Climate change models predict an increase in frequency and amplitude of extreme weather events, including heat waves. To better predict how the composition and distribution of plant assemblages might respond to these changes in temperature, it is important to understand how species currently respond to these extremes. Photosynthetic thermal tolerance (T25)and photosynthetic recovery (RT25) were quantified in 27 species. We also studied the relationships between T25, RT25 and leaf mass per area (LMA). Leaf temperature was also monitored in the field. Leaves used in this study were collected from two distinct environments representing desert and montane plant assemblages. T25 and RT25 were measured using a chlorophyll fluorescence protocol incorporating sub-saturating light and short duration heat stress. Mean T25and LMA were significantly different between environments. Mean RT25 was not significantly different between environments. There was a positive relationship between T25 and LMA in both environments. The ability to recover from heat stress does not differ between two biomes that experience vastly different mean maximum temperatures during the summer months. LMA is a predictive leaf trait for thermal tolerance.

Methods for rapid, safe and effective detection of unmapped buried ordnance are vital to the protection of humans and environmental quality throughout the world. This study aimed to investigate the use of phytosensing and to understand the physiological response of woody plants to 2,4,6-trinitrotoluene (TNT) contamination. Baccharis halimifolia were potted in soils containing various concentrations of TNT and physiological responses were observed over a 9-week experimental period. Measurements included the collection of remotely sensed data, such as hyperspectral reflectance and chlorophyll fluorescence, and traditional plant-level physiological data. In accordance with the hypothesis, low levels of TNT improved physiological response in plants due to the slight increase in nitrogen, while high levels of TNT induced stress. Key markers in stress responses were identified, specifically with reflectance indices and derivatives, which may separate TNT-contaminated plants from naturally stressed plants and would allow for accurate detection of buried ordnance at the landscape level.

Parmi les contraintes abiotiques la salinité est considérée comme un problème majeur, qui affecte le fonctionnement des plantes, en particulier leur croissance et leur rendement. Afin d’étudier le contrôle génétique de la tolérance à la salinité chez Medicago truncatula, plante modèle de la famille des légumineuses, deux expérimentations ont été réalisées. La première expérimentation visait à étudier l’effet de la contrainte saline sur différents paramètres morpho-physiologiques pour un panel de génotypes de M. truncatula afin de déterminer les traits de phénotypage pour la tolérance à la salinité. Les génotypes A17, TN1.11, DZA315.16, A20, TN1.12 et F83005.5 ont été sélectionnés parmi des lignées originaires de différents pays méditerranéens, qui ont été déjà séquencées (http://www1.montpellierinra.fr/BRC-MTR/mauguio/mauguio.php). Les génotypes ont été étudiés sous 6 traitements salins (0, 30, 60, 90,120 et 150 mM NaCl) dans un essai factoriel sous forme de blocs complets aléatoires en trois répétitions. L’analyse de la variance montre des différences significatives entre les niveaux de salinité et une interaction entre les génotypes et les traitements salins concernant la plupart des caractères étudiés. Le génotype « DZA315.16 » présente les valeurs les plus importantes concernant les effets principaux pour les caractères morphologiques alors que « TN1.11 » présente les valeurs les plus faibles. La projection verticale de la surface foliaire de la plante (Leaf Area=LA), significativement corrélée à la biomasse des plantes, apparaît comme un trait d’intérêt pour le phénotypage de la tolérance à la salinité. La concentration saline la mieux adaptée pour démontrer les différences parmi les lignes étudiées se situe entre 90 et 120 mM NaCl. Le génotype « TN1.11 » contrairement à « DZA315.16 » et à « Jemalong-A17 » présente un maintien de la surface foliaire de la plante en réponse à la salinité. Pour la deuxième expérimentation, une population de cent lignées recombinantes (Recombinant Inbred Lines=RILs) produite par le croisement entre « TN1.11 » et « Jemalong-A17 » a été retenue pour l’analyse du contrôle génétique de la tolérance à la salinité. Les RILs ont été développés par la méthode de descendant mono graines (Single Seed descent= SSD) jusqu’ à la génération F6 à l’INP-ENSAT, France. Le plan d’experimentation est « Spli plots » , sous forme de blocs randomisés avec trois répétitions et deux conditions : traitement salin (100 mM NaCl) et témoin (eau). L’expérience a été menée pour déterminer la variabilité génétique et pour identifier les QTLs contrôlant les caractères morphologiques et physiologiques chez la population des lignées recombinantes (RILs). L’analyse de la variance a montré une large variation génétique et une ségrégation transgressive pour les caractères étudiés. La différence entre la moyenne des RILs et la moyenne de leurs parents n’est pas significative concernant tous les caractères étudiés dans les deux conditions, ce qui montre que les RILs utilisées dans notre expérimentation sont représentatives de toutes les lignées recombinantes possibles du croisement « TN1.11 x Jemalong-A17 ». 21 QTLs ont été détectés dans la condition témoin et 19 QTLs ont été identifiés sous contrainte saline (100 mM NaCl). Le pourcentage de la variance phénotypique expliqué par les QTLs varie entre 4.60% et 23.01%. Certains de ces QTLs sont spécifiques à la condition saline, ce qui démontre l’existence du contrôle génétique de la tolérance à la salinité chez M. truncatula ; tandis que les autres ne sont pas spécifiques et contrôlent un même caractère dans les deux conditions. Des QTLs superposés concernant différents caractères ont été aussi observés. Les résultats fournissent des informations importantes en vue de futures analyses fonctionnelles de la tolérance à la salinité chez M.truncatula et pour d’autres espèces voisines. / Among abiotic stresses salinity is considered as a serious problem affecting plant functions especially growth and yield. In order to study the genetic control of salt stress in the model legume Medicago truncatula, two experiments were performed. The first experiment was conducted to study the effect of salt stress on some morpho-physiological parameters in M. truncatula genotypes and to determine the eventual use of some traits as tolerance criteria. Genotypes including A17, TN1.11, DZA315.16, A20, TN1.12 and F83005.5 are selected through a sequenced lines collection (http://www1.montpellierinra.fr/BRC-MTR/mauguio/mauguio.php) which are originated from different Mediterranean countries. Genotypes were studied under 6 salinity treatments (0, 30, 60, 90,120 and 150 mM NaCl) in a factorial design based on randomized complete blocks with three replications. Analysis of variance show significant differences among genotypes, salinity levels and interaction between genotypes and salt treatments for