Tolerance to zinc and lead of the suspension cells derived from metallophytes of the calamine soils

W niniejszej pracy badano poziom tolerancji komórek kultur zawiesinowych trzech wybranych gatunków metalofitów fakultatywnych z terenów metalonośnych: Arabidopsis halleri subsp. halleri, Armeria maritima subsp. halleri, Silene vulgaris subsp. humilis w celu odniesienia uzyskanych wyników do uzyskanych wcześniej, a opartych o analogiczne eksperymenty, nad kilkoma gatunkami metalofitów fakultatywnych, obligatoryjnych, niemetalofitów z rodzaju Viola. Komórki badanych metalofitów różniły się przeżywalnością po osobnym traktowaniu solami cynku i ołowiu (200 µM, 500 µM, 1000 µM) przez 24, 48 i 72 godziny oszacowaną testem alamarBlue®. Komórki A. maritima subsp. halleri, S. vulgaris subsp. humilis były wysoce tolerancyjne na cynk, w przeciwieństwie do A. halleri subsp. halleri. Wszystkie analizowane metalofity wykazywały niską tolerancję na ołów. W szczególności zaskakująca jest niska tolerancja na cynk i ołów A. halleri subsp. halleri, hiperakumulatora cynku i kadmu. Komórki wybranych metalofitów gleb galmanowych wykazały niższą tolerancję na cynk i ołów niż gatunki z rodzaju Viola. Uzyskane wyniki potwierdzają hipotezę, że gatunki z rodzaju Viola, niezależnie od ich statusu (metalofit vs. niemetalofit), mają wrodzoną, wysoką tolerancję na metale ciężkie.In this study, the tolerance level of suspended cells derived from three selected species of facultative metallophytes from metalliferous areas: Arabidopsis halleri subsp. halleri, Armeria maritima subsp. halleri, Silene vulgaris subsp. humilis was examined in order to compare the obtained results with analogous experiment on facultative,obligatory metallophytes and non-metallophytes of the Viola genus. The tested metallophyte cells differed in the survival rate after treatment with zinc and lead salts (200 µM, 500 µM, 1000 µM) for 24, 48 and 72 hours assessed by the alamarBlue test®. A. maritima subsp. halleri, S. vulgaris subsp. humilis cells were highly tolerant to zinc, in contrast to A. halleri subsp. halleri. All three analysed metallophytes showed low tolerance to lead. In particular, low tolerance to zinc and lead of A. halleri subsp. halleri, zinc and cadmium hyperaccumulator, was surprising. The cells of selected species of calamine metallophytes showed lower tolerance to zinc and lead than species of the genus Viola independently of their status (metallophyte vs. non-metallophyte). The obtained results support the hypothesis that species of the genus Viola have an innate, high tolerance to heavy metals

Konstytutywna tolerancja na metale ciężkie w rodzaju Viola L. (Violaceae Batsch.) na podstawie badań przeżywalności, parametrów fizjologicznych oraz gromadzenia metali w komórkach zawiesinowych.

Badania miały na celu potwierdzenie wrodzonej tolerancji na metale ciężkie u gatunków z rodzaju Viola L. (fiołki) W tej grupie występuje wiele gatunków, które rosną na glebach z metalami ciężkimi, od fakultatywnych metalofitów po hiperakumulatory. Nie wiadomo, czy jest to wynikiem stopniowych procesów mikroewolucyjnych prowadzących do adaptacji, czy cecha ta została odziedziczona po tolerancyjnym przodku (-ach).Wyprowadzono kultury zawiesinowe komórek z tkanki kalusa pięciu roślin: dwóch niemetalofitów; Arabidopsis thaliana (Col-0) i Viola x wittrockiana oraz trzech metalofitów; V. philippica, V. tricolor i Silene vulgaris subsp. humilis. Zmierzono żywotność komórek, zawartość nieenzymatycznych przeciwutleniaczy i akumulację metali ciężkich po traktowaniach komórek Zn lub Pb.Komórki gatunków z rodzaju Viola charakteryzowały się wysoką tolerancją na Zn i Pb (1000 μM), niezależnie od ich statutu ekologicznego (niemetalofity vs metalofity). Fiołki będące -metalofitami wykazywały wysoką zawartość przeciwutleniaczy, co świadczy o ich wysokiej reaktywności na metale ciężkie. U wszystkich gatunków ołów wykryto w protoplazmie komórek, nie w wakuoli lub ścianie komórkowej. Komórki wszystkich gatunków fiołków charakteryzowały się zdolnością do akumulacji ołowiu.Wyniki wskazują na wrodzoną (konstytutywną) tolerancję fiołków na metale ciężkie. Co więcej, wyniki uzyskane w badaniach fizjologicznych i biochemicznych na komórkach roślinnych przekładają się na tolerancję na metale ciężkie na poziomie całej rośliny.This research aimed to evaluate hypothesis of innate tolerance to heavy metals within Viola L. genus (violets). Violets enclose many species, which are heavy metal-tolerant, ranging from facultative metallophytes to hyperaccumulators. It is unknown, whether this tolerance is a re-sult of gradual microevolutionary adaptive processes or whether this trait was inherited from tolerant ancestors(s).Cell suspension cultures from callus tissue of five plants: two non-metallophytes; Arabidopsis thaliana (Col-0) and Viola x wittrockiana, and three metallophytes; V. philippica, V. tricolor and Silene vulgaris subsp. humilis were developed. Cell viability, non-enzymatic antioxidant content and the accumulation of heavy metals after cell treatment with Zn or Pb were exam-ined.Cells of violets were characterized by high tolerance, independently on their ecological status (non-metallophyte vs metallophyte), after exposure to Zn or Pb (1000 μM). Metallophytes belonging to Viola genus had high content of antioxidants, indicating their high responsivity to these metals. In all species, lead was detected in the protoplasm of the cells, not in the vac-uole or cell wall. All violets were characterized by the capacity to accumulate lead.The results indicate innate (constitutive) tolerance to heavy metals in violets. Furthermore, physiological and biochemical results obtained using plant cells translate into heavy metal tol-erance of the whole plant

