusaro)

According to results shown by Eccher et al. (1987), Bariteau (1992) and Ducci 273 and Guidi (1998), P. brutia showed higher homogeneity of performances, allowing 274 the deﬁnition of wider provenance regions which can be used as good seed sources: 275 predominantly the southern Turkey coastal range between 30° and 36° longitude 276 East. The easternmost provenances (B13 – Gamgolu to B16 – Zawita and E1) are the 277 most suitable for the warm deciduous oak range areas, whilst the western basic 278 material (B8 – Duzlerçani to B10 – Bozburun) performs better in the wet-cold 279 evergreen/deciduous oak transition range. The best stem quality can be found in the 280 oriental group of provenances. The Eldar pine performances could be interesting in 281 ecological areas on lime stones, higher elevations and a more continental climate, 282 whilst it must be avoided in lower sites and/or where there are larger amounts of clay. 283 In the two French sites, the lowest survival rates were observed for some 284 P. halepensis provenances from Greece (A2-Elea), Italy (A27-Vico del Gargano 285 and A26-Otricoli) and Morocco (A14-Ouardane Bouksane), while provenances 286 from France (A24-Gémenos), Spain and north-eastern Greece exhibited good sur- 287 vival rates, with no clear geographic pattern (Pichot and Vauthier 2007). The highest 288 M.R. Chambel et al.

Fig. 5.7 Location of provenances and experimental sites included in the Spanish Aleppo pine network of genetic trials. Black ﬂags represent provenance trial sites (6); blue ﬂags represent provenance/ progeny trial sites (2); dots represent sampled populations, 24 of them included in both trials

289 survival rates in this trial were those of the Pinus brutia provenances originating 290 from eastern Taurus (B15-Kisildag and B14-Baspinar).

291 5.4.1.2 Other Provenance Trial Networks

292 More recent experimental networks were implemented in Spain and France for 293 assessing respectively the variability of Aleppo pine national resources and evaluating 294 the adaptation of Brutia pine in less dry but colder Mediterranean (deeper soils or 295 higher elevation) conditions than those previously assessed. 296 In Spain, the network includes a six-site provenance trial with Spanish and 297 Mediterranean-wide provenances and a three-site trial with a progeny within prov- 298 enance structure, including only Spanish materials. The provenance trials were 299 planted in 1998 with 42 populations from Spain, one from France, three from 300 Greece, three from Italy and two from Tunisia. Additionally, three highly productive 301 plantations of unknown origin located in north continental Spain, a commercial 302 seed lot from eastern Spain and a seed lot from a Greek seed orchard (Fig. 5.7) were 303 also included in these trials. 304 Phenotypic plasticity (site effect) was by far the main source of variation for 305 growth variables in this trial, while provenance and g × e interaction had signiﬁcant 5 Mediterranean Pines (Pinus halepensis Mill. and brutia Ten.)

Fig. 5.8 Provenance/progeny trial with 9 years of age at a marginal site for the species (low winter temperatures, low rainfall and gypsum marls soil) in Central Spain, Valladolid province (R. López) but reduced effects. No clear geographic patterns were found for growth or phenotypic 306 plasticity (Chambel et al. 2007). The higher rankings across sites for growth were 307 obtained by the provenances from Greece, but their survival was generally low. 308 The provenance/progeny trial includes two sites with highly contrasting environ- 309 ments (Fig. 5.8), planted in 1995 with 148 open-pollinated families from 32 popula- 310 tions that cover the entire natural and planted range of peninsular Spain and Balearic 311 islands (Santos-del-Blanco et al. 2010); 24 of these populations are also included in 312 the provenance trial. Although also in this trial, site is still the most significant factor 313 influencing growth and survival; there were also significant differences among prov- 314 enances, while the effect of family within provenance was scarce. The estimated dif- 315 ference in biomass production between the best and the worst performing provenance 316 at age ten was as high as 145 % for the harsher site, highlighting the importance of an 317 adequate selection of the seed source used for afforestation (Climent et al. 2009). 318 In France, a “new” five-site comparative trial was settled between 1995 and 1997 319 within the Mediterranean Pine and Cedar (MPC) European project and the 320 FORADAPT INCO project (Plomion and Pichot 2001; Fig. 5.9; Table 5.4). Eighteen 321 Pinus brutia provenances from Turkey were planted in this trial along with four 322 P. halepensis populations to be used as controls (Table 5.5). An additional set of 323 seven P. brutia provenances commonly used by the French forest service were also 324 included in Bousquet d’Orb and Naucadery plantations. These plantations share 325 some provenances with trials established within the same project by Moroccan and 326 Tunisian partners. 327 M.R. Chambel et al.

