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In the Mediterranean basin, agriculture and other forms of human land use have shaped the environment since ancient times. Intensive and extensive agricultural systems managed with a few cultured plant populations of improved varieties are a widespread reality in many Mediterranean countries. Despite this, historical cultural landscapes still exist in interior and less intensively managed rural areas. There, ancient fruit tree varieties have survived modern cultivation systems, preserving a unique genetic heritage. In this study, we mapped and characterized 106 living fruit trees of ancient varieties in the Basilicata region of southern Italy. Tree ages were determined through tree ring measurements and radiocarbon analyses. We uncovered some of the oldest scientifically dated fruit trees in the world. The oldest fruit species were olive (max age 680 ± 57 years), mulberry (647 ± 66 years), chestnut (636 ± 66 years), and pear (467 ± 89 years). These patriarchs hold a unique genetic resource; their preservation and genetic maintenance through agamic propagation are now promoted by the Lucan Agency for the Development and Innovation in Agriculture (ALSIA). Each tree also represents a hub for biodiversity conservation in agrarian ecosystems: their large architecture and time persistence guarantee ecological niches and micro-habitats suitable for flora and fauna species of conservation significance. [ABSTRACT FROM AUTHOR] more...