Biochar in perennial crops: nutritional, agronomical and environmental implications

Biochar is the solid C-rich matrix obtained by pyrolysis of biomasses, currently promoted as a soil amendment with the aim to offset anthropogenic C emissions, while ameliorating soil properties and growth conditions. Benefits from biochar seem promising, although scientific understandings are beginning to be explored. In this project, I performed a suite of experiments in controlled and in field conditions with the aims to investigate the effect of biochar on: a) the interaction with minerals; b) Fe nutrition in kiwifruit; c) soil leaching, soil fertility, soil CO2 emissions partitioning, soil bacterial profile and key gene expression of soil nitrification-involved bacteria; d) plant growth, nutritional status, yield, fruit quality and e) its physical-chemical changes as affected by long-term environmental exposure. Biochar released K, P and Mg but retained Fe, Mn, Cu and Zn on its surface which in turn hindered Fe nutrition of kiwifruit trees. A redox reaction on the biochar surface exposed to a Fe source was elucidated. Biochar reduced the amount of leached NH4+-N but increased that of Hg, K, P, Mo, Se and Sn. Furthermore, biochar synergistically interacted with compost increasing soil field capacity, fertility, leaching of DOC, TDN and RSOC, suggesting a priming effect. However, in field conditions, biochar did not affect yield, nutritional status and fruit quality. Actinomadura flavalba, Saccharomonospora viridis, Thermosporomyces composti and Enterobacter spp. were peculiar of the soil amended with biochar plus compost which exhibited the highest band richness and promoted gene expression levels of Nitrosomonas spp., Nitrobacter spp. and enzymatic-related activity. Environmental exposure reduced C, K, pH and water infiltration of biochar which instead resulted in a higher O, Si, N, Na, Al, Ca, Mn and Fe at%. Oxidation occurred on the aged biochar surface, it decreased progressively with depth and induced the development of O-containing functional groups, up to 75nm depth.

1st and 2nd Generation Ethanol from Biomass Crops

In chapter 1 and 2 calcium hydroxide as impregnation agent before steam explosion of sugarcane bagasse and switchgrass, respectively, was compared with auto-hydrolysis, assessing the effects on enzymatic hydrolysis and simultaneous saccharification and fermentation (SSF) at high solid concentration of pretreated solid fraction. In addition, anaerobic digestion of pretreated liquid fraction was carried out, in order to appraise the effectiveness of calcium hydroxide before steam explosion in a more comprehensive way. In As water is an expensive input in both cultivation of biomass crops and subsequent pretreatment, Chapter 3 addressed the effects of variable soil moisture on biomass growth and composition of biomass sorghum. Moreover, the effect of water stress was related to the characteristics of stem juice for 1st generation ethanol and structural carbohydrates for 2nd generation ethanol. In the frame of chapter 1, calcium hydroxide was proven to be a suitable catalyst for sugarcane bagasse before steam explosion, in order to enhance fibre deconstruction. In chapter 2, effect of calcium hydroxide on switchgrass showed a great potential when ethanol was focused, whereas acid addition produced higher methane yield. Regarding chapter 3, during crop cycle the amount of cellulose, hemicellulose and AIL changed causing a decrease of 2G ethanol amount. Biomass physical and chemical properties involved a lower glucose yield and concentration at the end of enzymatic hydrolysis and, consequently, a lower 2G ethanol concentration at the end of simultaneous saccharification and fermentation, proving that there is strong relationship between structure, chemical composition, and fermentable sugar yield. The significantly higher concentration of ethanol at the early crop stage could be an important incentive to consider biomass sorghum as second crop in the season, to be introduced into some agricultural systems, potentially benefiting farmers and, above all, avoiding the exacerbation of the debate about fuel vs food crops.

RNA interference technology as a potential control method for fruit and horticultural crops pathogens Botrytis cinerea and Plasmopara viticola

Chapter 1, a general introduction on Botrytis cinerea and its threat to crop production is presented. What Botrytis looks like, its life cycle, why it is a threat to agricultural production, its worldwide pest status, and its current state of management is further elaborated on. Chapter 2, a general introduction on Plasmopara viticola, its threat to grape production and management strategies presented. Chapter 3, titled " RNA Interference Strategies for Future Management of Plant Pathogenic Fungi: Prospects and Challenges ", presents the rapid improvement and extensive implementation of RNA interference (RNAi) technology for the management of fungal pathogens. In this chapter, we describe the application of exogenous RNAi involved in plant pathogenic fungi and discuss dsRNA production, formulation, and RNAi delivery methods. Chapter 4, titled " Exogenous dsRNAs against chitin synthase and glucan synthase genes suppress the growth of the pathogenic fungus Botrytis cinerea " addresses two important questions: Is RNAi technology functional for B. cinerea control ? And which target genes can be exploited for RNAi-based B.cinerea disease control ? Upon target genes selections, an exogenous RNAi protocol was set up and we could effectively deliver a known dose of bacterially produced double stranded RNA (dsRNA) to induce RNAi in B. cinerea. Chapter 5, titled " Double-Stranded RNA Targeting Dicer-Like Genes Compromises the Pathogenicity of Plasmopara viticola on Grapevine “, which deals mainly on RNAi induction against Plasmopara viticola. This chapter addresses two main questions: Is RNAi technology functional in contrasting Plasmopara viticola? And which target genes can be exploited for RNAi-based disease control in Plasmopara viticola?. In the last Chapter (Chapter 6) titled “General discussions and perspectives for future research”, the major research findings from this thesis are discussed together with perspectives for future research.