EX SITU CONSERVATION OF RARE MEDICINAL PLANTS IN THE BOTANICAL GARDEN (INSTITUTE) OF ACADEMY OF SCIENCES OF MOLDOVA

Acta Horti Bot. Bucurest. 2013, 40: 43-48 DOI: 10.2478/ahbb-2013-0005 EX SITU CONSERVATION OF RARE MEDICINAL PLANTS IN THE BOTANICAL GARDEN (INSTITUTE) OF ACADEMY OF SCIENCES OF MOLDOVA CIOCÂRLAN Nina1 Abstract: This work refers to the native species of genus Astragalus L. (A. dasyanthus, A. ponticus), Adonis L. (A. vernalis, A. wolgensis) and Digitalis L. (D. lanata, D. grandiflora). The plants are cultivated in the Botanical Garden of Moldova in the field collection of the medicinal and aromatic plants. Investigation includes propagation aspects, research into cultivation techniques and conservation measures. The biological particularities and the phenologic rhythm are also registered. The obtained data shows the ecological flexibility of species and the possibility of preserving them in culture. Key words: cultivation, ex situ conservation, medicinal plants, propagation Received 15 March 2013 Revision accepted 9 October 2013 Introduction Medicinal and aromatic plants have been an important source for human health care from prehistoric times to the present day. According to the WHO (World Health Organization), the majority of the world s population depends on traditional medicine based on medicinal and aromatic plants (WHO, 2002). Today we find a renewed interest in traditional medicine. This interest in plant-derived drugs has necessitated an increased demand of medicinal plants leading to over-exploitation, unsustainable harvesting and to the virtual decimation of several valuable plant species in the wild. Because of the constant harvesting of these plants, many of them have become rare. Furthermore, the habitat degradation, illegal trade of rare and endangered medicinal plants and loss of regeneration potential of the degraded forests have accelerated the current rate of extinction of plants, particularly the medicinal ones. According to recent studies (Negru et al. 2002, 2005), a good few wild species, some of those medicinal and aromatic plants, are endangered in the Republic of Moldova, mainly due to habitat degradation, pollution, over-exploitation and unsustainable harvesting. Over the past few years, we have begun a research program into the preservation, propagation and re-introduction of indigenous medicinal and aromatic plants. More than 30 rare and threatened species, well known for their medicinal properties were included in ex situ collection of medicinal and aromatic 1 Botanical Garden (Institute) of Academy of Sciences of Moldova, 18, Pădurii St., 2002 - Chişinău, Republic of Moldova, e-mail: n_ciocarlan@mail.ru

44 Ciocârlan Nina plants in order to observe their reproductive behavior and accumulate experience on their agro-technical peculiarities. Material and methods The investigations were carried out in 2007-2011 at the experimental fields in the Botanical Garden (Laboratory Vegetal Resources) of Academy of Sciences of Moldova. Six medicinally important species were used in this experiment. An extensive survey of selected species was made according to their preliminary information of status, propagation and conservation strategy. The selected plant species were collected and identified with the help of researchers of Native Flora and Herbarium Laboratory of the Botanical Garden (Institute) of ASM. The nomenclature of the taxa is given according to some scientific papers (Chater 1968, Heywood 1972, Tutin & Akeroyd 1993, Negru 2007, Ciocârlan 2009). Investigations include propagation aspects, research into cultivation techniques and conservation measures. The biological particularities and the phenologic rhythm are also registered. Results and discussion Genus Digitalis L. (foxglove) (fam. Scrophulariaceae) comprises about 30 species of herbaceous biennials, perennials and shrubs. The genus is native to Europe, western and central Asia and north-western Africa (Ivanina 1981). Digitalis species and its derivates are critically important in the management of congestive heart failure and related cardiac disorders (Arnold et al. 1980, Gheorghiade et al. 2004). According to the literature, the foxglove (Digitalis) species which are used in the heart medications digitoxin and acetyldigitoxin have already become endangered and will need their conservation and multiplication (Hamkins 2008). There are two species native to the flora of the Republic of Moldova (Negru 2007). Digitalis lanata Ehrh. (Greek Foxglove) is a herbaceous perennial or biennial plant (30-100 cm tall) indigenous to Central Europe and the Balkan Fig. 1. Samples of D. lanata cultivated on Peninsula (Ivanina 1981, Heywood 1972). experimental plots (flowering plants, July The growing places in the Republic of 2009) Moldova are within the central area. It grows in glades and clearings in semi-shade (Negru et al. 2006). It is a critically endangered species and is included in the Red Book of Republic of Moldova (Nicolaev 2001) and the European Red List of Vascular Plants (Bilz et al. 2011). The state protection of the growing areas of the species includes the registration of the new growing places and the organization of its preservation (Nicolaev 2001). Digitalis grandiflora Mill. (Large Yellow Foxglove) is a biennial to perennial plant (30-100cm), native to central and south Europe and Caucasus (Ivanina 1981, Heywood 1972). In the Republic of Moldova it occurs in the central part of the country

