Photochemistry and photoprotection of 'Gem' avocado (Persea americana Mill.) leaves within and outside the canopy and the relationship with fruit maturity.

A reduction in photosynthesis results in a reduced CO2 assimilation rate and availability of carbohydrates essential for fruit growth and development. This study determined photosynthetic efficiency and photoprotection mechanisms within and outside leaf canopy positions in 'Gem' avocado orchards and their relationship with avocado fruit maturity. The study was conducted in a commercial orchard at Everdon Estate in KwaZulu-Natal, South Africa. A total of 15 eight-year-old avocado trees (cv. Gem) were selected in a completely randomised design with three replicates, with each replicate consisting of five trees. Data were collected bi-weekly on photosynthetic rate (A), effective quantum efficiency of photosystem II (ϕPSII), stomatal conductance (gs), transpiration rate (T), electron transport rate (ETR), minimum fluorescence (Fo'), maximum fluorescence (Fm'), variable fluorescence (Fv'), intrinsic water use efficiency (WUEi), instantaneous water use efficiency (WUEins), intercellular CO2 concentration (Ci) and photochemical quenching (qP) from full bloom to fruit physiological maturity (∼25 % dry matter content (DM)). The results showed that leaves from the outside position had higher A (29.46 mol CO2 m-2s-1); gs (0.078 mol CO2 m-2s-1); ΦPS II (0.32); and qP (0.52) compared to those within the canopy position with lower A (19.27 mol CO2 m-2s-1); gs (0.0037 mol CO2 m-2s-1); ΦPS II (0.044) and qP (0.075), respectively. Contrastingly, chlorophyll fluorescence and photoprotection parameters were higher within the canopy than on the outside, suggesting that the greater proportion of energy accumulated within the canopy was used for photoprotection other than photochemistry. Photosynthetic rate (A), gs, Ci, T, WUEi and WUEins, correlated significantly with mesocarp dry matter (DM), while all other parameters correlated poorly. The high photosynthetic efficiency of leaves from outside the canopy resulted in an average DM of 28.9 % compared to 26.9 % of fruit within the canopy. The present findings suggest that reduced photosynthetic efficiency of 'Gem' avocado within the canopy position does not compromise fruit DM by reserving more energy for photoprotection; however, it delays maturity by about two weeks.

Impact of Laurel Wilt, Caused by Raffaelea lauricola, on Leaf Gas Exchange and Xylem Sap Flow in Avocado, Persea americana.

Laurel wilt, caused by Raffaelea lauricola, is a destructive disease of avocado (Persea americana). The susceptibility of different cultivars and races was examined previously but more information is needed on how this host responds to the disease. In the present study, net CO2 assimilation (A), stomatal conductance of H2O (gs), transpiration (E), water use efficiency (WUE), and xylem sap flow rates were assessed in cultivars that differed in susceptibility. After artificial inoculation with R. lauricola, there was a close relationship between symptom development and reductions in A, gs, E, WUE, and mean daily sap flow in the most susceptible cultivar, 'Russell', and significantly greater disease and lower A, gs, E, WUE, and sap flow rates were usually detected after 15 days compared with the more tolerant 'Brogdon' and 'Marcus Pumpkin'. Significant differences in preinoculation A, gs, E, and WUE were generally not detected among the cultivars but preinoculation sap flow rates were greater in Russell than in Brogdon and Marcus Pumpkin. Preinoculation sap flow rates and symptom severity for individual trees were correlated at the end of an experiment (r=0.46), indicating that a plant's susceptibility to laurel wilt was related to its ability to conduct water. The potential management of this disease with clonal rootstocks that reduce sap flow rates is discussed.

Electrophysiological assessment of water stress in fruit-bearing woody plants.

