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Potassium (K) plays a crucial role in multiple physiological and developmental processes in plants. Its deficiency is a common abiotic stress that inhibits plant growth and reduces crop productivity. A better understanding of the mechanisms involved in plant responses to low K could help to improve the efficiency of K use in plants. However, such responses remain poorly characterized in fruit tree species such as pears (Pyrus sp). We analyzed the physiological and transcriptome responses of a commonly used pear rootstock, Pyrus betulaefolia, to K-deficiency stress (0 mM). Potassium deprivation resulted in apparent changes in root morphology, with short-term low-K stress resulting in rapidly enhanced root growth. Transcriptome analyses indicated that the root transcriptome was coordinately altered within 6 h after K deprivation, a process that continued until 15 d after treatment. Potassium deprivation resulted in the enhanced expression (up to 5-fold) of a putative high-affinity K+ transporter, PbHAK5 (Pbr037826.1), suggesting the up-regulation of mechanisms associated with K+ acquisition. The enhanced root growth in response to K-deficiency stress was associated with a rapid and sustained decrease in the expression of a transcription factor, PbMYB44 (Pbr015309.1), potentially involved in mediating auxin responses, and the increased expression of multiple genes associated with regulating root growth. The concentrations of several phytohormones including indoleacetic acid (IAA), ABA, ETH, gibberellin (GA3), and jasmonic acid (JA) were higher in response to K deprivation. Furthermore, genes coding for enzymes associated with carbon metabolism such as SORBITOL DEHYDROGENASE (SDH) and SUCROSE SYNTHASE (SUS) displayed greatly enhanced expression in the roots under K deprivation, presumably indicating enhanced metabolism to meet the increased energy demands for growth and K+ acquisition. Together, these data suggest that K deprivation in P. betulaefolia results in the rapid re-programming of the transcriptome to enhance root growth and K+ acquisition. These data provide key insights into the molecular basis for understanding low-K-tolerance mechanisms in pears and in other related fruit trees and identifying potential candidates that warrant further analyses.

Fruit size is a highly valued commercial trait in peach. Competition among fruit and among other sinks on a tree reduces potential growth rate of the fruit. Hence, crop-load management strategies such as thinning (removal of flowers or fruit) are often practiced by growers to optimize fruit size. Thinning can be performed at bloom or during early fruit development and at different intensities to optimize fruit growth responses. Responses to thinning may be cultivar and location specific. The objective of the current study was to fine-tune thinning strategies in the southeastern United States, a major peach producing region. Timing and intensity of thinning were evaluated across multiple cultivars over three years. Thinning at bloom or at 21 d after full bloom (DAFB) improved fruit size in comparison to unthinned trees in &lsquo, Cary Mac&rsquo, and &lsquo, July Prince&rsquo, respectively, in one year. Bloom-thinning reduced fruit yield (kg per tree) in the above cultivars in one year, suggesting that flower thinning alone may not be a viable option in this region. Intensity of thinning, evaluated as spacings of 15 cm and 20 cm between fruit, did not differentially affect fruit weight or yield. However, fruit diameter decreased quadratically with increasing fruit number per tree in &lsquo, &lsquo, Summer Flame&rsquo, Similarly, fruit weight decreased quadratically in response to increase in fruit number per tree in &lsquo, Further, yield-per-tree decreased with increasing fruit size in &lsquo, Importantly, these relationships were cultivar specific. Together, the data suggest that achieving a target fruit number per tree is an effective strategy for crop-load management to optimize fruit size in southeastern peach production. The target fruit number per tree may potentially be achieved through a combination of flower and fruit-thinning during early fruit development. Such an approach may provide flexibility in crop-load management in relation to adverse weather events.

Additional file 1. Spatiotemporal patterns of transcript abundance of ALUMINUM ACTIVATED MALATE TRANSPORTER 9 (MdALMT9) in apple fruit in response to fruit load reduction, measured using quantitative RT-PCR. CC: Control fruit load-Cortex; CP: Control fruit load-Pith; RC: Reduced fruit load-Cortex; RP: Reduced fruit load-Pith. Fruit load reduction treatment was performed at 11 d after full bloom. The mean and S.E. of the mean (n = 4) are displayed. Asterisk indicates significant difference between control and reduced fruit load treatments in the cortex (α = 0.05). The transcript abundance data are presented in reference to mean expression at 0 d after treatment in RC. Transcript abundance was normalized to that of the apple GAPDH (GLYCERALDEHYDE 3-PHOSPHATE DEHYDROGENASE) gene. Shaded regions in the background indicate early (dark grey), mid (light grey), and late fruit development (white) periods.

