Acta Silvae et Ligni 121 (2020), 49-58 Izvirni znanstveni članek / Original scientific paper MORPHOLOGICAL TRAITS OF SCOTS PINE (Pinus sylvestris L.) IN INTERNATIONAL PROVENANCE TESTS IN BOSNIA AND HERZEGOVINA MORFOLOŠKE LASTNOSTI RDEČEGA BORA (Pinus sylvestris L.) V MEDNARODNIH PROVENIENČNIH TESTIH V BOSNI IN HERCEGOVINI Mirzeta MEMIŠEVIĆ HODŽIĆ1, Semir BEJTIĆ2, Selma VEJZAGIĆ3, Dalibor BALLIAN4 (1) University of Sarajevo, Faculty of Forestry, Department for Silviculture and Urban Greenery, mirzeta.memisevic.hodzic@gmail.com (2) University of Sarajevo, Faculty of Forestry, semirb33@gmail.com (3) University of Sarajevo, Faculty of Forestry, selma.vejzagic.90@gmail.com (4) University of Sarajevo, Faculty of Forestry, Department for Silviculture and Urban Greenery, d.ballian@sfsa.unsa.ba ABSTRACT Scots pine (Pinus sylvestris L.) is one of the most important tree species in European forests. This study aims to determine whether there is inter-provenance variability in researched morphological traits in two international provenance tests of Scots pine in Bosnia and Herzegovina. We measured height, root collar diameter, and latest shoot length and counted branches on the latest branch whorl of Scots pine plants in two provenance tests. The provenance tests are located in Kupres and Žepče, in different climatic, edaphic, and orographic conditions. Kupres and Žepče contain 15 and 14 provenances, respectively, eleven of which are mutual to both sites. Descriptive statistics and analysis of variance showed differences among provenances in all investigated morphological traits. These differences were attributable to provenance test, provenance, and interaction between provenance test and provenance. The average values were higher in Žepče for all provenances and all studied traits. The Austria A1, Austria A2, Austria A3, and Poland P1 provenances showed the best growth in both tests, while the Italy I1 provenance showed good growth in Žepče but not in Kupres. Key words: Scots pine, provenance tests, morphological traits IZVLEČEK Rdeči bor (Pinus sylvestris L.) je ena izmed najpomembnejših drevesnih vrst v Evropi. Pričujoča raziskava ugotavlja, ali med proveniencami, uporabljenimi v dveh provenienčnih testih rdečega bora v Bosni in Hercegovini, obstaja variabilnost preučevanih morfoloških lastnosti. Izmerili smo višino dreves, premer koreninskega vratu in dolžino terminalnega poganjka ter prešteli veje na najmlajšem vencu vej v dveh provenienčnih testih. Provenienčna testa sta locirana v Kupresu (15 provenienc) in Žepčah (14 provenienc) v različnih podnebnih, edafskih in orografskih razmerah. Obema provinenčnima testoma je skupnih 11 provenienc. Deskriptivna statistika in analiza variance kažeta na razlike med proveniencami pri vseh preučevanih morfoloških lastnostih, ki jih pripisujemo provenienčnim testom, proveniencam in interakciji med provenienčnim testom in provenienco. Povprečne vrednosti so bile za vse provenience in vse preučevane lastnosti višje v Žepčah. Provenience Avstrija A1, Avstrija A2, Avstrija A3 in Poljska P1 so v obeh testih izkazale najboljšo rast, medtem ko je bila izmerjena rast Italije I1 dobra v provenienčnem testu v Žepčah, ne pa tudi v Kupresu. Ključne besede: rdeči bor, provenienčni testi, morfološke značilnosti GDK 164+174.7Pinus sylvestris L.(497.6)(045)=111 DOI 10.20315/ASetL.121.4 Licenced CC BY-SA 4.0 1 INTRODUCTION 1 UVOD Scots pine (Pinus sylvestris L.) is one of the most important tree species in European forests (Matyas et al., 2004) and is characterized by high genetic variability and complex population structure. In the Balkans, Scots pine is in a succession species, while populations of Scots pine in the Mediterranean region and Central Europe represent relict communities from the Pleistocene era (Mirov, 1967). Scots pine has a wide Prispelo / Received: 15. 11. 2019 Sprejeto / Accepted: 7. 2. 