Aluminium tolerance in bean traditional cultivars from Madeira

Common bean (Phaseolus vulgaris L.) is the most important legume crop in the world, providing low-cost, high quality protein, minerals and dietary fiber for human nutrition. The crop was originated from diversity centers in America and exhibits adaptation abilities to different environmental conditions, including soil with low pH. Acid soils occupy 30% of the agro ecosystem areas in the world. In Madeira, acid Andosols and unsatured Cambisols are the dominant groups of soils. Generally, under acidic and infertile conditions, besides of H+ toxicity, soluble aluminium (Al) is the most important abiotic factor limiting plant development and crop productivity. In the field, the hidden roots are also affected and the reduction of root growth under Al stress can be clearly observed in early stages. Seedlings of fifty bean accessions from the Archipelago of Madeira were tested under controlled conditions in the presence of 50 mM Al at pH 4.4. In general, the tested germplasm appeared to be sensitive or very sensitive to Al toxicity. However, fifteen traditional cultivars clearly exhibited elevated Al-tolerance, with an average root relative elongation (RRE) exceeding 50%, while top six accessions surpassed the 60% RRE mark. The Madeira bean germplasm is a valuable resource for sustainable crop production in acid soils and it could be used as parental lines in breeding programs aimed for Al tolerance in common beans.

Aluminium tolerance in bean traditional cultivars from Madeira

Common bean (Phaseolus vulgaris L.) is the most important legume crop in the world, providing low-cost, high quality protein, minerals and dietary fiber for human nutrition. The crop was originated from diversity centers in America and exhibits adaptation abilities to different environmental conditions, including soil with low pH. Acid soils occupy 30% of the agro ecosystem areas in the world. In Madeira, acid Andosols and unsatured Cambisols are the dominant groups of soils. Generally, under acidic and infertile conditions, besides of H+ toxicity, soluble aluminium (Al) is the most important abiotic factor limiting plant development and crop productivity. In the field, the hidden roots are also affected and the reduction of root growth under Al stress can be clearly observed in early stages. Seedlings of fifty bean accessions from the Archipelago of Madeira were tested under controlled conditions in the presence of 50 mM Al at pH 4.4. In general, the tested germplasm appeared to be sensitive or very sensitive to Al toxicity. However, fifteen traditional cultivars clearly exhibited elevated Al-tolerance, with an average root relative elongation (RRE) exceeding 50%, while top six accessions surpassed the 60% RRE mark. The Madeira bean germplasm is a valuable resource for sustainable crop production in acid soils and it could be used as parental lines in breeding programs aimed for Al tolerance in common beans.

Konstruktive und funktionale Entwicklung eines selbststeuernden Unterflurbewässerungsverfahrens

Unter Einfluss des gegenwärtigen Klimawandels sowie dem Anstieg der Erdbevölkerung ist der effiziente Umgang mit den vorhandenen Wasserressourcen ein zentrales Thema in der Bewässerungslandwirtschaft. Die Unterflurbewässerung ist ein seit Jahrtausenden bekanntes Verfahren, bei dem das Bewässerungswasser unterhalb der Erdoberfläche aufgebracht wird und der Pflanze direkt im Wurzelraum zur Verfügung steht. Die Gefäßbewässerung ist ein selbststeuerndes Verfahren wobei die Wassergabe aus dem unglasierten Tongefäß aufgrund der im umgebenden Boden anliegenden Saugspannung erfolgt. Diese wassersparende und dem Wasserbedarf der Pflanze angepasste Bewässerungsweise ist Grundlage der Überlegungen und Untersuchungen der vorliegenden Arbeit. Ausgehend von dem Gedanken der Kombination der Vorteile der Gefäßbewässerung mit denen moderner Materialien, die eine maschinelle Installation ermöglichen und eine lange Lebensdauer aufweisen, wurde die Entwicklung einer selbststeuernden Bewässerung verfolgt. Materialrecherchen und eine Auswahl als geeignet identifizierter Materialien führten zu Laboruntersuchungen an Hand derer die Darcy-Eigenschaften (linearer Zusammenhang zwischen Volumenstrom und anliegendem Druck) überprüft wurden. Membranen erwiesen sich danach als geeignet, so dass an einer ausgewählten Schlauchmembran weitergehende Untersuchungen zum Einsatz für die Bewässerung vorgenommen wurden. Diese umfassten Langzeitlaborversuche im Boden zur Untersuchung der Entwicklung der Durchlässigkeit der Schlauchmembran über die Zeit, sowie zum Nachweis der Selbststeuerung des Systems. Mit den gewonnenen Ergebnissen war es möglich großmaßstäbliche Versuche im Gewächshaus zu realisieren, bei der die Schlauchmembran im Vergleich zur Tropfbewässerung hinsichtlich Wasserverbrauch und Ernteertrag an Standorten in Deutschland und Algerien getestet wurde. Diese Gewächshausversuche zeigten vielversprechende Ergebnisse für die Membranschlauchbewässerung mit im Vergleich zur Tropfbewässerung erhöhten Erträgen bei gleichzeitig geringerem Wasserverbrauch. Dies wurde besonders bei Verwendung von Bewässerungswasser mit erhöhtem Salzgehalt deutlich. Sorgfältiger Beachtung bedarf die Wasserqualität, da die Membranschlauchbewässerung auf Wasserverunreinigungen mit Durchflussverminderung reagiert, die sich nachteilig auf die Pflanzenentwicklung auswirkt. Aufgabe weiterer Forschungen muss es demnach sein, technische Verfahren zu entwickeln, die die langfristige Leistungsfähigkeit der Membran im Bewässerungsbetrieb aufrechterhalten.