most of studied traits. “DZA315.16” genotype presents the highest main effect values for morphological traits whereas”TN1.11” has low values. Vertically projected leaf area (LA); show the highest variability through all studied salt concentrations. The best concentration to find differences between parental lines is 90 to 120 mM Nacl. A segregating population of recombinant inbred lines (100 RILs) of M.truncatula derived from a cross between TN1.11 and Jemalong-A17 was used for the second experiment. RILs were developed by single-seed descent until F6 generation at the INP-ENSAT, France. The experiment was undertaken to determine the genetic variability and to identify QTLs controlling several traits related to plant growth and physiology, in the population of recombinant inbred lines (RILs). Analyses of variance showed a large genetic variation and transgressive segregation for the traits studied. The difference between the mean of RILs and the mean of their parents was not significant for all of the traits in both conditions, showing that the RILs used in our experiment are representative of the possible recombinant lines from the cross TN1.11 x A17. A total of 21 QTLs were detected under control and 19 QTLs were identified under 100mM salt stress conditions. The percentage of total phenotypic variance explained by the QTLs ranged from 4.60% to 23.01%. Some of the QTLs were specific for one condition, demonstrating that the genetic control of a traits differed under control and salt stress conditions. Some others are non-specific and control a trait in both conditions. Overlapping QTLs for different traits were also observed. The results provide important information for further functional analysis of salt tolerance in M. truncatula

Pour étudier le rôle de la mitochondrie dans la tolérance à la sécheresse, la réponse à la contrainte hydrique a été comparée entre une lignée sauvage (WT) et un mutant CMSII (Cytoplasmic Male Sterile) de Nicotiana sylvestris. Chez le mutant CMSII, le complexe I mitochondrial est absent et la respiration est assurée par les NAD(P)H déshydrogénases alternes et elle est maintenue à un niveau supérieur de l’ordre de 20 à 30% à celui du WT. La différence observée entre les plantes WT et CMSII met en jeu non seulement le fonctionnement mitochondrial, mais également le fonctionnement des chloroplastes. En effet, l’activité photosynthétique du mutant est plus faible que celui du WT et elle est corrélée avec une plus faible conductance stomatique (gs) et mésophyllienne (gm).Après l’arrêt de l’arrosage, on observe que le contenu relatif en eau (RWC) diminue plus lentement chez les feuilles du CMSII. Ceci n’était pas le résultat d’une plus petite surface de transpiration ou d’une masse racinaire d’absorption plus élevé puisque le rapport partie aérienne/racine et la surface foliaire totale ont été similaires au début de l’expérience chez les deux génotypes. De plus la mutation n’a pas induit des changements au niveau des paramètres hydriques (P0, PTLP, RWCTLP) ni au niveau de la densité stomatique. La tolérance des plantes CMSII a été le plus probablement la conséquence de sa plus faible transpiration en conditions bien hydratées et aux premiers jours de déshydratation et non pas d’une meilleure efficacité d’absorption de l’eau puisque le contenu en eau du sol reste plus élevé chez CMSII après l’arrêt de l’arrosage. La plus faible conductance stomatique chez le CMSII bien hydraté a été expliquée par sa plus faible conductance hydraulique. De plus, contrairement au WT, le niveau des acides aminés totaux diminue au cours de la déshydratation lorsque le contenu en protéines solubles augmente chez les feuilles du CMSII, suggérant une accélération de la remobilisation des acides aminés. D’autre part, il a été aussi montré que le mutant CMSII est capable de s’acclimater mieux à la sécheresse que le WT lorsqu’ils ont été maintenus à un RWC de 80 % sur plusieurs jours. Sous ces conditions, la photosynthèse reste plus élevée chez le mutant que chez le WT. Cette meilleure acclimatation corrèle avec une plus forte photorespiration du CMSII sous conditions bien hydratées et sous conditions d’acclimatation. La photorespiration chez CMSII et le WT a été estimée par le transport électronique dévolu à l’oxygénation de RuBP et en plus par l’accumulation des métabolites impliqués dans la photorespiration. D’une part, l’acclimatation à la sécheresse diminue gm plus fortement chez le WT que chez le CMSII. D’autre part, le WT accumule la glycine ce qui laisse supposer que le glycine décarboxylase mitochondrial est plus affectée chez le WT que chez le CMS et inhibe ainsi la photorespiration. En effet, cette plus faible photorespiration chez le WT affecte les réactions primaires de la photosynthèse par une accumulation d’un gradient de protons estimé par le quenching non-photochimique (NPQ) de la fluorescence chlorophyllienne ce qui induit une diminution du transport électronique des réactions primaires de la photosynthèse. / To investigate the role of mitochondria in drought stress, the response to water deprivation was compared between Nicotiana sylvestris wild type (WT) plants and the CMSII respiratory complex I mutant. In CMSII, alternative NAD(P)H-dehydrogenases bypassing complex 1 allow respiration.. The difference of mitochondrial function between WT and CMSII plants affect also photosynthesis. The CMSII has lower photosynthetic actitvity than the WT and lower stomatal (gs) and internal (gm) conductances to CO2. When watering of plants with similar leaf surface and similar shoot/root ratio was stopped the relative water content (RWC) declined faster in WT as compared to CMSII leaves. Furthermore, CMSII and WT leaves had the same osmotic potential at leaf saturation (P0) and at leaf turgor lost pressure (PTLP) and the same stomatal density. The slower decline of RWC in CMSII, compared to WT leaves, was most likely the consequence of the lower stomatal conductance (gs) under well-watered conditions and during the first days after withholding watering, The lower stomatal conductance of well-watered CMSII leaves correlated with a lower hydraulic conductance of leaves. Remarkably, total free amino acid levels declined and total soluble protein content increased in CMSII leaves, while the opposite was observed in WT leaves. This suggests protein synthesis in CMSII but protein degradation in WT leaves during drought stress. We also show that CMSII leaves better acclimate to drought stress than the WT leaves. After several days at 80 % RWC , photosynthesis is higher in the mutant than in WT. As compared to the WT, the mutant shows higher rates of photorespiration before and after acclimation to drought.The strong accumulation of glycine in the WT suggests that photorespiration may be limited at the level of glycine decarboxylase. In addition, after acclimation to drought gm declined markedly in WT but not in CMSII leaves, thus further limiting CO2 supply for photosynthesis in the WT. The resulting lower photosynthesis and photorespiration in WT leaves affect also the primary reaction of photosynthesis by increasing the non-photochemical fluorescence quenching (NPQ) and decreasing linear electron transport.