Fig. 5.9 Location of provenances and experimental sites included in the ‘new’ French brutia pine provenance trial. Black triangles represent trial sites (5); orange dots (18) represent Pinus brutia provenances and red dots represent P. halepensis provenances used as control (4) t4.1 Table 5.4 List of the “new” French Pinus brutia provenance plantations t4.2 Number t4.3 Site Year plant Altitude Surface (ha) Soil of provenances t4.4 Bousquet d’Orb F 1995 580 2 Limestone 28 t4.5 Naucadery (Laure) S 1996 150 1.5 Limestone 27 t4.6 Toulourenc forest F 1996 600 2 Limestone 22 t4.7 Bedoin forest S 1997 600 2 Limestone 25 t4.8 Laquina S 1997 600 1.8 Schist 22

t5.1 Table 5.5 List of the provenances tested in the “new” 5-site trial t5.2 Species Provenance Country Species Provenance Country t5.3 P. brutia 050 Karsanti Turkey P. brutia 116 Eskibag Turkey t5.4 P. brutia 071 Kiyra Turkey P. brutia 120 Karabucak Turkey t5.5 P. brutia 073 Suçati Turkey P. brutia 121 Güzelcluk Turkey t5.6 P. brutia 076 Guzelbag Turkey P. brutia 122 Bigadic Turkey t5.7 P. brutia 079 Pinargözü Turkey P. brutia 124 Camkonagi Turkey t5.8 P. brutia 100 Karadag Turkey P. brutia 125 Göktepe Turkey t5.9 P. brutia 109 Gökçesu Turkey P. brutia 129 Koças Turkey t5.10 P. brutia 110 Findikpinari Turkey P. halepensis 001 St Etienne du Grès France t5.11 P. brutia 113 Melli Turkey P. halepensis 002 Port-Cros France t5.12 P. brutia 114 Merkez Turkey P. halepensis 003 Mejjou Morocco t5.13 P. brutia 115 Karaçay Turkey P. halepensis 011 Vilmorin France 5 Mediterranean Pines (Pinus halepensis Mill. and brutia Ten.)

5.4.2 Provenance Research in Other Countries 328

In Greece, a network of provenance trials has been established for both P. halepensis 329 (Fig. 5.10a) and P. brutia (Fig. 5.10b), to explore the levels of the existing variation 330 for adaptive traits. This network consisted of five trial sites for Pinus halepensis and 331 seven trial sites for Pinus brutia including both Greek and foreign materials. These 332 were, for P. halepensis: three Italian, one from Spain, three Moroccan, five Tunisian 333 and six other provenances form North African countries and, for Pinus brutia: two 334 Turkish provenances and one from Cyprus. 335 In Morocco, provenance research on the species of the halepensis group was very 336 active during the second half of the twentieth century, when there were a large number 337 of trials. A five-site Pinus brutia provenance test was planted in 1969; another trial 338 was installed in only one location in 1973–1974 with seven provenances of P. halepen- 339 sis and P. brutia; and in 1976–1977 a four-site provenance trial within the FAO/Silva 340 Mediterranea network including 51 provenances was installed. Unfortunately, most 341 of these plantations do not exist anymore and little information on the results is easily 342 available (Pichot 1995). In 1992 a new two-site provenance test was planted with 58 343 provenances (1 Eldarica pine, 8 Brutia pine and 49 Aleppo pine). In 1999, 18 P. brutia 344 provenances and 1 P. halepensis were planted in a two-site trial located in the Moroccan 345 dry area, implemented within the EU Mediterranean INCO-FORADAPT project. 346 In Turkey, a wide programme of provenance research was carried out by the 347 Turkish Forest Research Institute, with a set of 26 trial sites of Brutia pine planted 348 in 1988 and 1989 throughout Turkey and Northern Cyprus, with 47 provenances 349 from Turkey and 3 from Cyprus, mostly from selected stands. 350 In Tunisia, Aleppo pine populations cover 200,000 ha and P. halepensis is the first 351 species for afforestation (80,000 ha). The studies on genetic diversity of both Aleppo 352 and Brutia pines are based on two sets of provenance trials, a seven-site trial estab- 353 lished in the 1960s with 49 provenances, and a four-site trial planted in 1998, focusing 354 on drought tolerance, implemented within the EU Mediterranean INCO-FORADAPT 355 project and including 13 provenances of Aleppo pine and 27 of Brutia pine. 356

5.4.3 Geographic Patterns of Genetic Diversity 357

5.4.3.1 Biochemical and Molecular Variation 358

Discrimination between species and their hybrids was carried out by terpene and 359 spSSR analyses (Gallis et al. 1997; Korol et al. 1995; Tognetti et al. 1997; Bucci 360 et al. 1998). 361 Studies on Aleppo and Brutia pine genetic diversity and population genetics 362 based on isozymes (Schiller et al. 1986; Conkle et al. 1988; Teisseire et al. 1995, 363 Agúndez et al. 1997, 1999; Korol et al. 2002; Wahid et al. 2010), RAPDs and cpSSR 364 (Bucci et al. 1998; Gomez et al. 2001; Alrababah et al. 2011) generally indicate 365 lower levels of intra-population diversity for Aleppo pine and the existence of 366 genetic barriers for introgression between both species. 367 M.R. Chambel et al.

