Pasquale Losciale

A rapid, whole-tissue determination of the functional fraction of PSII after photoinhibition of leaves based on flash-induced P700 redox kinetics.

Assaying the number of functional PSII complexes by the oxygen yield from leaf tissue per saturating, single-turnover flash, assuming that each functional PSII evolves one oxygen molecule after four flashes, is one of the most direct methods but time-consuming. The ratio of variable to maximum Chl fluorescence yield (F(v)/F(m)) in leaves can be correlated with the oxygen yield per flash during a progressive loss of PSII activity associated with high-light stress and is rapid and non-intrusive, but suffers from being representative of chloroplasts near the measured leaf surface; consequently, the exact correlation depends on the internal leaf structure and on which leaf surface is being measured. Our results show that the average F(v)/F(m) of the adaxial and abaxial surfaces has a reasonable linear correlation with the oxygen yield per flash after varied extents of photoinactivation of PSII. However, we obtained an even better linear correlation between (1) the integrated, transient electron flow (Sigma) to P700+, the dimeric Chl cation in PSI, after superimposing a single-turnover flash on steady background far-red light and (2) the relative oxygen yield per flash. Leaves of C3 and C4 plants, woody and herbaceous species, wild-type and a Chl-b-less mutant, and monocot and dicot plants gave a single straight line, which seems to be a universal relation for predicting the relative oxygen yield per flash from Sigma. Measurement of Sigma is non-intrusive, representative of the whole leaf tissue, rapid and applicable to attached leaves; it may even be applicable in the field.

Changes in vascular and transpiration flows affect the seasonal and daily growth of kiwifruit (Actinidia deliciosa) berry.

BACKGROUND AND AIMS The kiwifruit berry is characterized by an early stage of rapid growth, followed by a relatively long stage of slow increase in size. Vascular and transpiration flows are the main processes through which water and carbon enter/exit the fruit, determining the daily and seasonal changes in fruit size. This work investigates the biophysical mechanisms underpinning the change in fruit growth rate during the season. METHODS The daily patterns of phloem, xylem and transpiration in/outflows have been determined at several stages of kiwifruit development, during two seasons. The different flows were quantified by comparing the diurnal patterns of diameter change of fruit, which were then girdled and subsequently detached while measurements continued. The diurnal courses of leaf and stem water potential and of fruit pressure potential were also monitored at different times during the season. KEY RESULTS Xylem and transpiration flows were high during the first period of rapid volume growth and sharply decreased with fruit development. Specific phloem import was lower and gradually decreased during the season, whereas it remained constant at whole-fruit level, in accordance with fruit dry matter gain. On a daily basis, transpiration always responded to vapour pressure deficit and contributed to the daily reduction of fruit hydrostatic pressure. Xylem flow was positively related to stem-to-fruit pressure potential gradient during the first but not the last part of the season, when xylem conductivity appeared to be reduced. CONCLUSIONS The fruit growth model adopted by this species changes during the season due to anatomical modifications in the fruit features.

Modulating the light environment with the peach ‘asymmetric orchard’: effects on gas exchange performances, photoprotection, and photoinhibition

The productivity of fruit trees is a linear function of the light intercepted, although the relationship is less tight when greater than 50% of available light is intercepted. This paper investigates the management of light energy in peach using the measurement of whole-tree light interception and gas exchange, along with the absorbed energy partitioning at the leaf level by concurrent measurements of gas exchange and chlorophyll fluorescence. These measurements were performed on trees of a custom-built ‘asymmetric’ orchard. Whole-tree gas exchange for north–south, vertical canopies (C) was similar to that for canopies intercepting the highest irradiance in the morning hours (W), but trees receiving the highest irradiance in the afternoon (E) had the highest net photosynthesis and transpiration while maintaining a water use efficiency (WUE) comparable to the other treatments. In the W trees, 29% and 8% more photosystems were damaged than in C and E trees, respectively. The quenching partitioning revealed that the non-photochemical quenching (NPQ) played the most important role in excess energy dissipation, but it was not fully active at low irradiance, possibly due to a sub-optimal trans-thylakoid ΔpH. The non-net carboxylative mechanisms (NC) appeared to be the main photoprotective mechanisms at low irradiance levels and, probably, they could facilitate the establishment of a trans-thylakoid ΔpH more appropriate for NPQ. These findings support the conclusion that irradiance impinging on leaves may be excessive and can cause photodamage, whose repair requires energy in the form of carbohydrates that are thereby diverted from tree growth and productivity.