Additional file 3 Spatiotemporal patterns of apple fruit tissue metabolite contents in response to fruit load reduction. Fruit metabolite concentration was multiplied with normalized tissue areas of the cortex and pith to obtain an estimate of tissue metabolite content. Tissue area was used as a proxy for tissue weight. The mean and standard error of the mean (n = 4) are displayed. CC: Control fruit load-Cortex; CP: Control fruit load-Pith; RC: Reduced fruit load-Cortex; RP: Reduced fruit load-Pith. Asterisks and dagger symbols indicate significant difference between control and reduced fruit load treatments in the cortex and pith, respectively (α = 0.05). Shaded regions indicate early (dark grey), mid (light grey), and late fruit development (white) periods.

Additional file 2. A correlation matrix among different fruit metabolites. Pearson correlation analysis was performed to determine relationships among all metabolites analyzed in this study. Fruit metabolite concentration data were used here. Correlations are presented as color-keyed correlation coefficients (numbers) and a heat map. Positive correlations are indicated in blue and negative correlations are in red. The intensity of the color represents the magnitude of the relationship between two metabolites.

Additional file 5. List of the apple genes and sequences of primers used in quantitative RT-PCR analyses.

Additional file 4. Fruit developmental stages of ‘Golden Delicious Smoothee’. Fruit developmental stages used in this study are displayed. Fruit represented here were longitudinally sliced and fixed in CRAF III fixative. Images were obtained using a flatbed scanner and processed with ImageJ. Bar indicates 1 cm. DAT: Days after treatment. Fruit load reduction was performed at 11 d after full bloom corresponding to 0 DAT.

Blueberry (Vaccinium sp.) is thought to display a preference for the ammonium (NH4+) form over the nitrate (NO3-) form of inorganic nitrogen (N). This N-source preference has been associated with a generally low capacity to assimilate the NO3- form of N, especially within the shoot tissues. Nitrate assimilation is mediated by nitrate reductase (NR), a rate limiting enzyme that converts NO3- to nitrite (NO2-). We investigated potential limitations of NO3- assimilation in two blueberry species, rabbiteye (Vaccinium ashei) and southern highbush (Vaccinium corymbosum) by supplying NO3- to the roots, leaf surface, or through the cut stem. Both species displayed relatively low but similar root uptake rates for both forms of inorganic N. Nitrate uptake through the roots transiently increased NR activity by up to 3.3-fold and root NR gene expression by up to 4-fold. However, supplying NO3- to the roots did not increase its transport in the xylem, nor did it increase NR activity in the leaves, indicating that the acquired N was largely assimilated or stored within the roots. Foliar application of NO3- increased leaf NR activity by up to 3.5-fold, but did not alter NO3- metabolism-related gene expression, suggesting that blueberries are capable of post translational regulation of NR activity in the shoots. Additionally, supplying NO3- to the cut ends of stems resulted in around a 5-fold increase in NR activity, a 10-fold increase in NR transcript accumulation, and up to a 195-fold increase in transcript accumulation of NITRITE REDUCTASE (NiR1) which codes for the enzyme catalyzing the conversion of NO2- to NH4+. These data indicate that blueberry shoots are capable of assimilating NO3- when it is directly supplied to these tissues. Together, these data suggest that limitations in the uptake and translocation of NO3- to the shoots may limit overall NO3- assimilation capacity in blueberry.

