Sistematic, population genetics, propagation and conservation of Prunus azorica (Rosaceae)

Tese de Doutoramento, Biologia (Ecologia Vegetal), 25 de Junho de 2013, Universidade dos Açores.

Transcriptional dynamics of the developing sweet cherry (Prunus avium L.) fruit: sequencing, annotation and expression profiling of exocarp-associated genes

The exocarp, or skin, of fleshy fruit is a specialized tissue that protects the fruit, attracts seed dispersing fruit eaters, and has large economical relevance for fruit quality. Development of the exocarp involves regulated activities of many genes. This research analyzed global gene expression in the exocarp of developing sweet cherry (Prunus avium L., 'Regina'), a fruit crop species with little public genomic resources. A catalog of transcript models (contigs) representing expressed genes was constructed from de novo assembled short complementary DNA (cDNA) sequences generated from developing fruit between flowering and maturity at 14 time points. Expression levels in each sample were estimated for 34 695 contigs from numbers of reads mapping to each contig. Contigs were annotated functionally based on BLAST, gene ontology and InterProScan analyses. Coregulated genes were detected using partitional clustering of expression patterns. The results are discussed with emphasis on genes putatively involved in cuticle deposition, cell wall metabolism and sugar transport. The high temporal resolution of the expression patterns presented here reveals finely tuned developmental specialization of individual members of gene families. Moreover, the de novo assembled sweet cherry fruit transcriptome with 7760 full-length protein coding sequences and over 20 000 other, annotated cDNA sequences together with their developmental expression patterns is expected to accelerate molecular research on this important tree fruit crop.

Genetic diversity assessment of the almond (Prunus dulcis (Mill.) DA Webb) traditional germplasm of Algarve, Portugal, using molecular markers

In this study, 123 almond (Prunus dulcis (Mill.) D. A. Webb) trees identified among traditional orchards in the Algarve region and 53 trees of the local field collection managed by the regional office of the Portuguese Ministry of Agriculture (DRAALG) were assessed using isozyme, inter- single sequence repeat and simple sequence repeat or microsatellite techniques for the evaluation of genetic diversity and genetic relatedness and identification of new accessions for the field collection. The isozyme analysis allowed the distribution of the 176 plants into 13 different classes of enzyme similarity, while the use of DNA markers increased the distribution of the analysed trees among 140 discriminating DNA patterns. Multiple cases of homonymy and synonymy were identified in the local germplasm. Some traditional varieties, such as Lourencinha, appeared to be relatively homogeneous, while other local denominations, e.g. Galamba, included diverse genotypes. Of the 13 commercial varieties analysed in this study, 11 assembled in one major cluster clearly differentiated from the majority of the local genotypes. These results reinforced the perception that the Algarve traditional germplasm constitutes an important repository of genetic diversity, eventually carrying alleles of high agricultural interest such as the recently identified Phomopsis resistance in the traditional variety Barrinho Grado.

Genetic diversity assessment of the almond (Prunus dulcis (Mill.) DA Webb) traditional germplasm of Algarve, Portugal, using molecular markers

In this study, 123 almond (Prunus dulcis (Mill.) D. A. Webb) trees identified among traditional orchards in the Algarve region and 53 trees of the local field collection managed by the regional office of the Portuguese Ministry of Agriculture (DRAALG) were assessed using isozyme, inter- single sequence repeat and simple sequence repeat or microsatellite techniques for the evaluation of genetic diversity and genetic relatedness and identification of new accessions for the field collection. The isozyme analysis allowed the distribution of the 176 plants into 13 different classes of enzyme similarity, while the use of DNA markers increased the distribution of the analysed trees among 140 discriminating DNA patterns. Multiple cases of homonymy and synonymy were identified in the local germplasm. Some traditional varieties, such as Lourencinha, appeared to be relatively homogeneous, while other local denominations, e.g. Galamba, included diverse genotypes. Of the 13 commercial varieties analysed in this study, 11 assembled in one major cluster clearly differentiated from the majority of the local genotypes. These results reinforced the perception that the Algarve traditional germplasm constitutes an important repository of genetic diversity, eventually carrying alleles of high agricultural interest such as the recently identified Phomopsis resistance in the traditional variety Barrinho Grado.

