Roots can produce mechanical and chemical alterations to building structures, especially in the case of underground historical artifacts. In archaeological sites, where vegetation plays the dual role of naturalistic relevance and potential threat, trees and bushes are under supervision. No customized measures can be taken against herbaceous plants lacking fast and reliable root identification methods that are useful to assess their dangerousness. In this study, we aimed to test the efficacy of DNA barcoding in identifying plant rootlets threatening the Etruscan tombs of the Necropolis of Tarquinia. As DNA barcode markers, we selected two sections of the genes rbcL and matK, the nuclear ribosomal internal transcribed spacer (nrITS), and the intergenic spacer psbA-trnH. All fourteen root samples were successfully sequenced and identified at species (92.9%) and genus level (7.01%) by GenBank matching and reference dataset implementation. Some eudicotyledons with taproots, such as Echium italicum L., Foeniculum vulgare Mill., and Reseda lutea L. subsp. lutea, showed a certain recurrence. Further investigations are needed to confirm this promising result, increasing the number of roots and enlarging the reference dataset with attention to meso-Mediterranean perennial herbaceous species. The finding of herbaceous plants roots at more than 3 m deep confirms their potential risk and underlines the importance of vegetation planning, monitoring, and management on archaeological sites.

The conservation of underground tombs is affected by several physical-chemical and biological factors, which could be reduced by insulating systems able to maintain the microclimatic stability also decreasing the biodeterioration risk. In Mediterranean areas, wild ephemeral plants, which reduce their cover during the hot season, seem unsuitable for reducing summer overheating. In this study, we wish to assess the influence of vegetation cover and of overlaying soil, after the establishment of an evergreen turf of a cultivar of Cynodon dactylon, on two tombs in the Etruscan Necropolis of Monterozzi, covered by linear-shaped tumuli. Therefore, we evaluated for 10 months the thermo-hygrometric values of these tombs, together with two tombs as controls. We also evaluated the different tumuli’s morphologies and the related received solar radiation. Results confirmed that late summer and early autumn as critical microclimatic periods for the risk factors of hypogeal paintings when peaks of superficial temperature occur. A positive influence of vegetation cover on maintaining constant humidity and internal temperatures was detected, but the mounds orientation, as well as soil depth, seems to have a relevant role. Considering the naturalistic features of the area and the related cultural ecosystem services, a careful selection of wild plants is suggested.