Drought stress reduces stem elongation and cell expansion. Since gibberellins (GAs) play an important role in controlling cell elongation, the objective of this study was to determine if the reduction in growth under drought stress is associated with altered GA metabolism or signaling. We exposed ‘Moneymaker’ tomato (Solanum lycopersicum) to drought stress to observe the effects on growth. Irrigation was automated using a data logger, which maintained volumetric water contents (VWC) of 0.35 and 0.15 m3·m−3 for well-watered and drought-stressed conditions, respectively. To further investigate the effect of GAs on elongation, paclobutrazol (PAC), a GA biosynthesis inhibitor, was applied to reduce endogenous GA production. Drought stress and PAC treatment reduced plant height. Internode length, cell size, and shoot dry weight displayed an interaction between the VWC and PAC treatments. The transcript levels of SlGA20ox1, -2, -3, and -4, SlGA3ox2, and SlGA2ox2, -4, and -5, corresponding to enzymes in GA metabolism, and LeEXP1, and -2, encoding expansin enzymes related to cell wall loosening necessary for cell expansion, were analyzed. Downregulation of transcript accumulation due to drought stress was observed for SlGA20ox4, SlGA2ox5, and LeEXP1, but not for any of the other genes. PAC increased expression of SlGA20ox-3, and SlGA3ox2, potentially through feedback regulation. These findings suggest that drought stress effects on elongation are at least partly mediated by altered GA metabolism.

BackgroundThe major fleshy tissues of the apple fruit are spatially separable into cortex and pith. These tissues display differential growth during development. Key features of such differential growth, and sink metabolic programs supporting it have not been investigated previously. We hypothesized that differential growth between these fruit tissues is supported by differential sink metabolic programs, particularly during early development. Growth, metabolite concentrations, and transcript abundance of metabolism-related genes were measured to determine characteristics of differential growth and their underlying metabolic programs.ResultsThe cortex displayed > 5-fold higher growth than the pith during early fruit development, indicating that differential growth was established during this period. Further, when resource availability was increased through sink-removal, cortex growth was preferentially enhanced. Greatest diversity in metabolic programs between these tissues was evident during early fruit development. Higher cortex growth during early development was facilitated by increased catabolism of imported carbon (C) resources, sorbitol and sucrose, and the nitrogen (N) resource, asparagine. It was also associated with enhanced primary C metabolism, and C storage as malate and quinate. The pith metabolic program during this period involved limited allocation of C and N to growth, but greater allocation to storage, and enhanced sucrose-sucrose cycling.ConclusionsTogether, these data indicate that the fruit cortex tissue displays a resource intensive metabolic program during early fruit development. This provides the C backbones, proteins, energy and osmolytes to support its higher growth.

Sustaining the fourfold increase in blueberry (Vaccinium sp.) production witnessed during the previous two decades requires better understanding of its mineral nutrient physiology. The primary goals of this review are to evaluate our current understanding of the physiology of nitrogen (N) and calcium (Ca) nutrition in blueberry. Nitrogen concentration in blueberry ranges from 0.4% to >2% across organs. Blueberry uses N in various forms (organic and inorganic), but it appears to display preference for ammonium (NH4+) over nitrate (NO3−). The roles of N acquisition, translocation and assimilation in determining N-source preference in blueberry are evaluated. Calcium plays important roles in determining fruit quality owing to its function in maintaining cell wall and membrane integrity. It is unique in its translocation characteristics being transported primarily via the xylem. Fruit [Ca2+] typically declines from around 0.2% during early development to 2+] are discussed. Areas where further research is warranted to improve our understanding of N and Ca physiology in blueberry are identified. Such knowledge is essential for sustainable nutrient management, improving productivity, and enhancing fruit quality in blueberry.