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No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tese_anderson_augusto_schock.pdf: 1520871 bytes, checksum: 3832517fddb50738dca235fa9b7cbf2e (MD5) Previous issue date: 2015-03-19 / Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul - FAPERGS / A pesquisa foi desenvolvida em dois experimentos com o objetivo de verificar o efeito da aplicação de piraclostrobina na mitigação do estresse causado pelo alagamento em plantas jovens de Jatropha curcas L.. Ambos os experimentos foram desenvolvidos na Universidade Federal de Pelotas, Campus Capão do Leão, Município de Capão do Leão, RS. Foram utilizadas sementes de Jatropha curcas L. (pinhão manso) oriundas do município de Janaúba- MG. A semeadura foi realizada em vasos plásticos com capacidade de 5 L, dispostos em espaçamento 50 x 50 cm, contendo uma mistura de solo com areia (proporção 2:1). O primeiro experimento consistiu em testar a influência da piraclostrobina sobre a fluorescência transiente da clorofila, trocas gasosas, curvas de resposta fotossintética, atividade da enzima redutase do nitrato e níveis de nitrogênio foliar. Para isso foram instalados quatro tratamentos com piraclostrobina com as seguintes quantidade de ingrediente ativo (i.a.) por litro de calda: T1 – 0 g i.a. L-1, T2 - 0,05 g i.a. L-1, T3 - 0,10 i.a. g L-1, T4 - 0,20 g i.a. L-1. Como fonte de piraclostrobina foi utilizado o fungicida Comet® (250 g i.a L-1). As plantas receberam duas aplicações nas referidas doses, a primeira aos 30 dias após a emergência e a segunda cinco dias após. Constatou-se que as diferentes concentrações de piraclostrobina ocasionaram respostas fisiológicas diferentes às plantas ao longo do experimento. Observou-se efeitos positivos nos índices de performance fotossintética nas plantas que receberam a piraclostrobina e as taxas assimilatórias líquidas aumentaram respectivamente com o aumento das concentrações. Por outro lado apenas concentração 0,1 g i.a. L-1 demonstrou correlação positiva nos índices de clorofila. O segundo experimento consistiu em testar a eficiência da aplicação de piraclostrobina na superação do estresse hipóxico causado pelo alagamento. Para tal foram avaliados os índices de fluorescência transiente da clorofila, trocas gasosas e parâmetros biométricos. Este ensaio foi constituído por quatro tratamentos nas seguintes condições: T1- normóxia sem piraclostrobina; T2- normóxia com piraclostrobina (0,1 g i.a. L-1); T3- hipóxia sem piraclostrobina e T4- hipóxia com piraclostrobina (0,1 g i.a. L-1). Os resultados demostram que a condição de hipóxia reduziu o crescimento das plantas, proporcionado também uma serie de outros efeitos fisiológicos negativos que prejudicaram o fluxo de elétrons na cadeia transportadora de elétrons cloroplastídica e também as trocas gasosas às plantas jovens de Jatropha curcas L.. Contudo, a aplicação de piraclostrobina causou menor redução dos índices de clorofila e biomassa e também proporcionou atraso dos efeitos negativos do estresse na cadeia transportadora de elétrons cloroplastídica. Diante do exposto pode-se dizer que os processos fisiológicos da Jatropha curcas L. respondem diferentemente as concentrações de piraclostrobina ao longo do tempo. Assim sendo, os resultados deste experimento podem fundamentar futuros estudos na utilização da piraclostrobina como uma alternativa a superação de estresses abióticos. / The research was conducted in two experiments in order to verify the application of pyraclostrobin effect in mitigating the stress caused by flooding in young plants of Jatropha curcas L. (physic nut ). Both experiments were conducted at Federal University of Pelotas, Campus Capão do Leão, City Capão do Leão, RS. Jatropha curcas L. seeds were used originated in the city of Janaúba- MG. The seeds were sown in plastic pots with 5 L capacity, arranged in spaced 50 x 50 cm, containing a soil mixture with sand (proportion 2: 1). The first experiment is to test the influence of pyraclostrobin on transient chlorophyll fluorescence, gas exchange, photosynthetic response curves reductase activity of the enzyme nitrate and leaf nitrogen levels. To this were four treatments with pyraclostrobin at the following amount of active ingredient (ai) per liter of water: T1 – 0 g i.a. L-1, T2 - 0,05 g i.a. L-1, T3 - 0,10 i.a. g L-1, T4 - 0,20 g i.a. L-1. As pyraclostrobin source was used COMET® fungicide (250 g ai L-1). Plants take two applications in these doses, the first at 30 days after emergence and the second five days. It was found that the different concentrations of pyraclostrobin caused different physiological responses to plants throughout the experiment. We observed positive effects on rates of photosynthetic performance on plants supplied pyraclostrobin and net assimilatory rates increased respectively with increasing concentrations. In contrast, only concentration 0.1 g ai L-1 showed a positive correlation chlorophyll indices. The second experiment is to test the pyraclostrobin application efficiency in overcoming the hypoxic stress caused by flooding. To this end we evaluated the transient fluorescence of chlorophyll indices, gas exchange and biometric parameters. This test consisted of four treatments as follows: T1 normoxia without pyraclostrobin; T2 normoxia with pyraclostrobin (0.1 g ai L-1); T3-T4 hypoxia without pyraclostrobin and hypoxia with pyraclostrobin (0.1 g ai L-1). The results demonstrate that hypoxia condition reduces plant growth, also provided a number of other negative physiological effects that hindered the flow of electrons in the electron transport chain cloroplastídica and also gas exchange in young plants of Jatropha curcas L .. However, the application of pyraclostrobin caused less reduction in chlorophyll contents and biomass and also caused a delay of the negative effects of stress on the conveyor chain cloroplastídica electrons. Given the above it can be said that the physiological processes of Jatropha curcas L. respond differently to pyraclostrobin concentrations over time. Therefore, the results of this experiment can support future studies on the use of pyraclostrobin as an alternative to overcome abiotic stresses.

