Two different experiments were carried out to study the production of protoplasts and the variation of protoplast yield from in vitro cultured shoot tips of tansy (Tanacetum vulgare L.) and pyrethrum (Tanacetum cinerariifolium (Trevir.) Schiltz-Bip). In the first experiment, light had more pronouced effect for tansy than for pyrethrum. When the donor tissues of tansy were cultured under high light intensity the leaves contained anthocyanin and became brown during enzyme maceration. In contrast, donor tissues cultured under low light intensity produced leaves without anthocyanin. Depending on the light intensity of donor tissues, on average 5.8 - 6.8 x 106 and 3.4 - 4.3 x 106 protoplasts were isolated from one gram of mesophyll leaves of tansy and pyrethrum, respectively. In the second experiment, the production of protoplasts from tansy and pyrethrum varied seasonally. The most successful season for the production of protoplasts from in vitro cultured shoot tips was between December and April, when also the highest number of protoplasts could be isolated. It was not possible to state whether Tanacetum species have rhythms, which could cause physiological or chemical changes for the in vitro grown shoot tips. However, some external or internal, possible seasonal-dependent stimuli may have caused variation in the number of protoplasts isolated from tansy and pyrethrum and favoured protoplast production during winter and spring.
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