2020 geographical distribution - from the Iberian Peninsula and Scotland to the Far East and from Scandinavia to Asia Minor (Boratinsky, 1991). Therefore, Scots pine is one of the most studied forest tree species in Europe (Giertych and Matyas (eds.), 1991). The first international experiment, a progeny test, was established in 1907 by the International Union of Forest Research Organizations (IUFRO). Scots pine has been the subject of numerous morphological studies in Europe (Alia et al., 2001; Ballian et al., 2009; Cvjetković et al., 2014; Barzdajn et al., 2016; Ballian and Šito, 2017; Gülcü and Bilir, 2017; Ballian and Lizdo, 2019). The territory of Bosnia and Herzegovina represents the southern border of the natural distribution of Scots pine. Provenance tests in Bosnia and Herzegovina, therefore, represent an opportunity to determine the adaptability and variability of different provenances of Scots pine from all over Europe, at the southern border of its natural distribution. Thus, two international provenance tests were established in Bosnia Herzegovina (Kupres and Žepče) in 2012. The study aims to determine whether there is inter-provenance variability in researched morphological traits in the Kupres and Žepče provenance tests in Bosnia and Herzegovina. The results of this study will enable the selection of Scots pine provenances with the best growth and adaptability and thus contribute to the in-situ and ex-situ conservation of the species. 2 MATERIAL AND METHODS 2 MATERIALI IN METODE In this study, we measured height, root collar diameter, and latest shoot length and counted the branches on the latest branch whorl on eight-year-old Scots pine Table 1: List of investigated provenances in the Kupres and Žepče provenance tests plants in two international provenance tests in Bosnia and Herzegovina. Height was measured to an accuracy of one cm using a wooden measuring stick. Root collar diameter was measured to an accuracy of 0.1 mm using a digital caliper. Descriptive analysis, analysis of variance (one- and two-way ANOVA), and Duncan's multiple interval test were performed for different morphological traits using IBM SPSS STATISTICS 20.0. The provenance tests were established in 2012 by planting two-year-old seedlings on two sites (Kupres and Žepče) with different climatic, edaphic, and oro-graphic conditions. The Kupres and Žepče provenance tests contain 15 and 14 provenances, respectively, from nine countries (Table 1, Figure 1). Seedlings were planted in holes measuring 30x30x30cm, with a planting distance of 2 x2m. In Kupres, 36 seedlings in five repetitions per randomized scheme were planted for each provenance (2700 seedlings in total), and in Žepče, 25 seedlings were planted in four repetitions (1400 seedlings in total). The provenances originate from areas with different climates. Provenances from Austria, Slovakia, Poland, and Romania are from a temperate continental climate and those from Bosnia and Herzegovina from a Preglednica 1: Seznam preučevanih provenienc v provenienčnih testih v Kupresu in Žepčah Provenance Test(s) Country Provenance Label Locality Latitude Longitude Kupres Austria A1 Kobersdorf/Lackenbach 47°53'12« 15°31'39« Zepče Kupres Austria A2 Panholz 47°07'14« 15°17'14« Zepče Kupres Austria A3 Lans/Tirol 47°13'49« 11°26'12« Zepče Kupres Bosnia & Herzegovina B1 Bugojno 44°03'00« 17°27'00« Zepče Kupres Germany NJ2 Trippstadt 49°21'35« 7°46' 29« Zepče Kupres Italy I1 Ca Del Lupo 44°45'25« 9°05'07« Zepče Kupres Italy I2 Fenestrelle (TO) 45°01'47« 7°03'38« Zepče Kupres Italy I3 Valda (TN) 46°13'00« 11°16'00« Zepče Kupres Poland P1 Ruciane - Nida 53°37'00« 21°29'00« Zepče Kupres Romania R1 Sacueni 47°21'09« 22°05'29« Zepče Kupres Slovakia SL1 Hanušovce 49°01'35« 21°30'01« Zepče Kupres Germany NJ1 Teisendorf 47°51'00« 12°49'00« Kupres Norway N1 Malvik 63°22' 22« 10°45'03« Kupres Norway N2 Arnes 60°07'20« 11°27'55« Kupres Ukraine U1 Delyatyn 48°32'41« 24°30'10« Zepče Norway N3 Narvik 68°25'14« 17°33'10« Zepče Scotland S1 Shieldaig 57°36'24« 5°55'17« Zepče Ukraine U2 Ivano Frankivsk 48°56'22« 24°31'52« Legend A uši ri a KoberstV irfLacktnbuch Austria Ponholz Austria Lan:;, fircl tJosma Ä Herzegovina Bugojro 1гл]у D?! I lipo liiiiy Fe nostra lie 0*01 riflly Valdft (TKi Ncmvay Malvik Norway Ames JJcrway Nan it Germany Teisjindorl' Germany Iripiwiadt Poiirtd RuctW Nida Romania 5ocucni Scotland Shiiridaig Slnvdkia HnitiJ^rtvcc T rkr^tbilii- DeLyfityn Ukraine Ivano Fnuifcjvsk Fig. 1: Map of the spatial distribution of the studied provenances subalpine temperate continental climate. Provenances from Italy are from a continental climate and those from Germany from a moderately warm and humid climate influenced by the Atlantic Ocean. Provenances from Ukraine are from a continental climate, but from one of the coldest regions in Ukraine with an average daily maximum temperature of only 12 °C and frequent rainfall and wind. The Shieldaig provenance from Scotland is from an oceanic climate. The Kupres provenance test is located at an altitude of 1140 m, on terrain characterized by sinkholes and numerous hills and valleys. The soil type is calcocam-bisol