Rootstock and Crop Load Effects on 'Honeycrisp' Photosynthetic Performance and Carbohydrate Accumulation.

Rootstock selection and crop load adjustment are key practices in apple orchard management; nevertheless, the effects of rootstocks and crop load levels on important physiological processes of the scions, such as photosynthetic performance and carbohydrate accumulation, are still unclear. To investigate the impact of different rootstocks and crop load levels on scion photosynthesis and carbohydrate buildup, in 2020, 'Honeycrisp' trees grafted on rootstocks 'G.41', 'G.935', and 'M.9-T337' were thinned to low and high crop load levels, and photosynthetic performance and carbohydrate accumulation in leaves and fruit were evaluated. Leaves from 'G.935' showed the highest net photosynthesis and electron use efficiency of photosynthesis and the lowest activity for non-net carboxylative processes, all together indicative of enhanced photosynthetic performance. High crop load determined an increase in gas exchange, suggesting a positive feedback of high fruit competition on carbon assimilation. While rootstock 'M.9-T337' showed a higher accumulation of starch in leaves, no pattern regarding the composition of leaf-soluble sugars among rootstocks could be identified. Conversely, by the end of the harvest season, leaves from low-cropping trees had higher fructose, glucose, and sorbitol than those from high-cropping trees, but differences in starch content were not significant. Fructose and sorbitol concentrations were affected by rootstock and crop load, respectively. Overall, this study showed that high cropping enhanced photosynthesis in 'Honeycrisp' apple and determined lower accumulation of some soluble carbohydrates (fructose, glucose, sorbitol) in leaves. This study also provided insights into how rootstocks affect photosynthetic performance of 'Honeycrisp', highlighting 'G.935' as the rootstock conferring the highest photosynthetic capacity under the present experimental conditions.

Detection and distribution of two dominant alleles associated with the sweet kernel phenotype in almond cultivated germplasm.

Almond [Prunus dulcis Miller (D. A. Webb), syn. Prunus amygdalus L.)] is the major tree nut crop worldwide in terms of production and cultivated area. Almond domestication was enabled by the selection of individuals bearing sweet kernels, which do not accumulate high levels of the toxic cyanogenic glucoside amygdalin. Previously, we showed that the Sweet kernel (Sk) gene, controlling the kernel taste in almond, encodes a basic helix loop helix (bHLH) transcription factor regulating the amygdalin biosynthetic pathway. In addition, we characterized a dominant allele of this gene, further referred to as Sk-1, which originates from a C1036→T missense mutation and confers the sweet kernel phenotype. Here we provide evidence indicating that the allele further referred to as Sk-2, originally detected in the cultivar "Atocha" and arising from a T989→G missense mutation, is also dominantly inherited and confers the sweet kernel phenotype in almond cultivated germplasm. The use of single nucleotide polymorphism (SNP) data from genotyping by sequencing (GBS) for population structure and hierarchical clustering analyses indicated that Sk-2 occurs in a group of related genotypes, including the widespread cultivar "Texas", descending from the same ancestral population. KASP and dual label functional markers were developed for the accurate and high-throughput selection of the Sk-1 and Sk-2 alleles, and the genotyping of a panel of 134 almond cultivars. Overall, our results provide further insights on the understanding of the almond cultivation history. In addition, molecular marker assays and genotypic data presented in this study are expected to be of major interest for the conduction of almond breeding programs, which often need to select sweet kernel individuals in segregant populations.

Susceptible and resistant olive cultivars show differential physiological response to Xylella fastidiosa infections.

Olive quick decline syndrome (OQDS) is a severe disease, first described in Italy in late 2013, caused by strains of Xylella fastidiosa subsp. pauca (Xfp) in susceptible olive cultivars. Conversely, resistant olive cultivars do not develop OQDS but present scattered branch dieback, which generally does not evolve to severe canopy decline. In the present study, we assessed the physiological responses of Xfp-infected olive trees of susceptible and resistant cultivars. Periodic measurements of stomatal conductance (gs) and stem water potential (Ψstem) were performed using a set of healthy and Xfp-infected plants of the susceptible "Cellina di Nardò" and resistant "Leccino" and "FS17" cultivars. Strong differences in Δgs and ΔΨstem among Xfp-infected trees of these cultivars were found, with higher values in Cellina di Nardò than in Leccino and FS17, while no differences were found among healthy plants of the different cultivars. Both resistant olive cultivars showed lower water stress upon Xfp infections, compared to the susceptible one, suggesting that measurements of gs and Ψstem may represent discriminating parameters to be exploited in screening programs of olive genotypes for resistance to X. fastidiosa.