Potassium (K) plays a crucial role in multiple physiological and developmental processes in plants. Its deficiency is a common abiotic stress that inhibits plant growth and reduces crop productivity. A better understanding of the mechanisms involved in plant responses to low K could help to improve the efficiency of K use in plants. However, such responses remain poorly characterized in fruit tree species such as pears (Pyrus sp). We analyzed the physiological and transcriptome responses of a commonly used pear rootstock, Pyrus betulaefolia, to K-deficiency stress (0 mM). Potassium deprivation resulted in apparent changes in root morphology, with short-term low-K stress resulting in rapidly enhanced root growth. Transcriptome analyses indicated that the root transcriptome was coordinately altered within 6 h after K deprivation, a process that continued until 15 d after treatment. Potassium deprivation resulted in the enhanced expression (up to 5-fold) of a putative high-affinity K+ transporter, PbHAK5 (Pbr037826.1), suggesting the up-regulation of mechanisms associated with K+ acquisition. The enhanced root growth in response to K-deficiency stress was associated with a rapid and sustained decrease in the expression of a transcription factor, PbMYB44 (Pbr015309.1), potentially involved in mediating auxin responses, and the increased expression of multiple genes associated with regulating root growth. The concentrations of several phytohormones including indoleacetic acid (IAA), ABA, ETH, gibberellin (GA3), and jasmonic acid (JA) were higher in response to K deprivation. Furthermore, genes coding for enzymes associated with carbon metabolism such as SORBITOL DEHYDROGENASE (SDH) and SUCROSE SYNTHASE (SUS) displayed greatly enhanced expression in the roots under K deprivation, presumably indicating enhanced metabolism to meet the increased energy demands for growth and K+ acquisition. Together, these data suggest that K deprivation in P. betulaefolia results in the rapid re-programming of the transcriptome to enhance root growth and K+ acquisition. These data provide key insights into the molecular basis for understanding low-K-tolerance mechanisms in pears and in other related fruit trees and identifying potential candidates that warrant further analyses.

The apple (Malus × domestica) fruit consists of two main fleshy tissues, cortex and pith, that are of diverse origins and contribute differentially to final fruit size. The contribution of these fruit spatial constituents to variation in fruit morphology has not been determined previously. We developed an image-analysis based method that uses two-dimensional images of the fruit sectioned in the proximo-distal plane to determine fruit, cortex, and pith volumes, and key shape descriptors. Fruit volume determined by image-analysis was compared to that determined using water displacement. These two methods were strongly correlated (R2 = 0.95) and displayed good agreement. The cortex volume constituted majority (> 86 %) of the fruit. Variation in cortex volume explained most of the variation in fruit volume (R2 = 0.99), while that of the pith explained a relatively minor portion (R2 = 0.32). Further, variation in pith volume explained only a minor part of cortex volume variation (R2 = 0.24), suggesting that growth in these tissues is regulated by a combination of shared and differential mechanisms. Proportion of the fruit constituted by cortex displayed limited variability across 28 apple cultivars. However, proportion of cortex was substantially lower, and that of pith higher, in a crabapple (Malus sp.) cultivar, Callaway, suggesting that change in tissue proportions contributes to increase in fruit size from that observed in crabapples. Limited variation in the fruit aspect ratio, a key shape descriptor, was noted in the apple cultivars analyzed. A weak relationship was observed between the fruit aspect ratio and that of the core (R2 = 0.29), indicating that differential mechanisms determine shapes of the fruit and its internal tissues. Together, data from this study suggest that growth of cortex and pith tissues is mediated in part by shared, and substantially by differential mechanisms.

The efficacy of auxinic herbicides, a valuable weed control tool for growers worldwide, has been shown to vary with the time of day in which applications are made. However, little is known about the mechanisms causing this phenomenon. Investigating the differential in planta behavior of these herbicides across different times of application may grant an ability to advise which properties of auxinic herbicides are desirable when applications must be made around the clock. Radiolabeled herbicide experiments demonstrated a likely increase in ATP-binding cassette subfamily B (ABCB)-mediated 2,4-D and dicamba transport in Palmer amaranth (Amaranthus palmeri S. Watson) at simulated dawn compared to mid-day, as dose response models indicated that many orders of magnitude higher concentrations of N-1-naphthylphthalamic acid (NPA) and verapamil, respectively, are required to inhibit translocation by 50% at simulated sunrise compared to mid-day. Gas chromatographic analysis displayed that ethylene evolution in A. palmeri was higher when dicamba was applied during mid-day compared to sunrise. Furthermore, it was found that inhibition of translocation via 2,3,5-triiodobenzoic acid (TIBA) resulted in an increased amount of 2,4-D-induced ethylene evolution at sunrise, and the inhibition of dicamba translocation via NPA reversed the difference in ethylene evolution across time of application. Dawn applications of these herbicides were associated with increased expression of a putative 9-cis-epoxycarotenoid dioxygenase biosynthesis gene NCED1, while there was a notable lack of trends observed across times of day and across herbicides with ACS1, encoding 1-aminocyclopropane-1-carboxylic acid synthase. Overall, this research indicates that translocation is differentially regulated via specific protein-level mechanisms across times of application, and that ethylene release, a chief phytotoxic process involved in the response to auxinic herbicides, is related to translocation. Furthermore, transcriptional regulation of abscisic acid involvement in phytotoxicity and/or translocation are suggested.

