Although genetic diversity has been recognized as a key component of biodiversity since the first Convention on Biological Diversity (CBD) in 1993, it has rarely been included in conservation policies and regulations. Even less appreciated is the role that ancient and historical DNA (aDNA and hDNA, respectively) could play in unlocking the temporal dimension of genetic diversity, allowing key conservation issues to be resolved, including setting baselines for intraspecies genetic diversity, estimating changes in effective population size (N e) , and identifying the genealogical continuity of populations. Here, we discuss how genetic information from ancient and historical specimens can play a central role in preserving biodiversity and highlight specific conservation policies that could incorporate such data to help countries meet their CBD obligations. Genetic biodiversityThree levels of biodiversity constitute the variation of life on our planet: diversity of ecosystems, species diversity (number and distribution of species), and genetic diversity (amount and distribution of genetic variation within species or populations). The need to monitor biodiversity at all three levels has been globally recognized in international policy since 1993 when the Convention on Biological Diversity i (CBD) came into effect. Today, we face dramatic biodiversity loss due to the combined effects of habitat damage, fragmentation and alteration, climate change, and other global change stressors. Most frequently, this loss is calculated in terms of the number of species, but relatively little is known about loss of diversity within species and populations at the genome level (but see [1]). Genetic diversity within species and populations is necessary for long-term survival as it allows resilience and adaptation not only for individuals, but also for populations, species, and entire ecosystems [2]. This diversity is particularly relevant in the Anthropocene, characterized by significant, rapid, and global changes to habitats and environmental conditions. Despite the importance of genetic diversity in biodiversity protection and management, it has rarely been included in policies and regulations [3]. But, with the ongoing development of the CBD post-2020 Global Biodiversity Framework (expected to be concluded in May 2022), there is an opportunity to address this significant blind spot by adopting genetic diversity targets and indicators.

A total of 132 mtDNA sequences from 10 Balkan donkey populations were analysed to ascertain their regional genetic structure and to contribute to the knowledge of the spreading of the species after domestication. The Balkan donkey sequences were compared with those from 40 Burkina Faso donkeys as an African outgroup to account for possible local Balkan scenarios. The 172 sequences gave 62 different haplotypes (55 in Balkan donkey). Virtually all the analysed populations had haplotypes assigned to either Clade 1 or Clade 2 even though the relative proportion of Clade 1 or 2 haplotypes differed across populations. Geographical maps constructed using factors computed via principal component analysis showed that the Balkan donkey populations are not spatially structured. AMOVA confirmed a lack of genetic structure in Balkan donkey mtDNA. Balkan populations were poorly differentiated (ΦST  = 0.071). Differentiation between the Balkan donkey and the African outgroup also was low. The lack of correspondence between geographical areas and maternal genetic structure is consistent with the hypothesis suggesting a very quick spread of the species after domestication. The current research illustrates the difficulties to trace routes of expansion in donkey, as the species has no geographical structure.

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Genetic polymorphisms in CSN3 gene in Pag (Croatia), Sarda (Italy) and Pramenka (Serbia) sheep breeds were investigated. A single nucleotide polymorphism (SNP) was localized by sequence analysis (sequence submitted to GenBank under accession AY237637) relying on an original primer pair. Primers for sequencing (kappa-casF and kappa-casR) were designed on the available CSN3 sequences to amplify the genomic region encoding the major part of the mature protein (exon 4). An SNP was detected at position 237 of the sheep kappa-casein mRNA (reference sequence: GenBank X51822), where a thymine was substituted for a cytosine. The SNP was typed by conventional PCR and SYBR Green I-based real-time PCR. C and T alleles were discriminated using a dedicated set of primers that consisted of one common forward primer (SNP-TC) and two reverse primers (SNP-T and SNP-C), the latter two differing in the 3' end base and in the presence of a 12 bp poly-G tail in SNP-C. The SNP was found in both the heterozygous and the homozygous state in Sarda and Pramenka breeds, and in the heterozygous state only in the Pag breed. The observed allelic frequencies of the SNP were 0.12 in Pag, 0.27 in Sarda and 0.45 in Pramenka.

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