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On the Taxonomical Identity of Abies Alba Ssp. Borisii- Regis (Mattf.) Koz

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On the Taxonomical Identity of Abies Alba Ssp. Borisii- Regis (Mattf.) Koz 34 (2): 127-132 (2010) Original Scientifi c Paper On the taxonomical identity of Abies alba ssp. borisii- regis (Mattf.) Koz. et Andr.– morphometry, fl avonoids and chorology in Bulgaria Malina Delcheva✳, Dimitar Peev and Milena Nikolova Institute of Botany, Bulgarian Academy of Sciences, 23 Acad. G. Bonchev St. bl. 23, 1113, Sofi a, Bulgaria ABSTRACT: Th irty four samples of Abies alba ssp. borisii-regis (Mattf.) Koz. et Andr, Abies alba Miller. and Abies cephalonica Loudon collected from Rila, Pirin, Western Rhodopes, Slavyanka, Black Sea coast and Greece have been subjected to variation and cluster analyses. Abies alba ssp. borisii-regis is mentioned in Bulgarian Red list under LC category. In the same time this species is a major species to the endemic habitat 9270 Hellenic beech forests with Abies borisii-regis distributed in Southern Bulgarian mountains and some other mountains of Balkan peninsula. Th e obtained results support the idea about intermediate character of Abies alba ssp. borisii-regis (between A. alba and A. cephalonica) and corresponding species rank - Abies borisii-regis. Key words: cluster analysis, Abies alba ssp. borisii-regis, Abies alba, Abies cephalonica, variation coeffi cient, fl avonoids. Received 12 October 2009 Revision accepted 18 June 2010 UDK 582.475(497.2) ; 519.237.8 INTRODUCTION MATERIALS AND METHODS Th e taxon A bies alba ssp. borisii-regis (Mattf.) Koz. et Andr. Plant material. Th irty four individual representative is given as subspecies in Bulgarian fl ora. Its distribution samples of Abies alba Mill. (16 samples), A. alba ssp. in Bulgaria is in the mountains in the South part of the borisii-regis (Mattf.) Koz. et Andr. (14 samples) and A. country (Slavyanka, Belasitsa, Southern Pirin, Middle and cephalonica Loudon. (3 samples) have been subjected to Western Rhodopes, Southwestern Rila) in area of elevation Variation and Cluster analyses. Th e origin of the samples between 900 and 1800 m. (Andreev 1992). Th e subspecies was from 5 fl oristic regions of Bulgaria: Rila, Pirin, Western take part in the threatened habitat of 9270 Hellenic beech Rhodopes, Slavyanka, Black Sea coast (north) and one forests with Abies borisii-regis. It is included in the Red sample from Greece. (Table 1) List in Bulgaria under the category of Least Concern (LC). (Petrova & Vladimirov 2009). In Flora Europaea the Variation and cluster analysis. Four m orphometric species rank status was adopted – A. borisii-regis Mattf. features were analyzed as follows: х1 – diameter of the It is distributed in the Balkan Peninsula from Southern circular-shaped base of the needle, х2 – needle length from Albania to the Th race region. Th e species has many varying the base, х3 – maximum needle width, х4 – thickness in the forms, b ut always more or less intermediate between A. middle of the needle. Th e morphometric features х1, х2 and alba Miller and A. cephalonica Loudon. It is most likely х3 were measured with accuracy of up to 0.05 mm and the of hybrid origin (Chater 1964). Our goal is to verify feature х4 - 0,025. Hundred needles of each sample have the statistical isolation and partially the phytochemical been measured. Th e degree of variation was measured isolation of Abies alba and Abies alba ssp. borisii-regis. according to the variation coeffi cient scale (Peev 1985): ✳correspondence: [email protected] © 2010 Institute of Botany and Botanical Garden Jevremovac, Belgrade 128 vol. 34 (2) Table 1. Origin and taxonomy of the analyzed samples Sample Floristic Date of m above sea level; Taxon Location (№) regions sampling longitude and latitude 1 A. alba ssp. borisii-regis Rila Hotel ‘Alen Mak’ 02.04. 2009 1716 m; 42o 12,718’ N/23 o 23,259’ E 2 A. alba ssp. borisii-regis Rila Hotel ‘Malyovitsa’ 02.04. 2009 1709 m; 42o 12,659’ N/ 23 o 23,266’ E 3 A. alba ssp. borisii-regis Rila Hotel ‘Malyovitsa’ 02.04. 2009 1709 m; 42o 12,659’ N/ 23 o 23,266’ E 4 A. alba ssp. borisii-regis Rila hut ‘Malyovitsa’ 02.04. 2009 1906 m; 42o 11,683’ N/ 23 o 22,732’ E 5 A. alba Rila Mecha polyana 02.04. 2009 1733 m; 42o 12,466’ N/ 23 o 23,250’ E By Yadenitsa river, at 15 km Western 6 A. alba ssp. borisii-regis northern from Golyamo 06.04. 