Análise funcional e potencial biotecnológico do gene DHN11 de macieira (Malus x domestica Borkh.)

A deficiência hídrica é o principal fator limitante à agricultura, causando significativas perdas de produtividade nas mais diversas culturas vegetais. Novas tecnologias agrícolas, tais como a geração de plantas geneticamente modificadas melhor adaptadas a condições adversas de cultivo, são necessárias. Em um estudo anterior do grupo caracterizando a família gênica das desidrinas (DHNs) em macieira (Malus x domestica Borkh.), foi verificado que o gene MdDHN11 possui níveis de expressão cerca de 6 mil vezes maior em sementes em relação a outros tecidos. Sugere-se que o acúmulo nos últimos estágios de desenvolvimento de sementes possua papel protetor na tolerância à dessecação. Além disso, as DHNs são proteínas caracterizadas por seu envolvimento na resposta a mudanças na disponibilidade de água, principalmente os causados por estresses abióticos.

Determinação da origem das lenticelas e sua relação com a lenticelose em maçãs (Malus domestica Borkh.).

2011

The Effects of Various Types of Feed additives on The Chemical Composition of Local Chicken Meat (Gallus domestica).

Abstract. This study aims to determine the effect of giving various types of feed additives to the chemical composition value of super chicken chicken (Gallus domestica). This research is an experimental research using 20 super chicken chickens that come from chicken growth research (growth study). The design used in this study was Completely Randomized Design (RAL), consisting of 4 treatments and 5 replications. The treatment given was (A0 = control (Vita chick 0.7 gram / liter; A1 = 20 ml / liter probio-FM; A2 = 0.08% MOS-oligosaccharide / kg of feed and A3 = herbal leuser KI 5 ml / liter). is a 90 day old super chicken breast Chicken Variable observed moisture content, protein content and fat content The data obtained were analyzed by using vocabulary and tested further by Duncan's Multiple Range Test The results showed that treatment (P> 0,05) to the value of water content and protein of super chicken fowl.Average value of water content at each treatment A0 (69,81%), A1 (70,74%), A2 (71,56%) and A3 (71,52%) while mean value of protein A0 (18,95%), A1 (19,61%), A2 (19,01%) and A3 (19,14%)) P <0,05) to the fat content of super chicken flesh, mean of fat content were A0 (2.02%), A1 (1.49%), A2 (1.37%) and A3 (2.0%).

Study of medicinal and food plants as a source of biologically active compounds.

In the last few decades, scientific evidence has pointed out the health-beneficial effects of phenolic compounds in foods, including a decrease in risk of developing degenerative and chronic diseases, known to be caused by oxidative stress. In this frame can be inserted research carried out during my PhD thesis, which concerns the phytochemical investigation of phenolic composition in sweet cherries (Prunus avium L.), apple fruits (Malus domestica L.) and quinoa seeds (Chenopodium quinoa Willd.). The first project was focused on the investigation of phytochemical profile and nutraceutical value of fruits of new sweet cherry cultivars. Their phenolic profile and antioxidant activity were investigated and compared with those of commonly commercialized cultivars. Their nutraceutical value was evaluated in terms of antioxidant/neuroprotective capacity in neuron-like SH-SY5Y cells, in order to investigate their ability to counteract the oxidative stress and/or neurodegeneration process The second project was focused on phytochemical analysis of phenolic compounds in apples of ancient cultivars with the aim of selecting the most diverse cultivars, that will then be assayed for their anti-carcinogenic and anti-proliferative activities against the hepato-biliary and pancreatic tumours. The third project was focused on the analysis of polyphenolic pattern of seeds of two quinoa varieties grown at different latitudes. Analysis of phenolic profile and in vitro antioxidant activity of seed extracts both in their free and soluble-conjugated forms, showed that the accumulation of some classes of flavonoids is strictly regulated by environmental factors, even though the overall antioxidant capacity does not differ in quinoa Regalona grown in Chile and Italy. During the internship period carried out at the Department of Organic Chemistry at Universidad Autónoma de Madrid (UAM), it was achieved the isolation of two pentacyclic triterpenoids, from an endemic Peruvian plant, Jatropha macrantha Müll. Arg., with bio-guided fractionation technique.