Fig. 5.10 Location of provenances and trial sites of the Pinus halepensis network (a) and of the Pinus brutia network (b) in Greece. Yellow stars represent trial sites and trees represent provenances (P. Alizoti) 5 Mediterranean Pines (Pinus halepensis Mill. and brutia Ten.)

The genetic variability of Pinus halepensis has a geographic structure that reﬂects 368 migration pathways from glacial remnants after the last glaciations (Gómez et al. 369 2005). Recent results conﬁrmed a marked loss of genetic diversity from Greek 370 populations towards the western range of the species, as well as molecular sig- 371 natures of intense bottlenecks (Grivet et al. 2009). Genetic analyses on P. brutia 372 indicated a high differentiation between populations but with no clear geographic 373 structure (Bucci et al. 1998). 374

5.4.3.2 Quantitative and Adaptive Variation 375

The previously mentioned trial networks and smaller-scale eco-physiological research 376 give evidence of both an adaptive divergence between the two species and differentia- 377 tion among provenances within species. A joint analysis of all extant data from different 378 trial sets is still lacking, nevertheless, there are several published works regarding 379 growth, mortality and frost resistance (Calamassi et al. 1984); needle morphology and 380 anatomy (Calamassi et al. 1980, 1987); drought tolerance (Calamassi et al. 1980; 381 Falusi et al. 1983; Grunwald and Schiller 1988; Atzmon et al. 2004; Voltas et al. 2008), 382 seed and cone morphology (Boulii et al. 2001, 2003) and pest resistance, stem form 383 and forking (Eccher et al. 1987; Weinstein 1989; Bariteau 1992; Ducci and Guidi 384 1998). The results obtained confirm that Pinus halepensis and Pinus brutia have 385 different ecological optima. Pinus brutia is less tolerant to dry environments, being 386 out-performed by most P. halepensis provenances at drier sites, but on the other hand, 387 it is less frost sensitive, therefore better adapted to continental areas and high altitudes. 388 Pinus brutia generally presents better form and it has also been proven to be more 389 resistant to the insect Pissodes notatus (Ducci and Guidi 1998). 390 In general, Aleppo pine tends to show a higher differentiation between prove- 391 nances than Brutia pine, but this varies considerably among studies due to prove- 392 nance sampling and test environments. 393 In Aleppo pine, the north-east–south-west cline for adaptive characters seems 394 very well established experimentally, with north-eastern provenances (Greece) 395 showing higher early growth rates and better forms than south Iberian and north 396 African provenances. Recently, a similar geographic trend for early reproductive 397 allocation has also been confirmed, with eastern and northern populations (particu- 398 larly Greece, Catalonia and Balearic Islands) far less precocious for cone bearing 399 than South Iberian and North African provenances (Climent et al. 2008). 400 For Brutia pine, despite its smaller range of distribution, there are also experi- 401 mental evidences of differences between populations and altitudinal gradients for 402 adaptive traits (i.e.: different resistance to cold and developmental rates, Isik 1986; 403 Kaya and Isik 1997; Pichot and Vauthier 2007). Provenances from the middle eleva- 404 tion of the Mediterranean region generally perform the best while provenances from 405 the peripheral areas of the distribution range of the species or isolated populations 406 generally grow more slowly. However, other reports postulate scarcely distinct 407 geographic patterns of diversity in this species as a consequence of human influence 408 and recurrent fires (Gülcü and Çelik 2009). 409 M.R. Chambel et al.

410 Besides this between-population variation, high intra-population diversity (both 411 through heritability and/or additive co-efﬁcient of variation) has been recorded in 412 Aleppo pine for growth (Matziris 2000) and particularly for reproductive traits 413 (Matziris 1997; Santos-del-Blanco et al. 2010). This is also true for Brutia pine, 414 with very similar heritability values for the different sets of traits (Gülcü and Çelik 415 2009; Isik et al. 2002; Kaya and Isik 1997). 416 In general, the formerly mentioned trial networks and smaller-scale eco-physio- 417 logical research (Grunwald and Schiller 1988) give evidence for both an adaptive 418 divergence between the two species and differentiation among provenances within. 419 The results obtained in different countries conﬁrm that Pinus halepensis and Pinus 420 brutia have different ecological optima species (Bariteau 1992; Ducci and Guidi 421 1998; Boulii et al. 2001, 2003). Pinus brutia is less tolerant to dry environments, 422 being out-performed by most P. halepensis provenances at more Mediterranean 423 sites, but on the other hand, it is less frost sensitive, presenting higher adaptability 424 to continental areas and high altitudes. Pinus brutia generally presents best stem 425 form and tree architecture and it has also proven to have better resistance to the 426 insect Pissodes notatus (Ducci and Guidi op. cit.). 427 Aleppo pine tends to show a high inter-provenance variability, while Brutia pine 428 tends to show lower differentiation between provenances, but the latter varies con- 429 siderably among studies (provenance sampling and environments). A joint analysis 430 of all extant data from different trial sets is still lacking. Therefore, the following 431 sections will quote the most consistent results and then some more details on the 432 results obtained in each country up until now.

