Serpentine environment prevails over geographic distribution in shaping the genetic diversity of Medicago lupulina L.

Edaphic conditions of serpentine soils, naturally rich in heavy metals, act as a strong selection pressure that shapes specific metal-tolerant ecotypes. Medicago lupulina L. (black medick) is not only a widespread plant species that prefers calcareous and dry soil types but also grows at the borders of serpentine formations. It can also be found in waste and disturbed habitats. This is a species with reported phytoremediation potential, however, there is no published data regarding the impact of the environment on the genetic distribution of this species. The aim of our research was to explore how selection pressure of serpentine soils affects genetic diversity of M. lupulina and to test heavy-metal accumulation capacity of this species. Specimens of 11 M. lupulina populations were collected from serpentine outcrops located in Central and Eastern Bosnia as well as from non-serpentine sites. Soil and plant samples were analyzed for the total contents of heavy metals using air-acetylene flame atomic absorption spectroscopy. Genetic diversity was analyzed using AFLP (Amplified Fragment Length Polymorphism) markers. Serpentine soils showed high nickel, cobalt, chromium and iron concentrations. Nickel and manganese concentrations in soil samples and plant material showed statistically significant correlation. Although plants in two populations show the ability to extract Ni, M. lupulina does not show hyperaccumulating properties. Despite severe selective pressure, genetic diversity in serpentine populations is not reduced. Analyses of intrapopulation and interpopulation genetic diversity showed significant genetic differentiation among populations which is not related to their geographic distance. Population from non-metalliferous soil showed clear separation from all other populations. Diversity data suggest that serpentine populations maintain genetic diversity by undetected mechanisms and that edaphic factors rather than geography influence genetic structure analyzed M. lupulina populations.

Geranium robertianum L. tolerates various soil types burdened with heavy metals.

Many heavy metals (HMs) are essential micronutrients for the growth and development of plants. However, human activities such as mining, smelting, waste disposal, and industrial processes have led to toxic levels of HMs in soil. Fortunately, many plant species have developed incredible adaptive mechanisms to survive and thrive in such harsh environments. As a widespread and ruderal species, Geranium robertianum L. inhabits versatile soil types, both polluted and unpolluted. Considering the ubiquity of G. robertianum, the study aimed to determine whether geographically distant populations can tolerate HMs. We collected soil and plant samples from serpentine, an anthropogenic heavy metal contaminated, and a non-metalliferous site to study the physiological state of G. robertianum. HMs in soil and plants were determined using flame atomic absorption spectrometry. Spectrophotometric methods were used to measure the total content of chlorophylls a and b, total phenolics, phenolic acids, flavonoids, and proline. Principal component analysis (PCA) was used to investigate the potential correlation between HMs concentrations gathered from various soil types and plant samples and biochemical data acquired for plant material. A statistically significant difference was observed for all localities regarding secondary metabolite parameters. A positive correlation between Ni and Zn in soil and Ni and Zn in plant matter was observed (p<0.0005) indicating higher absorption. Regardless of high concentrations of heavy metals in investigated soils, G. robertianum displayed resilience and was capable of thriving. These results may be ascribed to several protective mechanisms that allow G. robertianum to express normal growth and development and act as a pioneer species.