Ex situ conservation of rare medicinal plants in the Botanical Garden (Institute) 45 (the forested valleys of Dniester and Prut) (Negru et al. 2006). D. grandiflora as well as other foxglove species and their derivates possess cardiac, stimulant and tonic properties being used in the treatment of certain heart problems (Shchelokova 1980, Grinchevich et al. 1981). In the wild the species is becoming increasingly uncommon due to a combination of unfavorable factors including habitat destruction and anthropogenic activities (Negru et al. 2006). Under the conditions of the Republic of Moldova both species were successfully propagated by seeds and through seedlings (Fig. 1). The seeds were sown in open ground in late autumn or early spring. The seeds usually germinate in 2-3 weeks. When being propagated by seedlings, the seeds were sown in boxes with black soil and sand (1:1) in the glass house in the middle of February. The first plant appeared in 16-18 days after sowing. When the seedlings were 5-7cm high they were transferred to open field (in second decade of May). The planting was made in rows with a 50 cm distance between them and 30 cm distance between the plants. In the first year of growth both species produced only long basal leaves. In the second year, plants developed the erect leafy stem 0.5-1.2m high (Fig. 1). The flowering period lasted for 32-36 days, from the first decade of June to the second decade of July. The seeds ripened in the Mid-August. Genus Adonis L. (fam. Ranunculaceae) comprises approximately 20 annual and perennial plants, distributed in the temperate region of the northern hemisphere (Tutin & Akeroyd 1993, Sennicov 2001). There are five species native to the flora of the Republic of Moldova (Negru 2007). Adonis vernalis L. (False Hellebore) is a perennial, herbaceous plant native to Central and Southern Europe and eastwards to Western Siberia (Tutin & Akeroyd 1993, Sennicov 2001). In the Republic of Moldova it vegetates sporadically throughout the country (Negru et al. 2005, Teleuţă et al. 2008). It is a vulnerable species, protected by the environmental law (Parlamentul Republicii Moldova 1998). Artificial forestation of slopes in the steppe and over-collection of medicinal raw material negatively influenced the condition of A. vernalis populations. The species is included in the Red Data Book of Ukraine (Melnik & Peregrim 2009). Adonis wolgensis Steven is an herbaceous, perennial species native to Central Europe and eastwards to Western Siberia (Tutin & Akeroyd 1993, Sennicov 2001). In the Republic of Moldova is found in the south of the country (Negru et al. 2002). This species has a rare status, being protected by the Environmental law (Parlamentul Republicii Moldova 1998). It is included in the Red Data Book of Vascular Plants of Romania (Dihoru & Negrean 2009) and Red Data Book of Ukraine (Melnik & Peregrim 2009). The adult plants and seeds of A. vernalis and A. wolgensis were collected from nature. The first stage was transplanting adult individuals from their native populations. The second stage was propagation of plants from seeds collected from the wild as well as from seeds collected from the adult plants which set seeds successfully in the garden. The third stage was vegetative propagation through root Fig. 2. Samples of A. vernalis dividing (Fig. 2). The optimal time of seed (flowering plants, 2009)

