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Assessment of Genetic Diversity in Opuntia spp. Portuguese Populations Using SSR Molecular Markers

Publication . Reis, C.M.G.; Raimundo, Joana; Ribeiro, M.M.A.

The Opuntia spp., most likely few individuals, were introduced in the Iberian Peninsula in the beginning of the 16th century, after the discovery of America, spreading afterwards throughout the Mediterranean basin. We analysed, for the first time, the genetic diversity in a set of 19 Portuguese Opuntia spp. populations from the species O. ficus-indica, O. elata, O. dillenii and O. robusta using nuclear microsatellite (nuSSR) markers. The Italian cultivars ‘Bianca’, ‘Gialla’ and ‘Rossa’ were included in the study for comparison purposes. The nuSSR amplifications produced from five to 16 alleles, with an average of 9.2 alleles per primer pair, and average polymorphism information content of 0.71. The estimated Dice coefficient among populations varied from 0.26 to 1.0, indicating high interspecific genetic diversity but low genetic diversity at the intraspecific level. The hierarchical clustering analysis revealed four major groups that clearly separated the four Opuntia species. Among the O. ficus-indica populations, two sub-clusters were found, one including the white pulp fruits (with cv. Bianca) and the other with the orange pulp ones and including the cv. Gialla, the cv. Rossa, and one pale yellow pulp population. No genetic differences were found between the inermis form, O. ficus-indica f. ficus-indica, and the rewilded spiny one, O. ficus-indica f. amyclaea. The dendrogram indicated that the clustering pattern was unrelated to geographical origin. The results revealed a low level of genetic diversity among the Portuguese populations of O. ficus-indica.

2018Journal article

Open access

Rapid, simple and potentially universal method for DNA extraction from Opuntia spp. fresh cladode tissues suitable for PCR amplification

Publication . Raimundo, Joana; Reis, C.M.G.; Ribeiro, M.M.A.

In Opuntia spp., the cladode tissues contain many polysaccharides and secondary metabolites that interfere with obtaining high-quality deoxyribonucleic acid (DNA), using currently available methods. To circumvent this problem, three commercial kits, three modified versions of the conventional cetyltrimethylammonium bromide method (CTAB) method and one combined method were tested in Opuntia ficus-indica, O. robusta, O. dillenii and O. elata species. We obtained a rapid and simple protocol that allows the extraction of DNA from all the tested species with good DNA yield and purity, namely, the combined method. With this method (DNeasy® Plant Mini Kit combined with the CTAB method), DNA yields from 13.2 ± 7.8 to 15.9 ± 11.3 µg.g-1 of fresh tissue were obtained in the four Opuntia species. The purity, evaluated by the ratio A260/A280 ratio, ranged from 1.67 ± 0.12 to 2.01 ± 0.25, revealing low levels of problematic metabolites. The extracted DNA quality was confirmed by amplifying a set of nuclear microsatellites obtained for the genus. Reliable reproducible bands and electropherogram profiles were obtained. The combined method has potential to be universal for good-quality DNA extraction in cacti, particularly in the Opuntia genus and other difficult-to-extract species.

2018Journal article

Restricted access

Bioclimatic modeling in the Last Glacial Maximum, Mid-Holocene and facing future climatic changes in the strawberry tree (Arbutus unedo L.)

Publication . Ribeiro, M.M.A.; Roque, Natália; Ribeiro, Sílvia; Gavinhos, Catarina; Silva, Isabel Castanheira; Quinta-Nova, L.C.; Albuquerque, M.T.D.; Gerassis, Saki

Increasing forest wildfires in Portugal remain a growing concern since forests in the Mediterranean region are vulnerable to recent global warming and reduction of precipitation. Therefore, a long-term negative effect is expected on the vegetation, with increasing drought and areas burnt by fires. The strawberry tree (Arbutus unedo L.) is particularly used in Portugal to produce a spirit by processing its fruits and is the main income for forestry owners. Other applications are possible due to the fruit and leaves' anti-oxidant properties and bioactive compounds production, with a potential for clinical and food uses. It is a sclerophyllous plant, dry-adapted and fire resistant, enduring the Mediterranean climate, and recently considered as a possibility for afforestation, to intensify forest discontinuity where pines and eucalypts monoculture dominate the region. To improve our knowledge about the species' spatial distribution we used 318 plots (the centroid of a 1 km2 square grid) measuring the species presence and nine environmental attributes. The seven bioclimatic variables most impacting on the species distribution and two topographic features, slope and altitude, were used. The past, current and future climate data were obtained through WorldClim. Finally, the vulnerability of the strawberry tree to the effects of global climate change was examined in the face of two emission scenarios (RCP 4.5 and 8.5), to predict distribution changes in the years 2050 and 2070, using a species distribution models (MaxEnt). The reduction of suitable habitat for this species is significant in the southern regions, considering the future scenarios of global warming. Central and northern mountainous regions are putative predicted refuges for this species. Forest policy and management should reflect the impact of climate change on the usable areas for forestry, particularly considering species adapted to the Mediterranean regions and wildfires, such as the strawberry tree. The distribution of the species in the Last Glacial Maximum (LGM) and Mid-Holocene (MH) agrees with previous genetic and paleontological studies in the region, which support putative refuges for the species. Two in the southern and coastal-central regions, since the LGM, and one in the east-central mountainous region, considered as cryptic refugia.

2019Journal article

Open access

Bioclimatic modeling in the last glacial maximum, mid-holocene and facing future climatic changes in the strawberry tree (Arbutus unedo L.)

Publication . Ribeiro, M.M.A.; Roque, Natália; Ribeiro, Sílvia; Gavinhos, Catarina; Silva, Isabel Castanheira; Quinta-Nova, L.C.; Albuquerque, M.T.D.; Gerassis, Saki

2019Journal article

Open access

Future habitat suitability for species under climate change: lessons learned from the strawberry tree case study

Publication . Gerassis, Saki; Albuquerque, M.T.D; Roque, Natália; Ribeiro, Sílvia; Taboada, Javier; Ribeiro, M.M.A.

Climate change is already a reality, and habitat loss is affecting relentlessly tree species distributions. The strawberry tree (Arbutus unedo L., Ericaceae) is a Mediterranean evergreen tree used in this article as a case study to evince its expected threatening situation in the upcoming age. This research work seeks to identify the bioclimatic and physiographic variables that have the most impact on the strawberry tree’s spatial distribution in Portugal, acquiring vital knowledge for the design of accurate conservation and afforestation plans for the use and conservation of strawberry tree, working as a guide under a climate change scenario. For that, twenty-one bioclimatic variables, two physiographic attributes (altitude and slope), and the Emberger Index (EI) were used together with 318 observations of strawberry trees, to build a scalable Bayesian procedure, based on machine learning techniques, aimed to assess the species’ future habitat evolution through three temporal scenarios: (i) Control Run (1960–1990); (ii) 2050 and (iii) 2070. The results indicate for 2050 a 30% loss of the humid subregion and a 35% increase in the semi-arid sub-region towards the north. In 2070, it is expected a 2% recuperation for the sub-humid area, but an 8% loss of the humid sub-area. Under these extreme climate change conditions, it is anticipated an almost complete loss of habitat for the strawberry tree in the south of Portugal. The expected ecological evolvability may trigger future migration paths and new refuges’ settlement in the northern sub-region for the succeeding decades and suggesting after 2070 the possibility of habitat switch and species drifting.

2021Journal article

Restricted access