Initial growth of sunflower in soils with high concentrations of boron and heavy metals.

Phytoremediation studies have been conducted in an area contaminated by heavy metals, located in Piracicaba - SP, Brazil. This area was contaminated accidentally by the addition of auto scrap shredding to the soil and was limed later to reduce heavy metal mobility in the environment. Previous characterization showed that it also presents high concentration of boron, which has limited the initial plant development of some species. As sunflower plants require a high boron supply and the literature describes its use in the phytoremediation of soils contaminated with heavy metals under some conditions, the aim of this work was to evaluate its potential for the remediation of this area. In the present study, the results of preliminary tests are presented, aiming at the evaluation of sunflower plant germination and its initial development when cultivated in the contaminated soil described. Two sunflower hybrids were sown in soils treated with different rates of boron and in the soil from the contaminated area in study. The results showed that sunflower plants had a normal initial development, even in the soil from the contaminated area. Therefore, sunflower is a promising crop and further studies will be developed to evaluate the sunflower efficiency in phytoextraction or phytostabilization of heavy metals in areas where boron contamination also occurs, as is the case in the study area.

Genome-wide analysis of G-type lectin genes in Fragaria vesca and functional characterization of FaMBL1 gene in defense response of F. × ananassa to fungal pathogens

Strawberry (Fragaria × ananassa) is an important soft fruit but easily to be infected by pathogens. Anthracnose and gray mold are two of the most destructive diseases of strawberry which lead to serious fruit rot. The first chapter introduced strawberry anthracnose caused by Colletotrichum acutatum. The infection strategy, disease cycle and management of C. acutatum on strawberry were reported. Likewise, the second chapter summarized the infection strategy of Botrytis cinerea and the defense responses of strawberry. As we already know white unripe strawberry fruits are more resistant to C. acutatum than red ripe fruits. During the interaction between strawberry white/red fruit and C. acutaum, a mannose binding lectin gene, FaMBL1, was found to be the most up-regulated gene and induced exclusively in white fruit. FaMBL1 belongs to the G-type lectin family which has important roles in plant development and defense process. To get insight into the role of FaMBL1, genome-wide identification was carried out on G-type lectin gene family in Fragaria vesca and the results were showed in chapter 3. G-type lectin genes make up a large family in F. vesca. Active expression upon biotic/abiotic stresses suggested a potential role of G-lectin genes in strawberry defenses. Hence, stable transgenic strawberry plants with FaMBL1 gene overexpressed were generated. Transformed strawberry plants were screened and identified. The results were showed in chapter 4, content of disease-related phytohormone, jasmonic acid, was found decreased in overexpressing lines compared with wild type (WT). Petioles inoculated by C. fioriniae of overexpressing lines had lower disease incidence than WT. Leaves of overexpressing lines challenged by B. cinerea showed remarkably smaller lesion diameters compared with WT. The chitinase 2-1 (FaChi2-1) showed higher expression in overexpressing lines than in WT during the interaction with B. cinerea, which could be related with the lower susceptibility of overexpressing lines.

Novel insights into plant vascular development: disclosing a mechanism to maintain thermospermine homeostasis in the xylem

Dissertation presented to obtain the Ph.D degree in Biology

An integrated approach to Physcomitrella patens transcriptomics reveals importantclues of the evolutionary development of plant organs and sexual reproduction

Land plant evolution required the generation of a new body plan that could resist the harsher and fluctuating environmental conditions found outside of aquatic environments. Unraveling the genetic basis of plant developmental innovations is not only revealing in terms of an evolutionary point of view, but it is also important for understanding the emergence of agronomically important traits. Comparative genetic studies between basal and modern land plants, both at the genome and trancriptome levels, can help in the generation of hypotheses related to the genetic basis of plant evolutionary development.(...)