Quantifying and monitoring functional photosystem II and the stoichiometry of the two photosystems in leaf segments: approaches and approximations

Given its unique function in light-induced water oxidation and its susceptibility to photoinactivation during photosynthesis, photosystem II (PS II) is often the focus of studies of photosynthetic structure and function, particularly in environmental stress conditions. Here we review four approaches for quantifying or monitoring PS II functionality or the stoichiometry of the two photosystems in leaf segments, scrutinizing the approximations in each approach. (1) Chlorophyll fluorescence parameters are convenient to derive, but the information-rich signal suffers from the localized nature of its detection in leaf tissue. (2) The gross O2 yield per single-turnover flash in CO2-enriched air is a more direct measurement of the functional content, assuming that each functional PS II evolves one O2 molecule after four flashes. However, the gross O2 yield per single-turnover flash (multiplied by four) could over-estimate the content of functional PS II if mitochondrial respiration is lower in flash illumination than in darkness. (3) The cumulative delivery of electrons from PS II to P700+ (oxidized primary donor in PS I) after a flash is added to steady background far-red light is a whole-tissue measurement, such that a single linear correlation with functional PS II applies to leaves of all plant species investigated so far. However, the magnitude obtained in a simple analysis (with the signal normalized to the maximum photo-oxidizable P700 signal), which should equal the ratio of PS II to PS I centers, was too small to match the independently-obtained photosystem stoichiometry. Further, an under-estimation of functional PS II content could occur if some electrons were intercepted before reaching PS I. (4) The electrochromic signal from leaf segments appears to reliably quantify the photosystem stoichiometry, either by progressively photoinactivating PS II or suppressing PS I via photo-oxidation of a known fraction of the P700 with steady far-red light. Together, these approaches have the potential for quantitatively probing PS II in vivo in leaf segments, with prospects for application of the latter two approaches in the field.

The positive effect of skin transpiration in peach fruit growth

The effect of fruit transpiration on the mechanisms driving peach (Prunus persica (L.) Batsch) daily growth was investigated. In peach, fruit water losses increase during the season and might play a key role in determining fruit growth. Skin transpiration was reduced during the cell expansion stage by enclosing fruit in plastic bags fitted with holes. In the first year, diameter changes of bagged and control fruit were precisely monitored for 15 days, and percentage dry matter and soluble solids content were determined during the experiment and at harvest. In the second year, midday fruit water potential, daily patterns of fruit growth and of vascular and transpiration flows were monitored. Bagging reduced fruit daily growth on some days, and negatively affected both fruit dry matter percentage and soluble solids content. Fruit transpiration rate was reduced during the midday hours, thus increasing midday fruit water potential and lowering xylem inflows. In accordance with the Münch hypothesis on traslocation, these conditions likely decreased the necessary gradient needed for the transport of phloem sap to sink organs, as in the afternoon, bagged fruit showed lower phloem inflows. These data suggest that skin transpiration in peach has a positive effect on fruit growth, as it enhances fruit phloem import.

Whole-tissue determination of the rate coefficients of photoinactivation and repair of photosystem II in cotton leaf discs based on flash-induced P700 redox kinetics

Using radioactively labelled amino acids to investigate repair of photoinactivated photosystem II (PS II) gives only a relative rate of repair, while using chlorophyll fluorescence parameters yields a repair rate coefficient for an undefined, variable location within the leaf tissue. Here, we report on a whole-tissue determination of the rate coefficient of photoinactivation ki, and that of repair kr in cotton leaf discs. The method assays functional PS II via a P700 kinetics area associated with PS I, as induced by a single-turnover, saturating flash superimposed on continuous background far-red light. The P700 kinetics area, directly proportional to the oxygen yield per single-turnover, saturating flash, was used to obtain both ki and kr. The value of ki, directly proportional to irradiance, was slightly higher when CO2 diffusion into the abaxial surface (richer in stomata) was blocked by contact with water. The value of kr, sizable in darkness, changed in the light depending on which surface was blocked by contact with water. When the abaxial surface was blocked, kr first peaked at moderate irradiance and then decreased at high irradiance. When the adaxial surface was blocked, kr first increased at low irradiance, then plateaued, before increasing markedly at high irradiance. At the highest irradiance, kr differed by an order of magnitude between the two orientations, attributable to different extents of oxidative stress affecting repair (Nishiyama et al., EMBO J 20: 5587–5594, 2001). The method is a whole-tissue, convenient determination of the rate coefficient of photoinactivation ki and that of repair kr.

A multivariate approach for assessing leaf photo-assimilation performance using the IPL index.

