Abstract
Sideritis raeseri Boiss & Heldr. subsp. raeseri, known in Greece as Mountain tea of Parnassus or Velouchi is a range restricted medicinal plant of the Balkan peninsula. Conventional propagation methods did not allow the mass production of plant material in a short period of time due to both low seed germination rate and rooting of cuttings. Therefore, the aim of this study was to establish a reliable, reproducible and efficient regeneration protocol for mass and large-scale micropropagation, germplasm and ex situ conservation of S. raeseri Boiss & Heldr. using Imazalil and Carbendazim fungicides. After 9 weeks, 2.5-10 mg/l Imazalil stimulated root length by 1 cm but diminished root number and rooting percentage. Optimum shoot number (5.58), shoot length (24.91 mm), shoot multiplication (100%), root number (20.63) and rooting (66.67%) were recorded with 0.5 mg/l kinetin (KN) + 0.05 mg/l ?-napthaleneacetic acid (NAA) (Imazalil-free). After 4 ½ weeks, 1 mg/l Carbendazim + 0.5 mg/l 6-benzyladenine (BA) yielded 5.77 shoots/explant, 16.1 mm shoot length and 100% shoot multiplication. However, Carbendazim did not result in rooting. The ex vitro survival percentage of rooted shoot-tip explants derived from Imazalil experiment was 95%. Carbendazim proved useful in shoot proliferarion and Imazalil in root elongation of S. raeseri Boiss & Heldr. subsp. raeseri micropropagation system.
Highlights
Sideritis L. (Family Lamiaceae) comprises approximately 150 species of annuals and perennials distributed throughout North Africa, the Iberian Peninsula, the Mediterranean countries, and the Middle East region [1]
In S. raeseri Boiss & Heldr, Imazalil exerted an inhibitory effect on shoot regeneration
From the results obtained it is evident that the imidazole fungicide Imazalil cannot be used in tissue culture systems as a promoting shoot proliferation agent, its use is recommended only for enhancing root elongation of S. raeseri shoot tip explants
Summary
Sideritis L. (Family Lamiaceae) comprises approximately 150 species of annuals and perennials distributed throughout North Africa, the Iberian Peninsula, the Mediterranean countries, and the Middle East region [1]. (Mountain tea of Velouchi or Parnassus) is distributed in central, north-central and northern parts of the country (typically in the range of Pindos) or in neighbouring areas with Albania and the FYR of Macedonia according to Strid and Kit Tan [2]. A number of interesting side effects of the imidazole fungicide Imazalil can be observed in diverse tissue cultured plant species, i.e. excessive shoot formation in Araceae, inhibition of bushiness in Gerbera, restoration of normal embryo development in Citrus and histogenic instability of Ficus benjamina L. When screening fungicides as potential tissue culture medium additives to control fungal development, it was found that the imidazole fungicide Imazalil exuberantly enhanced the shoot (bud)-inducing effect of BA in Spathiphyllum floribundum. According to Werbrouck and Debergh [8], TDZ mediated shoot proliferation improved by the inclusion of Imazalil in S. floribundum culture
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