Low and high elevation Heliosperma species (Caryophyllaceae) : insight based on chromosome number, pollen characters and seed micromorphology

Miszczak, Szymon, Shuka, Donald, Shuka, Lulëzim, Migdałek, Grzegorz, Słomka, Aneta (2022): Low and high elevation Heliosperma species (Caryophyllaceae)-insight based on chromosome number, pollen characters and seed micromorphology. Phytotaxa 554 (1): 32-46, DOI: 10.11646/phytotaxa.554.1.2, URL: http://dx.doi.org/10.11646/phytotaxa.554.1.

Innate, high tolerance to zinc and lead in violets confirmed at the suspended cell level

Many species of the Viola L. genus (violets) colonize areas with high concentrations of trace elements in the soil, e.g., nickel, cadmium, zinc, and lead. Although tolerance to heavy metals is a common phenomenon in violets, it is not clear whether this is the result of gradual microevolutionary processes as a part of the adaptation to the specific conditions, or whether the tolerance was inherited from the ancestor(s). We developed cell suspension cultures of five plant species: two non-metallophytes—Arabidopsis thaliana (Col-0) and Viola · wittrockiana, and three metallophytes—V. philippica, V. tricolor, and Silene vulgaris subsp. humilis for tolerance tests. The aim of the study was to measure the level of tolerance of violets in comparison with species from the other genera to verify the hypothesis of the high, innate tolerance of the former. We measured cell viability, non-enzymatic antioxidant content, and the accumulation of heavy metals after cell treatment with Zn or Pb. The results indicate they are innate and independent on the ecological status (metallophyte vs. non-metallophyte) and high in comparison with other species tolerance to Zn and Pb in violets. Viability of the cells after Zn and Pb (1000 μM) exposure for 72 h was the highest in violets. Antioxidant content, after heavy metal treatment, increased significantly, particularly in metallophyte violets, indicating their high responsivity to metals. In all species, lead was detected in the protoplasm of the cells, not in the vacuole or cell wall. All violets were characterized by the accumulation capacity of lead. Here, we clearly show that the physiological and biochemical studies conducted with the use of heavy metals on plant cells translate into the heavy metal tolerance of the species

Stable Artificial Autopolyploids of the Zn/Cd Accumulator Arabidopsis arenosa—A Promising Genetic Resource for Phytoremediation

Arabidopsis arenosa is a good candidate for phytoremediation due to its high tolerance to Zn and Cd as well as its accumulation ability. However, its small size and low biomass are the largest obstacles to applying it on a broad scale. The aim was to obtain polyploid specimens, which tend to have higher biomass to increase the accumulation and translocation capacity of heavy metals in this metal-tolerant plant. Doubled polyploids (octaploids) were obtained via indirect organogenesis on a ½ MS medium supplemented with 1 mg L−1 TDZ, followed by rooting on the same medium without growth regulators. Callus tissue of a high endopolyploidy level (the (Ʃ>2C)/2C ratio over 2.5) obtained on seedling fragments on ½ MS supplemented with 2 mg L−1 2,4-D + 2 mg L−1 BAP served as a source material. Among the regenerants successfully obtained (without using antimitotic agents), over half, regardless of the stage of regenerant development, were octaploid (54–78%; 2C DNA = 1.642 pg). Octaploids were not affected by ploidy or in vitro culture conditions; they were fully fertile, produced normal pollen (~97% of viability), and set seeds capable of germinating (78%). Their cell and organ size was affected by genome doubling resulting in longer stomata, bigger pollen grains, and flowers with a larger area and width in comparison with tetraploid regenerants and initial plants. The promising results of measurements of morpho-anatomical, physiological, and reproductive parameters indicate that, in the future, after passing tolerance tests, the obtained polyploids could be used in phytoremediation of metal-contaminated areas