433 5.4.4 Seed and Plant Transfer: History and Current Trends

434 Today, in Italy, France and Spain, seed imports from Turkey are allowed for Pinus 435 brutia. In Italy, imports have been programmed with reference to indications issued 436 from FAO Silva Mediterranean field tests. At present, most needs can be covered by 437 a seed orchard derived from these programmes. 438 By contrast, Aleppo pine seeds must come from the national provenance regions 439 and seed stands in the three countries (Fig. 5.11). In France, however, significant [AU3] 440 quantities of Aleppo pine seeds were imported in the past from some foreign countries 441 for historical afforestation programmes in the South of France. This reproduc- 442 tive material often came from stands exhibiting warmer climatic conditions than 443 those observed in France and its susceptibility to frost led to high mortality during the 444 coldest winters (1956 and 1985) within forest stands planted respectively with 445 Algerian and Italian provenances (Tabeaud and Simon 1993; Bedel 1986). 446 The situation was different in Italy and Spain, where the importation of foreign 447 Aleppo pine seeds, if any, can be considered negligible due to the high production 448 of seeds from local basic materials. Traceability of former seed transfer between 449 regions within Spain is unrealistic, so both successful and unsuccessful plantations 450 outwith the natural range are difficult to trace in terms of seed origin. 5 Mediterranean Pines (Pinus halepensis Mill. and brutia Ten.)

Fig. 5.11 Aleppo pine seeds clean and ready for the market (1966, courtesy of the Italian Forest Service)

In Italy, wide forestation programmes were carried out in Tuscany for coastal 451 dunes and abandoned interior and Mediterranean areas. This work was initiated by 452 the Dukes of Tuscany before 1860 and then continued by the Kingdom of Italy after 453 the Uniﬁcation. The amount of Aleppo pine seed distributed documented by the 454 Italian Forest Service was nearly 2,000 kg in the early 1960s, reaching about 455 20,000 kg in the early 1970s with a peak in 1989 when about 40,000 kg were pro- 456 duced. These amounts were estimated to cover 60 % of the Aleppo pine seeds used 457 at national level. Thereafter, production decreased progressively and today only a 458 few hundred kilos are being produced and only a proportion are sold. That is due to 459 the end of forestation programmes in the early 1990s (Mencaccini 2010). 460

5.4.5 Approved Forest Reproductive Material 461 Within the TreeBreedex Countries 462

A total of 54 seed stands registered according to rules under the European Directive 463 1999/105/CE are officially approved in the three TreeBreedex countries for the pro- 464 duction of Pinus halepensis seeds (Table 5.6). 465 In Spain there are 14 selected stands, belonging to 7 out of the 20 provenance 466 regions defined for the species (Fig. 5.12). These selected stands cover a total of 467 495 ha. All except one were homologated in February 2002 (the remaining one was 468 homologated in July 2009). There is also one clonal seed orchard which was 469 homologated in July 2007 with material selected from three provenance regions. 470 The selection of parent material for this seed orchard was based on growth and 471 form. The seed from this seed orchard is currently catalogued as qualified FRM, but 472 M.R. Chambel et al. t6.1 Table 5.6 Homologated FRMs in TreeBreedex countries t6.2 Country Selected Qualified Tested t6.3 Spain Number 14 selected seed stands 1 seed orchard (P. halepensis) – t6.4 Area 495 ha 4.31 ha – t6.5 France Number 25 selected seed stands – – t6.6 Area 228 ha – – t6.7 Italy Number 15 selected seed stands 1 seed orchard (P. brutia) – t6.8 Area About 2,600 ha 2.5 ha –

Fig. 5.12 Regions of provenance deﬁned in Spain for Aleppo pine: 1. Alta Cataluña, 2. Cataluña Litoral, 3. Cataluña Interior, 4. Bárdenas-Ribagorza, 5. Ibérico Aragonés, 6. Monearos-Depresión del Ebro, 7. Alcarria, 8. La Mancha, 9. Maestrazgo-Los Serranos, 10. Levante Interior, 11. Litoral Levantino, 12. Pitiusas, 13. Sudeste, 14. Bética, 15. Bética Meridional, 16. Cazorla, 17. Sur, 18. Mallorca y Menorca, 19. Repoblaciones de la Meseta Norte. Region 19, in blue, consists of plantations outwith the natural range of the species

473 there are already comparative trials which have been planted with the aim of raising 474 the category up to tested FRM. 475 In France, there are 25 selected stands for the production of Pinus halepensis 476 seed, totalling 228 ha, and all of them are included in the only provenance 477 region deﬁned for the species (Fig. 5.13) with all having been homologated in 478 November 2003. 479 In Italy, there are 15 selected seed stands, broadly covering the national range of 480 the species (Fig. 5.14). All of these seed stands were homologated before 1975 481 (Morandini and Magini 1975; Ducci et al. 2010). 5 Mediterranean Pines (Pinus halepensis Mill. and brutia Ten.)