Aspectos moleculares del desarrollo de biofilms, la colonización de la planta y la emisión de compuestos orgánicos volátiles en bacterias rizosféricas. Molecular aspects of biofilm development, plant colonization and microbial volatile organic emission in rhizospheric bacteria.

Las bacterias que habitan la rizosfera y que poseen la capacidad de provocar un efecto positivo sobre las plantas son denominadas en su conjunto como Rizobacterias Promotoras del Crecimiento Vegetal (PGPR). Estas bacterias han desarrollado diferentes estrategias para adaptarse a diversas condiciones ambientales. La capacidad para responder a variaciones en la disponibilidad nutricional permite la persistencia de la bacteria en el suelo y mejora sus posibilidades para colonizar la planta hospedadora. En la naturaleza, a menudo las bacterias se encuentran en estructuras de comunidades de microorganismos interconectados denominados biofilms, con un estilo de vida diferente al de la vida en forma planctónica. La formación del biofilm podría representar una estrategia de supervivencia de la rizobacteria a condiciones adversas del suelo. Por Microscopía Confocal de Barrido Láser (CLSM), hemos observado que Rhizobium leguminosarum desarrolla un biofilm característico sobre una superficie abiótica. Hemos identificado algunos de los factores genéticos que influyen en su formación. El presente proyecto propone avanzar en el conocimiento de los factores ambientales y genéticos que influyen sobre la capacidad de las rizobacterias para formar biofilms y su impacto en la interacción con las plantas. A través de enfoques genéticos (mutacionales y de expresión génica) y análisis por CLSM nos proponemos acercarnos a un modelo de los factores de superficie, extracelulares y regulatorios propios de la bacteria que influyen en las propiedades de adhesión y la formación de biofilms. Por último, se intentará correlacionar la emisión de compuestos orgánicos volátiles por las bacterias rizosféricas con ciertos aspectos de la promoción del crecimiento de las plantas.

Development of a rehabilitation system for the red mud waste generated at the Aughinish Alumina Bayer Plant

Aughinish Alumina Limited (AAL) have an obligation by terms of their Integrated Pollution Control Licence (IPCL) and Planning Permission to establish vegetation on the red mud stack at their plant at Aughinish, Co. Limerick. High pH and high exchangeable sodium percentage are the main known factors limiting the establishment of vegetation on red mud. Gypsum addition has been known to assist in alleviating these problems in other countries. However, there is no experience or published information on red mud rehabilitation under Irish conditions. Red mud with organic and inorganic waste-derived ameliorants as well as selected grassland species were examined under laboratory controlled environment conditions as well as in field plot trials. Also, in order that it would be economically achievable, the research utilised locally available waste products as the organic amendments. Screening trials found that physical constraints severely limit plant germination and growth in red mud. Gypsum addition effectively lowers pH, exchangeable sodium percentage and the availability of A1 and Fe in the mud. A strong relationship between pH, ESP and A1 levels was also found. Gypsum addition increased germination percentages and plant growth for all species investigated. Greenhouse trials demonstrated that organic wastes alone did not greatly improve conditions for plant growth but when used in conjunction with gypsum plant performances for all species investigated was significantly increased. There was a high mortality rate for grasses in non-gypsum treatments. An emerging trend of preferential iron uptake and calcium deficiency in non-gypsum treatments was found at pot screening stage. Species also displayed manganese and magnesium deficiencies.

The endodermis--development and differentiation of the plant's inner skin.

Controlling external compound entrance is essential for plant survival. To set up an efficient and selective sorting of nutrients, free diffusion via the apoplast in vascular plants is blocked at the level of the endodermis. Although we have learned a lot about endodermal specification in the last years, information regarding its differentiation is still very limited. A differentiated endodermal cell can be defined by the presence of the "Casparian strip" (CS), a cell wall modification described first by Robert Caspary in 1865. While the anatomical description of CS in many vascular plants has been very detailed, we still lack molecular information about the establishment of the Casparian strips and their actual function in roots. The recent isolation of a novel protein family, the CASPs, that localizes precisely to a domain of the plasma membrane underneath the CS represents an excellent point of entry to explore CS function and formation. In addition, it has been shown that the endodermis contains transporters that are localized to either the central (stele-facing) or peripheral (soil-facing) plasma membranes. These features suggest that the endodermis functions as a polar plant epithelium.

