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2. Dirr M. A and Heuser C. W. 2006. The Reference Manual of Woody Propagation. Timber Press, pp.43-45. 3. Fira Alexandru, Doina Clapa. 2009. Ex-Vitro Acclimation of some Horticultural in Hydroculture. Bulletin UASVM, nr. 66 (1), Horticulture, Print ISSN 1843-5254; Electronic ISSN 1843-5394, pp 44-51. 4. Fira Al, D. Clapa, C. Plopa. 2010. New Aspects Regarding the Micropropagation of Blackberry Cultivar ‘Thornless ’. Bulletin UASVM Horticulture, 67(1)/2010, Print ISSN 1843-5254; Electronic ISSN 1843-5394. 5. B. A. Sheikh, 2006, Hydroponics: Key to Sustain Agriculture in Water Stressed and Urban Environment, Pak. J. Agri., Agril. Engg., Vet. Sc. 22 (2). 6. Tyson, R. V., White J. M and King K. V. 1999. Outdoor Floating Hydroponic Systems for Leafy Salad crop and Herb Production. Proc. Fla. State. Hort. Soc. 112:313-315. 7. Zaharia, D., 1994, Floricultura, Tipo Agronomia Cluj-Napoca, pp82-88. 8. A. H. S. 1999. Plant Propagation. DK Publishing, Inc.

FLOWER TRAITS VARIABILITY OF DATE PLUM ( LOTUS L.) GENOTYPES GROWN IN SLOVAKIA FROM SEEDS Olga Grygorieva1 – Ján Brindza2 – Dezider Tóth2 – Vlasta Abrahám2 – Radovan Ostrovský2 1M. M. Grishko National Botanical Garden of Ukraine National Academy of Sciences, Kiev,Ukraine 2Institute of Conservation and Biosafety, Slovak University of Agriculture, Nitra, Slovak Republic

Summary. This work aimed at the variability determination of date plum (Diospyros lotus L.) male and female fl owers. In the experimental collection were included genotypes of date plum grown from seeds in Arboretum Mlynany (Slovakia). During the bloom stage fl owers were withdrawn from the tested genotypes. The male and female fowers were evaluated separately. Morphological traits determination was based on image analysis. rom these analyses resulted data for the male/female fl owers as follows: number of sepals 3-5/3-7, sepal height 1.97/5.45 (mm), sepal width 2.13/4.98 (mm), crown height 6.00/6.47 (mm), crown diameter 4.21/5.33 (mm), number of crown sepals 2-6/3-7, number of stamens in the 1st row of male fl owers 6-9/7-9 and number of stamens in the 2nd row of male fl owers 6-9/0. In the evaluation process dealing with the tested traits dependences has been found a partial agreement among the correlation coeffi cients of the male and female fl owers only in the cases of calyx sepals height and crown height (0.476 – 0.481), calyx sepals height and stamen length in the 1st row (0.137 – 0.152) and crown height and stamen length in the 1st row (0.360 – 0.301), respectively. The rest of traits the when compared bilaterally, exerted different values of correlation coeffi cients. It means, that the dependencies between the male and female fl owers 34 are variable. For male fl owers between the calyx sepals height (mm) and the crown diameter (-0.285) as well as between the crown height (mm) and crown diameter (-0.400) were statistically detected signifi cant negative correlations.