Fig. 5.13 Regions of provenance deﬁned in France for Aleppo pine: a single provenance region named PHA 700: Region Méditerranéenne

Fig. 5.14 (a) The range of Pinus halepensis in Italy and the location of selected seed stands: 125 – Vallecrosia (Imperia-Liguria), 110 – Chiavari-Le Grazie (Genova-Liguria), 15 –Cerchiara (Terni-Umbria), 18 – Arrone (Terni-Umbria), 19 – Otricoli (Terni-Umbria), 60 – Gargano-Marzini (Foggia-Puglia), 78 – Gargano-Monte Pucci (Foggia-Puglia), 59 – Patemisco (Taranto-Puglia), 52, 53, 54, 55, 56, 57, 58 – Litorale Tarantino (Taranto-Puglia). Capital letters represent FAO trial sites (see Table 5.2). (b) The provisional ofﬁcial map of provenance regions in Italy (Partitionist method 2010) agreed by all the Regional and State Forest Services. A – Alpine Region, B – Planitial Po valley Region, C – Central–Northern Mediterranean Region, D – Southern Mediterranean region, E – Sardinian (Mediterranean) Region, F – Sicily (Mediterranean) Region M.R. Chambel et al.

482 There are no speciﬁc provenance regions deﬁned for P. brutia within TreeBreedex 483 countries, but a seed orchard of this species was recently registered in Italy (Ducci 484 et al. 2010).

485 1. Among all the TreeBreedex countries, 70 Aleppo pine and 77 of Brutia pine selected 486 seed stands were inventoried by FAO Silva Mediterranea (Topak 1997). Seed orchards 487 recorded were 4 for Pinus halepensis, 55 for Brutia pine and 2 for Pinus eldarica, 488 The majority are located in Cyprus (6) and Turkey (49) and only one in Egypt.

489 5.4.6 Web Links to Approved FRMs in TreeBreedex Countries

490 Spain 491 http://www.mma.es/portal/secciones/biodiversidad/montes_politica_forestal/recur- 492 sos_geneticos_forestal/programas_mejora_genetica/catalogo_materiales_base/ 493 index.htm 494 https://sgw.mma.es/ (this page can only be access by authorized users)

495 France 496 http://agriculture.gouv.fr/graines-et-plants-forestiers 497 http://agriculture.gouv.fr/IMG/pdf/pin_alep-2.pdf

498 Italy 499 http://www.infc.it 500 http://www.ricercaforestale.it/ 501 http://www.ricercaforestale.it/modules.php?op=modload&name=BoschiDaSeme& 502 ﬁle=index

503 5.5 Current Breeding Programmes, Description and Results

504 5.5.1 Breeding Within TreeBreedex Countries

505 In Spain, a seed orchard programme was carried out in the 1980s and 1990s, from a 506 selection of plus trees in Eastern continental Spain (three regions of provenance). 507 Progeny trials were established, both with open-pollinated seeds from the selected 508 mother trees and, more recently, with the seeds collected from seed orchards. 509 A more recent set of comparative trials was planted to promote the FRM of the most 510 successful seed orchard to the tested category (Nieto et al. 2009), even though the 511 demand for improved seed of this species in Spain (justifying a higher price) is very 512 low. Thanks to the precocious and proliﬁc cone-bearing in seedlings, progeny tests 513 can easily be converted into seedling seed orchards of 1.5 generations. 514 In France, controlled pollinations inter- and intra-species and provenances were 515 done in order to study the relationships and genetic control of main adaptive traits 516 (survival, growth, phenology, architecture and fertility) within the halepensis group. 5 Mediterranean Pines (Pinus halepensis Mill. and brutia Ten.)

