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O género Opuntia spp. pertence à família Cactaceae e tem origem na América Central. A O. ficus-indica é a espécie mais importante sob o ponto de vista económico e é cultivada em vários países para a produção de fruto e de cladódios. Esta espécie foi introduzida na Península Ibérica provavelmente no início do séc. XVI e encontra-se naturalizada na bacia Mediterrânica. Neste estudo foram utilizados seis marcadores SSR, previamente desenvolvidos para O. echios, na análise da diversidade genética de 19 ecótipos Portugueses de Opuntia spp. pertencentes a quatro espécies, O. ficus-indica (16 ecótipos), O. dillennii (1), O. elata (1) e O. robusta (1). Três cultivares italianas de O. ficus-indica, “Bianca”, “Gialla” e “Rossa”, foram utilizadas como termo de comparação e estudaram-se 15 plantas por ecótipo. Devido à existência de diferentes níveis de ploidia, os dados foram tratados como marcadores dominantes. Foi construída uma matriz binária, a partir da qual foram determinados os índices de similaridade de Dice e seguidamente procedeu-se à análise de cluster pelo método UPGMA. Com base na matriz de distância genética de Nei foi feita a análise de coordenadas principais (PCoA). Adicionalmente, foi realizada uma AMOVA para estimar os componentes da variância e a variação genética entre populações dentro de grupos e entre grupos. Detetou-se um total de 55 alelos, com média de 9,2 alelos por cada par de primers. A análise de cluster revelou quatro grupos que permitem diferenciar as quatro espécies de Opuntia spp estudadas. No caso de O. ficus-indica os ecótipos foram separados apenas em dois subgrupos, verificando-se baixo nível de variação genética intraespecífica. Um dos subgrupos continha os frutos de polpa branca (incluindo a cv. “Bianca”) e o ecótipo OFI-01 (que corresponde à forma espinescente O. ficus-indica f. amyclaea) e o outro continha os frutos de polpa laranja (incluindo a cv. “Gialla”), a cv. 'Rossa', de polpa vermelha, e um ecótipo de polpa amarela (OFI-04). Em O. ficus-indica, não foi possível realizar o DNA fingerprinting claro de ecótipos, de cultivares e das formas espinescentes relativamente às formas inermes. A AMOVA confirmou elevada diferenciação entre grupos e variação reduzida dentro das populações.

Existe um interesse crescente pelas espécies do género Opuntia spp. devido à sua rusticidade e às várias possibilidades de utilização económica que estas representam, nomeadamente na produção de fruto, forragem e nas indústrias farmacêutica e cosmética. A caracterização da diversidade genética por marcadores moleculares bem como a avaliação de germoplasma é importante no melhoramento de plantas e no desenvolvimento de novas cultivares. As plantas do género Opuntia spp. caraterizam-se por conter muitos polissacarídeos e metabolitos secundários que interferem na obtenção de DNA de alta qualidade. Por esta razão, vários métodos descritos na literatura e ‘kits’ comerciais disponíveis são, frequentemente, pouco eficazes na extração de DNA genómico. De modo a contornar este problema, foram testadas várias metodologias de extração de DNA a partir de cladódios de quatro espécies do género Opuntia (O. ficus-indica, O. robusta, O. dillenii e O. elata). Utilizaram-se três ‘kits’ comerciais, três versões modificadas do método padrão de brometo de cetil trimetil amónio (CTAB) e um método, por nós melhorado, que se baseia no método CTAB combinado como ‘mini kit’ comercial Plant DNeasy® (Qiagen). O protocolo combinado, permite a rápida extração de quantidades razoáveis de DNA genómico de alta qualidade em cladódios de todas as espécies utilizadas, destacando-se a O.dillenii e a O.elata como espécies onde o teor de polissacarídeos e metabolitos secundários é muito elevado. Foram obtidos em O. ficus-indica, O. robusta, O. dillenii e O. elata rendimentos em DNA de 10.5, 12.0, 13.2 e 15.9 μg.g-1 de tecido fresco, respetivamente. Com este protocolo, a pureza, avaliada pela razão A260/A280, foi respectivamente de 1.89, 1.75, 1.67 e 2.01. No entanto, quando utilizado isoladamente, o mini kit Plant DNeasy® permite extrair facilmente DNA de O. ficus-indica e O. robusta com rendimentos de 18.3 e 12.5 μg.g-1 de tecido fresco, respetivamente. Neste caso, a pureza, avaliada pela referida razão, foi de 1.84 e 1.74, respetivamente. A qualidade do DNA extraído foi confirmada pela amplificação de um conjunto de microsatélites nucleares obtidos para o género Opuntia spp. Da análise dos produtos amplificados, por electroforese em gel de agarose e eletroforese capilar em sequenciador, verificaram-se perfis de bandas e eletroferogramas reprodutíveis.

