Martino Orru

Morphological characterisation of Vitis vinifera L. seeds by image analysis and comparison with archaeological remains

In archaeobotanical studies, the taxonomic classification of diaspores has usually been done by simple morphological observation and visual comparison with ex situ collections of seeds, although the use of biometric indices has often proved to be a powerful approach in the taxonomic studies of the genus Vitis as well as for the species attribution of archaeological remains. Using image analysis techniques, seeds from two Sardinian archaeological sites, the pre-Nuragic and Nuragic complex of Sa Osa in central-western Sardinia, attested as the oldest Sardinian archaeological site with remains of Vitis seeds, and the Isola di Coltellazzo in southwest Sardinia, were selected and characterized on the basis of morphological features and Elliptic Fourier Descriptors. Moreover, seeds of five modern populations of V. vinifera ssp. sylvestris collected from southwest Sardinia and the seeds of 41 cultivars of V. vinifera ssp. vinifera mainly from southern and central-western Sardinia were also analysed by computer image analysis. The obtained data were used to implement a database of biometric parameters and to compare the unknown archaeological seeds with the characterized recent seeds, using Linear Discriminant Analysis. The similarity of the archaeological seeds to V. vinifera ssp. vinifera cultivars rather than to V. vinifera ssp. sylvestris populations could allow it to be stated that, between the Middle and Final Bronze Age, varieties very close to modern V. vinifera ssp. vinifera were already being used to produce wine and/or to be preserved for foodstuffs. Moreover, the better matching of the archaeological seeds to white grapes rather than black grape cultivars could indicate the origins of the traditional cultivation of white grapes in these regions of Sardinia.

Thermal thresholds as predictors of seed dormancy release and germination timing: altitude-related risks from climate warming for the wild grapevine Vitis vinifera subsp. sylvestris.

BACKGROUND AND AIMS The importance of thermal thresholds for predicting seed dormancy release and germination timing under the present climate conditions and simulated climate change scenarios was investigated. In particular, Vitis vinifera subsp. sylvestris was investigated in four Sardinian populations over the full altitudinal range of the species (from approx. 100 to 800 m a.s.l). METHODS Dried and fresh seeds from each population were incubated in the light at a range of temperatures (10-25 and 25/10 °C), without any pre-treatment and after a warm (3 months at 25 °C) or a cold (3 months at 5 °C) stratification. A thermal time approach was then applied to the germination results for dried seeds and the seed responses were modelled according to the present climate conditions and two simulated scenarios of the Intergovernmental Panel on Climate Change (IPCC): B1 (+1·8 °C) and A2 (+3·4 °C). KEY RESULTS Cold stratification released physiological dormancy, while very few seeds germinated without treatments or after warm stratification. Fresh, cold-stratified seeds germinated significantly better (>80 %) at temperatures ≥20 °C than at lower temperatures. A base temperature for germination (T(b)) of 9·0-11·3 °C and a thermal time requirement for 50 % of germination (θ(50)) ranging from 33·6 °Cd to 68·6 °Cd were identified for non-dormant cold-stratified seeds, depending on the populations. This complex combination of thermal requirements for dormancy release and germination allowed prediction of field emergence from March to May under the present climatic conditions for the investigated populations. CONCLUSIONS The thermal thresholds for seed germination identified in this study (T(b) and θ(50)) explained the differences in seed germination detected among populations. Under the two simulated IPCC scenarios, an altitude-related risk from climate warming is identified, with lowland populations being more threatened due to a compromised seed dormancy release and a narrowed seed germination window.

Computer vision as a method complementary to molecular analysis: Grapevine cultivar seeds case study

Despite different breeding events, as well as the domestication phenomena which contributed to enrich the grape varietal heritage in Sardinia, many local varieties simply are the product of linguistic distorting due to the wide heterogeneity historic-cultural of the island. This phenomenon generated a great assortment of grape names, that, together with the huge real number of cultivars, is the cause of the incredible current grapevine Sardinian panorama. The goal of this article is to compare the published molecular data of 40 Sardinian autochthonous cultivars with the results achieved by the germplasm phenotypical characterization, on the basis of morpho-colorimetric features and Elliptic Fourier Descriptors (EFDs), measured by image analysis. Statistical classifiers were implemented to discriminate dissimilar seeds and carry out hypothetical synonymy groups to compare with those proposed on the basis of Simple Sequence Repeat (SSR) markers. This work represents the first trial to validate a morpho-colorimetric characterization method by direct comparison with molecular data, proving that the 113 measured features of the germplasm resulted adequate to achieve a clear discrimination among the synonymy groups.

The Aichi Biodiversity Target 12 at regional level: an achievable goal?

The Aichi Biodiversity Target 12 aims to prevent the extinction risk of known threatened species and to improve their conservation status by 2020. We present the integrated strategy implemented in the last 10 years for the keystone plant species of Sardinia (Italy, W. Mediterranean Basin), which includes the following activities: conservation status assessment (following the IUCN protocol), ex situ conservation, in situ monitoring and active protection measures. To date, an average of 51.8% of keystone plant species have been subjected to the latter first three activities while, due to the higher costs, only few active conservation measures have been carried out. Considering the activities realised since 2004, we have also predicted the conservation effectiveness towards 2020 and have elaborated an index to evaluate it. Halfway through the strategic plan, we argue that more efforts are needed to guarantee the effective conservation of all threatened plants in Sardinia.

Predictive method for correct identification of archaeological charred grape seeds: Support for advances in knowledge of grape domestication process

The identification of archaeological charred grape seeds is a difficult task due to the alteration of the morphological seeds shape. In archaeobotanical studies, for the correct discrimination between Vitis vinifera subsp. sylvestris and Vitis vinifera subsp. vinifera grape seeds it is very important to understand the history and origin of the domesticated grapevine. In this work, different carbonisation experiments were carried out using a hearth to reproduce the same burning conditions that occurred in archaeological contexts. In addition, several carbonisation trials on modern wild and cultivated grape seeds were performed using a muffle furnace. For comparison with archaeological materials, modern grape seed samples were obtained using seven different temperatures of carbonisation ranging between 180 and 340ºC for 120 min. Analysing the grape seed size and shape by computer vision techniques, and applying the stepwise linear discriminant analysis (LDA) method, discrimination of the wild from the cultivated charred grape seeds was possible. An overall correct classification of 93.3% was achieved. Applying the same statistical procedure to compare modern charred with archaeological grape seeds, found in Sardinia and dating back to the Early Bronze Age (2017–1751 2σ cal. BC), allowed 75.0% of the cases to be identified as wild grape. The proposed method proved to be a useful and effective procedure in identifying, with high accuracy, the charred grape seeds found in archaeological sites. Moreover, it may be considered valid support for advances in the knowledge and comprehension of viticulture adoption and the grape domestication process. The same methodology may also be successful when applied to other plant remains, and provide important information about the history of domesticated plants.