Five seed trees within each of four provenances (two of P. brutia: Baspinar and 517 Marmaris, and two of P. halepensis: Cehegin and Gémenos) were pollinated with 518 mixtures of pollen collected from ten pollen trees within each of the four prove- 519 nances. The seeds produced made it possible to plant an experimental trial in 2000, 520 in the South of France, near Perpignan. Preliminary results reveal signiﬁcant differ- 521 ences among families for survival but no species effect. Pinus halepensis families 522 grow faster than P. brutia and P. brutia × P. halepensis hybrids are signiﬁcantly 523 more vigorous than pure P. brutia trees. In order to estimate genetic parameters in 524 P. brutia, one provenance/progeny test with 90 half-sib families (3 stands × 30 families) 525 was planted in southern France in 2000. 526 In Italy, two progeny trials with half-sib families of the best performing prove- 527 nances from Greece and Italy, selected by stem form, were established in 1985, 528 covering 4.5 ha. 529

5.5.2 Breeding in Other Countries 530

Long-lasting breeding programmes of both Aleppo and Brutia pines have been car- 531 ried out in other countries outwith the TreeBreedex consortium. 532 In Greece, the initial breeding effort that was launched in 1962 mainly involved 533 hybridization efforts between Pinus brutia and Pinus halepensis. Organized breed- 534 ing programmes were initiated in 1970, with selection and collection of material on 535 a provenance and mother-tree basis. Both P. halepensis and P. brutia provenances 536 exhibited significant genetic variation for all the adaptive traits studied, while their 537 performances across sites indicated the high potential for effective across-site selec- 538 tion (Matziris 1997; Alizoti et al. 2000). P. brutia provenances of Greek origin out- 539 grew the provenances originating from Turkey and Cyprus when tested in two 540 different sites (Alizoti et al. 2001). The outstanding performance of P. brutia × P. 541 halepensis hybrids is worth mentioning, as when tested together with their parental 542 species in harsh environmental conditions they significantly outperformed the only 543 parent that was able to survive (P. brutia). The results indicate the existence of 544 ample genetic variation among and within the natural populations of both species 545 for adaptive traits (Matziris 2000), the potential for selection and breeding and the 546 need for conservation of their unique genetic resources in the face of global change. 547 A first-generation seed orchard exists for Pinus halepensis covering 20 ha and 548 including 76 genotypes (2,630 grafted individuals in total). There are two progeny 549 trials associated with this seed orchard, established in 1987, with 70 open-pollinated 550 families that correspond to 70 out of the 76 clones from the seed orchard. 551 In Israel, a three-site provenance trial with provenances of Pinus halepensis Mill. 552 and related species, P. brutia Ten. and P. eldarica Medw. was established within the 553 framework of the FAO/Silva Mediterranea programme FAO/SCM/CRFM/4 bis, as 554 for the other partners of Silva Mediterranea. The results obtained 10 years after 555 planting were discussed by Weinstein (1989). According to these results, the most 556 promising provenances of P. halepensis are from the lower altitudes of Greece and 557 M.R. Chambel et al.

Fig. 5.15 The Yatir (edges of Negev desert) plantations (G. Schiller)

558 some native stands within Israel, while the most promising seed sources of P. brutia 559 are from the lower elevation on the Mediterranean coast of Turkey. In addition, 560 P. eldarica from Iran showed good growth potential for Israel. Research projects 561 continued within the Aleppo pine forest plantations and new provenance trials at 562 Yatir, near the northern edge of the Israeli Negev desert (Schiller and Atzmon 2009), 563 were established, with the aim of improving the sustainability of plantations with 564 this species (Fig. 5.15). 565 These aims imply that seed sources to be used (provenances, ecotypes) must be 566 selected according to their genetic diversity, drought resistance and water use 567 efﬁciency. Harsh environmental site conditions at the edge of the desert provide the 568 opportunity of exerting heavy selection pressure among and within provenances of 569 a priori drought-tolerant species. Survivors should be used as a seed source for new 570 forest plantations aiming to combat desertiﬁcation. 571 In Morocco, as already mentioned, intense provenance research for both Aleppo 572 and Brutia pine was undertaken in the second half of the twentieth century, includ- 573 ing several trials based on the Silva Mediterranea initiative, and a P. halepensis seed 574 orchard was established in 1978. 575 In Tunisia, as well as the provenance trial sets of Aleppo and Brutia pines planted 576 since the 1960s, a P. brutia provenance/progeny test was planted in 2000 with 90 577 half-sib families (three stands × 30 families). 578 In Turkey, again in parallel with wide provenance research formerly described 579 for Brutia and Aleppo pines, a wide programme of seed orchard plantation has been 580 carried out. As many as 67 seed orchards of Brutia pine covering 472 ha and 2 seed 5 Mediterranean Pines (Pinus halepensis Mill. and brutia Ten.)

