Biochar as a soil amendment in the tree establishment phase: What are the consequences for tree physiology, soil quality and carbon sequestration?

Trees play a pivotal role in the urban environment alleviating the negative impacts of urbanization, and for this reason, local governments have promoted strongly tree planting policies. However, poor soil quality and neglect tree maintenance (e.g., irrigation and fertilization) can seriously mine the plant health status during the tree establishment phase. The use of biochar to provide long-lasting C to the soil and, at the same time, improving soil properties (e.g., improved water holding capacity), soil enzymes activities and NPK concentrations, is a promising research field. Therefore, with a two-step experiment, the study aimed to assay the physiological responses of a commonly used urban tree species (Tilia × europaea L.) to 1.5 % (w/w) biochar amendment (B), and secondly, to assess the ability of trees, grown in biochar amended soil, to tolerate a period of drought. Biochar amendment increased P and K availability in the soil, resulting in higher P and K concentrations in B than control leaves, according to the leaf stage. This induced B trees, higher values in both total biomass than controls (+22 %) in well-watered plants. Moreover, the higher water availability in soil amended with biochar helped B trees to tolerate water stress, with better leaf photosynthetic performances and a faster recovery than stressed controls after the re-watering. This study highlights the dual function of the biochar, improving CO2 sequestration and soil properties, and at the same time, enhancing plant physiological responses to environmental constraints. The use of biochar at the tree planting, especially in an urban environment, is a feasible and environmentally sustainable strategy to improve the success during the tree establishment phase.

First Characterization of the Formation of Anthocyanin-Ge and Anthocyanin-B Complexes through UV-Vis Spectroscopy and Density Functional Theory Quantum Chemical Calculations.

The occurrence of anthocyanin (ACN) and metal (Me) complexes has been widely supported by many research works while the possibility that ACNs bind to metalloids (Mds) is yet to be proven. Here, metalloids (H3BO3 for B; GeO2 for Ge) were added to cyanidin-based solutions at pH 5, 6, and 7 and ACN-Md stoichiometric ratios of 1:1, 1:10, 1:100, and 1:500, and UV-vis transmittance spectroscopy as well as density functional theory (DFT) calculations were performed to test this hypothesis. Ge and B addition caused bathochromic and hyperchromic shifts on ACN UV-vis spectra, particularly pronounced at pH 5 and a 1:500 (ACN:Md) ratio. ACN-Me complexation reactions have been evaluated where Ge showed a higher capability to bind to ACNs than B. Among the complexes envisioned, those labeled as b1, b2, and b3 feature UV-vis spectra compatible with experiments. The combination of experimental and computational data offers for the first time evidence of the formation of ACN-Md complexes.

Girled-induced anthocyanin accumulation in red-leafed Prunus cerasifera: Effect on photosynthesis, photoprotection and sugar metabolism.

The feedback regulation of photosynthesis depends on the cooperation of multiple signals, including sugars. Herein, the effect of shoot girdling was monitored on a daily basis for three days in green- and red-leafed Prunus cerasifera plants (GLP and RLP, respectively). The effect of anthocyanin presence was investigated in terms of photosynthesis, sugar metabolism and photoprotection. Net photosynthesis (A390) and stomatal conductance were reduced on the first day at 12:00 only in the girdled GLP (29 and 33 %, respectively). Moreover, the girdled GLP displayed at 12:00 higher sucrose, glucose and fructose concentrations than control leaves. Conversely, girdled RLP showed the first reduction of A390 at 18:00, with no significant differences at 12:00 in sucrose and glucose concentrations. The increased biosynthesis of anthocyanins that was only detected in girdled RLP contributed to lowering the accumulation of hexoses. Overall, these results revealed a sugar-buffering role exerted by anthocyanins that positively influence the feedback regulation of photosynthesis. Moreover, non-photochemical quenching, namely pNPQ, revealed the ability of anthocyanins to photoprotect photosystem II from supernumerary photons reaching the chloroplast, whose function was compromised by girdling. The present study provides a starting point to understand the possible link between photosynthesis regulation through sugar signalling and anthocyanin upregulation.

Red versus green leaves: transcriptomic comparison of foliar senescence between two Prunus cerasifera genotypes.

The final stage of leaf ontogenesis is represented by senescence, a highly regulated process driven by a sequential cellular breakdown involving, as the first step, chloroplast dismantling with consequent reduction of photosynthetic efficiency. Different processes, such as pigment accumulation, could protect the vulnerable photosynthetic apparatus of senescent leaves. Although several studies have produced transcriptomic data on foliar senescence, just few works have attempted to explain differences in red and green leaves throughout ontogenesis. In this work, a transcriptomic approach was used on green and red leaves of Prunus cerasifera to unveil molecular differences from leaf maturity to senescence. Our analysis revealed a higher gene regulation in red leaves compared to green ones, during leaf transition. Most of the observed DEGs were shared and involved in transcription factor activities, senescing processes and cell wall remodelling. Significant differences were detected in cellular functions: genes related to photosystem I and II were highly down-regulated in the green genotype, whereas transcripts involved in flavonoid biosynthesis, such as UDP glucose-flavonoid-3-O-glucosyltransferase (UFGT) were exclusively up-regulated in red leaves. In addition, cellular functions involved in stress response (glutathione-S-transferase, Pathogen-Related) and sugar metabolism, such as three threalose-6-phosphate synthases, were activated in senescent red leaves. In conclusion, data suggests that P. cerasifera red genotypes can regulate a set of genes and molecular mechanisms that cope with senescence, promoting more advantages during leaf ontogenesis than compared to the green ones.