La violenza di genere: analisi della casistica autoptica di femminicidio e degli ammonimenti del Questore per stalking e per violenza domestica della citta metropolitana di Bologna

Il presente lavoro tratta il tema della violenza di genere, in particolare femminicidio, stalking e violenza domestica per la connessione tra questi intercorrente nell’escalation violenta. Spesso, i femminicidi sono preceduti da stalking o ripetuti episodi di violenza fisica prettamente consumata in ambito domestico e relazioni affettive pregresse/attuali. La prima parte del lavoro descrive ambito scientifico e giuridico, internazionale e nazionale, della violenza di genere, dimensione del fenomeno ed evoluzione normativa a tutela delle vittime, come richiesto dalla Convenzione di Istanbul. La seconda parte affronta aspetti medico-legali della violenza di genere (attività di patologia forense, genetica forense e tossicologia forense nei femminicidi, assistenza medico-legale alla vittima di maltrattamenti e violenza sessuale, valutazione medico-legale del danno alla persona per stalking). La terza parte presenta lo studio su casistica autoptica di femminicidi del 1950-2019 e provvedimenti di ammonimento del Questore per stalking e per violenza domestica del 2009-2020 nella provincia bolognese. Secondo i risultati il femminicidio è un fenomeno di “vecchia data”, in un quadro normativo-culturale che “tollerava” violenza a danno di vittime femminili. L’andamento del fenomeno è costante fino a oggi con modifiche di cause e mezzi del decesso. L’aumentata età media delle vittime richiama il crescente fenomeno dell’elder abuse. Negli ammonimenti per stalking e per violenza domestica analizzati prevalgono vittime di genere femminile e autori di genere maschile, in vicende affettive e familiari. Lo studio approfondito delle caratteristiche di autori e vittime di femminicidio, stalking e violenza domestica permette di individuare indicatori di rischio per implementare strategie di prevenzione mirate. Il punto di vista privilegiato medico-legale può assumere un ruolo centrale insieme alle altre figure coinvolte in prevenzione, repressione e contrasto alla violenza di genere. Solo un rigoroso approccio metodologico multidisciplinare può aiutare nella prevenzione. La ricerca in tale ambito è il punto di forza della gestione multidisciplinare della vittima.

Edilizia domestica e cultura materiale nelle colonie di Ariminum e Bononia (III-I sec. a.C.)

Il tema affrontato nella presente ricerca sono le trasformazioni intercorse nella vita quotidiana tra il III e il I secolo a.C. in due colonie latine, Ariminum e Bononia, attraverso le evidenze archeologiche. Vengono indagate su scala locale le conseguenze di un fenomeno di grande portata, la colonizzazione romano-latina, mettendo a fuoco le forme dell’abitare, le tradizioni artigianali e le pratiche alimentari. La principale base documentaria sono le testimonianze archeologiche di edilizia domestica e le ceramiche, rinvenute nelle aree di abitato di Rimini e Bologna e nei territori limitrofi. Per cogliere a pieno le trasformazioni intercorse, vengono passate in rassegna le principali caratteristiche del popolamento, dell'architettura domestica e delle ceramiche precedenti la colonizzazione romano-latina. Le due colonie, le abitazioni e le ceramiche sono considerate, inoltre, nel contesto territoriale più ampio, volgendo lo sguardo anche all'area medio-adriatica e alla Cispadana. Allo stesso tempo, sono continui i riferimenti all'Italia medio-tirrenica, poiché permettono di comprendere molte delle evidenze archeologiche e dei processi storici in esame. Il primo capitolo tratta della colonizzazione romano-latina, calata nelle realtà di Rimini e Bologna. La domanda a cui si vuole rispondere è: chi erano gli abitanti delle due colonie? A questo proposito, si affronta anche la questione degli insediamenti precoloniali. Nel secondo capitolo si analizzano le abitazioni urbane. Quali furono le principali innovazioni nell'architettura domestica introdotte dalla colonizzazione? Come cambiarono le forme dell’abitare ad Ariminum e Bononia in età repubblicana? Il terzo capitolo si concentra sulla ceramica per la preparazione e il consumo del cibo nei contesti di abitato. Come cambiarono nelle due città le pratiche alimentari e le tradizioni artigianali utilizzate nella produzione di ceramiche? L'ultimo capitolo discute alcuni quadri teorici applicati ai fenomeni descritti nei capitoli precedenti (romanizzazione, acculturazione, identità, globalizzazione). L'ultimo paragrafo entra nel merito delle trasformazioni avvenute nella vita quotidiana di Ariminum e Bononia.