Development of specific ITS markers for plant DNA identification within herbivorous insects

DNA based techniques have proved to be very useful methods to study trophic relationships 17 between pests and their natural enemies. However, most predators are best defined as omnivores, 18 and the identification of plant-specific DNA should also allow the identification of the plant 19 species the predators have been feeding on. In this study, a PCR approach based on the 20 development of specific primers was developed as a self-marking technique to detect plant DNA 21 within the gut of one heteropteran omnivorous predator (Macrolophus pygmaeus) and two 22 lepidopteran pest species (Helicoverpa armigera and Tuta absoluta). Specific tomato primers 23 were designed from the ITS 1-2 region, which allowed the amplification of a tomato DNA 24 fragment of 332 bp within the three insect species tested in all cases (100% of detection at t = 0) 25 and did not detect DNA of other plants nor of the starved insects. Plant DNA half-lives at 25ºC 26 ranged from 5.8h, to 27.7h and 28.7h within M. pygmaeus, H. armigera and T. absoluta, 27 respectively. Tomato DNA detection within field collected M. pygmaeus suggests dietary mixing 28 in this omnivorous predator and showed a higher detection of tomato DNA in females and 29 nymphs than males. This study provides a useful tool to detect and to identify plant food sources 30 of arthropods and to evaluate crop colonization from surrounding vegetation in conservation 31 biological control programs.

Light-regulated plant growth and development.

Plants are sessile and photo-autotrophic; their entire life cycle is thus strongly influenced by the ever-changing light environment. In order to sense and respond to those fluctuating conditions higher plants possess several families of photoreceptors that can monitor light from UV-B to the near infrared (far-red). The molecular nature of UV-B sensors remains unknown, red (R) and far-red (FR) light is sensed by the phytochromes (phyA-phyE in Arabidopsis) while three classes of UV-A/blue photoreceptors have been identified: cryptochromes, phototropins, and members of the Zeitlupe family (cry1, cry2, phot1, phot2, ZTL, FKF1, and LKP2 in Arabidopsis). Functional specialization within photoreceptor families gave rise to members optimized for a wide range of light intensities. Genetic and photobiological studies performed in Arabidopsis have shown that these light sensors mediate numerous adaptive responses (e.g., phototropism and shade avoidance) and developmental transitions (e.g., germination and flowering). Some physiological responses are specifically triggered by a single photoreceptor but in many cases multiple light sensors ensure a coordinated response. Recent studies also provide examples of crosstalk between the responses of Arabidopsis to different external factors, in particular among light, temperature, and pathogens. Although the different photoreceptors are unrelated in structure, in many cases they trigger similar signaling mechanisms including light-regulated protein-protein interactions or light-regulated stability of several transcription factors. The breath and complexity of this topic forced us to concentrate on specific aspects of photomorphogenesis and we point the readers to recent reviews for some aspects of light-mediated signaling (e.g., transition to flowering).

Development of a two-step screening ESI-TOF-MS method for rapid determination of significant stress-induced metabolome modifications in plant leaf extracts: the wound response in Arabidopsis thaliana as a case study.

To study the stress-induced effects caused by wounding under a new perspective, a metabolomic strategy based on HPLC-MS has been devised for the model plant Arabidopsis thaliana. To detect induced metabolites and precisely localise these compounds among the numerous constitutive metabolites, HPLC-MS analyses were performed in a two-step strategy. In a first step, rapid direct TOF-MS measurements of the crude leaf extract were performed with a ballistic gradient on a short LC-column. The HPLC-MS data were investigated by multivariate analysis as total mass spectra (TMS). Principal components analysis (PCA) and hierarchical cluster analysis (HCA) on principal coordinates were combined for data treatment. PCA and HCA demonstrated a clear clustering of plant specimens selecting the highest discriminating ions given by the complete data analysis, leading to the specific detection of discrete-induced ions (m/z values). Furthermore, pool constitution with plants of homogeneous behaviour was achieved for confirmatory analysis. In this second step, long high-resolution LC profilings on an UPLC-TOF-MS system were used on pooled samples. This allowed to precisely localise the putative biological marker induced by wounding and by specific extraction of accurate m/z values detected in the screening procedure with the TMS spectra.

Plant polyamines in stress and development: an emerging area of research in plant sciences.

Compelling evidence indicates the participation of polyamines in abiotic and biotic stress responses in plants. Indeed, genetic engineering of polyamine levels in plants has successfully improved biotic and abiotic stress resistance in model plants and crops. We anticipate that many of the current challenges in agriculture to cope with climate change and maintain nutritional quality of fruits and vegetables can be approached by considering the polyamine pathway...