The Arbuscular mycorrhiza (AM) is an endosymbiont of higher plant roots. Most land plants and cultivated crops are concerned to AM symbiosis. This endosymbiosis is based on the mutual exchange of nutrients between plant and fungus. Therefore, AM symbiosis leads to an increased demand for photosynthetic products. The aim of this study was to investigate the pathway used by plants during AM symbiosis to increase photosynthetic performance. Therefore, we have carried out a systematic characterization of photosynthesis in Medicago truncatula (M. truncatula), which is a model legume. We observed colonization by the fungus in roots and that AM symbiosis increases the fresh and dry plant biomass. This could be attributed to an increase in both photosynthetic efficiency and capacity in AM plants. Consistent with these observations, AM symbiosis enhanced phosphorus uptake from the soil into roots, stems and leaves, as based on analyses of phosphorus content. Based on equal chl loading, no differences were found regarding D1, Lhcb1 and Lhcb2 protein content in four plant groups. This indicates similar ratio between chl and PSII proteins. Furthermore, AM symbiosis increases the amount of chlorophyll, steady state oxygen evolution activities, maximum quantum yield (Fv/Fm), and photosynthetic electron transport rate (about 5 fold). Nevertheless, photoprotection was not affected by AM symbiosis. We observed an increase in weight of seed/fruit and weight of seed/plant in AM plants (about 2 fold). Based on these results, we propose that AM symbiosis increases both the efficiency and the capacity of photosynthetic apparatus in the M. truncatula.

Η υδατική καταπόνηση αποτελεί έναν από τους κυριότερους περιβαλλοντικούς παράγοντες περιορισμού της φυτικής ανάπτυξης και απόδοσης στα ξηρά και ημίξηρα οικοσυστήματα, συμπεριλαμβανομένων κι αυτών με μεσογειακού τύπου κλίμα. Σκοπός της παρούσας ερευνητικής εργασίας ήταν η εκτίμηση των επιδράσεων της υδατικής καταπόνησης στη φωτοχημική ικανότητα του PSII και τις φωτεινές αντιδράσεις της φωτοσύνθεσης, σε τυπικά μεσογειακά είδη. Η υδατική καταπόνηση επιτεύχθηκε τόσο εργαστηριακά εντός διαστήματος λίγων ωρών, όσο και σε ημι-φυσικές συνθήκες περιβάλλοντος σε διάστημα περίπου δύο εβδομάδων. Μετά τις δύο εβδομάδες τα φυτά επανυδατώθηκαν, με στόχο την εκτίμηση της ικανότητας ανάκαμψης από το έντονο υδατικό stress. Η ανάλυση της κινητικής της επαγωγής του φθορισμού της χλωροφύλλης a μέσω του JIP-test αποκάλυψε την άμεση επίδραση της έλλειψης νερού στη ροή των ηλεκτρονίων, όπως αυτή εκφράζεται από τις παραμέτρους ψΕο, φΕο, δRo και φRo. Επιπλέον, παρατηρήθηκε αρνητική συσχέτιση του δείκτη RWC με το συνολικό απόθεμα των ενεργών κέντρων του PSI και τους τελικούς υποδοχείς του PSI, καθώς και με τους δείκτες φωτοσυνθετικής απόδοσης PIABS & PItotal. Η μέγιστη φωτοχημική ικανότητα του PSII, φPo(=Fv/Fm) μειώθηκε μόνο σε πολύ χαμηλές τιμές RWC στις εργαστηριακές μετρήσεις που πραγματοποιήθηκαν, ενώ στις ημι-φυσικές συνθήκες η μείωση του φPo την δεύτερη εβδομάδα της υδατικής καταπόνησης συνοδεύτηκε από αυξημένη θερμική απόσβεση της απορροφούμενης ενέργειας διεγέρσεως. Η ανάπτυξη του υδατικού stress σε ημι-φυσικές συνθήκες οδήγησε επίσης στη μεταβολή των ειδικών ενεργειακών ροών ανά δραστικό ενεργό κέντρο του PSII, πιθανώς λόγω της μερικής αδρανοποίησης ενεργών κέντρων στις συνθήκες αυτές. Επιπλέον, καταγράφηκε αύξηση του φθορισμού στα 300 μsec (Σημείο Κ), ένα φαινόμενο που συνδέεται με καταστολή της λειτουργίας του συμπλόκου έκλυσης οξυγόνου. Οι εν λόγω μεταβολές δεν παρατηρήθηκαν στον ίδιο βαθμό στις εργαστηριακές μετρήσεις, γεγονός που πιθανώς οφείλεται στη σταδιακή ανάπτυξη της ξηρασίας και τη συνδυαστική δράση της έλλειψης νερού, της υψηλής θερμοκρασίας και των υψηλών εντάσεων φωτός κάτω από φυσικές συνθήκες περιβάλλοντος. Από την πρώτη κιόλας ημέρα μετά την επανυδάτωση των φυτών, όλες οι παράμετροι του JIP-test άρχισαν να επανακάμπτουν. Ωστόσο, εντοπίστηκαν διαφορές στην ταχύτητα επαναφοράς μεταξύ των διαφορετικών ειδών. Συνολικά, τα ευρήματα μας υποδεικνύουν καταστολή των φωτεινών αντιδράσεων της φωτοσύνθεσης σε συνθήκες υδατικής καταπόνησης, μεταβολή η οποία αντιστράφηκε γρήγορα στα πειράματά μας. Θα μπορούσε λοιπόν, να θεωρηθεί ως ένας μηχανισμός προσαρμογής που εξασφαλίζει στα μεσογειακά είδη που μελετήθηκαν την διατήρηση της λειτουργικότητας της φωτοσυνθετικής συσκευής, ακόμη και σε συνθήκες έντονης υδατικής καταπόνησης. / Water stress is one of the most important environmental factors limiting plant growth and yield in arid and semi-arid ecosystems, including those with Mediterranean climate. The objective of this study was to evaluate the effects of water stress on the photosynthetic capacity of PSII and on the light reactions of photosynthesis of typical Mediterranean species. Water stress experiments were conducted both under laboratory conditions, through dehydration of leaf-disks within hours and under semi-natural conditions by withholding water for about two weeks. Drought period was followed by six days of re-watering, in order to assess the recovery of the photosynthetic apparatus. Analysis of the polyphasic OJIP fluorescence transient through JIP-test, revealed direct limitations in electron transport, as expressed by the parameters ψΕο, φΕο, δRo and φRo. Furthermore, a negative correlation was found between RWC and the pool of the RCs of PSI, the pool of