Investigating the causes of late fruit drop in ‘Regina’ sweet cherry (Prunus avium)

The presented study aimed to correctly describe the (late) fruit drop pattern of sweet cherry cv. Regina grafted on ‘Gisela 5’ and investigate its internal causes. In the first season, a method to describe the fruit drop pattern was defined and validated. The second season was devoted to a province-based screening of the phenomenon to identify potential influences of environmental, physiological and management factors. The multisite trial involved 6 commercial orchards located at different elevations, from 225 up to 1175m a.s.l. The third season was dedicated to find confirmation of the hypothesis formulated during the previous year. The multisite comparison was maintained but reduced to only two orchards to allow more frequent samplings. It emerged that late fruit drop is a complex phenomenon showing variable intensity: the percentage of late fruit drop ranged from 7 to 76% of the fruitlets set, depending on the orchard and on the year considered. Two main waves of fruitlets drop have been observed: the first one was composed by unfertilized parthenocarpic fruitlets, probably caused by late or missing fertilization, that immediately after bloom already showed smaller diameters and symptoms of senescence; the second one (the focus of this study) was composed by fully developed fruits that at a certain point decreased their growth rate and got senescent. All the late dropped cherries showed an aborted embryo. This sudden change has been observed to be concomitant both with prolonged periods of low temperatures (or sudden severe decreases in the daily Growing Degree Hours accumulation) and with extraordinary high temperatures close to or above 30°C. Other factors, such as the position of the limb within the canopy, its orientation (sunny vs. shady side) or nutrition played only a marginal role. Excessive vigor can increase late fruit drop intensity but is not its main cause.

Ultra-sonografia do fígado, aparelho renal e reprodutivo da jibóia (Boa constrictor)

Atualmente os animais silvestres têm despertado o interesse particular na criação domestica. Na medicina de animais selvagens, os exames ultra-sonográficos podem ser considerados como ferramenta para diagnosticar e prevenir doenças. Deste modo, realizou-se um estudo em 20 jibóias (Boa constrictor), a fim de caracterizar a morfologia e aparência ultra-sonográfica das estruturas presentes da cavidade celomática desses animais. Ultra-sonograficamente, o fígado apresentou-se variando de hipoecóica a levemente hiperecogênica, com margens ecogênicas e ecotextura homogênea em toda sua extensão. Os rins mostraram formato elipsóide, com cápsula fina, regular e hiperecóica. Os folículos ovarianos apresentaram formato ovóide, margens finas, regulares e discretamente hiperecóicas. As estruturas do sistema reprodutor do macho não foram evidenciadas com precisão, devido a sua ecogenicidade similar em relação às estruturas adjacentes e pela presença do "corpo gorduroso" localizado nessa região. A ultra-sonografia da cavidade celomática em jibóias demonstrou ser uma técnica rápida e de fácil acesso, permitindo identificar a morfologia, sintopia e aparência ultra-sonográfica de estruturas como o fígado, rins e de folículos vitelogênicos nas fêmeas.

Condições de atmosfera controlada para pêssegos "maciel" colhidos em dois estádios de maturação

O objetivo deste trabalho foi avaliar o efeito de duas temperaturas e condições de atmosfera controlada (AC) sobre a conservação de pêssegos da cultivar Maciel, colhidos em dois estádios de maturação. Os tratamentos avaliados foram: armazenamento refrigerado (AR) na temperatura de +0,5°C; AR na temperatura de -0,5°C; 2,0kPa O2 + 4,0kPa CO2 em -0,5°C; 1,0kPa O2 + 3,0kPa CO2 em -0,5°C; 2,0kPa O2 + 6,0kPa CO2 em -0,5°C. As avaliações foram realizadas após 60 dias de armazenamento e mais dois e quatro dias de exposição dos frutos à temperatura de 20ºC. Na análise realizada após dois meses de armazenamento, mais dois dias a 20°C, verificou-se que os frutos submetidos a 2,0kPa de O2 + 4,0 kPa de CO2 apresentaram maior firmeza de polpa em relação aos demais tratamentos, sendo que a mesma não foi influenciada pelo estádio de maturação. Os sólidos solúveis totais foram maiores em frutos com estádio de maturação maduro independente da condição de armazenamento. A ocorrência de podridões e escurecimento interno da polpa não foi influenciada pelo estádio de maturação. No entanto, a condição de AC de 1,0 kPa de O2 + 3,0kPa de CO2 proporcionou o menor percentual de podridões e escurecimento interno da polpa em relação aos demais tratamentos. Na avaliação realizada aos quatro dias de exposição a 20°C, os frutos colhidos no estádio maduro estavam completamente podres, independente da condição de armazenamento praticada.