Introduction Natural copses of date plum (Diospyros lotus L.) are widely occurring in Japan, China, India and Iran. In cultural form this species is spread in Korea, Pakistan, Afghanistan, Turkey, Albania, Spain, France and Poland (Kuliyeva, 1962). Date plum is a typical dioecious with . In natural conditions these achieve the height up to 5 m (Ayaz and Kadioglu, 1999). Date plum are of globular form with a diameter around 8-16 mm (Brezhnev and Korovina, 1981). Generally interesting by this species is the biology of blooming and the fl owers morphological structure. There are joined together 1-3 staminate fl owers, pedicel lenght is up to 6 mm, number of calyx lobes 4 or 5, corolla is reddish to pale yellow, urceolate, ca. 4 mm; number of corolla lobes 4 and stamens 16. Pistillate fl owers are subsessile, pale green to reddish; they have 4 calyx lobes, corolla urceolate, ca. 6 mm, corolla lobes 4 or rarely 5, staminodes 8, ovary 8-locular, glabrous except for apex, styles 4. Fruiting calyx lobes 4, ovate, apex obtuse. Berry pale yellow, becoming bluish black with a glaucous bloom, subglobose to ellipsoid, 1-2 cm in diameter. Seeds are brown, compressed, approximately 10 × 6 mm in diameter (Shu-kang et al., 1996). Female fl owers subsessile, solitary on short, pubescent, bracteate pedicels 2-3 mm. long; calyces green, 7-8 mm. long, densely rufous- pubescent on the adaxial surface of the calyx tube, accrescent in and sometimes persistent on the branchlets, the 4 or 5 lobes foliaceous, 6-7 mm. long at anthesis; corolla reddish-brown, broadly urceolate, ca. 5 mm. long, the lobes 2-3 mm. long, ± recurved; staminodia 8, curved over the surface of the ovary, pubescent with long, silvery hairs; ovary ± globose, glabrous or pubescent at apex, the 4 (or 5) styles glabrous or fi nely pubescent, connate basally (Spongberg, 1977). Male fl owers produced on the current year’s growth, held ± nodding beneath the leaves. Staminate fl owers 6-7 mm. Long at anthesis, short- pedicellate, 3-5 together (or fewer through abortion) in rufous-pubescent, short-pedunculate, the bracts caducous; calyces green, fi nely pubescent, the 4 (or 5) deltoid lobes 1.5-2 mm. long; corolla 4.5-6 mm. long, white, ± campanulate and weakly 4- (or 5-) ribbed, the lobes pinkish or yellowish, recurved, ca. 2 mm, long; stamens 16, rarely fewer, epipetalcus in 2 whorls, the largest stamens ca. 4 mm. long; gynoecia abortive or rudimentary (Spongberg, 1977). 35 Materials and methods In the experiments was studied the male and female fl owers variability of date plum (Diospyros lotus L.). In the tested collection were included genotypes of date plum grown from seeds in Arboretum Mlynany of the Slovak Republic. As all genotypes were cultivated from the seeds in one locality, the tested collection of trees was perfectly adapted on the local conditions. In the blossom-time were from the genotypes withdrawn samples of fl owers and then determined their morphological traits – number of calyx petals, height and width of calyx petals (in mm), height and width of crown (mm), height and width of crown petals (mm), number of crown petals, number of stamens in the 1st/2nd rows, length of stamens in the 1st/2nd rows. To evaluate the determined morphological traits, the image analysis was applied using the software AxioVs40 V 4.8.2.0.

Results and discussions Plant populations cultivated from seeds are well adapted on different conditions including that of Botanical gardens in Slovakia (Grygorieva et al., 2009). The gained data and those reported in literature are signalling, that the date plum owing to its biological, nutritional and phytotherapeu- tic value of fruits and ather plant parts is a suitable species to be used in Slovakia as well, and therefore were made the experiments described in this paper.Four acids: palmitic acid (16:0), palmitoleic acid (16:1), linoleic acid (18:2), and linolenic acid (18:3) were identifi ed as the major acids in signifi cant amounts (P = 0.05) during development. Ripe fruits of date plum are consumed in fresh state, sometimes sprin- kled with lemon juice. These fruits are widely exploited for preparation of salads, cakes, marmalades, syrups, jams and distilled spirits (Kremer, 1995). In some countries are given as feed to utility animals (Holdeman, 1998). In China medicine the date plum is traditionally applied due to neuro-protective effect of this fruit to patients suffering apoplexy (Bei et al., 2007). Loizzo et al., 2009 studied their extract and their antioxidant and antiproliferative properties. antioxidative and antiproliferative ef- fects. Eight compounds were isolated from D. In extracts from fruit were determined the gallic and elagic acids,lotus and identifi ed as gallic acid, methylgallate, ellagic acid, kaempferol, quercetin, myricetin, myricetin 3- O - β -glucuronide, and myricetin-3- O - α -rhamnoside. D caempferol, quercetin, myricetin and other important biologically active components. Azadbakhta et al. (2010) experimentally proved the hypo-glycemic activ- ity in water extract of date plum fruits. Around with the fruits are the date 36 plum fl owers interesting as well from the view of their economic exploita- tion. To gain more complex data, we have taken into account both, the male and female fl ower traits (Table 1). From the morphometric analyses resulted the following outcomes: number of sepals 3.91/4.34, sepal height 1.97/5.45 (mm), sepal width 2.13/4.98 (mm), crown height 6.00/6.47 (mm), crown diameter 4.21/5.33 (mm), number of crown sepals 4.17/4.05, number of stamens in the 1st/2nd row of male fl owers 7,91/7,42, length of stamens in the 1st/2nd row of male fl owers 3.36/2.79 (mm).

