Micropropagation and conservation of threatened endemicTripleurospermum conoclinium and T. hygrophilum (asteraceae) species

In this thesis, ex situ conservation of the threatened endemic species Tripleurospermum conoclinium (Boiss. & Bal.) Hayek and Tripleurospermum hygrophilum (Bornm.) Bornm. was carried out by micropropagation. The stability tests of the micropropagated plants were performed by flow cytometry (FCM), chromosome counting and stomatal analysis. In micropropagation of T. conoclinium and T. hygrophilum, seedlings obtained from achenes germinated in vitro were used as explant source. MS suplemented with the combination of 6-benzylaminopurine (6-BA, 0,25, 0,5 and 1 mg L−1), kinetin (KIN, 0,25, 0,5 and 1 mg L−1), 6- y,dimethylallylamino)-purine (2iP, 0,25, 0,5 and 1 mg L−1) with indole-3-butyric acid (IBA, 0,1 mg L−1) and naphthaleneacetic acid (NAA, 0,1 mg L−1) were used at shoot multipication. The highest number of shoots per explant for T. conoclinium was obtained from plants in MS medium suplemented with 0,25 mg L−1 6-BA + 0,1 mg L−1 NAA. For T. hygrophilum, the highest shoot number was obtained from plants in MS medium suplemented with 0,25 mg L−1 6-BA + 0,1 mg L−1 IBA. MS medium suplemented with indole-3-butyric acid (IBA; 0,25, 0,5, and 1 mg L−1) and naphthalene acetic acid (NAA; 0,25, 0,5 and 1 mg L−1) was used for rooting. The highest number of roots per explant for T. conoclinium was obtained from plants in MS medium suplemented with 1 mg L−1 NAA. For T. hygrophilum, the highest root number was obtained from plants in MS medium suplemented with 0,25 mg L−1 NAA. Nuclear DNA of micropropagated plants did not change when compared to the nuclear DNA amounts in natural populations of these plants. It was determined that the chromosome number of T. conoclinium and T. hygrophilum reproduced in vitro was 2n = 2x = 18. The chromosome number was found to be similar to the number of chromosomes in plants in the natural environment. It was observed that the stomatal length of the acclimatized plants was the same as the stomatal length of the plants growing in the natural environment. The survival rate of T. conoclinium transferred to ex vitro conditions was 95%, and the survival rate of T. hygrophilum was determined 98%. Plants transferred to the ex vitro flowered and formed achene in the second year. These data have contributed to conservation biology.