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GIAMPIERO CAI – curriculum vitae METRICS Total Impact Factor: 230 (95 in the last 5 years) Publications: 65 (30 in the last 5 years) H-index (Scopus): 20 (with 1242 citations); last five years: 10 (with 225 citations) H-index (Google Scholar): 22 (with 1702 citations); last five years: 15 (with 714 citations) SCIENTIFIC ACTIVITY 1) The cytoskeleton and organelle motility in pollen tubes Microtubule motors are a group of proteins that cooperate with microtubules in delivering cell components to specific destinations. Microtubule motors are classified as two superfamilies, the kinesins (further subdivided into 14 subfamilies) and dyneins. In previous years, major undertaken efforts were directed to identify proteins similar to kinesins in pollen tubes. This effort led to the characterization of a 90-kD protein, which showed several features of the kinesin family and it was found capable of transporting organelles in the cortical region of pollen tubes. In particular, we obtained evidence that kinesin is involved in the slow movement of mitochondria, as opposed to the fast movement along actin filaments. In the last period, we focused on the association between kinesins and HSP70 (70-kDa heat shock proteins), a family of proteins ubiquitously expressed and important for protein folding. We have identified a novel isoform of HSP70 in tobacco pollen tubes capable of binding to microtubules in an ATP-dependent manner. The HSP70 was identified in association with the 90-kD kinesin. The identity of the HSP70 was validated by immunoassays and mass spectrometry. Two-dimensional electrophoresis analysis showed that a specific isoform of HSP70 is associated with the 90-kD kinesin and is also associated with mitochondria of pollen tubes. Several assays suggested that the binding (but not the release) of the microtubule-HSP70 is facilitated by the 90-kD kinesin. 2) Activity and dynamics of transglutaminase (TGase) in pollen tubes TGases are a family of calcium-dependent acyl-transferases that catalyze the transfer of an acyl group from glutamine residue to primary amines, including polyamines (PA) or lysine residues. The purpose of this research is to determine the roles of TGase in pollen tubes. In particular, after analyzing the interaction between TGase and cytoskeleton, we proceeded with the identification of different subcellular compartments in association with TGase (cytosol, membranes and cell walls). Membrane TGase is probably associated with the Golgi and plasma membrane, suggesting that TGase might be exported by exocytosis. The immunolocalization of TGase showed that the enzyme is present in the growth region of pollen tubes and that the enzyme co-fractionates with specific markers of cell wall. Analysis by gel electrophoresis showed that different isoforms of TGase are associated with subcellular compartments, suggesting both different roles or different regulatory mechanisms. Different subcellular distribution of TGase might be also regulated by the dynamics of membranes and by the integrity of the cytoskeleton. These data indicate that a cytoplasmic TGase interacts with the cytoskeleton because a different isoform of TGase is secreted in the pollen tube cell wall, where it could play a role in the regulation of apical growth. 3) Dynamics of cellulose synthase and callose synthase in pollen tubes Like other plant cells, the pollen tube is characterized by a cell wall composed of several components. We examined the distribution of callose synthase and cellulose synthase in tobacco pollen tubes in relation to the dynamics of actin filaments, microtubules and in relation to the endomembrane system. Both enzymes are associated with the plasma membrane, but cellulose synthase is present along the entire length of pollen tubes (with a higher concentration at the apex) while callose synthase is located at the apex and in distal regions. In longer pollen tubes, callose synthase accumulates around the callose plugs, suggesting the involvement of the enzyme in plug synthesis. Actin filaments and endomembranes are critical for the proper distribution of callose synthase and cellulose synthase, suggesting that enzymes are transported through actin filaments by vesicles. Conversely, microtubules appear to be critical in the distal positioning of callose synthase and around callose plugs. In contrast, cellulose synthase only partially aligned with cortical microtubules and seemed independent of deposition of callose plugs. In addition, callose synthase comigrates with tubulin in Blue Native PAGE analysis. 