Development and evaluation of a real-time plant water stress sensor, based on the electrophysiological behavior of fruit-bearing woody plants is presented. Continuous electric potentials are measured in tree trunks for different irrigation schedules, inducing variable water stress conditions; results are discussed in relation to soil water content and micro-atmospheric evaporative demand, determined continuously by conventional sensors, correlating this information with tree electric potential measurements. Systematic and differentiable patterns of electric potentials for water-stressed and no-stressed trees in 2 fruit species are presented. Early detection and recovery dynamics of water stress conditions can also be monitored with these electrophysiology sensors, which enable continuous and non-destructive measurements for efficient irrigation scheduling throughout the year. The experiment is developed under controlled conditions, in Faraday cages located at a greenhouse area, both in Persea americana and Prunus domestica plants. Soil moisture evolution is controlled using capacitance sensors and solar radiation, temperature, relative humidity, wind intensity and direction are continuously registered with accurate weather sensors, in a micro-agrometeorological automatic station located at the experimental site. The electrophysiological sensor has two stainless steel electrodes (measuring/reference), inserted on the stem; a high precision Keithley 2701 digital multimeter is used to measure plant electrical signals; an algorithm written in MatLab(®), allows correlating the signal to environmental variables. An electric cyclic behavior is observed (circadian cycle) in the experimental plants. For non-irrigated plants, the electrical signal shows a time positive slope and then, a negative slope after restarting irrigation throughout a rather extended recovery process, before reaching a stable electrical signal with zero slope. Well-watered plants presented a continuous signal with daily maximum and a minimum EP of similar magnitude in time, with zero slope. This plant electrical behavior is proposed for the development of a sensor measuring real-time plant water status.

Decreased photochemical efficiency of photosystem II following sunlight exposure of shade-grown leaves of avocado: because of, or in spite of, two kinetically distinct xanthophyll cycles?

This study resolved correlations between changes in xanthophyll pigments and photosynthetic properties in attached and detached shade-grown avocado (Persea americana) leaves upon sun exposure. Lutein epoxide (Lx) was deepoxidized to lutein (L), increasing the total pool by ΔL over 5 h, whereas violaxanthin (V) conversion to antheraxanthin (A) and zeaxanthin (Z) ceased after 1 h. During subsequent dark or shade recovery, de novo synthesis of L and Z continued, followed by epoxidation of A and Z but not of L. Light-saturated nonphotochemical quenching (NPQ) was strongly and linearly correlated with decreasing [Lx] and increasing [L] but showed a biphasic correlation with declining [V] and increasing [A+Z] separated when V deepoxidation ceased. When considering [ΔL+Z], the monophasic linear correlation was restored. Photochemical efficiency of photosystem II (PSII) and photosystem (PSI; deduced from the delivery of electrons to PSI in saturating single-turnover flashes) showed a strong correlation in their continuous decline in sunlight and an increase in NPQ capacity. This decrease was also reflected in the initial reduction of the slope of photosynthetic electron transport versus photon flux density. Generally longer, stronger sun exposures enhanced declines in both slope and maximum photosynthetic electron transport rates as well as photochemical efficiency of PSII and PSII/PSI more severely and prevented full recovery. Interestingly, increased NPQ capacity was accompanied by slower relaxation. This was more prominent in detached leaves with closed stomata, indicating that photorespiratory recycling of CO(2) provided little photoprotection to avocado shade leaves. Sun exposure of these shade leaves initiates a continuum of photoprotection, beyond full engagement of the Lx and V cycle in the antenna, but ultimately photoinactivated PSII reaction centers.

From ecophysiology to phenomics: some implications of photoprotection and shade-sun acclimation in situ for dynamics of thylakoids in vitro.

Half a century of research into the physiology and biochemistry of sun-shade acclimation in diverse plants has provided reality checks for contemporary understanding of thylakoid membrane dynamics. This paper reviews recent insights into photosynthetic efficiency and photoprotection from studies of two xanthophyll cycles in old shade leaves from the inner canopy of the tropical trees Inga sapindoides and Persea americana (avocado). It then presents new physiological data from avocado on the time frames of the slow coordinated photosynthetic development of sink leaves in sunlight and on the slow renovation of photosynthetic properties in old leaves during sun to shade and shade to sun acclimation. In so doing, it grapples with issues in vivo that seem relevant to our increasingly sophisticated understanding of ΔpH-dependent, xanthophyll-pigment-stabilized non-photochemical quenching in the antenna of PSII in thylakoid membranes in vitro.

Lutein from deepoxidation of lutein epoxide replaces zeaxanthin to sustain an enhanced capacity for nonphotochemical chlorophyll fluorescence quenching in avocado shade leaves in the dark.