or brown soil, and the geological substrates are limestones and dolomites. According to the classification of climate types in BiH (Milosavljević, 1973), this area belongs to the mountain climate, with short summers and long, cold, and very snowy winters. The mean annual air temperature is 6.2 °C. The minimum temperature was measured in January (-26.8 °C) and the maximum in September (34.9 °C). The annual average of registered frosts is 155 days. The average annual precipitation is 1221 mm, and its distribution varies slightly by season. The Žepče provenance test is located at an altitude of 600 m in the oak and beech forest belt. The soil type is dystric cambisol. The climate (Milosavljević, 1973) in this area is temperate continental, with warm sum- Slika 1: Zemljevid prostorske razporeditve preučevanih provenienc mers and harsh winters. The average annual air temperature is 10.7 °C, with the minimum temperature measured in January (-27.5 °C) and the maximum in August (39.5 °C). The annual average of registered frosts is 85 days. The average annual precipitation is 1040 mm. 3 RESULTS 3 REZULTATI 3.1 Plant height 3.1 Višina rastlin Average plant height per provenance and provenance test is presented in Figure 2. The average height of all measured plants in Kupres and Žepče was 181.0 cm and 294.9 cm, respectively. Average height was higher in Žepče for all provenances. The Italy I1 provenance in Žepče had the highest average height. The analysis of variance for height (Table 2) showed a statistically significant difference between provenances in both tests, with a probability of 95 %. Duncan's multiple interval test showed grouping of provenances in Kupres and Žepče into seven and eight overlapping groups, respectively. Two-way analysis of variance (Table 3) showed statistically significant differences for provenance test (Sig. = 0.000), provenance (Sig. = 0.000), and provenance test x provenance interaction (Sig. = 0.000). \vtrage litlevl sluwt length (cm) per provenance ami provmanc* tfsl W ца Ml j.i ■ Kill*?* ei.tot* Fig. 2: Average height per provenance and provenance test Slika 2: Povprečna višina glede na provenienco in provenienčni test Table 2: Analysis of variance for height Preglednica 2: Analiza variance višine Source of Variation Sum of Squares Df Mean Squares F Sig. Duncan's Test Number of Groups Kupres Provenance Test Between Groups 1961118.082 14 140079.863 60.743 0.000 7 Within Groups 3701289.841 1605 2306.100 Total 5662407.923 1619 Žepče Provenance Test Between Groups 878310.996 13 67562,384 10.434 0.000 8 Within Groups 4668816.369 721 6475.473 Total 5547127.366 734 Table 3: Two-way analysis of variance for height Preglednica 3: Dvofaktorska analiza variance višine Source Type III Sum of Squares Df Mean Square F Sig. Partial Eta Squared Provenance Test 5084878.343 1 5084878.343 1393.634 0.000 0.438 Provenance 802327.846 10 80232.785 21.990 0.000 0.110 Provenance Test * Provenance 791165.236 10 79116.524 21.684 0.000 0.108 Table 4: Analysis of variance for root collar diameter Preglednica 4: Analiza variance premera koreninskega vrat Source of Variation Sum of Squares Df Mean Squares F Sig. Duncan's Test Number of Groups Kupres Provenance Test Between Groups 155866.958 14 11133.354 51.175 0.000 7 Within groups 349177.103 1605 217.556 Total 505044.061 1619 Žepče Provenance Test Between Groups 46328.465 13 3563.728 9.166 0.000 6 Within groups 280328.020 721 388.804 Total 326656.484 734 Average root collar diameter (mm) per provenance aud provenance test 0 ^ & ^ ^ ^ ^ ^ ^ ^ ^ ^ c>> ^ ^ I eS" «У «У Л?' tP cP //////7// I Kupres ■ Zepče Fig. 3: Average root collar diameter per provenance and pro- Slika 3: Povprečen premer koreninskega vratu glede na venance test provenienco in provenienčni test Table 5: Two-way analysis of variance for root collar diame- Preglednica 5: Dvofaktorska analiza variance premera koter reninskega vratu Source Type III Sum of Squares Df Mean Square F Sig. Partial Eta Squared Provenance Test 105152.836 1 105152.836 377.244 0.000 0.174 Provenance 54523.937 10 5452.394 19.561 0.000 0.099 Provenance Test * Provenance 40859.133 10 4085.913 14.659 0.000 0.076 3.2 Root collar diameter 3.2 Premer koreninskega vratu Average root collar diameter per provenance and provenance test is presented in Figure 3. Average root collar diameter was 51.6 mm in Kupres and 67.9 mm in Žepče. Average root collar diameter was larger in Žepče compared to Kupres for all provenances. The Italy I1 provenance in Žepče had the largest average root collar diameter. The analysis of variance for root collar diameter (Table 4) showed a highly statistically significant difference between provenances in both provenance tests, with a probability of 95 %. Duncan's multiple interval test showed grouping of provenances in Kupres and Žepče into seven and six overlapping groups, respectively. Two-way analysis of variance (Table 5) showed statistically significant differences for provenance tests (Sig. = Average root coLLar diameter (mm) per provenance and provenance test 32 ES 72 62 52 42 32 22 10 2 # J* -Č» ^ * * * * ^ A** ^ .