The energy cost of repairing photoinactivated photosystem II: an experimental determination in cotton leaf discs.

Photosystem II (PSII), which splits water molecules at minimal excess photochemical potential, is inevitably photoinactivated during photosynthesis, resulting in compromised photosynthetic efficiency unless it is repaired. The energy cost of PSII repair is currently uncertain, despite attempts to calculate it. We experimentally determined the energy cost of repairing each photoinactivated PSII in cotton (Gossypium hirsutum) leaves, which are capable of repairing PSII in darkness. As an upper limit, 24 000 adenosine triphosphate (ATP) molecules (including any guanosine triphosphate synthesized at the expense of ATP) were required to repair one entire PSII complex. Further, over a 7-h illumination period at 526-1953 µmol photons m-2 s-1 , the ATP requirement for PSII repair was on average up to 4.6% of the ATP required for the gross carbon assimilation. Each of these two measures of ATP requirement for PSII repair is two- to three-fold greater than the respective reported calculated value. Possible additional energy sinks in the PSII repair cycle are discussed.

Wah Soon Chow, a teacher, a friend and a colleague.

To finish this special issue, some friends, colleagues and students of Prof. Chow (Emeritus Professor, the Research School of Biology, the Australian National University) have written small tributes to acknowledge not only his eminent career but to describe his wonderful personality.

Red and Blue Netting Alters Leaf Morphological and Physiological Characteristics in Apple Trees.

There is little information about the role of red and blue light on leaf morphology and physiology in fruit trees, and more studies have been developed in herbaceous plants grown under controlled light conditions. The objective of this research was to evaluate the effect of red and blue screens on morpho-anatomy and gas exchange in apple leaves grown under ambient sunlight conditions. Apple trees cv. Fuji were covered by 40% red and blue nets, leaving trees with 20% white net as control. Light relations (photosynthetic photon flux density, PPFD; red to far-red light ratio, R/FR and blue to red light ratio, B/R), morpho-anatomical features of the leaf (palisade to spongy mesophyll ratio, P/S, and stomata density, SD) and leaf gas exchange (net photosynthesis rate, An; stomatal conductance, g s; transpiration rate, E; and intrinsic water use efficiency, IWUE) were evaluated. Red and blue nets reduced 27% PPFD, reducing by 20% SD and 25% P/S compared to control, but without negative effects on An and g s. Blue net increased g s 21%, leading to the highest E and lowest IWUE by increment of B/R light proportion. These findings demonstrate the potential use of red and blue nets for differential modulation of apple leaf gas exchange through sunlight management under field conditions.

Almond diversity and homozygosity define structure, kinship, inbreeding, and linkage disequilibrium in cultivated germplasm, and reveal genomic associations with nut and seed weight.

Almond [Prunus dulcis Miller (D.A. Webb)] is the main tree nut species worldwide. Here, genotyping-by-sequencing (GBS) was applied to 149 almond cultivars from the ex situ collections of the Italian Council for Agricultural Research (CREA) and the Spanish National Research Council (CSIC), leading to the detection of 93,119 single-nucleotide polymorphisms (SNPs). The study of population structure outlined four distinct genetic groups and highlighted diversification between the Mediterranean and Californian gene pools. Data on SNP diversity and runs of homozygosity (ROHs) allowed the definition of kinship, inbreeding, and linkage disequilibrium (LD) decay in almond cultivated germplasm. Four-year phenotypic observations, gathered on 98 cultivars of the CREA collection, were used to perform a genome-wide association study (GWAS) and, for the first time in a crop species, homozygosity mapping (HM), resulting in the identification of genomic associations with nut, shell, and seed weight. Both GWAS and HM suggested that loci controlling nut and seed weight are mostly independent. Overall, this study provides insights on the almond cultivation history and delivers information of major interest for almond genetics and breeding. In a broader perspective, our results encourage the use of ROHs in crop science to estimate inbreeding, choose parental combinations minimizing the risk of inbreeding depression, and identify genomic footprints of selection for specific traits.