electron end-acceptors, and the photosynthetic efficiency, as expressed by the parameters PIABS & PItotal. The maximum quantum yield of primary photochemistry φPο (=Fv/Fm) was affected only under very low RWC values during the laboratory experiments. On the other hand, a reduction was found during the second week of drought stress treatment, along with an increased conversion rate of absorbed light into thermal energy. Under semi-natural conditions, water stress also caused a shift in the specific energy flows per RC of PSII, caused possibly by inactivation of RCs. Moreover, there was an increase in fluorescence intensity at 300 microseconds (K-band), a phenomenon associated with inhibition of the oxygen evolving complex. These alterations were not observed to the same extent in the laboratory experiments, so they can be attributed to the gradual onset of drought and the combined effect of dehydration, high temperature and high light intensities under natural conditions. Within one day after rehydration, all the parameters of the JIP-test began to recover. However, differences in the recovery rate were observed between different species. In conclusion, our findings indicate down-regulation of light reactions of photosynthesis under water stress. This phenomenon was quickly reversed in our experiments. Therefore, it could be regarded as an adaptive response, which maintains the functionality of the photosynthetic apparatus of the Mediterranean species studied, even under severe water deficit.

Cultivars of Agrostis stolonifera showed weak and strong responsiveness to the systemic acquired resistance (SAR) activator, benzothiadiazole (BTH), or the induced systemic resistance (ISR) activator, 2R, 3R-butanediol (BD). Next Generation RNA sequencing was used to identify 2163 putative transcripts with increased expression in BTH versus water-treated A. stolonifera. Among three BTH-induced genes, AsASP-2 and AsHIR-1 were induced faster, while AsLOX-1 had stronger transient induction, in one out of two strongly BTH-responsive cultivars. Three ISR-responsive genes, AsGNS-5, AsOPR-4 and AsAOS-1, showed no greater induction or priming in the strongly versus weakly BD-responsive cultivars. Cultivars of A. stolonifera vary significantly in their response to defense activators, however this is not consistently related to defense gene expression. To quantify disease severity, chlorophyll fluorescence imaging of the maximum quantum efficiency of photosystem II (Fv/Fm) was tested on Nicotiana benthamiana infected with Colletotrichum orbiculare. Leaf areas of healthy, non-necrotic affected and necrotic tissue could be individually quantified, which demonstrated that BD delayed symptom development by approx. 24-hour and reduced non-necrotic affected tissue compared to controls. Chlorophyll fluorescence imaging can quantify and reveal novel features about induced disease resistance.

Στα φύλλα κάποιων φυτικών ειδών παρουσιάζεται παροδική ερυθρότητα, λόγω συσσώρευσης ανθοκυανινών, η οποία μπορεί να είναι τόσο έντονη που να καλύπτεται το χρώμα της χλωροφύλλης. Δεδομένου ότι απορροφούν και στο ορατό φάσμα (κυρίως στην πράσινη και ελαφρώς στη μπλε και κίτρινη περιοχή) οι ανθοκυανίνες πρακτικώς λειτουργούν ως φίλτρα, αφού απορροφούν μέρος της προσπίπτουσας φωτοσυνθετικά ενεργής ακτινοβολίας και ανταγωνίζονται τις χλωροφύλλες στην απορρόφηση του φωτός. Εξ αιτίας αυτής της ιδιότητάς τους, ένας από τους ρόλους που τους έχει αποδοθεί είναι η φωτοπροστατευτική δράση, ενώ παράλληλα (και ανεξαρτήτως ρόλου που επιτελούν) έχει προταθεί ότι διαμορφώνουν ένα ιδιαίτερο μικροπεριβάλλον σκιάς στο εσωτερικό του φύλλου. Προκειμένου να διερευνηθεί η πιθανή συνεισφορά των ανθοκυανινών στη φωτοπροστασία αλλά και στην επαγωγή σκιόφιλων χαρακτήρων, στην παρούσα διατριβή παρακολουθήσαμε την πορεία των φωτοσυνθετικών και φωτοπροστατευτικών χαρακτηριστικών, παράλληλα με τη μεταβολή της ερυθρότητας, σε φύλλα του ίδιου είδους που διαφέρουν ως προς το ανθοκυανικό περιεχόμενο. Εξετάστηκαν πέντε είδη, τρία ως κύρια πειραματόφυτα (Cistus creticus L., Photinia x fraseri Dress και Quercus coccifera L.) και δύο ως συμπληρωματικά στον προσδιορισμό της στοιχειομετρίας των δύο φωτοσυστημάτων (Ricinus communis L., Rosa sp. L.). Στο C. creticus, η παροδική ερυθρότητα εμφανίζεται στα ώριμα φύλλα και σχετίζεται με αβιοτικούς περιβαλλοντικούς παράγοντες, συγκεκριμένα το συνδυασμό χαμηλών θερμοκρασιών και υψηλών εντάσεων φωτός. Στα υπόλοιπα, η παροδική συσσώρευση ανθοκυανινών επάγεται από οντογενετικούς παράγοντες (νεαρά κόκκινα φύλλα που πρασινίζουν με την ωρίμανση). Στα P. x fraseri και R. communis, η χρωματική ποικιλομορφία αφορά φύλλα διαφορετικής ηλικίας, που ανήκουν όμως στο ίδιο άτομο (νεαρά κόκκινα-ώριμα πράσινα). Tα υπόλοιπα τρία είδη εμφανίζουν ενδο-ειδική ποικιλομορφία ως προς τον ανθοκυανικό χαρακτήρα, και επομένως συγκρίνονται φύλλα ίδιας κάθε φορά φυσιολογικής ηλικίας, που ανήκουν σε άτομα διαφορετικών φαινοτύπων. Στην περίπτωση που οι ανθοκυανίνες λειτουργούν φωτοπροστατευτικά, αναμένεται τα φύλλα που τις περιέχουν να αντιμετωπίζουν μικρότερο κίνδυνο φωτοαναστολής και να έχουν μικρότερη ανάγκη για μη-φωτοχημική απόσβεση σε σύγκριση με τα αντίστοιχα πράσινα. Ωστόσο, από τις μετρήσεις του in vivo φθορισμού της χλωροφύλλης σε προ-σκοτεινιασμένα δείγματα (OJIP-ανάλυση) δεν διαπιστώθηκε τα ανθοκυανικά φύλλα να διαθέτουν κάποιο συγκριτικό πλεονέκτημα