„■ ... ^ y J- ^ # I Kupres BZepie Fig. 4: Average latest shoot length per provenance and provenance test Slika 4: Povprečna dolžina terminalnega poganjka glede na provenienco in provenienčni test 0.000), provenances (Sig. = 0.000), and provenance test x provenance interaction (Sig. = 0.000). 3.3 Latest shoot length 3.3 Dolžina terminalnega poganjka Average latest shoot length per provenance and provenance test is presented in Figure 4. Average latest shoot length for all plants was 42.3 cm in Kupres and 66.8 cm in Žepče. Similar to height and root collar diameter, average latest shoot length was higher in Žepče compared to Kupres for all provenances. The Italy I1 provenance in Žepče had the highest average latest shoot length. The analysis of variance for latest shoot length (Table 6) showed a highly statistically significant difference between provenances in both provenance tests, with a probability of 95 %. Duncan's test for latest shoot length showed grouping of provenances in Kupres and Žepče into eight and five overlapping groups, respectively. Two-way analysis of variance (Table 7) showed statistically significant difference by provenance tests (Sig. = 0.000), provenances (Sig. = 0.000), and provenance test x provenance interaction (Sig. = 0.000). 3.4 Number of branches on the latest branch whorl 3.4 Število vej na najmlajšem vencu vej The average number of branches on the latest branch whorl is presented in Figure 5. The average number of branches on the latest whorl was six in Table 6: Analysis of variance for latest shoot length Table 7: Two-way analysis of variance for latest shoot length Kupres and eight in Žepče. The average number of branches on the latest branch whorl was higher in Žepče for all provenances. The analysis of variance for the number of branches on the latest whorl (Table 8) showed a highly statistically significant difference between provenances in Kupres and a statistically significant difference between provenances in Žepče, with a probability of 95 %. Duncan's multiple interval test showed a grouping of provenances in Kupres and Žepče into five and four overlapping groups, respectively. Two-way analysis of variance (Table 9) showed statistically significant differences for provenance tests (Sig. = 0.000), provenances (Sig. = 0.008), and provenance test x provenance interaction (Sig. = 0.000). 4 DISCUSSION 4 RAZPRAVA Scots pine has been the subject of numerous studies in Europe, both morphological and genetic, and there is a need for continued research, as the species is of very high value in modern forestry (Pintarić, 2002). Although molecular methods are dominant in population studies, morphometric methods still play a significant role in illustrating patterns of intra-provenance and inter-provenance variability (Zebec et al., 2010; Brus et al., 2011; Galvan et al., 2012; Jasinska et al., 2012; Paridari et al., 2013; Poljak et al., 2014; Zebec et al., 2014; Popović and Kerkez, 2016). This study included 15 provenances in the Kupres Preglednica 6: Analiza variance dolžine terminalnega poganjka Preglednica 7: Dvofaktorska analiza variance dolžine terminalnega poganjka Source Type III Sum of Squares Df Mean Square F Sig. Partial Eta Squared Provenance Test 229601.577 1 229601.577 883.389 0.000 0.331 Provenance 69108.271 10 6910.827 26.589 0.000 0.130 Provenance Test * Provenance 40897.904 10 4089.790 15.735 0.000 0.081 Source of Variation Sum of Squares Df Mean Squares F Sig. Duncan's Test Number of Groups Kupres Provenance Test Between Groups 146373.477 14 10455.248 48.283 0.000 Within Groups 347328.328 1604 216.539 8 Total 493701.805 1618 Žepče Provenance Test Between Groups 43040.534 13 3310.810 9.371 0.000 Within Groups 254734.032 721 353.307 5 Total 297774.566 734 \vwigt iniiuhrrnf hriini'he« (in Ihi1 bloi lniimh wlmM j i L~k' ] ll'4> V L"ll il hC L1 J It [I |H4Vi'h:LJU'c list 5 3 6 i 4 j 1 1 p H _ i> ЈУ č ,<У O J.4 rfv s> J> Л /VV