Leaves of avocado (Persea americana) that develop and persist in deep shade canopies have very low rates of photosynthesis but contain high concentrations of lutein epoxide (Lx) that are partially deepoxidized to lutein (L) after 1 h of exposure to 120 to 350 µmol photons m(-2) s(-1), increasing the total L pool by 5% to 10% (ΔL). Deepoxidation of Lx to L was near stoichiometric and similar in kinetics to deepoxidation of violaxanthin (V) to antheraxanthin (A) and zeaxanthin (Z). Although the V pool was restored by epoxidation of A and Z overnight, the Lx pool was not. Depending on leaf age and pretreatment, the pool of ΔL persisted for up to 72 h in the dark. Metabolism of ΔL did not involve epoxidation to Lx. These contrasting kinetics enabled us to differentiate three states of the capacity for nonphotochemical chlorophyll fluorescence quenching (NPQ) in attached and detached leaves: ΔpH dependent (NPQ(ΔpH)) before deepoxidation; after deepoxidation in the presence of ΔL, A, and Z (NPQ(ΔLAZ)); and after epoxidation of A+Z but with residual ΔL (NPQ(ΔL)). The capacity of both NPQ(ΔLAZ) and NPQ(ΔL) was similar and 45% larger than NPQ(ΔpH), but dark relaxation of NPQ(ΔLAZ) was slower. The enhanced capacity for NPQ was lost after metabolism of ΔL. The near equivalence of NPQ(ΔLAZ) and NPQ(ΔL) provides compelling evidence that the small dynamic pool ΔL replaces A+Z in avocado to "lock in" enhanced NPQ. The results are discussed in relation to data obtained with other Lx-rich species and in mutants of Arabidopsis (Arabidopsis thaliana) with increased L pools.

Insights into carotenoid dynamics in non-foliar photosynthetic tissues of avocado.

Leaves are the main photosynthetically active tissues in most plants. However, stems and fruits are also important for the overall carbon balance of the plant because of their contribution to fixation of the CO(2) released by respiration. Photosynthesis could not be possible without a complete set of photoprotection mechanisms, which include the ubiquitous violaxanthin (V) cycle and the taxonomically restricted lutein epoxide (Lx) cycle. In this work, we characterise carotenoid stoichiometry in photosynthetic stems and fruits of avocado in comparison with that of leaves and specifically whether Lx is present in these tissues and also whether it is involved in a light-driven cycle. Avocado was selected as model species to study whether both cycles were functional in non-foliar photosynthetic structures (stems and fruits). An unusual pigment composition was observed in avocado fruit, with a high content of cis-V and cis-Lx, suggesting a different photosynthetic function. In stems, both xanthophylls de-epoxidated upon illumination, but only V recovered in the dark, indicating the existence of a possible 'truncated' Lx cycle. Lx in fruits was de-epoxidated only when its pool was higher than a threshold of 30 mmol mol(-1) chlorophyll, indicating a high non-photoconvertible pool of Lx. We conclude that, at least in stems, the dynamic regulation of photosynthetic activity could also depend on the Lx cycle.

De novo synthesis and degradation of Lx and V cycle pigments during shade and sun acclimation in avocado leaves.

The photoprotective role of the universal violaxanthin cycle that interconverts violaxanthin (V), antheraxanthin (A), and zeaxanthin (Z) is well established, but functions of the analogous conversions of lutein-5,6-epoxide (Lx) and lutein (L) in the selectively occurring Lx cycle are still unclear. We investigated carotenoid pools in Lx-rich leaves of avocado (Persea americana) during sun or shade acclimation at different developmental stages. During sun exposure of mature shade leaves, an unusual decrease in L preceded the deepoxidation of Lx to L and of V to A+Z. In addition to deepoxidation, de novo synthesis increased the L and A+Z pools. Epoxidation of L was exceptionally slow, requiring about 40 d in the shade to restore the Lx pool, and residual A+Z usually persisted overnight. In young shade leaves, the Lx cycle was reversed initially, with Lx accumulating in the sun and declining in the shade. De novo synthesis of xanthophylls did not affect alpha- and beta-carotene pools on the first day, but during long-term acclimation alpha-carotene pools changed noticeably. Nonetheless, the total change in alpha- and beta-branch carotenoid pools was equal. We discuss the implications for regulation of metabolic flux through the alpha- and beta-branches of carotenoid biosynthesis and potential roles for L in photoprotection and Lx in energy transfer to photosystem II and explore physiological roles of both xanthophyll cycles as determinants of photosystem II efficiency.