Discerning between Two Tuscany (Italy) Ancient Apple cultivars, 'Rotella' and 'Casciana', through Polyphenolic Fingerprint and Molecular Markers.

Ancient apple cultivars usually have higher nutraceutical value than commercial ones, but in most cases their variability in pomological traits does not allow us to discriminate among them. Fruit of two Tuscany ancient apple cultivars, 'Casciana' and 'Rotella', picked from eight different orchards (four for each cultivar) were analyzed for their pomological traits, organoleptic qualities, polyphenolic profile and antiradical activity. The effectiveness of a polyphenol-based cluster analysis was compared to molecular markers (internal transcribed spacers, ITS1 and ITS2) to unequivocally discern the two apples. 'Casciana' and 'Rotella' fruit had a higher nutraceutical value than some commercial cultivars, in terms of phenolic abundance, profile and total antiradical activity. Although pedo-climatic conditions of different orchards influenced the phenolic profile of both apples, the polyphenolic discriminant analysis clearly separated the two cultivars, principally due to higher amounts of procyanidin B2, procyanidin B3 and p-coumaroylquinic acid in 'Casciana' than in 'Rotella' fruit. These three polyphenols can be used proficiently as biochemical markers for distinguishing the two apples when pomological traits cannot. Conversely, ITS1 and ITS2 polymorphism did not allow us to distinguish 'Casciana' from 'Rotella' fruit. Overall, the use of polyphenolic fingerprint might represent a valid tool to ensure the traceability of products with a high economic value.

Ancient apple cultivars from Garfagnana (Tuscany, Italy): A potential source for 'nutrafruit' production.

Ancient apple cultivars are known for their organoleptic properties over a small geographic area, but little is known of their nutraceutical properties, which might be useful in large-scale breeding programmes. Nine ancient apple cultivars from Tuscany (Italy) were characterized for their organoleptic properties, phenolic profiles, and antioxidant activity. These cultivars had high polyphenol concentrations (principally flavanols and phenolic acids) and high total antioxidant capacity compared with most commercial apple cultivars. Fruits from the cultivars 'San Michele' and 'Del Debbio' showed a good compromise between fruit size and solid soluble content, and might be suitable for fresh consumption, while fruit from 'Benito', 'Della Piastra', 'Lugliese Grisanti', 'Del Sangue' and 'Ruggine' had high polyphenol contents and excellent antioxidant capacity, and may be suitable for breeding programmes. 'Ruggine' fruit could also be used for sweet juices with good nutraceutical properties due to their high soluble solid content and high flavanol concentration.

Chlorophyll Fluorescence, Photoinhibition and Abiotic Stress: Does it Make Any Difference the Fact to Be a C3 or C4 Species?

Chlorophyll fluorescence analysis is one of the most powerful and widely used techniques to study the effect of stresses on the photosynthetic process. From the first utilization, the F v/F m ratio has been largely used as a sensitive indicator of plant photosynthetic performance. Decreases of this index are indicative of the reduction of photosystem II (PSII) efficiency, namely photoinhibition. In the last 20 years, application of chlorophyll fluorescence has been largely improved, and many other informative parameters have been established to detect PSII photochemical efficiency and the partitioning of light energy to alternative dissipative mechanisms (qE, energy-dependent quenching; qZ, zeaxanthin-dependent quenching and qI, photoinhibitory quenching; qH, sustained photoprotective antenna quenching; qM, quenching dependent to chloroplast movement; qT, light harvesting complexes II-I state-transition) such as the recently developed "photoprotective power" of non-photochemical quenching (pNPQ). This review reports a brief description of the main chlorophyll fluorescence parameters and a wide analysis of the current bibliography on the use of different parameters which are useful to detect events of PSII photoinhibition. In addition, in view of the inherent differences in morpho-anatomical, physiological and biochemical features between C3 and C4 metabolism, possible differences in terms of photoinhibition between C3 and C4 plant species under stress conditions are proposed. The attempt is to highlight the limits of their comparison in terms of susceptibility to photoinhibition and to propose direction of future research which, assisted by chlorophyll fluorescence, should improve the knowledge of the different sensitivity of C3 and C4 to abiotic stressors.

Multiple Consequences Induced by Epidermally-Located Anthocyanins in Young, Mature and Senescent Leaves of Prunus.

Anthocyanic morphs are generally less efficient in terms of carbon gain, but, in turn, are more photoprotected than anthocyanin-less ones. To date, mature leaves of different morphs or leaves at different developmental stages within the same species have generally been compared, whereas there is a lack of knowledge regarding different stages of development of red vs. green leaves. Leaves (1-, 7-, and 13-week-old) of red- (RLP) and green-leafed (GLP) Prunus in terms of photosynthetic rate, carbon metabolism and photoprotective mechanisms were compared to test whether anthocyanin-equipped leaves perform better than anthocyanin-less leaves and whether photoprotection is the primary role of epidermally-located anthocyanins, using for the first time a recently-developed parameter of chlorophyll fluorescence (qPd). GLP leaves had a higher photosynthetic rate in 1- and 7-week-old leaves, but RLP leaves performed better at an early stage of senescence and had a longer leaf lifespan. Anthocyanins contributed to leaf photoprotection throughout the leaf development, but were tightly coordinated with carotenoids. Besides photoprotecting, we propose that epidermal anthocyanins may be principally synthetized to maintain an efficient carbon-sink strength in young and senescent leaves, thus extending the RLP leaf lifespan.