Innovative Soil Management and Micro-Climate Modulation for Saving Water in Peach Orchards.

Microclimatic and soil management studies emphasize that roofing above the canopy or soil mulching contributes to reduce water losses from horticultural cropping systems and, at the same time, to increase water use efficiency. The aim of this 2-year on-farm study, carried out on a late ripening peach (cv. California) orchard, was to investigate the combined effect of water supply (full or deficit irrigation, DI), incoming light (hail or shading net), and soil management (tilling or mulching) on: microclimate; fruit growth; yield; irrigation water use productivity (WPI); and soil water stress coefficient (Ks). Shading hail net reduced air temperature (-1°C), wind speed (-57%), solar radiation (-32%), while increased relative air humidity (+9.5%). Compared to the control treatment (hail net coverage, soil tillage, and full irrigation), the innovative management (DI + shading hail net + mulching) reduced seasonal volumes of irrigation water (-25%) and increased both final yield (+36%) and WPI (+53%). Saving water resources without losing yield is an achievable goal by peach orchards growing under the Mediterranean climate if the DI agro-technique is adopted conjointly with shading hail net and soil mulching.

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.

Frequently asked questions about chlorophyll fluorescence, the sequel.

Using chlorophyll (Chl) a fluorescence many aspects of the photosynthetic apparatus can be studied, both in vitro and, noninvasively, in vivo. Complementary techniques can help to interpret changes in the Chl a fluorescence kinetics. Kalaji et al. (Photosynth Res 122:121-158, 2014a) addressed several questions about instruments, methods and applications based on Chl a fluorescence. Here, additional Chl a fluorescence-related topics are discussed again in a question and answer format. Examples are the effect of connectivity on photochemical quenching, the correction of F V /F M values for PSI fluorescence, the energy partitioning concept, the interpretation of the complementary area, probing the donor side of PSII, the assignment of bands of 77 K fluorescence emission spectra to fluorescence emitters, the relationship between prompt and delayed fluorescence, potential problems when sampling tree canopies, the use of fluorescence parameters in QTL studies, the use of Chl a fluorescence in biosensor applications and the application of neural network approaches for the analysis of fluorescence measurements. The answers draw on knowledge from different Chl a fluorescence analysis domains, yielding in several cases new insights.

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).

Increasing water stress negatively affects pear fruit growth by reducing first its xylem and then its phloem inflow.

Drought stress negatively affects many physiological parameters and determines lower yields and fruit size. This paper investigates on the effects of prolonged water restriction on leaf gas exchanges, water relations and fruit growth on a 24-h time-scale in order to understand how different physiological processes interact to each other to face increasing drought stress and affect pear productive performances during the season. The diurnal patterns of tree water relations, leaf gas exchanges, fruit growth, fruit vascular and transpiration flows were monitored at about 50, 95 and 145 days after full bloom (DAFB) on pear trees of the cv. Abbé Fétel, subjected to two irrigation regimes, corresponding to a water restitution of 100% and 25% of the estimated Etc, respectively. Drought stress progressively increased during the season due to lower soil tensions and higher daily vapour pressure deficits (VPDs). Stem water potential was the first parameter to be negatively affected by stress and determined the simultaneous reduction of fruit xylem flow, which at 95 DAFB was reflected by a decrease in fruit daily growth. Leaf photosynthesis was reduced only from 95 DAFB on, but was not immediately reflected by a decrease in fruit phloem flow, which instead was reduced only at 145 DAFB. This work shows how water stress negatively affects pear fruit growth by reducing first its xylem and then its phloem inflow. This determines a progressive increase in the phloem relative contribution to growth, which lead to the typical higher dry matter percentages of stressed fruit.

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 k i , and that of repair k r 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 k i and k r . The value of k i , 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 k r , sizable in darkness, changed in the light depending on which surface was blocked by contact with water. When the abaxial surface was blocked, k r first peaked at moderate irradiance and then decreased at high irradiance. When the adaxial surface was blocked, k r first increased at low irradiance, then plateaued, before increasing markedly at high irradiance. At the highest irradiance, k r 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 k i and that of repair k r .

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 O(2) yield per single-turnover flash in CO(2)-enriched air is a more direct measurement of the functional content, assuming that each functional PS II evolves one O(2) molecule after four flashes. However, the gross O(2) 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.

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 DeltapH. 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 DeltapH 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.

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.