Summary Thlaspi caerulescens (Brassicaceae) is a promising plant model with which to study heavy metal hyperaccumulation. Population genetics studies are necessary for a better understanding of its history, which will be useful for further genomic studies on the evolution of heavy metal hyperaccumulation. The genetic structure of 24 natural Swiss locations was investigated using nuclear and plastid loci. Population genetics parameters were estimated and genetic pools were identified using Bayesian inference on eight putatively neutral nuclear loci. Finally, the effect of cadmium (Cd) and zinc (Zn) soil concentrations on genetic differentiation at loci located in genes putatively involved in heavy metal responses was examined using partial Mantel tests in Jura, western Switzerland. Four main genetic clusters were recognized based on nuclear and plastid loci, which gave mostly congruent signals. In Jura, genetic differentiation linked to heavy metal concentrations in soil was shown at some candidate loci, particularly for genes encoding metal transporters. This suggests that natural selection limits gene flow between metalliferous and nonmetalliferous locations at such loci. Strong historical factors explain the present genetic structure of Swiss T. caerulescens populations, which has to be considered in studies testing for relationships between environmental and genetic variations. Linking of genetic differentiation at candidate genes with soil characteristics offers new perspectives in the study of heavy metal hyperaccumulation.

Genetic polymorphism was investigated in Thlaspi caerulescens J. & C. Presl at 15 gene regions, of which seven have been identified to putatively play a role in heavy-metal tolerance or hyperaccumulation. Single nucleotide and length polymorphisms were assessed at four cleaved amplified polymorphic sequences (CAPS) and 11 simple sequence repeat (microsatellite) loci, respectively. The utility of these loci for genetic studies in T. caerulescens was measured among seven natural populations (135 individuals). Fourteen loci rendered polymorphism, and the number of alleles per locus varied from 2 to 5 and 1 to 27 for CAPS and microsatellites, respectively. Up to 12 alleles per locus were detected in a population. The global observed heterozygosity per population varied between 0.01 and 0.31. Additionally, cross-species/genera amplification of loci was investigated on eight other Brassicaceae (five individuals per population). Overall, 70% of the cross-species/genera amplifications were successful, and among them, more than 40% provided intraspecific polymorphisms within a single population. This indicates that such markers may, as well, allow comparative population genetic or mapping studies between and within several Brassicaceae, particularly for genes involved in traits such as heavy-metal tolerance and/or hyperaccumulation.

During the last decade, the metal hyperaccumulating plants have attracted considerable attention because of their potential use in decontamination of heavy metal contaminated soils. However, in most species, little is known regarding the function, the ecological and the evolutionary significances of hyperaccumulation. In our study, we investigated the parameters influencing the Cd concentration in plants as well as the biological implications of Cd hyperaccumulation in nine natural populations of Thlaspi caerulescens. First, we showed that Cd concentration in the plant was positively correlated with plant Zn, Fe, and Cu concentrations. This suggested that the physiological and/or molecular mechanisms for uptake, transport and/or accumulation of these four heavy metals interact with each other. Second, we specified a measure of Cd hyperaccumulation capacity by populations and showed that T. caerulescens plants originating from populations with high Cd hyperaccumulation capacity had better growth, by developing more and bigger leaves, taller stems, and produced more fruits and heavier seeds. These results suggest a tolerance/disposal role of Cd hyperaccumulation in this species.

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Abstract. In order to fully understand the hyperaccumulation process and to increase the potential of plants for phytoextraction purposes, there is a need for more investigation of hyperaccumulating species or populations. Five Swiss populations of Thlaspi caerulescens J. & C. Presl originating from non-metalliferous but naturally Cd-rich soils (1.1-9.2 mg Cd kg −1 ) were compared with Ganges and Prayon populations and a non-accumulating species, Thlaspi perfoliatum (L.) F.K. Meyer, for their tolerance (shoot and root dry weight and root length) and Cd hyperaccumulation in hydroponics (0, 1, 5, 10, 20 and 50 µM Cd). In the field, the Swiss populations accumulated Zn and clearly hyperaccumulated Cd (up to 505 mg Cd kg −1 dry weight). The general response was significantly different between populations but in general an increasing Cd concentration in solution led to a decrease in dry weight production and an increase in Cd concentration in shoots. The shoot dry weight was a more discriminating parameter for tolerance than root dry weight and total root length. The Swiss populations behaved similarly to the Ganges population but differently from the Prayon population. Cadmium concentrations in shoots were above 100 mg kg −1 when plants were grown in 1 µM Cd, except for the Prayon population and T. perfoliatum. In addition, as 1 µM Cd did not induce any visible toxicity symptoms, it was found to be adequate to test Cd hyperaccumulation. However, the most striking feature was the positive linear relationship observed between the transfer factor (TF) calculated in the field and the response of a population to increasing Cd concentrations in solution, indicating that plant uptake in the field had an influence on the plant response in solution.