10th Symposium on the Flora of Southeastern Serbia and ...

10 th <strong>Symposium</strong> on the Flora of Southeastern Serbia and Neighbouring regions,Vlasina 17 to 20 June 2010carantion plantlets were left to grow in natural places where they bloomed. In future,these ''in vitro'' carnation plantlets will be reintroduced in nautural enviroment.Important of in vitro horse chestnut androgenic embryosproductionĆalić, D. 1 , Radojević, Lj. 11 Institut za biološka istraživanja „Siniša Stanković“, Beograd, Srbijadusica.calic@gmail.comThe aim of this research was to study influence of activated charcoal (AC),abscisic acid (ABA) and polyethylene glycol (PEG) on the maturation andconversion of horse chestnut androgenic embryo for the diversity protection andconservation of horse chestnut. Horse chestnut (Aesculus hippocastanum L.,Hippocastanaceae) represent a relict species of the tertiary flora and endemit ofBalkan peninsula. The common name horse chestnut is reported as having originatedfrom the erroneous belief that the tree was a kind of chestnut, together with theobservation that eating them cured horses of chest complaints. Horse chestnut treesare native to the Balkan peninsula, but grow as ornamental trees in parks andavenues throughout the Northern Hemisphere. Because of the slow and difficultreproduction of great importance to be fast and cheap in vitro multiplication.Possible solution is regenerated by androgenesis. Anther culture has been used inrecent years as a tool for producing haploid plants in a varyety of higher plants, butthe low frequencies of microspore-derived plants restrict the use of the technique inplant breeding. There are several factors affecting androgenesis in horse chestnut,such as genotypes, growth of donor plants, pretreatments of anthers, composition ofmedium and culture conditions. Androgenic embryos originating from microsporesand anther culture were maturated over 90 days. Androgenic embryos on mediacontaining PEG (50 g l-1), in combination with AC (1 g l-1) showed a rapiddevelopment of embryos in the cotyledonary stage and lowered percentage ofabnormal structures. The best results of androgenic microspore embryo germinationwas observed on media supplemented with AC alone (99%), and in combinationwith PEG (100%). Also, the greatest number of androgenic microspore plants (18%)and androgenic anther plants (12%) were formed on media enriched with 1 % AC.Lowest germination percentages, 37 % and 39 % in microspore culture and 33 %and 38 % in anther culture were obtained on maturation media with ABA 20 mg l-1alone and in combination with AC 1g l-1. Flow cytometric analysis showed thatmost of the androgenic embryos were haploid, corresponding to their microsporeorigin, while half of these became diploid, after maturation for 90 days. Allregenerants originating from microspore culture were haploid immediately after76