Fig. 5.16 Dr. Murat Alan follows Brutia pine progeny tests in Turkey (Ducci et al. 2010)

orchards of Aleppo pine totalling 8.2 ha were established. Progeny testing has also 581 been regularly undertaken and has been the object of intense research (Kaya and 582 Isik 1997; Isik et al. 2002; Fig. 5.16). 583

5.6 Additional Factors Affecting Future Breeding of Aleppo 584 and Brutia Pines 585

5.6.1 Aleppo and Brutia Pines and Climate Change 586

Past predictions of enhanced growth in Mediterranean countries due to the fertilizing 587 effect of increased CO2 (Sabaté et al. 2002) are largely surpassed today because of 588 the crucial role of water availability (Scarascia-Mugnozza 1986; Schroter et al. 589 2005). Nevertheless, both experimental evidences and predictions of the effects of 590 climate change on Aleppo and Brutia pine growth and distribution are highly diver- 591 gent, depending on the particular region. 592 In northern Mediterranean regions, such as southern France and north-eastern 593 Spain, growth increase and/or enlargement of the potential area for the species have 594 [AU4] been postulated, following temperature rising and increased CO2 (Rathgeber et al. 595 2000; Thuiller 2003). A putative decrease of total and summer rainfall (Fig. 5.17) 596 would favour Aleppo pine’s spreading and colonizing potential at the expense of 597 other more mesic species, such as evergreen oaks (Zavala and Zea 2004). 598 By contrast, a marked decline in growth has been demonstrated by dendro- 599 chronological methods on the edge of distributions, such as very dry coastal areas 600 of south-eastern Spain for Aleppo pine (De Luis et al. 2007) or some Greek Islands 601 for Brutia pine (Körner et al. 2005; Sarris et al. 2007). In these cases, extreme 602 drought will put some populations at risk of extinction. One main aim of future 603 M.R. Chambel et al.

Fig. 5.17 Annual (a) and summer (b) changes in precipitation, expressed in % as simulated by ENSEMBLES regional climate models for the IPCC SRES A1B emission scenario (Source: DMI ensembles-eu.metofﬁce.com, www.nature.com. http://ensemblesrt3.dmi.dk/ENSEMBLE-MEAN_ A1B_GCM_MM_25km-CRU_pr.nc)

604 breeding programmes is to evaluate and select provenances and genotypes adapted 605 to drought and efficient for water and nutrients use. In recent decades, innovative 606 methods and new indices have been introduced to quantify the impact of stress on 607 carbon gas exchanges between plants and the atmosphere and the efficiency of uti- 608 lization of light and water resources (Farquhar et al. 1989; Tognetti et al. 1997; 609 Scarascia-Mugnozza et al. 2011; Table 5.7). Good examples of physiologically- 610 based selection of drought-tolerant genetic materials of Aleppo pine have been 611 made in recent decades in the Negev desert in Israel (Schiller 2011). 612 A less obvious outcome of climate change is the higher risk of frost damage in 613 mountain or continental areas, derived from a lack of hardening and more sudden 614 than normal temperature extremes accompanying global warming (Kozlowski and 615 Pallardy 2002; Fernández et al. 2003). This effect can be more relevant for Aleppo 616 pine, given its lower general tolerance to cold temperatures (Calamassi et al. 2001; 617 Climent et al. 2009). 618 The increase in fire recurrence and virulence as experienced in recent decades 619 over southern France (Fig. 5.18), Spain and Greece has been shown to favour pyro- 620 phites like Aleppo and Brutia pines at the expense of less adapted species such as 621 P. nigra or even P. pinaster (Tapias et al. 2001, 2004). However, even when some 622 degree of fast adaptation through reducing the age or size needed for reproduction 623 can be expected according to recent research (Santos-del-Blanco et al. 2010) where 624 there is a short recurrence interval between forest fires the resilience of these species 625 can be challenged (Rigolot 2004). Table 5.7 Possible relationships between some physiological and growth-related traits and t7.1 drought resistance among Mediterranean and related conifers (Scarascia-Mugnozza et al. 2011) t7.2

Traits Xeric ecotypes Mesic ecotypes References t7.3 Photosynthetic High Low P. taeda: t7.4 capacity Teskey et al. (1986) t7.5 Stomatal High Low P. halepensis: t7.6 conductance Tognetti et al. (1997) t7.7 Stomatal Low High P. halepensis: t7.8 sensitivity Tognetti et al. (1997) t7.9 [AU5] Mesophyll High Low ? t7.10 conductance t7.11 Transpiration rate High Low P. halepensis: t7.12 Tognetti et al. (1997) t7.13 Intrinsic water-use Low High P. pinaster: t7.14 [AU6] efﬁciency Tognetti et al. (2000) t7.15 Hydraulic High Low P. halepensis: t7.16 conductance Tognetti et al. (1997) t7.17 Above ground Slow Fast ? t7.18 juvenile growth t7.19 Above ground adult Fast Slow ? t7.20 growth t7.21 Adult tree shape Vigorous Tall P. pinaster: t7.22 Guehl et al. (1995) t7.23 Carbon isotope High Low P. pinaster: t7.24 discrimination Tognetti et al. (2000) t7.25 Drought adaptive Find water Save water t7.26 strategy and use it t7.27

Fig. 5.18 Number of fires and burnt areas from 1980 to 2002 in France, Greece, Italy, Portugal and Spain (Source: European Commission, Topic Centre for Terrestrial Environment (ETC./TE). Forest fires in Europe – 2002 fire campaign) M.R. Chambel et al.