όσον αφορά στη φωτοσυνθετική τους λειτουργία. Αντιθέτως, τα κόκκινα φύλλα των ειδών P. x fraseri και C. creticus εκτός του ότι παρουσιάζουν ελαφρώς, αλλά στατιστικώς σημαντικά, χαμηλότερες τιμές μέγιστης φωτοχημικής απόδοσης του PSII (Fv/Fm) συναντούν και σαφώς μεγαλύτερους περιορισμούς στα μετέπειτα στάδια ηλεκτρονιακής ροής και μετατροπής της ενέργειας σε σχέση με τα αντίστοιχα πράσινα. Και στις δυο περιπτώσεις είναι πιθανό να συμβαίνει απενεργοποίηση κάποιων λειτουργικών ενεργών κέντρων του PSII. Στο Q. coccifera, τα όμοιας ηλικίας φύλλα των δύο φαινοτύπων δεν παρουσιάζουν διαφορές στις ανωτέρω παραμέτρους. Οι μετρήσεις φθορισμού της χλωροφύλλης κάτω από συνθήκες ακτινικού φωτός έδειξαν ότι τα κόκκινα φύλλα έχουν όμοιες (P. x fraseri και C. creticus) ή χαμηλότερες (Q. coccifera) τιμές τρέχουσας φωτοχημικής απόδοσης (YII) και αντίστοιχα όμοιες ή υψηλότερες τιμές μη-φωτοχημικής απόσβεσης (YNPQ). Ωστόσο, λόγω της απορρόφησης των ανθοκυανινών, οι χλωροπλάστες των κόκκινων φύλλων δέχονται πραγματική ένταση φωτός χαμηλότερη της προσπίπτουσας, και επομένως τα ανωτέρω αποτελέσματα είναι αντίθετα των αναμενόμενων. Διαφωτιστικές προς αυτή την κατεύθυνση είναι οι μετρήσεις φθορισμού της κάτω (πράσινης) φυλλικής επιφάνειας σε φωτιζόμενα φύλλα των δύο φαινοτύπων του C. creticus. Σ’ αυτήν την περίπτωση, έχοντας παρακάμψει το πρόβλημα της απορρόφησης των ανθοκυανινών, τα φύλλα του κόκκινου φαινοτύπου εμφανίζουν υψηλότερη ανάγκη μη-φωτοχημικής απόσβεσης (YNPQ) και υψηλότερη ευαισθησία έναντι της φωτοαναστολής (YNΟ), παρ’ όλη τη θεωρούμενη προστασία των υπερκείμενων ανθοκυανινών της άνω επιφανείας. Η μεγαλύτερη ανάγκη μη φωτοχημικής απόσβεσης των κόκκινων φύλλων διαφαίνεται και από την υψηλότερη επένδυση σε συστατικά του κύκλου των ξανθοφυλλών (P. x fraseri) ή την καλύτερη λειτουργικότητα του κύκλου (C. creticus), σε συνθήκες έντονου φωτισμού. Μάλιστα, κατά την καταπονητική περίοδο του χειμώνα ο κόκκινος φαινότυπος του C. creticus εμφανίζει μείωση των περιεχομένων χλωροφυλλών, πιθανόν σε μια προσπάθεια περαιτέρω ελάττωσης της απορροφούμενης ενέργειας διεγέρσεως, παρ’ όλη την ύπαρξη του ανθοκυανικού φίλτρου. Με βάση τα ανωτέρω ευρήματα, η υπόθεση του φωτοπροστατευτικού ρόλου των ανθοκυανινών δεν ενισχύεται, εκτός εάν υποτεθεί είτε ότι ο στόχος για την προστασία δεν συνδέεται με τη φωτοσύνθεση είτε ότι τα κόκκινα φύλλα εάν δεν τις διέθεταν θα ήταν σε πολύ πιο δυσμενή θέση, σε σύγκριση πάντα με τα πράσινα. Όσον αφορά στη συνεισφορά των ανθοκυανινών στην επαγωγή σκιόφιλων χαρακτήρων βρέθηκαν κάποιες ισχυρές ενδείξεις προς αυτήν την κατεύθυνση, ανεξάρτητα από τον παράγοντα που επάγει τη συσσώρευσή τους. Έτσι, και στα πέντε είδη που εξετάστηκαν, παρατηρήθηκε αύξηση της σχετικής αναλογίας PSII/PSI στα κόκκινα φύλλα, η οποία είναι στατιστικά σημαντική (εκτός από την περίπτωση του Q. coccifera) και βαίνει μειούμενη με τη μείωση των περιεχομένων ανθοκυανινών. Αύξηση της αναλογίας PSII/PSI έχει συσχετισθεί με την ποιοτική αλλοίωση του φωτός σε συνθήκες σκιάς και στοχεύει στην εξισορροπημένη διέγερση των δύο φωτοσυστημάτων. Από τον τρόπο προσδιορισμού της (ως F686/F735), η ανωτέρω αναλογία δεν κάνει διάκριση μεταξύ φωτοσυλλεκτικής κεραίας και ενεργών κέντρων των δύο φωτοσυστημάτων. Μία άλλη παράμετρος, που θεωρείται τυπικός δείκτης σκιοφιλίας και μετρήθηκε στο σύνολο των πειραματοφύτων, είναι η αναλογία Chl a/b. Μειωμένη αναλογία Chl a/b σε συνθήκες σκιάς υποδεικνύει αύξηση των φωτοσυλλεκτικών κεραιών σε σχέση με τα ενεργά κέντρα. Στατιστικά σημαντική μείωση της ανωτέρω αναλογίας βρέθηκε στα κόκκινα φύλλα των ειδών C. creticus, Q. coccifera και Rosa sp.. Στα P. x fraseri και R. communis δεν παρατηρήθηκαν στατιστικά σημαντικές διαφορές μεταξύ κόκκινων και πράσινων φύλλων. Τέλος, στα είδη C. creticus, P. x fraseri και Q. coccifera προσδιορίσθηκε και η παράμετρος ABS/RC (όπως προκύπτει από την OJIP-ανάλυση), όπου παρατηρήθηκε στατιστικώς σημαντική αύξηση στα κόκκινα φύλλα των δύο πρώτων ειδών. Στο Q. coccifera η ίδια τάση δεν είναι στατιστικά σημαντική. Αύξηση της αναλογίας ABS/RC, η οποία αποτελεί θα λέγαμε έναν λειτουργικό περισσότερο δείκτη, μπορεί να αποδοθεί σε αυξημένο μέγεθος της φωτοσυλλεκτικής κεραίας ή/και σε μείωση του αριθμού των λειτουργικών ενεργών κέντρων του PSII. Από το συνδυασμό των παραπάνω αποτελεσμάτων διαφαίνεται ότι στα είδη C. creticus, Rosa sp. και Q. coccifera η επίδραση των ανθοκυανινών εντοπίζεται κυρίως σε αύξηση του μεγέθους της φωτοσυλλεκτικής κεραίας. Στα P. x fraseri και R. communis, η αύξηση του λόγου PSII/PSI αντανακλά πιθανώς μια αύξηση των ενεργών κέντρων του PSII. / Leaves of some plant species appear transiently red because of the accumulation of anthocyanins, at levels sufficient to mask the green chlorophyll color. Given that leaf anthocyanins absorb also in the visible spectrum (strongly in the green and less in the blue-violet and yellow region), they act as sunscreens attenuating part of the photosynthetically active radiation and hence they compete with chlorophylls for photon capture. Due to this attribute, one of the ascribed roles of leaf anthocyanins is the photoprotection of the photosynthetic mesophyll. Additionally (and regardless of the proposed role) it has been suggested that they shape a particular shade microenvironment in the leaf interior. In order to investigate these two hypotheses, in the present study we monitored the course of photosynthetic and