Table 1 Variability of tested traits in the collection of date plum (Diospyros lotus L.) seedlings grown up in the Slovakia Male fl owers Female fl owers Tested fl ower traits n min max x V% n min max x V% Calyx sepals height(mm) 66 1.23 2.96 1.97 49.83 74 2.35 8 5.45 25.45 Calyx sepals width 66 1.32 3 2.13 16.59 74 3.29 7.42 4.98 21.55 (mm) Calyx sepals number 66 3 5 3.91 10.71 74 3 7 4.34 20.09 Crown height (mm) 66 4.44 8.16 6.00 18.05 74 4.21 8.26 6.47 14.55 Crown diameter (mm) 66 3.66 4.98 4.21 7.15 74 3.73 8.51 5.33 17.30 Number of crown sepals 66 2 6 4.17 17.65 74 3 7 4.05 13.47 Stamen numbers in the 66 6 9 7.91 6.55 74 7 9 7.84 5.98 1st row Stamen numbers in the 66 7 9 7.42 7.11 - - - - - 2nd row Stamen length in the 1st 66 2.08 4.06 3.36 8.98 - - - - - row (mm) Stamen length in the 2nd 66 1.83 3.68 2.79 14.25 - - - - - row (mm)

There were found signifi cant differences between the tested genotypes in amount of outgrowing male buds (Figure 1), number of crown sepals in the female (Figure 2) and male fl owers (Figure 3). Signifi cant differences were detected in the shapes of calyx and crown of female (Figure 4) and male fl owers (Figure 5).

37 Figure 1. Number of buds growing in male infl orescences of date plum (Diospyros lotus L.) genotypes cultivated from the seeds. Photo: Olga Grygorieva, 2010

Comparison of our results with those in literature (Spongberg, 1977; Shu-kang et al., 1996) showed for several traits striking differences. These discrepancies could be primarily connected with testing different genotypes collections.

Figure 2. Number of sepals in female fl owers of date plum (Diospyros lotus L.) genotypes cultivated from the seeds. Photo: Olga Grygorieva, 2010

Figure 3. Number of sepals of male fl owers of date plum (Diospyros lotus L.) genotypes cultivated from the seeds. Photo: Olga Grygorieva, 2010

Figure 4. Comparison of the female fl ower calyces and crowns of date plum (Diospyros lotus L.) genotypes cultivated from the seeds. Photo: Olga Grygorieva, 2010 38 Figure 5. Comparison of the male fl ower calyces and crowns of date plum (Diospyros lotus L.) genotypes cultivated from the seeds. Photo: Olga Grygorieva, 2010 Among the tested traits has been described high positive dependency as documented by the correlation analysis results in Table 2. In the evaluation process dealing with the tested traits dependences has been found a partial agreement among the correlation coeffi cients of the male and female fl owers only in the cases of calyx sepals height and crown height (0.476 – 0.481), calyx sepals height and stamen length in the 1st row (0.137 – 0.152) and crown height and stamen length in the 1st row (0.360 – 0.301), respectively. The rest of traits the when compared bilaterally, exerted different values of correlation coeffi cients. It means, that the dependencies between the male and female fl owers are variable. For male fl owers between the calyx sepals height (mm) and the crown diameter (-0.285) as well as between the crown height (mm) and crown diameter (-0.400) were statistically detected signifi cant negative correlations.

Conclusions From the study of morphological traits variability of date plum (Diospyros lotus L.) fl owers resulted the following knowledge: a) For all tested male and female fl owers traits was confi rmed a high variability degree. b) In the genotypes population grown up in Slovakia from the seeds were detected signifi cant genotypic differences for all evaluated traits. c) Date plum (Diospyros lotus L.) could be declared for an economically usable species in several branches for food and feed production as well as for pharmaceutical purposes. d) Further possibilities of date plum exploitation: as ornamental tree species with motley fl ower colours, and an additional nectar and pollen source for the .

Acknowledgments This work was supported by the Operational Programme Research and Development of the European Regional Development Fund in the frame 39 of the project „Support of technologies innovation for special bio-food products for human healthy nutrition“, ITMS 26220220115. Authors are indebted to Mrs. Eva Chovancova and Mr. Alexej Oravec for their technical assistance by the collection and evaluation of fl owers.

