Berry shrivel in grapevine: a review considering multiple approaches.

Grapevine berry shrivel, a ripening disorder, causes significant economic losses in the worldwide wine and table grape industries. An early interruption in ripening leads to this disorder, resulting in shriveling and reduced sugar accumulation affecting yield and fruit quality. Loss of sink strength associated with berry mesocarp cell death is an early symptom of this disorder; however, potential internal or external triggers are yet to be explored. No pathogens have been identified that might cause the ripening syndrome. Understanding the underlying causes and mechanisms contributing to berry shrivel is crucial for developing effective mitigation strategies and finding solutions for other ripening disorders associated with climacteric and non-climacteric fruits. This review discusses alterations in the fruit ripening mechanism induced by berry shrivel disorder, focusing primarily on sugar transport and metabolism, cell wall modification and cell death, and changes in the phytohormone profile. The essential open questions are highlighted and analyzed, thus identifying the critical knowledge gaps and key challenges for future research.

Understanding calcium functionality by examining growth characteristics and structural aspects in calcium-deficient grapevine.

This study characterized growth characteristics and cellular details employing microscopy techniques in hydroponically-grown Ca2+-sufficient and Ca2+-deficient grapevines (Vitis vinifera) in a glasshouse. The Ca2+-deficient vines exhibited significant reductions in shoot length, shoot and trunk fresh weights, leaf area, chlorophyll, which eventually led to drooping, yellowing, and chlorosis of leaves. Roots were less dense and primarily dark and necrotic. Furthermore, their xylem vessels were small, polygonal, and appeared to be collapsed yet increased in number and developed lateral roots. Despite such alterations, the anatomical organization of leaves was not affected, yet they developed with more xylem vessels with thick walls and lignin in their mesophyll and vascular tissues. The chloroplasts in internodes' chlorenchyma, phloem, and cambium underwent significant ultrastructural modifications. The concentrations of macro and micronutrients varied significantly among the roots, trunk, canes, and leaves, including the growth characteristics. These structural and growth modifications of calcium deficiency enable us to understand better the link between the symptoms and functions and for a holistic understanding of Ca2+ functionalities.

The Molecular Regulation of Carbon Sink Strength in Grapevine (Vitis vinifera L.).

Sink organs, the net receivers of resources from source tissues, provide food and energy for humans. Crops yield and quality are improved by increased sink strength and source activity, which are affected by many factors, including sugars and hormones. With the growing global population, it is necessary to increase photosynthesis into crop biomass and yield on a per plant basis by enhancing sink strength. Sugar translocation and accumulation are the major determinants of sink strength, so understanding molecular mechanisms and sugar allocation regulation are conducive to develop biotechnology to enhance sink strength. Grapevine (Vitis vinifera L.) is an excellent model to study the sink strength mechanism and regulation for perennial fruit crops, which export sucrose from leaves and accumulates high concentrations of hexoses in the vacuoles of fruit mesocarp cells. Here recent advances of this topic in grape are updated and discussed, including the molecular biology of sink strength, including sugar transportation and accumulation, the genes involved in sugar mobilization and their regulation of sugar and other regulators, and the effects of hormones on sink size and sink activity. Finally, a molecular basis model of the regulation of sugar accumulation in the grape is proposed.

In-Depth Aroma and Sensory Profiling of Unfamiliar Table-Grape Cultivars.

We present an in-depth analysis of aroma profiles and sensory attributes, employing solid-phase microextraction gas chromatography/mass spectrometry (SPME-GC-MS) to identify the key compounds driving consumer preference in 19 unfamiliar cultivars. In combination with popular cultivars, we identified a total of 100 compounds in all table grapes, of which 26 key volatiles were correlated with consumer liking. Based on this relationship, five aroma combinations (AC) were formulated, wherein 33 compounds contributed to aroma intensity, and thus, were viewed as active volatiles. The fruity, floral, and sweet aromas were further divided into secondary aromatic series, of which the apple, citrus, orange, rose, geranium, violet, and honey aromas constituted the predominant series in unfamiliar cultivars. Xiangyue and Heikuixiang emerged as the preferred table grapes according to our analysis. By comparison, the popular cultivars showed relatively fewer volatiles, but their contents were much greater than the large number of volatiles identified in the unfamiliar cultivars.

Plant hydraulic conductance adapts to shoot number but limits shoot vigour in grapevines.