4) Traceability, characterization and valorization of agro-food chains This researches line (in collaboration with other members of the department) are focused on the protection, management and conservation of the local agro-biodiversity, on genetic traceability and on chemical and nutraceutical characterization of food resources. Activities are applied on specific issues: development and validation of molecular methods for the authenticity and control of labelled products (e.g. Panforte and Ricciarelli di Siena PGI); traceability of high-quality products, fresh and processed, using DNA markers (e.g. truffles and saffron); genetic and proteomic characterization and development of molecular database of herbaceous plants and fruit trees (e.g. olive, chestnut, goji and ancient fruits); characterization of antioxidant activity, polyphenols, amino acids, sugars and vitamins within fresh or processed foods; cytological, physiological, biochemical, and palynological aspects of pollen and honey through bio-molecular and chemical-physical analysis; analysis of environmental stress (e.g. temperature variation) on the productivity of commercial plants (vegetables and fruit trees). GRANTS 1. Research project within the framework of the European Community (Biotechnology Division) to characterize the transfer cells during development of maize endosperm (1997-2000, 4th Framework Programme). 2. Grant of the University of Siena under the project of the University research (PAR) to study the movement of pollen organelles along microtubules (1999). 3. European Community Funding for a project on the development of genetic and molecular tools to reconstruct the functional domains in maize seeds (2000-2003, fifth framework programme). 4. Grant under the project of the University research (PAR) at the University of Siena for studying organelle motility along microtubules and actin filaments pollen (2002). 5. Grant in the University Research Project (PAR) at the University of Siena for studying motility of pollen organelle along microtubules and filaments of actin (2004). 6. Grant of the University of Siena under the project of the University research (PAR) for characterization of kinesins in the pollen tube (2007). 7. Grant from the Ministry of University within the PRIN research national program for the characterization of the interactions between the cell wall and cytoskeleton in pollen tubes (2009) 8. Grant by the Tuscany regional authority for the project measure 124 (PSR 2007-2013) entitled as MODOLIVI concerning the effects of olive wastes on cytological and molecular aspects of cultivated plants. 9. Grant by the Tuscany regional authority for the project Measure the RDP 2007-2013 entitled APISALUS - development of new technologies for health protection of honey bees (Apis mellifera). 10. Grant by the Tuscany regional authority for the TOSCOLATA project "Valorization of Tuscan products in innovative nutraceutical food based on cocoa" 2012-2015 under the Tuscany Measure 1.2.e. 11. Grant by the Tuscany regional authority for the project TOSCO MAGNATUM TRACE "Molecular and digital tracking of Tuscan white truffle-based products" as part of 2012-2015 Tuscany Measure 1.2.e. EDUCATION 1. High School "Liceo Scientifico" (Colle Val d'Elsa, Italy, 1982) 2. Degree in Biology at the University of Siena, Faculty of Mathematical, Physical and Natural Sciences (Italy, 1987). Vote 110 cum laude. 3. Doctorate in Environmental Biology at the University of Siena (Siena, Italy, 1993) WORK EXPERIENCE 1993-94; Chiron Biocine (Siena, Italy): research contract. 1994-99; University of Siena (Siena, Italy): position of permanent laboratory technician. 2000-01; University of Siena (Siena, Italy): Research Specialist. 2001 – 2015; University of Siena (Siena, Italy): position of full-time researcher. 2015 – present; University of Siena (Siena, Italy): Associate Professor of Botany. TEACHING ACTIVITY (University of Siena) A. Teaching as pre-researcher 1. 1992-93 to 1996-97: course in cellular technologies for the "school for technicians in biotechnology”. 2. 1999-2000 and 2000-01: course in plant biotechnology for the Interfaculty graduate course in Biotechnology. B. Teaching activities as researcher 1. 2001-02: laboratory of Systematic Botany for the degree course in Biological Sciences. 2. 2002-03 to 2005-06: cellular technologies-cell biology laboratory for the degree course in Biological Sciences. 3. 2005-06: cell biology for the graduate course LS-TAIE. 4. 2001-02 to 2008-09: Plant Biology and Plant Biotechnology within the Interfaculty Bachelor degree in Biotechnology. 5. 2007-08 and 2008-09: Applied Botany for the degree course in Biotechnology. 6. 2007-08 and 2008-09: Biochemistry of receptors for the graduate course LS-TAIE. 7. 2009-10, 2010-11, 2013-14 and 2014-15: Interactions between plant cells and environment for master LM-ESA. 8. 2009-10 to 2014-15: Botany for the degree in Biotechnology. 9. 2012-13 and 2013-14: Plant Biotechnology in the three-year degree course in Biotechnology. C. Teaching as associate professor 1. 2015-16: Plant Biotechnology in the master of science in Molecular and Cellular Biology. 2. 2015-16: Plant Biotechnology for the three-year degree course in Biotechnology. 3. 2015-16: Interactions between plant cells and environment for the MSC course LM-ESA. PARTICIPATION IN EDUCATIONAL COMMITTEES 1. Council of the PhD school in Science and Technology applied to Environment coordinated by the Department of Environmental Sciences (University of Siena) until 2013. 2. Academic Committee of the three-year degree course in Biotechnology (Faculties of Physical, Mathematical and Natural Sciences – Pharmacy – Medicine) from 2009 to 2013. 3. Academic Committee in the degree of Environmental and Natural Sciences and LM-ESA (Department of Physical, Land and Environmental Sciences, Department of Life Sciences). 4. Joint Commission of the Department of Life Sciences