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Lupinus mariae-josephae  is a recently discovered endemism that is only found in alkaline-limed soils, a unique habitat for lupines, from a small area in Valencia region (Spain). In these soils,  L. mariae-josephae  grows in just a few defined patches, and previous conservation efforts directed towards controlled plant reproduction have been unsuccessful. We have previously shown that  L. mariae-josephae  plants establish a specific root nodule symbiosis with bradyrhizobia present in those soils, and we reasoned that the paucity of these bacteria in soils might contribute to the lack of success in reproducing plants for conservation purposes. Greenhouse experiments using  L. mariae-josephae  trap-plants showed the absence or near absence of  L. mariae-josephae -nodulating bacteria in “terra rossa” soils of Valencia outside of  L. mariae-josephae  plant patches, and in other “terra rossa” or alkaline red soils of the Iberian Peninsula and Balearic Islands outside of the Valencia  L. mariae-josephae  endemism region. Among the bradyrhizobia able to establish an efficient symbiosis with  L. mariae-josephae  plants, two strains, LmjC and LmjM3 were selected as inoculum for seed coating. Two planting experiments were carried out in consecutive years under natural conditions in areas with edapho-climatic characteristics identical to those sustaining natural  L. mariae-josephae  populations, and successful reproduction of the plant was achieved. Interestingly, the successful reproductive cycle was absolutely dependent on seedling inoculation with effective bradyrhizobia, and optimal performance was observed in plants inoculated with LmjC, a strain that had previously shown the most efficient behavior under controlled conditions. Our results define conditions for  L. mariae-josephae  conservation and for extension to alkaline-limed soil habitats, where no other known lupine can thrive.

Conservation of Endangered Lupinus mariae-josephae in Its Natural Habitat by Inoculation with Selected, Native Bradyrhizobium Strains