Zastosowanie kultur in vitro w zintegrowanej ochronie gatunków zagrożonych w Polsce: Hacquetia epipactis L., Primula farinosa L. i Echium russicum J.F. Gmel

Barbara Nowak

Abstract

The aim of the investigation was to elaborate micropropagation protocols of three endangered species of Polish flora: Ilacquetia epipactis L., Primula farinosa L., and Echium russicum L. to meet the needs of genotype protection and to satisfy the commercial demand. For this reason, for each species, the protocols of micropropagation from seeds or shoot tips, and morphogenesis from differentiated tissues were developed. The cultures were started from seeds of E. russicum obtained from Okolog Botanik Garten in Bayreuth (Germany), seeds of P farinosa harvested at the only Polish population in Beskid S4clecki and shoot tips of H. epipactis from private garden. The MS medium supplemented with auxins NAA, IBA and IAA in various concentrations combined with cytokinins BAP and kinetin, solidified with 0.8% agar, pH 5.8 were used for propagation. Rooting was carried out on MS medium with addition of 0.1 mg • dm-3 IAA, NAA or IBA. Rooted H. epipactis plants were planted into pots with mixture of penile and peat substrate (1 : 2), P farinosa into mixture of peat and peat substrate (1 : 1), and E. russicum into mixture of perlite and peat substrate (1 : 1), and kept for six weeks in mini glass-houses for acctima¬tiza lion. The highest multiplication coefficient for FL epipactis was 3.8 on medium with 0.2 mg - dm-3 IBA and 3.0 mg • dm-3 BAP but growth of the species was slow and the share of small and adventitious buds was high (41.3 and 50.1% respectively). Among rooting media the best contained 0.1 tug - dm-3 of NAA, on which 50.9% shoots tooled, however 68.3% shoots developed roots also on multiplication medium with 0.1 mg dm-3 NAA and 0.5 mg • dm-3BAP. Multiplication coefficient of P. farinosa ranged from 1.5 to 5.0 on multiplication and rooting media. There were few multiplication media that supported efficient (4.0 and higher) multiplication: 0.1 mg • dm-3 IAA + 0.2 mg • dm-3 BAP, 0.1 mg • dm-3 IAA + 0.05 mg - dm-3BAP + 0.5 mg • dm-3 GA3, 0.05 mg • dm-31BA + 0.1 mg • dm-' BAP, and 0.05 mg • dm-3 NAA + 0,2 mg • dm-3 BAP, and two rooting media with 0.1 mg • dm-3 IAA and 0.1 mg • din-3 IAA. Roots developed on all rooting media in more than 82.0%, but also on proliferating media with 0.1 mg • dm-3 IAA + 0.1 mg • dm-3 BAP or 0.05 tug • dm-3 IBA + 0.1 mg • dm-3 BAR Multiplication coefficient of E. russicum was the highest (11.0) on medium supplemented with 0.05 mg • elm-3 IBA and 0.2 mg • dm-3 BAP, however the percentage of vitrified shoots was also high (32.8%). Percentage of rooted shoots was more than 57% and was similar on all rooting media. In two years, 95%, on average, of planted IL epipactis plants survived acclimatization. The survival of E. russicum and P farinosa was respectively 85 and 82%. The callus culture of H. epipactis was started on fragments of leaf petioles excised from in vitro grown plants: kept in photoperiod conditions and etiolated during storage in darkness in +4°C. The explants were placed on MS medium supplemented with 2.5 mg • dm-3 NAA and 10 mg • dm-3 TDZ in darkness or photoperiod conditions. Callus developed only on etiolated explants cultured in darkness and after 3-5 months single adventitious buds developed. Leaf and inflorescence shoots of in vivo growing plants were source of explants for callus culture of P farinosa. Explants were placed on A; medium supplemented with 8.0 mg • dm-3 of picloram or 4.0 mg • dm-3 of 2,4-D and kept in darkness or photoperiod conditions. More explants developed callus on medium containing picloram, darkness favored caulogenesis, and leaf explants were better than inflorescence shoot fragments; the growth of fresh weight of callus in the best combina¬tion was 180.9%. The supplementation of media with. TDZ suppressed the callus growth. It was possible to obtain morphogenesis in callus of P farinosa after four weeks of culture on MS medium supplemented with 2.5 mg • dm-3 NAA and 10 mg - dnr3 TDZ in darkness, followed by four weeks of cultivation on MS medium without growth regulators. The differentiated structures were able to develop leaves and roots on medium MS with 0.05 mg • dm-3 IBA and 0.1 mg • dm-3 BAP. Adventitious buds of E. russicum were obtained through indirect organogcnesis on leaf explants excised from plants grown in vitro. The percentage of regenerating explants ranged depending on medium from 0-30% in photoperiod conditions and from 29.9-85.4% in darkness. The light conditions of cultivation had crucial meaning for the organogensis efficiency. The ploidy level assessed for sampled plants revealed that two out of ten plants of micropropagated E. russicum were tetraploids, and two plants of H. epipactis developed after organogenesis were triploids.
Other language title versionsApplication of in vitro techniques in integrated conservation of endangered species in Poland: !lamella epipactis 1., Primula farinosa L. and Echium russicum J.F. Gmei
Book typeMonograph
Other categories ROZPRAWA_HABILITACYJNA
Author Barbara Nowak (FoBaH / IoPBaB / DoBaPP)
Barbara Nowak,,
- Department of Botany and Plant Physiology
PublisherUniwersytet Rolniczy im. Hugona Kołłątaja w Krakowie, MNiSW [80]
Publishing place (Publisher address)Kraków
Issue year2013
Book series /Journal (in case of Journal special issue)Zeszyty Naukowe Uniwersytetu Rolniczego im. Hugona Kołłątaja w Krakowie. Rozprawy, ISSN 1899-3486, (0 pkt)
Vol513
No513
Pages100
Publication size in sheets6
Languagepl polski
Score (nominal)25
ScoreMinisterial score = 25.0, 30-03-2019, BookNotMainLanguagesAuthor
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