46 Ciocârlan Nina sowing in open ground in conditions of our country is in late autumn (November). The set of seedlings of A. vernalis was produced in the glasshouse to be transferred in the open ground of the experimental plot of the Botanical Garden. Seeds were sowed in the last decade of February. They were incorporated into the soil at 1.5-2 cm depth. First seedlings were registered in 18-20 days. The seed germination coefficient was 98%. At the first decade of May, the seedlings at the 3-6 true leaf stage (after being hardened) were transferred into experimental plot. The planting scheme is 50 20 cm. Artificial watering of the seedlings was applied periodically. Genus Astragalus L. is the largest genus in the family Fabaceae. It includes about 2000 species distributed in Europe, Asia and North America (Chater 1968, Vasiljeva 1987). In our flora it is represented by 16 species (Negru 2007). The present studies refer to A. dasyanthus and A. ponticus, both species being rare and vulnerable. Astragalus dasyanthus Pall. is a herbaceous perennial found in the EastEuropean Plain and on the Balkan Peninsula (Chater 1968, Vasiljeva 1987). In the Republic of Moldova it vegetates in the central and south districts (Negru et al. 2005). A. dasyanthus is a vulnerable species, included in the Red Book of the Republic of Moldova (Șabanov 2001), Red Data Book of Ukraine (Korotcenco & Kritzka 2009) and European Red List of Vascular Plants (Bilz et al. 2011). In some Balkan countries it is Critically Endangered (Stoyanov & Tzonev 2009). Astragalus ponticus Pall. is a herbaceous perennial plant indigenous to Southern Europe and East-Mediteranean region (Chater 1968, Vasiljeva 1987). In the Republic of Moldova it is found in the south of country. It is threatened species, becoming rare because of habitat loss (Negru et al. 2002). The adult plants of A. dasyanthus transplanted from their native populations in 2007 developed well. A. ponticus developed especially well, looked more vigorous and the blooming period was two weeks earlier in comparison to the native habitats. The 91.3% of the A. dasyanthus transplants survived until the next season but in the first two years viable seeds were not obtained, although it bloomed abundantly in the ex situ conditions (Fig. 3). In 2009 seeds collected from the adult plants Fig. 3. A. dasyanthus (multibranched adult transplanted in the field in 2007 were tested. More than a half of them were plant, 2010) damaged by insects, the rest of the seeds demonstrated poor germination, even being stratified and scarified. We started more investigations and germination experiments for the optimization of the propagation process. In the collection of medicinal and aromatic plants there have been initiated introduction studies on some other rare medicinal plants: narrow-leaved Bindweed (Convolvulus lineatus L.), Sweet Flag (Acorus calamus L.), asparagus (Asparagus officinalis L.), Rupturewort (Herniaria glabra L), Martagon lily (Lilium martagon L.), Tatarian Sea-kale (Crambe tataria Sebeok), dwarf everlast (Helichrysum arenarium

Ex situ conservation of rare medicinal plants in the Botanical Garden (Institute) 47 (L.) Moench.), liverwort (Hepatica nobilis Mill.). In the future it is planned to increase the number of rare and threatened medicinally important plant species, in order to ensure their ability for ex situ propagation. Conclusions Digitalis lanata developed very well and set seed for the following seasons. Astragalus ponticus developed especially well, looked more vigorous and the blooming period was two weeks earlier in comparison to the native habitats. It should be propagated by seed and some more investigations are necessary for the optimization of the process. Good germination was observed for Adonis vernalis (87%) and Digitalis lanata (96%). Astragalus dasyanthus seed propagation was difficult enough, so vegetative propagation for this threatened species could be a successful strategy for cultivation. All obtained data shows the ecological flexibility of species and the possibility of preserving them in culture. References Arnold, S., Byrd, R. & Meister, W. (1980). Long-term digitalis therapy improves left ventricular function in heart failure. N. Engl. J. Med., 303, 1443-1448. Bilz, M., Kell, S. P., Maxted, N., Langdown, R. V. (2011). European Red List of Vascular Plants. Luxembourg: Publications Office of the European Union. Chater, A. O. (1968). Astragalus L. In T. G. Tutin, V. H. Heywood, N. A. Burges, D. M. Moore, D. H. Valentine, S. M. Walters & D. A. Webb (Eds.) Flora Europaea. Vol. 2. (pp. 108-124). Cambridge: Cambridge University Press. Ciocârlan, V. (2009). Flora ilustrată a României. Pteridophyta et Spermatophyta. Bucureşti: Edit. Ceres. Didukh, Y. A. (2009). Astragalus ponticus Pall.. In Y.A. Didukh (Ed.). Red Data Book of Ukraine. Vegetable Kingdom (pp. 445). Kyiv: Globalconsulting. Dihoru, G. & Negrean, G. (2009). Cartea roşie a plantelor vasculare din România. Bucureşti: Edit. Academiei Române. Gheorghiade, M., Kirkwood, F. & Wilson, S. (2004). Digoxin in the management of Cardiovascular Disorders. Circulation, 109, 2959-2964. Grinkevich, N. T., Sorokina, A. A. & Kucherov E. V. (1981). Contents of cardiac glycosides and microelements in Digitalis grandiflora Mill. Rastit. Resur., 17(3), 406-410. Hamkins, B. (2008). Plants for Life: Medicinal plant conservation and Botanic gardens. Richmond: Botanic Gardens Conservation International. Heywood, V. H. 1972. Digitalis L.. In T. G. Tutin, V. H. Heywood, N. A. Burges, D. M. Moore, D. H. Valentine, S. M. Walters & D. A. Webb (Eds.) Flora Europaea. Vol. 3. (pp. 239-241). Cambridge: Cambridge University Press. Ivanina, L. (1981). Digitalis L. In A. Fedorov (Ed.). Flora Partis Europaea URSS. Tom. 5. (pp. 239-240). Leningrad: Edit. Nauka. Korotcenko, I. & Kritzka, L. (2009). Astragalus dasyanthus Pall.. In Y. A. Didukh (Ed.). Red Data Book of Ukraine. Vegetable Kingdom (pp. 438). Kyiv: Globalconsulting.

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