Use of tissue culture in prezervation and conservation of endemic plants in Herzegovina

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1 AGROMEDITERANSKI FAKULTET-MOSTAR UNIVERZITET DŽEMAL BIJEDIĆ - MOSTAR AGROMEDITERANSKI FAKULTET Adresa:USRC Midhad Hujdur-Hujka, Mostar Studentska služba: Tel/Fax: af@unmo.ba Use of tissue culture in prezervation and conservation of endemic plants in Herzegovina Alisa Hadžiabulić Doc.dr. Semina Hadžiabulić

2 The position of Herzegovina as the southern part of Bosnia and Herzegovina is influenced by Mediterranean, continental, and mountain climates. The Euro-Siberian and Meditterranean floral regions have created such a diverse bio system that well over half the total number of flowering plants on the Balkan Peninsula can be found here. These plants are either rare, endemic and as such their protection should be of our interest.

3 Its richness is only comparable to the abundance of plants in tropical and sub-tropical regions. There are over 3,700 identified species of flowering plants in BiH and hundreds of endemic species.

4 During the last couple of years, the activities for preservation of genetic resources were intensified The first step in preservation and conservation of endemic plants is their inventarisation in the area of Bosnia and Herzegovina. The aim of this work was to register and protect as many endemic plants as possible, so they can be used for further multiplication and conservation.

5 Endemic plants of Herzegovina

6 Aquielgia dinarica Dinarska kandilika Endemic flowering rocky plant Dinarian kandilka Territory: endem of Dinaric range, Prenj, Čvrsnica, Plasa, Čabulja, Vran, Velež Description: distinctive flowers, generally bellshaped, with each petal modified into an elongated nectar spur. Size: app. 25cm Colour: lilac, internal petals white Bloom: summer (VII VIII) Cultivation: Species lives in rock cervices in mountain region from 1200 to 2100 m,limestone rocks mostly, sunny, dry, rock crevices,

7 Campanula hercegovina Hercegovački zvončić Endemic flowering rocky plant Herzegovinian bellflower Territory: Herzegovina, mountains of Prenj and Velez, Neretva canyon, Diva Grabovica valley Description: leaves ovate, green, denticulate, corolla erect, openly campanulate, 15-20mm long Size: 5-15cm Colour: deep lilac Bloom: summer Cultivation: sunny, dry, rock crevices, protection against winter wet drained soil,

8 Postal marks Flora: Dinarian kandilka Aquilegia dinarica Herzegovinian bellflower Campanula hercegovina

9 Endemic flowering rocky plant Dianthus, pinks Territory: Herzegovina, mainly Prenj mountain Description: grasslike, gray-green leaves, finally toothed, flower with fingered petals Size: 1-7cm Colour: pink Bloom: late spring Cultivation: sunny, limestone and dolomite soil, good drainage Dianthus freynii Frajnov karanfil

10 Endemic flowering rocky plant Mountain Mint Territory: Herzegovina mountains Description: leaves ovate, green, denticulate, corolla erect, openly campanulate, 15-20mm long Size: 5-15cm Colour: white Bloom: summer/autumn (VII-X) Cultivation: sunny, dry, rock crevices, mainly on karst, but also on serpentine, ranging from 30 m to 2000 m. Micromeria thymifolia Vrisić

11 Primula kitaibeliana Jaglac Endemic flowering rocky plant Kitajbelian primula Territory: Herzegovina, mountains from Velez to Visocica, National park Blidinje Description: leaves ovate, green, denticulate, corolla erect, openly campanulate, 15-20mm long Size: 1,5-18 cm Colour: pink Bloom: summer (VII) Cultivation: It prefers rock cervices in mountain region from 1600 to 2100 m, sunny positions, dry, limestone soil,

12 Gentiana lutea Dinarska enciana Perrenial medicinal plant Territory: Herzegovina, Dinaric range, National park Blidinje Description: broad lanceolate to elliptic leaves10-30 cm long and 4-12 cm broad. Size: 1-2 m Colour: yellow Bloom: summer (VI VIII) Cultivation: sunny, calcareous soils drained soil, in the mountain areas from 1500 to 1800 m.

13 Measures for preservation and propagation In situ - conventional methods Ex- situ in vitro culture

14 Culture of tissue is fascinating and useful tool which allows the rapid production of many genetically identical plants using relatively small amounts of space, supplies and time. This is a useful approach for ex situ conservation of rare, endemic or threatened plant species.

15 Taking of an explant in this cases (such as a stem tip, node, meristem, embryo, or even a seed), since none made it before, we should take them all and the experiment will result with the most suitable one We are not sure on the formulation of the growth medium, using the literature sources we found some which might be used.

16 Growth mediums Murashige and Skoog (MS) formula MS Salts mg/liter MAJOR SALTS Ammonium nitrate (NH4NO3) 1650 Calcium chloride (CaCl2 2H2O) 440 Magnesium sulfate (MgSO4 7H2O) 370 Potassium nitrate (KNO3) 1900 Potassium phosphate (KH2PO4) Subtotal 4530 MINOR SALTS Boric acid (H3BO3) 6.2 Cobalt chloride (CoCl2 6H2O) Cupricus sulfate (CuSO4 5H2O) Manganese sulfate (MgSO4 H2O) 16.9 Potassium iodide (KI) 0.83 Sodium molybdate (Na2MoO4 2H2O) 0.25 Zinc sulfate (Zn SO4 7H2O) 8.6 Subtotal32.83 IRON Ferrous sulfate (FeSO4 7H2O) 27.8 Na2EDTA 37.3 Subtotal 65.1 Total mg/liter

17 Dianthus media Compound Stage I II III mg/lite Ammonium nitrate 400 Boric acid 1.6 Calcium nitrat 144 Magnesium sulfate 72 Manganese sulfate 6.5 Potassium chloride 65 Potassium iodide 0.75 Potassium nitrate 80 Potassium phosphate 12.5 Zinc sulfate 2.7 MS salts Ferrous sulfate 27.8 Na2EDTA 32.8 trans-cinnamic acid 1.5 1,5 1,5 D-Pantothenic acid calcium salt Inosital Thiamine HCl Nicotinic acid Pyridoxine HCl NAA Kinetin Glycine Casein hydrolysate 3, Sucrose 30,000 30,000 30,000 Agar 6,000 6,000 6,000 ph 5.0

18 Saintpaulia media Compound Stage I & II IIII LeafPetiole mg/liter MS salts 4,628 4,628 Sodium phosphate Inositol 100-Thiamine HCl Nicotinic acid Pyridoxine HCl0.4-IAA NAA-0.1BA Sucrose 30,000 30,000 Agar 8,000 8,000 ph 5.5

19 Primula media Compound Stage I & II IIImg/liter Ammonium nitrate Boric acid Calcium chloride Cobalt chloride Cupricus sulfate Magnesium sulfate Manganese sulfate Potassium iodide Potassium nitrate Sodium molybdate Sodium phosphate Zinc sulfate Ferrous sulfate Na2EDTA Adenine sulfate 80 - Inositol Thiamine HCl IAA1.50.5BA2.5-S ucrose30,00030,000agar6,0006,000 ph 5.7