Fig. 5.19 Experimental heteroblastic grafting of black pine (Pinus nigra subsp. salzmannii) on Aleppo pine rootstocks (left) and Scots pine on Pinus brutia rootstocks at Alaquas and Puerta de Hierro breeding centres, Spain (Ministry of Environment), completely outwith the climatic range of these species (J. Climent)

626 5.6.2 Vegetative Propagation and Somatic Embryogenesis

627 Somatic embryogenesis of both Aleppo pine and Brutia pine has been achieved suc- 628 cessfully over the last decades (Diamantoglou and Banilas 1996; Lambardi et al. 629 1993; Scaltsoyiannes et al. 1994). However, this technique has been applied only at 630 an experimental level, and not for breeding purposes. 631 Operational propagation has been carried out using cuttings and grafting, with a 632 previous emphasis on the multiplication of P. brutia × P. halepensis hybrids (Panetsos 633 et al. 1994). Interestingly, Aleppo and Brutia pines have been used as dwarﬁng 634 rootstocks for enhancing cone production in heteroblastic grafts with scions of other 635 species, like Pinus pinea or even Pinus nigra and P. sylvestris (Climent et al. 1997; 636 Gil et al. 1990; Parra 1980). Both Aleppo and Brutia pine rootstocks induce a dwarﬁng 637 effect and a neat increase of reproductive allocation, and can be specially indicated 638 for ex-situ conservation clonal banks or seed orchards (Fig. 5.19).

639 5.6.3 Knowledge Gaps and Research Needs

640 Aleppo pine provides a very ﬁne example of colonization from eastern to western 641 Mediterranean and, as such, population genetics of this species are still under-inves- 642 tigated. It is surprising how fast adaptive radiation at the western extreme of the 5 Mediterranean Pines (Pinus halepensis Mill. and brutia Ten.) natural distribution range has occurred despite a westward reduction in neutral 643 genetic variation. Separating founder effects from local adaptation processes driven 644 by climate, soil and perturbation regimes (mostly ﬁres) is the object of ongoing 645 research that should be reinforced in the near future. 646 A joint analysis of all extant data from different trial sets would be extremely 647 useful, provided that advanced, suitably powerful statistical tools are used. 648 The knowledge on the vegetative and reproductive phenology of this species and 649 the effects of climate change would be extremely enhanced if the genetic variation 650 (both between and within provenances) and g × e interaction were considered (Girard 651 et al. 2011); there are interesting data sets still under-exploited for this purpose. In 652 all these aspects, integrating the fast progresses of molecular genetics will foster 653 high-quality research with the potential application for any level of genetic and 654 breeding activities. 655

5.6.4 European-Wide Breeding Perspectives 656

A collaborative, joint analysis of the extant field trial networks would form the basis 657 for a putative European-wide breeding programme for Aleppo and Brutia pines. 658 However, the interested countries would be different for the two species. From this 659 review, it seems that breeding of Aleppo pine for wood production would be more 660 rewarding for North African countries than for western European countries, whose 661 wood production is more focused on other species. However, some interest could be 662 found in increasing growth for maximizing biomass production (hence carbon 663 capture) under limiting edapho-climatic conditions. Brutia pine could be a valuable 664 resource for plantation programmes in areas where both moderate drought and 665 frosts are combined limiting factors. Mixed plantations with broadleaves could 666 enhance resilience and maximize carbon fixation in durable wood products for 667 mitigation purposes. 668 Maintaining the adaptive capacity of Mediterranean forests through optimal 669 deployment of the available genetic resources of these species, or enhancing some 670 particular traits through artificial selection, already counts with a wide knowledge 671 base which could be used in a collaborative way through actions such as FAO’s 672 Silva Mediterranea or other support tools. 673

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Queries Details Required Author’s Response AU1 Please provide department name for all the authors. AU2 Thibault (1992), Bariteau et al. (1992), Agúndez (1997, 1999), Schiller et al. (2011) have been changed to Thibault et al. (1992), Bariteau (1992), Agúndez et al. (1997, 1999), Schiller (2011), respectively as per the reference list. Please check. AU3 Please confrm the inserted citation for Fig. 5.11.

2 AU4 CO it should be changed to CO2. Please confrm. AU5 Please provide signifcance for “?” in Table 5.7. AU6 Partitionist Method (2010), Teskey et al. (1986), Tognetti et al. (2000), Guehl et al. (1995) are cited in text but not given in the reference list. Please check. AU7 Please provide complete details in Anonymous (2010), Pichot (1995), Plomion and Pichot (2001).