Evaluation of Antihyperglycemia and Antihypertension Potential of Native Peruvian Fruits Using In Vitro Models

Local food diversity and traditional crops are essential for cost-effective management of the global epidemic of type 2 diabetes and associated complications of hypertension. Water and 12% ethanol extracts of native Peruvian fruits such as Lucuma (Pouteria lucuma), Pacae (Inga feuille), Papayita arequipena (Carica pubescens), Capuli (Prunus capuli), Aguaymanto (Physalis peruviana), and Algarrobo (Prosopis pallida) were evaluated for total phenolics, antioxidant activity based on 2, 2-diphenyl-1-picrylhydrazyl radical scavenging assay, and functionality such as in vitro inhibition of alpha-amylase, alpha-glucosidase, and angiotensin I-converting enzyme (ACE) relevant for potential management of hyperglycemia and hypertension linked to type 2 diabetes. The total phenolic content ranged from 3.2 (Aguaymanto) to 11.4 (Lucuma fruit) mg/g of sample dry weight. A significant positive correlation was found between total phenolic content and antioxidant activity for the ethanolic extracts. No phenolic compound was detected in Lucuma (fruit and powder) and Pacae. Aqueous extracts from Lucuma and Algarrobo had the highest alpha-glucosidase inhibitory activities. Papayita arequipena and Algarrobo had significant ACE inhibitory activities reflecting antihypertensive potential. These in vitro results point to the excellent potential of Peruvian fruits for food-based strategies for complementing effective antidiabetes and antihypertension solutions based on further animal and clinical studies.

The gene transformer-2 of Anastrepha fruit flies (Diptera, Tephritidae) and its evolution in insects

Background: In the tephritids Ceratitis, Bactrocera and Anastrepha, the gene transformer provides the memory device for sex determination via its auto-regulation; only in females is functional Tra protein produced. To date, the isolation and characterisation of the gene transformer-2 in the tephritids has only been undertaken in Ceratitis, and it has been shown that its function is required for the female-specific splicing of doublesex and transformer pre-mRNA. It therefore participates in transformer auto-regulatory function. In this work, the characterisation of this gene in eleven tephritid species belonging to the less extensively analysed genus Anastrepha was undertaken in order to throw light on the evolution of transformer-2. Results: The gene transformer-2 produces a protein of 249 amino acids in both sexes, which shows the features of the SR protein family. No significant partially spliced mRNA isoform specific to the male germ line was detected, unlike in Drosophila. It is transcribed in both sexes during development and in adult life, in both the soma and germ line. The injection of Anastrepha transformer-2 dsRNA into Anastrepha embryos caused a change in the splicing pattern of the endogenous transformer and doublesex pre-mRNA of XX females from the female to the male mode. Consequently, these XX females were transformed into pseudomales. The comparison of the eleven Anastrepha Transformer-2 proteins among themselves, and with the Transformer-2 proteins of other insects, suggests the existence of negative selection acting at the protein level to maintain Transformer-2 structural features. Conclusions: These results indicate that transformer-2 is required for sex determination in Anastrepha through its participation in the female-specific splicing of transformer and doublesex pre-mRNAs. It is therefore needed for the auto-regulation of the gene transformer. Thus, the transformer/transfomer-2 > doublesex elements at the bottom of the cascade, and their relationships, probably represent the ancestral state ( which still exists in the Tephritidae, Calliphoridae and Muscidae lineages) of the extant cascade found in the Drosophilidae lineage ( in which tra is just another component of the sex determination gene cascade regulated by Sex-lethal). In the phylogenetic lineage that gave rise to the drosophilids, evolution co-opted for Sex-lethal, modified it, and converted it into the key gene controlling sex determination.