Effect of water quality on the spatial distribution of charophytes in the Peshawar Valley, Khyber Pakhtunkhwa, Pakistan

The presented research was conducted in 2018–2019 in the Peshawar Valley, Pakistan, to study for the first time the effect of water quality on the spatial distribution of charophytes. A total of six taxa of charophytes were found at 41 sites in the Peshawar Valley along the banks of seven rivers, 16 streams and two wetlands: Chara braunii C.C.Gmelin, C. connivens Salzmann ex A. Braun, C. contraria A. Braun ex Kützing, C. globularis Thuiller, C. vulgaris Linnaeus, and Nitellopsis obtusa (Desvaux) J. Groves. Chara vulgaris was the most abundant species, followed by C. globularis, and C. contraria. Water pH, electrical conductivity (EC), total dissolved solids (TDS), salinity and dissolved oxygen (DO) were within the permissible limits for Pakistan, while water temperature, oxidation reduction potential (ORP) and resistivity showed deviations. Canonical Correspondence Analysis (CCA) revealed that DO affected Chara vulgaris, pH and resistivity affected C. braunii, C. connivens and C. globularis, temperature and ORP affected C. contraria and Nitellopsis obtusa. Furthermore, CCA showed that TDS, EC, and salinity had no effect on the spatial distribution of Chara contraria, C. vulgaris and Nitellopsis obtusa. Chara contraria and Nitellopsis obtusa should be protected under VU (Vulnerable) status (IUCN) along with their habitats.

Impact of Nitellopsis obtusa (Desv.) J. Groves, a regionally alien and invasive charophyte, on macrophyte diversity in the species native range

This study aimed to determine the relationships between the abundance of Nitellopsis obtusa, a controversial charophyte, locally red-listed in its native Eurasian range but invasive in North America, and the species diversity of macrophyte stands dominated by N. obtusa. Three lakes of different morphology, productivity and catchment were surveyed in the species native range. In each lake, the species composition and cover of three N. obtusa-dominated stands were determined monthly from spring to autumn and illustrated by the Shannon–Wiener diversity index. Water chemistry supplemented vegetation study. The species diversity turned out to be lake-specific and declined with the increasing share of N. obtusa, which developed mass occurrence in less mineralised and less fertile waters, leaving no space and limiting light and nutrient availability for large and branchy macrophytes. We postulate that this mechanism makes N. obtusa a superior competitor in less fertile waters and seems common to both native and invaded territories, as is the pool of macrophyte species most frequently co-occurring with N. obtusa.

Low Benthic Oxygen and High Internal Phosphorus-Loading are Strongly Associated With the Invasive Macrophyte Nitellopsis obtusa (starry stonewort) in a Large, Polymictic Lake

Lake Scugog is an important headwater to the Trent Severn-Waterway in Ontario, Canada. In recent years, notable ecosystem-level changes have occurred coinciding with the emergence of the non-native invasive charophyte Nitellopsis obtusa. Despite N. obtusa arriving in North America in the early 1970s, studies documenting the impact of N. obtusa on invaded ecosystems are scarce. Given the increasing dominance of N. obtusa in inland waters of the Great Lakes basin, we investigated the ecosystem-level impacts of N. obtusa in Lake Scugog over a 3-year period. We show for the first time a strong association between N. obtusa occurrence and biomass with benthic anoxia in this shallow, polymictic lake. Benthic dissolved oxygen concentrations were significantly lower (p-value < 0.001) at sites with N. obtusa compared to sites without N. obtusa. Additionally, N. obtusa biomass was a negative predictor of near-bed oxygen concentration (R2 = 0.59, p-value < 0.001). Knowing that anoxia can promote the internal loading of phosphorus, we measured soluble reactive phosphorus (SRP) in the pore-water of sediments at each site, and found N. obtusa biomass explained 90% of sediment pore-water SRP (R2 = 0.90, p-value < 0.001). These notable associations between N. obtusa and key lake elements indicates that N. obtusa may be acting as an ecosystem engineer in invaded lakes by altering the biogeochemical fate of oxygen and phosphorus.