The rate of shoot growth (vigour) in grapevines tends to decrease as the number of shoots per plant increases. Because the underlying causes of this relationship remain unclear, they were studied by variable pruning of field-grown, deficit-irrigated Merlot grapevines (Vitis vinifera L.). Shoot number ranged from 11 to 124 per vine and was inversely correlated with shoot growth rate, leaf appearance rate, axillary bud outgrowth, internode length, leaf size, shoot leaf area, carbon partitioned to the fruit (Cfruit) per shoot, average daily maximum photosynthesis (Amax), stomatal conductance (gmax), and leaf-specific hydraulic conductance (Kl). Shoot number was positively correlated with canopy leaf area, whole-vine Cfruit, whole-plant hydraulic conductance (Kv), and canopy conductance (Kc). Higher shoot vigour was associated with higher Amax, gmax, predawn leaf water potential (Ψpd), shoot hydraulic conductance (Ks), Kl, and Kv. Vigorous shoots supported both more vegetative growth and more reproductive growth; thus fruit growth did not compete with shoot growth for photosynthates. These results indicate that the hydraulic capacity of grapevines adapts to varying shoot numbers to support leaf physiology, growth, and carbon partitioning, but adaptation may be limited, putting upper bounds on the growth of individual shoots and fruit.

Sugar demand of ripening grape berries leads to recycling of surplus phloem water via the xylem.

We tested the common assumption that fleshy fruits become dependent on phloem water supply because xylem inflow declines at the onset of ripening. Using two distinct grape genotypes exposed to drought stress, we found that a sink-driven rise in phloem inflow at the beginning of ripening was sufficient to reverse drought-induced berry shrinkage. Rewatering accelerated berry growth and sugar accumulation concurrently with leaf photosynthetic recovery. Interrupting phloem flow through the peduncle prevented the increase in berry growth after rewatering, but interrupting xylem flow did not. Nevertheless, xylem flow in ripening berries, but not berry size, remained responsive to root or shoot pressurization. A mass balance analysis on ripening berries sampled in the field suggested that phloem water inflow may exceed growth and transpiration water demands. Collecting apoplastic sap from ripening berries showed that osmotic pressure increased at distinct rates in berry vacuoles and apoplast. Our results indicate that the decrease in xylem inflow at the onset of ripening may be a consequence of the sink-driven increase in phloem inflow. We propose a conceptual model in which surplus phloem water bypasses the fruit cells and partly evaporates from the berry surface and partly moves apoplastically to the xylem for outflow.

Loss of rachis cell viability is associated with ripening disorders in grapes.

Rachises of grape (Vitis vinifera L.) clusters that appeared healthy or displayed symptoms of the ripening disorders berry shrivel (BS) or bunch-stem necrosis (BSN) were treated with the cellular viability stain fluorescein diacetate and examined by confocal microscopy. Clusters with BS and BSN symptoms experienced a decrease of cell viability throughout the rachis, and their berries contained 70-80% less sugar than healthy berries. The xylem-mobile dye basic fuchsin, infiltrated via the cut base of shoots with one healthy and one BS cluster, moved to all berries on the healthy cluster but generally failed to move into the peduncle of the BS cluster. Peduncle girdling did not interrupt dye movement in the xylem, but stopped solute accumulation in berries and led to berry shrinkage. In contrast, surgically destroying the peduncle xylem at the onset of ripening did not affect berry growth and solute accumulation. These results indicate that cessation of sugar and water accumulation in BS and BSN is associated with phloem death in the rachis. Although xylem flow to the berries may also cease, a functional xylem connection to the shoot may not be required for normal ripening, while water loss from berries by transpiration and xylem efflux may explain the characteristic berry shrinkage that is associated with these ripening disorders. The similarity of internal tissue breakdown in BS and BSN rachises and the correlation observed here between the proportion of shrinking berries on a cluster and the severity of rachis necrosis suggest that there may be a gradual transition between the two ripening disorders. Seeds from healthy and BS clusters showed no differences in colour, morphology, weight, viability, and ability to germinate, which indicates that the disorder may not appear until seeds are mature.

Ripening grape berries remain hydraulically connected to the shoot.

