In vitro spore germination and phytoremediation of Hg and Pb using gametophytes of Pityrogramma calomelanos

Few pteridophytes have proven the capacity to accumulate and remediate heavy metals from contaminated soils. Pityrogramma calomelanos, a non-seasonal fast-growing, a cosmopolitan fern, is a good indicator of environmental conditions, was used in the present study. The life cycle of ferns alternates with haploid gametophyte and diploid sporophyte. The present study was undertaken to access the effect of mercury, in form of mercury (II) chloride [(HgCl₂)] and lead as lead nitrate [Pb(NO₃)₂] in developmental studies using in vitro spore germination. Periodic recording of the germination, protonemal growth, rhizoid formation and differentiation of sex organs in different concentrations of heavy metals were conducted for a period of 6 weeks. It was found that the percentage of spore germination and the number of protonemal cells reduced significantly causing developmental defects in the presence of HgCl₂ as compared to Pb(NO₃)₂ (p < 0.0001). A significant decrease in the number of archegonial count and chlorophyll content was observed in different concentrations of the heavy metals tested. Gametophytes of P. calomelanos recorded lead uptake of 646.51 ± 0.93 mg/kg in treatments of 25 ppm of lead and high mercury accumulation up to 1,885 ± 1.98 mg/kg at 10 ppm of mercury, indicating successful uptake of heavy metals. Novelty statement: Pityrogramma calomelanos is gaining interest amongst pteridologists upon proving its exclusive capacity of phytoremediation. It is superior in comparison to the most popular, patented fern Pteris vittata. Our study demonstrates the effective use of the promising fern in its simplest, nonvascular form of gametophytes as a phytoremediation agent in controlled conditions.

Keywords: Pityrogramma calomelanos; Gametophytes; heavy metals; phytoremediation.