Ultramafic areas are critical for nickel (Ni) phytomining due to the high concentration of this element in their soils and the number of hyperaccumulators they harbor. The aim of the present study was to evaluate the potential of the Morais massif, an ultramafic area in Portugal, for phytomining using the hyperaccumulator species Alyssum serpyllifolium subsp. lusitanicum. Soil samples and A. serpyllifolium specimens were collected in four locations of the Morais massif. After determination of Ni concentrations in the samples, the results show that soil pseudo-total Ni concentrations in sites number 1 and 2 are significantly higher than in the soil samples collected in the other two locations, with 1918 and 2092 mg kg −1 , respectively. Nickel accumulation is significantly greater in the aerial parts of plants collected at sites 1, 2, and 4, presenting Ni harvestable amount means of 88.36, 93.80, and 95.56 mg per plant, respectively. These results suggest that the sites with highest potential for phytomining are sites 1, 2, and 4. A nickel agromining system in these locations could represent an additional source of income to local farmers, since ultramafic soils have low productivity for agriculture and crop production. and Bragança massifs localized in the Trás-os-Montes region and harbor the nickel hyperaccumulator species Alyssum serpyllifolium subsp. lusitanicum [11]. Hyperaccumulators are remarkable plants that accumulate extreme amounts of metals (such as cadmium, cobalt, magnesium, and zinc) in their tissues while remaining sufficiently healthy to maintain a self-sustaining population [12]. It has been hypothesized that high metal accumulation in these plants may serve various ecological functions such as metal tolerance, resistance to environmental stress, competitive strategy (allelopathy), and defense against pathogens and herbivores [13][14][15][16]. In the case of A. serpyllifolium, there is evidence that the high concentrations of Ni protect the species against herbivores [13]. The first discovery of a Ni hyperaccumulator was made in 1948 by Minguzzi and Vergnano with the species Alyssum bertolonii [17]. In 1977, Brooks et al. defined a threshold for nickel hyperaccumulation of 1000 mg kg −1 in the dry matter of plant shoots [18]. Currently, there are approximately 500 plant species identified that hyperaccumulate nickel, making up 70% of all known hyperaccumulators [19]. Nickel hyperaccumulators are widely dispersed, reflecting the distribution of serpentine soils from which they absorb this element [20]. The plant A. serpyllifolium subsp. lusitanicum T. R. Dudley & P. Silva (or Alyssum pintodasilvae) was described by Dudley in 1967 [21,22]. It belongs to the Brassicaceae family, and it is classified as an herbaceous and perennial plant that presents a ramified stem that can reach 10 to 30 cm and yellow flowers. It is endemic to Morais and Bragança massifs and can accumulate up to 8000 mg kg −1 (dry weight) of nickel [11].Phytomining is a recent phytotechnology based on the use of hyperaccumulators t...