Berry diameter was monitored during dry-down and rewatering cycles and pressurization of the root system of Vitis vinifera (cv. Merlot) and Vitis labruscana (cv. Concord) to test changes in xylem functionality during grape ripening. Prior to veraison (onset of ripening), berries maintained their size under declining soil moisture until the plants had used 80% of the transpirable soil water, began to shrink thereafter, and recovered rapidly after rewatering. By contrast, berry diameter declined slowly but steadily during post-veraison water stress and did not recover after rewatering; irrigation merely prevented further shrinking. Preconditioning vines with a period of water stress after flowering did not influence the berries' reaction to subsequent changes in transpirable soil water. Pressurizing the root system led to concomitant changes in berry diameter only prior to veraison, although some post-veraison Concord, but not Merlot, berries cracked under root pressurization. The xylem-mobile dye basic fuchsin, infused via the shoot base, moved throughout the berry vasculature before veraison, but became gradually confined to the brush area during ripening. When the dye was infused through the stylar end of attached berries, it readily moved back to the plant both before and after veraison. Our work demonstrated that berry-xylem conduits retain their capacity for water and solute transport during ripening. It is proposed here that apoplastic phloem unloading coupled with solute accumulation in the berry apoplast may be responsible for the decline in xylem water influx into ripening grape berries. Instead, the xylem may serve to recycle excess phloem water back to the shoot.

Functional xylem in the post-veraison grape berry.

A number of studies have shown a transition from a primarily xylem to a primarily phloem flow of water as fleshy fruits develop, and the current hypothesis to explain this transition, particularly in grape (Vitis vinifera L.) berries, is that the vascular tissue (tracheids) become non-functional as a result of post-veraison berry growth. In most studies, pedicels have been dipped in a vial containing an apoplastic dye, which was taken up into the entire peripheral and axial xylem vasculature of pre-veraison, but not post-veraison berries. The pressure plate/pressure membrane apparatus that is commonly used to study soil moisture characteristics was adapted and the pre- to post-veraison change in xylem functionality in grape berries was re-evaluated by establishing a hydrostatic (tension) gradient between the pedicel and a cut surface at the stylar end of the berry. Under the influence of this applied hydrostatic gradient, movement of the apoplastic tracer dye, basic fuchsin, was found in the pedicel and throughout the axial and peripheral xylem of the berry mesocarp. A similar movement of dye could be obtained by simply adjoining the stylar cut surface to a dry, hydrophilic wicking material. Since both pre- and post-veraison berries hydrate when the pedicel is dipped in water, it is hypothesized that the absence of dye movement into the vasculature of post-veraison berries indicates not a loss of xylem function, but rather the loss of an appropriate driving force (hydrostatic gradient) in the berry apoplast. Based on this hypothesis, and the substantial decrease in xylem flows that occur in intact grape berries at veraison, it is suggested that there may be significant changes in the pattern of solute partitioning between the fruit symplast and apoplast at veraison. It is further suggested that diurnal patterns in symplast/apoplast solute partitioning in grapes and other fleshy fruit, may explain the observed minimal xylem contribution to the water budgets of these fruits.

Absorption of foliar-applied arsenic by the arsenic hyperaccumulating fern (Pteris vittata L.).

The fact that heavy metals can enter various domains of the plant system through foliar pathways spurred us to explore if the fronds of the Chinese brake fern (Pteris vittata L.), a hyperaccumulator of arsenic, a carcinogenic metalloid, was proficient in absorbing arsenic in the form of sprays. The specific objective of this study was to investigate the impact of frond age, form of arsenic, and time of application on the absorption of foliar-applied arsenic by the brake fern; also examined were the effects of foliar sprays on surface ultrastructure and arsenic speciation in the frond following absorption. Foliar sprays of different arsenic concentrations (0, 50, 100, 200, and 400 ppm) were applied to young and fertile fronds. A positive linear relationship existed between arsenic concentration and absorption; the arsenic concentration of fronds increased from 50 to 200 ppm. Time-course analysis with excised pinnae indicated an initial linear increase followed by a plateau at 48 h. The young fronds with immature sori absorbed more arsenic (3100 ppm) than the fertile mature fronds (890 ppm). In the frond, the arsenic absorption was greatest in the lamina of the pinnae followed by the sori and the rachis. Applying arsenic during night (20:00-22:00 h) or afternoon (12:00-14:00 h) resulted in greater absorption of arsenic than the application in the morning (08:00-10:00 h). The arsenic absorption was greater through abaxial surfaces than through adaxial surfaces. The brake fern absorbed more arsenic when it was applied in the form of arsenite. Regardless of the form of arsenic and the surface it was applied to, arsenic occurred as arsenite, the reduced and the most toxic form of arsenic, after having been absorbed by the fronds. Scanning electron microscopy revealed no surface morphological alterations following all arsenic sprays. The study unequivocally illustrated that the Chinese brake fern absorbed foliar-applied arsenic with great efficiency. Consequently, the arsenic concentrations in the fronds transcended the levels of hyperaccumulation; such a characteristic could be exploited in the phytoremediation of groundwater contaminated with arsenic.