2009 1129 m; 42,10493 o N/ 23,90043 o E Rhodopes Belovo By Yadenitsa river, at 15 km Western 7 A. alba ssp. borisii-regis northern from Golyamo 06.04. 2009 1129 m; 42,10493 o N/ 23,90043 o E Rhodopes Belovo 8 A. alba Rila Hotel ‘Alen Mak’ 02.04. 2009 1716 m; 42o 12,718’ N/ 23 o 23,259’ E 9 A. alba Rila Hotel ‘Alen Mak’ 02.04.2009 1716 m; 42o 12,718’ N/ 23 o 23,259’ E Western 10 A. alba By Yadenitsa river 06.04. 2009 1129 m; 42,10493 o N/ 23,90043 o E Rhodopes Western 11 A. alba By Yadenitsa river 06.04. 2009 1129 m; 42,10493 o N/ 23,90043 o E Rhodopes 12 A. cephalonica Greece Town of Th e tree planted in the Town 13 A. cephalonica 12.06.2009 596 m; 42о 28,686’ N/ 23 о 44,831’ E Zemen Hall yard 14 A. alba ssp. borisii-regis Slavyanka Hambar dere 1.07.2009 1000 m Western 15 A. alba Direction to Sarnitsa Town 16.05.2009 1632 m; 41,89093 о N/ 23,93705 о E Rhodopes Western 16 A. alba By Yadenitsa river 16.05.2009 1129 m; 42,10493 о N/ 23,90043 о E Rhodopes Western At 10 km from Velingrad 17 A. alba 16.05.2009 988 m; 42,03136 о N/ 23,90343 о E Rhodopes Town, to Yundola By the road between the Western 18 A. alba Chairite Dam and the 21.06.2009 1050 m; 42о 11,215’ N/23 о 52,952’ E Rhodopes Sestrimo village Western 19 A. alba Krastava place 16.05.2009 1023 m; 41,93547о N/ 23,92017о E Rhodopes Western 20 A. alba Sofan dere 16.05.2009 1056 m; 41,93153о N/ 23,92623о E Rhodopes Western 5 km before Yundola on the 21 A. alba 19.06.2009 1270 m; 42о 05,442’ N/ 23о 53,302’ E Rhodopes road from Belovo Western 22 A. alba Grashevo Village 16.05.2009 1295 m; 42,08445о N/ 23,88552о E Rhodopes 23 A. alba Pirin hut ‘Gotse Deltchev’ 21.06.2009 1479 m; 41о 45,608’ N/ 23 о 32,814’ E Above Papaz Chayir, near 24 A. alba South Pirin 6.07.2009 1524 m; 41о 33,101’ N/ 23 о 37,722’ E Orelek peak 25 A. alba South Pirin Popovi livadi 18.07.2009 1432 m; 41,33152о N/ 23,38138о E Malina Delcheva et al.: On the taxonomical identity of Abies alba ssp. borisii-regis (Mattf.) Koz. et Andr.– morphometry, fl avonoids and chorology in Bulgaria 129 Sample Floristic Date of m above sea level; Taxon Location (№) regions sampling longitude and latitude Northern Specimen planted in the 26 A. cephalonica Black Sea University Botanic Garden in 24.07.2009 13 m; 43о 24,251’ N/ 28 о 8,917 E coast town of Balchik On the trekking course 27 A. alba ssp. borisii-regis Rila between the Rila Monastery 17.07.2009 1544 m; 42,07614 оN/ 23,19240 о E and hut ‘Iv.Vazov’. On the trekking course 28 A. alba Rila between the Rila Monastery 17.07.2009 1307 m; 42,07283 о N/ 23,19747 о E and Hut ‘Iv.Vazov’. 9 km off village of Pastra, on 29 A. alba ssp. borisii-regis Rila 1.08.2009 1616 m; 42о 09,489 ‚N/ 23 о 13,579’ E the way to Kalin Dam. 9 km off village of Pastra, on 30 A. alba ssp. borisii-regis Rila 1.08.2009 1616 m; 42о 09,489 ‚N/ 23 о 13,579’ E the way to Kalin Dam. 3 km off Kirilova polyana, on 31 A. alba ssp. borisii-regis Rila 1.08.2009 1527 m; 42о 09,334 ‚N/ 23 о 24,582’ E the way to Ribni lakes. 32 A. alba ssp. borisii-regis Rila Kirilova polyana 1.08.2009 1475 m; 42о 09,301 ‚N/ 23 о 24,007’ E 33 A. alba ssp. borisii-regis Rila Kravarsko dere 1.08.2009 1587 m; 42о 16,371 ‚N/ 23 о 19,994’ E 34 A. alba ssp. borisii-regis Slavyanka Above Parilska saddle 6.08.2009 1625 m; 41о24’ 37,3’’N/23о 38’ 54,3’’E Znepole At 3 km Eastern from Tran 35 A. cephalonica 5.01.2005 800 m; 42.83479 о N/22.69361 о E Region Town V<7% - very low, 7,1<V<12% - low, 12,1<V<20% - medium, TLC analysis of fl avonoid components. Th e 20,1<V<40% - high, V>40,1% - very high. Th e obtained identifi cation of fl avonoid aglycones was determined by data were subject to cluster analysis (Nearest Neighbor co-chromatography with reference compounds. Two TLC Method, Euclidean distance), whereby the clusters were systems confi rmed the identifi cation of the fl avonoids. formed by calculating the degree of similarity, based upon System 1: silica gel, toluene–dioxane–acetic acid 95 : 25 : 4 the average values. v/v/v; 2: cellulose plates, acetic acid–water 60–30 v/v. Flavonoid analysis. RESULTS AND DISCUSSION Preparation of plant extracts. Ground plant material (1g) was extracted twice with 30 ml 80% metanol. Variation analysis Th e extract was then evaporated and transferred to a Morphometric features.
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