Bibliography 1. Ayaz, F. A., Kadioglu, A. 1999. Fatty acid compositional changes in developing (Diospyros lotus L.) fruit. In New Zealand Journal of Crop and Horticultural Science, vol.27, 199, p. 257-261. 2. Azadbakhta, M., Safapour, S., Ahmadi, A., Ghasemi, M. and Shokrzadeh, M. 2010. Anti-diabetic effects of aqueous fruits extract of Diospyros lotus L. on streptozotocin-induced diabetic rats and the possible morphologic changes in the liver, kidney and heart. In Journal of Pharmacognosy and Phytotherapy, vol. 2, 2010, no. 2, p. 010–016. 3. Bei, W., Peng, W., Ma, Y., Xu, A. 2005. Flavonoids from the Leaves of Reduce Hydrogen Peroxide-induced Injury of NG108-15 Cells. In Life Sciences, LXXVI., vol. 11, 2005, p. 1975 -1988. 4. Brezhnev D.D., Korovina O. N. 1981. Wild Relatives of Cultivated in Soviet Union Flora. Kolos, Leningrad. 376 p. (in Russian). 5. Grygorieva, O., Klymenko, S., Brindza, J., Kochanova, Z., Toth, D., Derevjanko, V. 2009. Introduction, breeding and use of persimmon species (Diospyros spp.) in Ukraine. Acta Hort. (ISHS), 2009, no. 833, p. 57-62. 6. Holdeman, Q. L. 1998. for Louisiana´s Children – Young and Old. 1998.16 s. 7. Kremer, B. 1995. Stromy. Bratislava : Ikar, 1995. s. 250-251. ISBN 80- 7118-177-3. 8. Kuliyeva, G. 1962. Morfologicheskije osobennosti kaukazskoj churmy (Diospyros lotus L.). In Botanicheskij zhurnal, vol. 47, 1962, no. 10, p. 1446 – 1454. 9. Shu-kang, L., Gilbert, M. G., White, F. 1996. . 15: p. 215– 234. 10. Loizzo, M. R., Said, A., Tundis, R., Hawas, U.W., Rashed, K., Menichini, F., Frega, N.G., Menichini, F. 2009. Antioxidant and Antiproliferative activity of Diospyros lotus L. Extract and Isolatid Compounds. In Plant foods for human nutrition, vol. 94, 2009, no. 4. 11. Sponberg, S. A. 1977. hardy in temperate North America. In Journal of the Arnold Arboretum, vol. 58, 1977, p. 146-160.

40 Table 2 Table L.) 0.0226 0.0055 0.1935 0.0031 0.0090 0.0000 0.0000 of t (p) -0.0000 Probability Probability t Diospyros lotus 95% owers fl ndence r ndence fi row (mm) row row (mm) row st st row (mm) row st row (mm) row st traits of date plum ( ower fl r Interval of con 0.319 0.0986<= r >= 0.5110 2.86 0.152 -0.0784<= r >= 0.3685 1.31 0.301 0.0786<= r >= 0.4960 2.68 0.481 0.2845<= r >= 0.6398 4.66 0.495 0.3004<= r >= 0.6500 4.83 0.0009 0.2706 0.0030 0.0003 0.0206 of t (p) Probability Probability cients of evaluated fi Crown height (mm) x Crown diameter (mm) diameter height (mm) x Crown Crown Calyx sepals height (mm) x Crown height (mm) Calyx sepals height (mm) x Crown Calyx sepals width (mm) x Crown height (mm) Calyx sepals width (mm) x Crown Calyx sepals width (mm) x Crown diameter (mm) diameter Calyx sepals width (mm) x Crown Calyx sepals height (mm) x Crown diameter (mm) diameter Calyx sepals height (mm) x Crown t Calyx sepals height (mm) x width 0.81 0.4181 0.264 0.0387<= r >= 0.4652 2.33 1.424.33 0.1582 0.0001 0.339 0.794 0.1203<= r >= 0.5271 0.6916<= r >= 0.8658 3.06 11.10 Crown height (mm) x Stamen length in the 1 Crown Crown diameter (mm) x Stamen length in the 1 diameter Crown Calyx sepals width (mm) x Stamen length in the 1 Calyx sepals height (mm) x Stamen length in the 1 95% owers Female fl Male ndence r ndence fi Linear dependency correlation coef dependency correlation Linear r Interval of con 0.137 -0.1080<= r >= 0.36740.476 1.11 0.2652<= r >= 0.6444 0.360 0.1294<= r >= 0.5539 3.08 0.427 0.2070<= r >= 0.6068 3.78 -0.096 -0.3304<= r >= 0.1495 0.77 0.4430 0.301 0.0781<= r >= 0.4956 2.68 0.0091 -0.101 -0.3352<= r >= 0.1442 -0.400 -0.5855<= r >= -0.1751 3.49 -0.175 -0.4007<= r >= 0.0693 -0.060 -0.2980<= r >= 0.1844 0.48 0.6305 0.494 0.2995<= r >= 0.6493 4.82 0.0000 -0.284 -0.4927<= r >= -0.0457 2.37

41