Global high-throughput genotyping of organellar genomes reveals insights into the origin and spread of invasive starry stonewort (Nitellopsis obtusa)

Aquatic invasive species are damaging to native ecosystems. Preventing their spread and achieving comprehensive control measures requires an understanding of the genetic structure of an invasive population. Organellar genomes (plastid and mitochondrial) are useful for population level analyses of invasive plant distributions. In this study we generate complete organellar reference genomes using PacBio sequencing, then use these reference sequences for SNP calling of high-throughput, multiplexed, Illumina based organellar sequencing of fresh and historical samples from across the native and invasive range of Nitellopsis obtusa (Desv. in Loisel.) J.Groves, an invasive macroalgae. The data generated by the analytical pipeline we develop indicate introduction to North America from Western Europe. A single nucleotide transversion in the plastid genome separates a group of five samples from Michigan and Wisconsin that either resulted from introductions of two closely related genotypes or a mutation that has arisen in the invasive range. This transversion will serve as a useful tool to understand how Nitellopsis obtusa moves across the landscape. The methods and analyses described here are broadly applicable to invasive and native plant and algae species, and allow efficient genotyping of variable quality samples, including 100-year-old herbarium specimens, to determine population structure and geographic distributions.

Impact of Mammalian Two-Pore Channel Inhibitors on Long-Distance Electrical Signals in the Characean Macroalga Nitellopsis obtusa and the Early Terrestrial Liverwort Marchantia polymorpha

Inhibitors of human two-pore channels (TPC1 and TPC2), i.e., verapamil, tetrandrine, and NED-19, are promising medicines used in treatment of serious diseases. In the present study, the impact of these substances on action potentials (APs) and vacuolar channel activity was examined in the aquatic characean algae Nitellopsis obtusa and in the terrestrial liverwort Marchantia polymorpha. In both plant species, verapamil (20–300 µM) caused reduction of AP amplitudes, indicating impaired Ca2+ transport. In N. obtusa, it depolarized the AP excitation threshold and resting potential and prolonged AP duration. In isolated vacuoles of M. polymorpha, verapamil caused a reduction of the open probability of slow vacuolar SV/TPC channels but had almost no effect on K+ channels in the tonoplast of N. obtusa. In both species, tetrandrine (20–100 µM) evoked a pleiotropic effect: reduction of resting potential and AP amplitudes and prolongation of AP repolarization phases, especially in M. polymorpha, but it did not alter vacuolar SV/TPC activity. NED-19 (75 µM) caused both specific and unspecific effects on N. obtusa APs. In M. polymorpha, NED-19 increased the duration of repolarization. However, no inhibition of SV/TPC channels was observed in Marchantia vacuoles, but an increase in open probability and channel flickering. The results indicate an effect on Ca2+ -permeable channels governing plant excitation.

Biomarker identification of isolated compartments of the cell wall, cytoplasm and vacuole from the internodal cell of characean Nitellopsis obtusa

Cells of characean algae are attractive for plant cell physiologists because of their large size and their close relation to higher plant cells. The objective of our study was to evaluate the purity of the compartments (cell wall, cytoplasm with plastids, mitochondria, nuclei and endomembrane system, and vacuole) separated mechanically from the internodal cells of Nitellopsis obtusa using enzymatic markers. These included α-mannosidase and malate dehydrogenase, vacuolar and cytoplasmic enzymes, respectively. The biomarkers applied revealed the degree of compartment contamination with the material from unwanted cell parts. The cell wall was contaminated slightly by vacuole and cytoplasm residuals, respectively by 12.3 and 1.96% of corresponding biomarker activities. Relatively high activity of vacuolar marker in the cell wall could be associated with the cell vacuoles in the multicellular structure of the nodes. The biomarkers confirmed highly purified vacuolar (99.5%) and cytoplasmic (86.7%) compartments. Purity estimation of the cell fractions enabled reevaluating nCuO related Cu concentrations in the compartments of charophyte cell. The internalisation of CuO nanoparticles in N. obtusa cell occurred already after 0.5h. In general, the approach seems to be useful for assessing the accumulation and distribution of various xenobiotics and/or metabolites within plant cell. All this justifies N.obtusa internodal cells as a model organism for modern studies in cell biology and nanotoxicology.