Aldrovanda vesiculosa is a carnivorous water plant which is endangered by extinction worldwide. The number of natural stands and populations has decreased; therefore, there is a need for its active protection. The best method would be an in vitro culture. One of the main problems is disinfection of the explants. Therefore, it was decided that we should treat the explants with nano-silver particles. The explants were shoot fragments which were disinfected with sodium hypochlorite and then placed in a liquid 1/5 MS medium, supplemented with silver nanoparticles (AgNPs) at a concentration of 5 mg·dm⁻³. It was observed that AgNPs reduced the number of contaminations but also led to necrosis of the shoots. The shoots, which undertook regeneration in presence of AgNPs, were smaller and did not form traps; however, after being moved to fresh media twice, they started to develop normal leaves. Taking into consideration both disinfection and regeneration rates, it might be advisable to disinfect aldrovanda shoots in sodium hypochlorite only, without AgNPs. The results of the research might indicate a toxic activity of AgNPs towards water plants, which seems a big problem, as nanoparticles are commonly used in all the fields of life. However, the matter should be studied further.

To obtain healthy and good quality plants from in vitro cultivation, it is necessary to produce plantlets with well-developed rooting systems because they must undergo acclimatization, a final and a very difficult stage of micropropagation. In the present research, the effect of auxins NAA, IAA and IBA in concentrations of 0.5; 1; 2.5 and 5 mg·dm⁻³ on the Paphiopediluminsigne in vitro rooting was studied, and it was noted that 1 mg·dm⁻³ of IAA or IBA enabled the obtaining of a lot of rooted and good quality plantlets. The subsequent influence of the two most advantageous auxins on the acclimatization of plantlets in different substrates (sphagnum moss, sphagnum moss + substrate for orchids, substrate for orchids, substrate for orchids + acid peat) was tested, in the means of morphological features of plants and their physiological parameters, i.e., chlorophyll fluorescence (FV, Fm, Fv/Fm), stress enzyme activity (catalase, ascorbate peroxidase), and water balance. Considering all the tested features, it might be stated that the best results were obtained when explants were rooted in vitro in the presence of 1 mg·dm⁻³ of IAA and then planted ex vitro in substrate for orchids.

Initiation of tissue culture of many plant species is a very difficult stage due to appearance of many contaminations. The other problem might be a choice of media for regeneration. Initiation of grass species tissue cultures are thought to be very difficult. Therefore, a research was undertaken to evaluate the use of nano-silver particles for plant material disinfection and to estimate a medium Pennisetum alopecuroides. The plant material were buds and nodal explants that were disinfected in 2% NaOCl for 30 min or 0.1% HgCl2 for 1 min. Half of the explants disinfected with NaOCl were soaked in 50, 100 or 250 mg·dm Ag–3 NPs for 1 hour. Explants not soaked in nano-silver were placed on media with Ag NPs at concentrations of 4, 8 or 16 mg·dm–3. An influence of growth regulators on Pennisetum alopecuroides was evaluated in vitro. Regenerated shoots were placed on MS media with: 3 mg·dm–3 BA + 0.3 mg·dm–3 IBA, 3 mg·dm–3 KIN + 0.3 mg·dm–3 IAA, 1 mg·dm–3 BA + 0.1 mg·dm–3 IBA. It was observed that the use of nano-silver particles lowered the level of contamination. The best results were obtained when Ag NPs was used at concentration of 100–250 mg·dm–3 alone or as a supplementation of the media, at concentration of 4 mg·dm–3 for nodes and 16 mg·dm–3 for adventitious buds. The use of nodal explants allowed to obtain less contamination. Regeneration depended on a media content. The most regenerated shoots were obtained on the MS media supplemented with 1 mg·dm–3 BA and 0.1 mg·dm–3 IBA.

Salix lapponum L. is a boreal relict species, threatened with extinction in Poland. An 80% decrease in the number of its stands was confirmed in the last half-century, so that to prevent the loss of downy willow, attempts were made to reintroduce this species in natural habitats. Micropropagation was chosen as a first stage of its active conservation. S. lapponum shoots were collected and disinfected with NaOCl, AgNO₃, or HgCl₂ or with a two-step disinfection with NaOCl and then placed on MS medium with BA 1 mg·dm⁻³ and IBA 0.1 mg·dm⁻³. Regenerated shoots were cultivated with addition of BA, KIN, or 2iP, alone or in combination with auxins, to find the highest multiplication rate. Inter-simple sequence repeat (ISSR) analysis and flow cytometric analyses were conducted on in vitro regenerated plants to check their genetic stability. Disinfection was quite difficult and the use of HgCl₂ was the most efficient. The highest multiplication rate was obtained in presence of KIN at 0.5 mg·dm⁻³ + IAA at 0.5 mg·dm⁻³. The analysis confirmed the genome size stability, which is in agreement with the results obtained by ISSR, revealing no somaclonal variation in plantlets and therefore allowing the use of the obtained plants for reintroduction.

Salix myrtilloides L. is a relict species, threatened with extinction in many European countries. To prevent the loss of the species, tissue culture was established to produce plant material for reintroduction in natural habitats. Micropropagation was chosen as a method to obtain new plants. S. myrtilloides shoots were disinfected with NaOCl, AgNO₃, or with a two-step disinfection with NaOCl, and then placed on MS medium supplemented with BA at 1 mg·dm⁻³ and IBA at 0.1 mg·dm⁻³. Regenerated shoots were cultivated in presence of BA, KIN, and 2iP to select the treatment with the highest multiplication rate. The obtained plants were acclimatized to ex vitro conditions. Inter-simple sequence repeat (ISSR) and flow cytometric analyses were conducted on in vitro regenerated plants to check their genetic stability. The best disinfection results were obtained when explants were treated with 1.5% NaOCl for 20 min. The highest multiplication rate and good quality plants were noted in the control media, without growth regualtors and in presence of kinetin at 0.5 mg·dm⁻³. Flow cytometry and ISSR analyses confirmed genetic stability in plantlets, which indicated the possibility to use the in vitro obtained plants for reintroduction.

Effect of activated charcoal and ascorbic acid on in vitro morphogenesis and o-dihydroxyphenols content in Paphiopedilum insigne | Monika Poniewozik, Marzena Parzymies, Paweł Szot | Agricultural Journals