Leaf chlorophyll, net gas exchange and chloroplast ultrastructure in citrus leaves of different nitrogen status.

One-year-old 'Cleopatra mandarin' (Citrus reticulata Blanco) seedlings were raised in a greenhouse and fertilized with nitrogen (N) at four application frequencies. Nitrogen-deficient leaves (86 mmol N m-2) had less chlorophyll per unit area, but a greater chlorophyll a:b ratio than N-fertilized leaves (> 187 mmol N m-2). Leaf dry mass per area (DM area-1) and total chlorophyll concentration increased linearly with increasing leaf N, whereas chlorophyll a:b ratio declined. Net assimilation of CO2 (A(CO2)) and leaf water-use efficiency (WUE) reached maximum values in leaves with approximately 187 mmol N m-2. Nitrogen-deficient leaves exhibited small chloroplasts with no starch granules; grana and stroma lamellae that coincided with the accretion of numerous large plastoglobuli in the stroma disintegrated. High-N leaves had large chloroplasts with well-developed grana, stroma lamellae and starch granules that enlarged with increasing N concentration. The lack of an increase in A(CO2) capacity at leaf N concentrations above 187 mmol N m-2 appeared to be correlated with the presence of numerous large starch granules.

Comparative anatomy and morphology of Vitis vinifera (Vitaceae) somatic embryos from solid- and liquid-culture-derived proembryogenic masses.

Ontogeny of somatic embryos of grapevine (Vitis vinifera) produced from solid- and liquid-culture-derived proembryogenic masses (PEM) was compared using light and scanning electron microscopy. Somatic embryos produced from solid-medium-derived PEM (SPEM) had large cotyledons, little or no visible suspensor structure, and a relatively undeveloped concave shoot apical meristem, whereas those from liquid-medium-derived PEM (LPEM) had smaller cotyledons, a distinct suspensor, and a flat-to-convex shoot apical meristem. The convex shoot apical meristem in LPEM-derived somatic embryos formed as early as the heart stage of development; it was 4-6 cell layers deep and rich in protein. Suspensors persisted in fully developed and mature LPEM-derived somatic embryos. The SPEM-derived somatic embryos exhibited dormancy, as do mature zygotic embryos, which also have a rudimentary suspensor, whereas LPEM-derived embryos were not dormant. We hypothesize that the presence of a persistent suspensor in LPEM-derived somatic embryos modulates development, ultimately resulting in rapid germination and a high plant-regeneration rate.

Arsenic accumulation in the hyperaccumulator Chinese brake and its utilization potential for phytoremediation.

The unique property of arsenic hyperaccumulation by the newly discovered Chinese brake (Pteris vittata L.) fern is of great significance in the phytoremediation of arsenic-contaminated soils. The objectives of this study were to (i) examine arsenic accumulation characterized by its distribution pattern in Chinese brake, and (ii) assess the phytoextraction potential of the plant. Young ferns with five or six fronds were transferred to an arsenic-contaminated soil containing 98 mg As kg-1 and grown for 20 wk in a greenhouse. At harvest, the Chinese brake produced a total dry biomass of 18 g plant-1. Arsenic concentration in the fronds was 6000 mg kg-1 dry mass after 8 wk of transplanting, and it increased to 7230 mg kg-1 after 20 wk with a bioconcentration factor (ratio of plant arsenic concentration to water-soluble arsenic in soil) of 1450 and a translocation factor (ratio of arsenic concentration in shoot to that in root) of 24. The arsenic concentrations increased as the fronds aged, with the old fronds accumulating as much as 13,800 mg As kg-1. Most (approximately 90%) of the arsenic taken up by the Chinese brake was transported to the fronds, with the lowest arsenic concentrations in roots. About 26% of the initial soil arsenic was removed by the plant after 20 wk of transplanting. Our data suggest that the arsenic hyperaccumulating property of the Chinese brake could be exploited on a large scale to remediate arsenic contaminated soils.