VARIABILIT Y OF T HE SWE ET CHESTNUT ( Castanea sativa Mill.

) BASED ON T HE MORPHOL OGICAL PROPERT IE S OF THE NUT AND CUPUL E IN NATURA L POPUL ATIONS OF BOSANSKA KR AJINA Aida Mujagi-Pai , Dalibor B allian
1

Faculty of Biotechnology of the University of Biha, Bosnia and Herzegovina (aida.mujagic@gmail.com) 2 Faculty of Forestry of the University of Sarajevo, Bosnia and Herzegovina

Abstract The purpose of this study is to establish an intra-population and inter-population variability of the sweet chestnut (Castanea sativa Mill.) based on the morphological properties of the nut and cupule within the natural populations in the region of Bosanska Krajina (Bosnia and Herzegovina). The material has been collected from 16 subpopulations in the region of Bosanska Krajina (Cazin, Buim and Velika Kladua). A total of 3,631 nuts and 1,315 cupules have been collected, and the following variables have been analysed: nut length, nut width, nut thickness, hilum width, hilum length, nut mass, cupule width and cupule height, calculated nut shape, relative hilum size and cupule size, nut shape, nut embryony, penetration of the pericarp into the cotyledon, number of nuts in a cupule and length of spines in a cupule. An analysis of the morphological properties of the nuts and cupules of the sweet chestnut (Castanea sativa Mill.) has established an intra-population and interpopulation variability of the sweet chestnut. Key words: sweet chestnut, Bosanska Krajina, nut, cupule, variability

Introduction The Castanea (Fagaceae) genus features seven species which are mainly distributed in temperate regions of the Northern Hemisphere, more specifically in the south-eastern parts of North America, Southern Europe, Northeast Africa and Western and Eastern Asia (Fernndez-Lpez and Alia, 2003). The priority topics which will have to be addressed in the country in the future, when this species is in question, are the integrated and coordinated taxonomic approach to chestnut forests at European level, the impact of the conditions and different management systems on the growth, selection and conservation of the sweet chestnut, optimal methods of pest and disease control, and the use of the species within the climate change context (Mujagi-Pai and Ballian, 2012).

Material and methods A total of 16 subpopulations and 140 sweet chestnut trees were selected among the natural populations of the sweet chestnut in Bosanska Krajina (Cazin, Buim and Velika Kladua) in 2009. The selection of subpopulations made sure that scientifically interesting, minor and local subpopulations are equally represented. A total of 3,631 nuts and 1,315 cupules were collected. The measured variables of the nut and cupule were: nut length in mm (DPL), nut width in mm (PL), nut thickness in mm (DEP), hilum width in mm (H), hilum length in mm (DH), nut mass in g (MP), cupule width in mm (K) and cupule height in mm (VK). The following relations were calculated on the basis of known values according to Serdar et al. (2011): calculated nut shape in mm (DP/P), relative hilum size in mm (HxDH) and cupule size in mm (KxVK), the derived properties: nut shape (OP), nut embryony (EP), penetration of the pericarp into the cotyledon (PSO), number of nuts in a cupule (BPL) and length of spines in a cupule (DI) were determined according to the descriptor for the sweet chestnut (UPOV 1989). The morphological data from the measurements was processed statistically by means of the XLSTAT 2011 and STATISTICA 10 packages.
Table 2. Eigenvalues and cumulative variability in the first five PC factors

Results and discussion An analysis of the morphological properties of the nut and cupule of sweet chestnut in the region of Bosanska Krajina within its natural populations was used to establish with statistical methods the interaction between the genotype and environment and to raise awareness about the need to preserve the natural populations of the sweet chestnut, because they represent the main source of the variability of genetic material of this species. There are several studies (Aravanopoulos et al., 2005, Pandit et al., 2009, Zarafshar et al., 2010) which use different morphological analyses to estimate the variability of sweet chestnut populations and which confirm that the parameters of the nut, leaf and cupule can be appropriate variables for establishing the level of phenotype variability among natural populations of the sweet chestnut. An analysis of variance for the studied properties of the nut and cupule imply the existence of statistically significant differences (Sig. <0.0001) among the groups, which shows a pronounced inter-population variability of the studied properties in comparison to intra-population variability.

The results of a principal component analysis (PCA) of the morphological properties of the sweet chestnut nut and cupule show that the cumulative variability in the first three PC factors amounts to 81.95 %, with the expected biggest participation of the first PC1 factor of 59.64 %, while the PC2 and PC3 factors contribute a lot less to total variability 13.27 % and 9.029 %, respectively (Table 2).

An agglomerative hierarchical clustering (AHC) analysis was used to detect possible similarities and differences between the 16 studied subpopulations of the sweet chestnut in terms of morphological properties of the nut and cupule. The topology of the dendrogram clearly shows the existence of four main groups of the studied subpopulations relating to the 16 studied properties of the nut and cupule (Picture 1).

Picture 2. Principal component methods (PC1 vs. PC2) for the morphological properties of the nut and cupule

The PCA (Picture 2) confirms the conclusion resulting from the cluster analysis, in which four distinctive groups have been established based on the pomological properties of the sweet chestnut

Picture 1. Dendrogram of grouping of studied subpopulations by morphological properties of the sweet chestnut (Castanea sativa Mill.)

The first group features four subpopulations with the average mean values of the studied properties. The second group features three subpopulations with distinctly small nuts and the smallest number of nuts in a cupule. The third group features five subpopulations with the average mean values of the studied properties of the nut and cupule and different geographical origin, while the fourth group features subpopulations originating from the same geographical area (Cazin) and having the biggest mean values for a majority of the studied properties of the nut. Genotypes from these subpopulations can serve as a good basis for the selection, plant breeding and preservation of the genetic diversity, and in the differentiation of the lower taxa of the Castanea genus, because they have displayed the best pomological properties in comparison to the genotypes of other subpopulations.

Conclusion The results point to the existence of variability within and among the studied subpopulations, which means that the studied morphological properties of the nut and cupule can be appropriate variables for determining phenotype variability in sweet chestnut populations. In the coming period, the results should be confirmed at the molecular and genetic level.
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