The detection of leaf functionality is of pivotal importance for plant scientists from both theoretical and practical point of view. Leaves are the sources of dry matter and food, and they sequester CO2 as well. Under the perspective of climate change and primary resource scarcity (i.e. water, fertilizers and soil), assessing leaf photo-assimilation in a rapid but comprehensive way can be helpful for understanding plant behavior under different environmental conditions and for managing the agricultural practices properly. Several approaches have been proposed for this goal, however, some of them resulted very efficient but little reliable. On the other hand, the high reliability and exhaustive information of some models used for estimating net photosynthesis are at the expense of time and ease of measurement. The present study employs a multivariate statistical approach to assess a model aiming at estimating leaf photo-assimilation performance, using few and easy-to-measure variables. The model, parameterized for apple and pear and subjected to internal and external cross validation, involves chlorophyll fluorescence, carboxylative activity of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCo), air and leaf temperature. Results prove that this is a fair-predictive model allowing reliable variable assessment. The dependent variable, called IPL index, was found strongly and linearly correlated to net photosynthesis. IPL and the model behind it seem to be (1) reliable, (2) easy and fast to measure and (3) usable in vivo and in the field for such cases where high amount of data is required (e.g. precision agriculture and phenotyping studies).

A Universal Correlation Between Flash-Induced P700 Redox Kinetics and Photoinactivation of Photosystem II in All Leaves?

Assaying functional Photosystem II (PSII) complexes by flash-induced oxygen yield from leaf tissue after photoinhibition is most direct but time-consuming, while measurement of chlorophyll fluorescence in leaves is only representative of chloroplasts near the leaf surface. To circumvent these deficiencies, we obtained an excellent linear correlation between (a) the integrated, transient electron flow (Σ) to PS I, after superimposing a single-turnover flash on steady far-red light, and (b) the relative oxygen yield per flash during progressive photoinactivation of Photosystem II. Leaves of C3 and C4 plants, woody and herbaceous species, wild type and a chlorophyll b-less mutant, and monocot and dicot plants gave a single linear correlation, which seems to be a universal relation for predicting the relative oxygen yield per flash from Σ.

Photosynthetic Performance and Vegetative Growth in a New Red Leaf Pear: Comparison of Scion Genotypes Using a Complex, Grafted-Plant System

Leaf photosynthetic performance of a new red-skinned inter-specific hybrid pear variety called ‘PremP009’ (PIQA®BOO®) is presently unknown and therefore was compared to the Asian pear variety ‘Hosui’. The seasonal growth patterns and the final dry matter accumulation of all tree components were also investigated for both genotypes in their first year of growth after grafting. Leaf gas exchange and tree growth comparisons were assessed using an innovative grafted plant system, which involved a bi-axis tree with the presence of combinations of identical or mixed (one of each genotype) ‘PremP009’ and ‘Hosui’ scion genotypes grafted onto a single clonal rootstock (‘Buerre Hardy’ BA29). This experimental grafted plant system allowed a technique for comparing leaf photosynthesis of two scion genotypes on the same root system, thereby avoiding between-plant differences in plant water relations. ‘PremP009’ had higher leaf photosynthesis and higher leaf mass compared with ‘Hosui.’ However, by the end of the first year of growth, primary shoots of ‘PremP009’ were shorter with fewer nodes, corresponding to less dry weight gain in primary shoot leaves and stems. This vegetative behavior of ‘PremP009’ is likely a response to the smaller individual leaf area in the early season affecting light capture that greatly limits dry matter accumulation of young trees. HIGHLIGHTS - The bi-axis grafting technique never showed before in a scientific paper presents a strategic system for a comparative study of red/green leaf photosynthetic performance and related dry matter partitioning.

Erratum to: Frequently asked questions about chlorophyll fluorescence, the sequel

Hazem M. Kalaji1 · Gert Schansker2 · Marian Brestic3 · Filippo Bussotti4 · Angeles Calatayud5 · Lorenzo Ferroni6 · Vasilij Goltsev7 · Lucia Guidi8 · Anjana Jajoo9 · Pengmin Li10 · Pasquale Losciale11 · Vinod K. Mishra12 · Amarendra N. Misra13 · Sergio G. Nebauer14 · Simonetta Pancaldi6 · Consuelo Penella5 · Martina Pollastrini4 · Kancherla Suresh15 · Eduardo Tambussi16 · Marcos Yanniccari16 · Marek Zivcak3 · Magdalena D. Cetner1 · Izabela A. Samborska1 · Alexandrina Stirbet17 · Katarina Olsovska18 · Kristyna Kunderlikova18 · Henry Shelonzek19 · Szymon Rusinowski20 · Wojciech Bąba21