Propagating of bryum capillare hedw., Sphagnum palustre L. ve thamnobryum alopecurum (Hedw.) gangulee with tissue culture methods, identifying biological activity and phenolic content

In this study, it is aimed to find out the quantitative amount of Bryum capillare Hedw., Sphagnum palustre L. ve Thamnobryum alopecurum (Hedw.) Gangulee species, which have important roles in the ecosystem, via HPLC-UV analysis by using the in vitro tissue culture, antioxidant activity, total phenol content and 12 phenolic compound standart. For in vitro cultures, spores of the mature capsules and apical shoots of gametophyte of B. capillare and T. alopecurum species were used, whereas for S. palustre L. species only the capitulums parts of gametophyte were used due to the fact that its sporophytes stage of was not seen. Through in vitro tissue culture, the regeneration of these three species which were cultivated (by adding various concentrations of plant growth regulators and/or sugar) in 8 different modified MS (Murashige and Skoog, 1962) nutrient medium was successfully achieved. In B. capillare spores culture and gametophyte culture, the medium which showed the best growth was MS3 (30 g sucrose, 0.5 mg/l kinetin, 0.5 mg/l 2,4 D). In S. palustre gametophyte culture, the best growth was observed in MS1 (plant growth regulator and sugar free) medium. While the best growth in T. alopecurum spores culture was observed in MS3 (30 g sucrose, 0.5 mg/l kinetin, 0.5 mg/l 2,4 D) medium, in gametophyte culture it was MS1 (plant growth regulator and sugar free). The antioxidant activity of the fresh samples of these three species that have been collected from nature was done with the DPPH (2,2 diphenyl- 1, picryl hydrazyl) free radical scavenging activity method, while their total phenol amount was identified with the Folin-Ciocaltae method. When the antioxidant activity of these three species was compared withBHT (Butylated hydroxytoluene), BHA (Butylated hydroxyanisole) and ascorbic acid, the highest free radical scavenging effect, nearly about 85%, was seen in B. capillare. In terms of total phenol content, according to gallic acid equivalents, it was defined that the highest B. capillare extract had an approximate content of 0.034 ± 0.002 mg/g, S. palustre 0.031 ± 0.004 mg/g and T. alopecurum had the content of 0.0065 ± 0.0015 mg/g on the average. On the fresh samples of three species collected from nature in November and May (2012), the B. capillare and T. alopecurum herbarium samples collected at different times and the other samples which were grown with in vitro tissue culture, 12 different phenolic compounds (gallic acid, caffeic acid, p-coumaric acid, taxifolin, rosmarinic acid, kaempferol, genistein, quercetin, biochanin A, daidzein, formononetin, shikimic acid) were determined quantitatively by means of HPLC-UV. Among the species in 12 different compounds, B. capillare had the highest amount. In B. capillare spores culture, the highest amount of shikimic acid was in MS4 medium (261.65 ± 21.84 mg/g); in gametophyte culture, the highest amount of the same acid was in MS6 medium (220.4 ± 17.3 mg/g), while in herbarium samples the highest amount of gallic acid collected from the 9-month-old extracts was 1.20 ± 0.4 mg/g. The highest amounts of shikimic acid in S. palustregametophyte culture, T. alopecurum spores culture and in gametophyte culture were determined in MS4 medium (251.88 ± 88.3 mg/g), in MS8 medium (120.92 ± 41.4 mg/g) and in MS8 medium (120.92 ± 41.4 mg/g) respectively. However, in herbarium samples the highest amount of shikimic acid collected from the 4-month-old extracts was seen as 3.52 ± 1.84 mg/g. By means of the obtained results, these three species were found to be rich in phenolic compounds and also with antioxidant activity.