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.

The cork oak (Quercus suber L.) has remarkable ecological, social and economic value in the Mediterranean region. Due to the growing economic interest in cork, human intervention in the plant production and renewal of this species is crucial. Thus, the optimization of the propagation methodologies to produce selected and improved trees for high quality cork production is a key factor in the species improvement program. Rooting and survival of cuttings are greatly affected by several external and internal factors. To circumvent this problem, experiments were made using young seedlings as a source of cork oak cuttings for two consecutive years. We studied the influence of the application of synthetic auxins, indole-3-butyric acid (IBA) and α-naphthalene acetic acid (NAA), the IBA concentration and the basal bark removal in rooting and survival of cork oak cuttings. The IBA improved the rooting, the survival and the mean length of the longest root per rooted cutting, but not the mean number of primary roots produced. The basal bark removal associated with 0.5% of IBA gave the highest percent of rooting and plant survival (60% and 54%, respectively). The application of 0.1% of NAA, the origin of the cutting (whether basal or apical) had no significant influence on the rooting and survival. Thus, it is possible to draw the conclusion that wounding along with 0.5 % IBA produced the highest percentage of rooting and survival with cuttings planted in April, however complementary studies involving different physical and chemical conditions are required.

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.

Cistus ladanifer has a well-defined taxonomic identity. 2,2,6-trimethylcyclohexanone may be an authenticity and taxonomic marker. Its traits and applications make it a possible economic resource fitted for Mediterranean areas. Cistus ladanifer is a dominant shrub species endemic to the western Mediterranean region. Due to its dominant nature and its potential ecological, aromatic or pharmacological applications, C. ladanifer has been the object of numerous studies. In this review current knowledge on different aspects of this species is summarized, from its taxonomy to its chemical characterisation or its competitive traits. There are no doubts about the taxonomic entity of C. ladanifer, although the recognition of infraspecific taxa deserves more attention. Given that the fragrant exudate of C. ladanifer holds a very specific composition, one species specific carotenoid, 2,2,6-trimethylcyclohexanone, derivative is proposed as an authenticity marker for uses of C. ladanifer in pharmacological or aromatic industries. Evidence is also gathered on the extreme adaptation of C. ladanifer to stressful conditions in the Mediterranean region, such as the ability to survive in low hydric and high solar exposition conditions, presistence in poor and contaminated soils, and growth inhibition of several other plants through the release of allelochemicals. Thus, the finding of potential applications for this plant may contribute to enhance the economic dimension of derelict lands, such as mine tailings or poor agricultural Mediterranean areas.

Main conclusion: The combination of genotypic selection, targeted and improved cultivation, and processing techniques for specific applications gives C. ladanifer the potential to be used as a valuable resource in Mediterranean areas with poor agronomic advantages. Cistus ladanifer (rockrose) is a perennial shrub, well adapted to the Mediterranean climate and possibly to upcoming environmental changes. As a sequence to a thorough review on taxonomic, morphological, chemical and competitive aspects of C. ladanifer, the research team focuses here on the economic potential of C. ladanifer: from production to applications, highlighting also known biological activities of extracts and their compounds. The use of this natural resource may be a viable solution for poor and contaminated soils with no need for large agricultural techniques, because this species is highly resistant to pests, diseases and extreme environmental factors. In addition, this species reveals interesting aptitudes that can be applied to food, pharmaceutical, phytochemical and biofuel industries. The final synthesis highlights research lines toward the exploitation of this neglected resource, such as selection of plant lines for specific applications and development of agronomic and processing techniques.