photoprotective characteristics in leaves of the same species with different anthocyanic content. Five species have been used, three of them as main experimental material (Cistus creticus L., Photinia x fraseri Dress και Quercus coccifera L.) and two additional (Ricinus communis L., Rosa sp. L.) for the assessment of the stoichiometry of the two photosystems. In Cistus creticus (L.) transient redness is induced in mature leaves by abiotic environmental factors, i.e. the combination of low temperatures and high light intensity during winter. In the rest species redness is developmentally determined (young red leaves which become red upon maturation). In P. x fraseri and R. communis the color variation concerns leaves of different age from the same individual (comparison of young red and mature green). The rest three species show intra-species variation of the anthocyanic trait. Thus, leaves of the same physiological age from different individuals and phenotypes (occupying the same habitat) were compared. The photoprotective hypothesis of leaf anthocyanins entails that red leaves would be less sensitive against photoinhibition and have a reduced need of non-photochemical quenching, compared to their green counterparts. However, according to our chlorophyll fluorescence measurements with dark adapted samples (OJIP-analysis), anthocyanic leaves did not display a comparative advantage in their photosynthetic function. On the contrary, red leaves of P. x fraseri and C. creticus, apart from their slightly, yet statistically significant, lower maximum PSII yield (Fv/Fm), they also confront higher limitations in the further steps of excitation energy processing and its transformation compared to greens. In both species, a transformation of active PSII centers to non-QA -reducing ones, could be inferred. In Q. coccifera, no differences were observed between leaves of the same physiological age from the two phenotypes in the above parameters. Moreover, chlorophyll fluorescence measurements in light acclimated samples have shown that red leaves have similar (P. x fraseri και C. creticus) or lower (Q. coccifera) PSII effective yield (YII) and similar or higher corresponding non-photochemical energy quenching (YNPQ). It has to be noted, however, that the actual PAR levels penetrating to the red leaf mesophyll are in fact lower compared to their green counterparts, due to anthocyanin absorption of the white and blue actinic light used during these measurements. Accordingly, for the same incident PAR, the opposite trend was expected in red leaves. In C. creticus, measurements performed in the lower leaf side by-pass this problem, since anthocyanins accumulate only in the palisade mesophyll (of the upper side) and the abaxial leaf surface is always green. In that case, red leaves showed a higher need for non-photochemical quenching (YNPQ) and a higher vulnerability to photoinhibition (YNΟ), in spite of the supposed photoprotective function of the overlying anthocyanins. The trend for an enhanced need of non-photochemical dissipation in red leaves is indicated also by the higher investment in “xanthophyll cycle” components (P. x fraseri) and/or the lower EPS values (P. x fraseri and C. creticus) during midday. In addition, during the superimposed winter stress, red leaves of C. creticus proceed to a Chl loss, possibly as an attempt to decrease further the absorbed excitation energy, in spite of the anthocyanic screen. The above results weaken the photoprotective hypothesis for leaf anthocyanins, unless it is assumed either that photosynthesis is not the target for protection or that red leaves would be in a worse position, compared to greens, if anthocyanins were absent. Some of our results support the shade acclimation hypothesis in red leaves, irrespectively of the factor that induces the accumulation of anthocyanins. In all tested species, red leaves display a higher, statistically significant, relative PSII/PSI ratio compared to greens (except of Q. coccifera, where the same was observed as a trend). PSII/PSI ratio declines in parallel with the decrease of anthocyanin accumulation. A higher PSII/PSI ratio is considered as an acclimation to the altered light quality under shade conditions in order to adjust the excitation pressure of two photosystems. Chl a/b was statistically lower in red leaves of C. creticus, Q. coccifera και Rosa sp., while in P. x fraseri and R. communis no differences were observed. A decrease in Chl a/b ratio is a typical shade acclimation feature, indicating an increased ratio of light harvesting antennae per reaction center. Finally, another parameter indicative of shade acclimation is the mean antenna size (calculated as ABS/RC ratio), which was considerably higher in red leaves of C. creticus and P. x fraseri (again in Q. coccifera only a similar trend was observed). Yet, according to the JIP-analysis the antenna size is expressed per active RC, hence the increased ABS/RC ratio could be the combined effect of an increase in the antenna size and a decrease in active PSII centers. The combination of the above results indicates that anthocyanin accumulation in C. creticus, Rosa sp. and Q. coccifera contributes mainly to a higher relative size of LHC. In the case of P. x fraseri and R. communis, the enhanced PSII/PSI ratio of red leaves reflects rather an increase of PSII centers.

This study aimed to characterize the physiological responses to drought of two sugarcane (Saccharum spp.) varieties in order to contribute information to study the physiological processes involved in plant adaptation to drought. To this end, an experiment was installed in a completely randomized design in a greenhouse at the Laboratory of Plant Physiology of the Center for Agrarian Sciences, Federal University of Alagoas, located in Rio Largo - AL (9° 28 S, 35° 49 W, with 127 m of altitude), and where the varieties RB855536 and RB867515 with contrasting responses to drought, were subjected to two, four, six and eight days of water stress and rehydration. The physiological and biochemical variables were analyzed, leaf water potential (w), solute potential (s), chlorophyll fluorescence (Fv/Fm and ФPSII), soluble sugars (glucose, fructose, sucrose) and starch and antioxidant enzymes, catalase (CAT), superoxide dismutase (SOD) and ascorbate peroxidase (APX). Water stress resulted in significant reduction in the w and s in two varieties studied, which did not support a drought of more than six days. After rehydration, the varieties studied, showed the ability to restore in the w and s. The RB867515 had greater reductions in the w and s in relation to RB855536. Both varieties studied presented chronic and dynamic photoinhibition gradually with the increase in time of drought stress and a strong reduction in the quantum potential efficiency (Fv/Fm) and effective (ΦPSII). The variety RB867515 showed higher capacity of recovery of the ΦPSII and rapair the effects of photoinhibition. The drought caused the decrease in the levels of sucrose and starch in two varieties, with greater reduction of recovery sucrose and starch compared to control, presented by RB867515. Increases in the levels of glucose and fructose were also observed in both varieties under water restriction. Changes in levels of sugars, indicating that rising levels of glucose and fructose occurred through the degradation of sucrose and starch, likely to hold the osmotic adjustment. The variety RB867515 showed higher activity of CAT and APX, in relation to RB855536, and SOD activity increased with increasing time of drought. In this work, the variety RB867515 showed greater efficiency in the activation of mechanisms of tolerance to drought stress. / Conselho Nacional de Desenvolvimento Científico e Tecnológico / O presente estudo teve por objetivo, caracterizar fisiologicamente as respostas à seca de duas variedades de cana-de-açúcar (Saccharum spp.), afim de contribuir com informações para o estudo dos processos fisiológicos envolvidos na adaptação das plantas ao déficit hídrico. Para tanto, instalou-se um experimento em delineamento inteiramente ao acaso, em casa de vegetação no Laboratório de Fisiologia Vegetal do Centro de Ciências Agrárias da Universidade Federal de Alagoas, localizado no município de Rio Largo - AL (9° 28 S, 35° 49 W, a 127 m de altitude), onde as variedades RB855536 e RB867515 com respostas contrastantes à seca, foram submetidas a dois, quatro, seis e oito dias de estresse hídrico e posterior reidratação. As variáveis fisiológicas e bioquímicas analisadas foram, potencial hídrico foliar (w), potencial de solutos (s), fluorescência da clorofila a (Fv/Fm e ФPSII), açúcares solúveis (glicose, frutose, sacarose) e amido, e atividade de enzimas antioxidantes, catalase (CAT), superóxido dismutase (SOD) e ascorbato peroxidase (APX). O estresse hídrico ocasionou em redução significativa no w e s nas duas variedades estudadas, as quais não suportaram um período de seca superior a seis dias. Após a reidratação, as variedades pesquisadas, mostraram capacidade de restabelecimento do w e s. A RB867515 teve maior decréscimo no w e s em relação a RB855536. Ambas as variedades pesquisadas apresentaram fotoinibição crônica e dinâmica gradativa com o aumento do tempo de estresse hídrico além de forte redução na eficiência quântica potencial (Fv/Fm) e efetiva (ΦPSII). A variedade RB867515 mostrou maior capacidade de recuperação do ΦPSII e de reparação aos efeitos da fotoinibição. O déficit hídrico causou a diminuição dos teores de sacarose e amido nas duas variedades, com maior nível de redução de sacarose e amido em relação ao controle, apresentado pela RB867515. Incrementos nos níveis de glicose e frutose também foram observados nas duas variedades sob restrição hídrica. As mudanças nos níveis de açúcares, indicam que a elevação nos níveis de glicose e frutose se deu através da degradação de sacarose e amido, provavelmente, no sentido de realizar o ajuste osmótico. A variedade RB867515 mostrou maior atividade de CAT e APX, em relação a RB855536, além de atividade crescente da SOD com aumento do tempo de seca. No presente trabalho, a variedade RB867515 revelou maior eficiência na ativação de mecanismos de tolerância ao estresse hídrico.