Fruit abscission in blueberry (Vaccinium sp.) occurs at the pedicel/peduncle junction (PPJ). Growth regulators such as methyl jasmonate (MeJa) and ethephon accelerate the progression of abscission at this zone. It is not known whether the abscission zone at the PPJ is sufficient to perceive and respond to these growth regulator applications or if the fruit and leaf tissues are required to elicit these responses. Furthermore, the effects of injury to the fruit and leaves on fruit detachment responses have not been previously reported in blueberry. In this study, the requirement of the fruit and leaves to respond to MeJa and ethephon applications was investigated through organ removal treatments in rabbiteye blueberry. Removal of the fruit or the fruit and leaves on the branch followed by MeJa application delayed the progression of abscission at the PPJ suggesting that the fruit tissue is required only to accelerate the progression of fruit detachment in response to MeJa. Interestingly, the extent of fruit/pedicel detachment in response to ethephon applications was higher in the organ removal treatments compared with the control indicating that the PPJ was sufficient to perceive and respond to ethephon and that wounding caused by organ removal synergistically enhanced fruit abscission in response to ethephon. Mechanical wounding of the fruit by removing the distal half of the berry resulted in accelerated fruit detachment at the PPJ. Detachment of non-injured fruit was unaffected by mechanical wounding of adjacent fruit. These data suggest that wounding generates a local signal capable of accelerating fruit abscission at the PPJ. This information may have implications for fruit retention or drop in response to injury to the fruit as caused by herbivore feeding or as a result of insects and pathogens.

Carbohydrate availability is a key factor determining fruit growth in apple (Malus ×domestica) and other fruits. However, the molecular mechanisms regulating fleshy fruit growth in response to changes in carbohydrate availability are not well understood. In this study, carbohydrate availability was manipulated by reducing fruit load, and its effects on apple fruit growth, cell production and expansion, and the expression of genes associated with these processes was investigated. Reduction of fruit load during early fruit development led to a rapid increase in early fruit growth. The increase in fruit growth was associated with a transient increase in sorbitol and fructose concentrations and altered expression of sorbitol dehydrogenase and sucrose synthase genes. Increase in early fruit growth was mediated primarily by an increase in cell production. The aintegumenta gene, MdANT1, an AP2-domain transcription factor associated with the regulation of cell production and fruit growth, displayed an increase in expression by up to 5-fold during early fruit development in response to the reduction in fruit load. Additionally, multiple cell cycle genes positively associated with cell production such as the cyclins, MdCYCA2;1, MdCYCA2;3, MdCYCB1;1, and MdCYCB2;2, and the B-type cyclin-dependent kinases, MdCDKB1;1, MdCDKB1;2, and MdCDKB2;2, displayed higher expression by up to 5-fold under reduced fruit load conditions during early fruit growth. These data indicate that carbohydrate availability affects the expression of key transcription factors and cell proliferation genes, thereby regulating cell production during early fruit growth. Several genes associated with cell expansion such as the expansins, and cobra and cobra-like genes, also displayed altered expression in response to the reduction in fruit load. The expression of three expansin genes was higher under reduced fruit load conditions at maturity, a stage at which a minor increase in cell size was apparent. Together, data from this study indicate that fruit load reduction induces changes in carbohydrate availability and metabolism, which in turn affect cell production-related mechanisms, thereby enhancing early fruit growth.

A better understanding of fruit detachment and the processes mediating it is essential to improve the efficiency of mechanical harvesting in blueberry (Vaccinium sp.). In blueberry, fruit detachment may occur either at the point of attachment of the pedicel to the peduncle [peduncle–pedicel junction (PPJ)] or at the point of attachment of the pedicel to the fruit [fruit–pedicel junction (FPJ)]. The fruit detachment responses of the PPJ and the FPJ to different conditions are not entirely clear. Additionally, whether fruit detachment at these junctions is mediated by the physiological process of abscission or through physical separation of the organ from the parent plant is not well understood. In this study, a series of experiments were performed to determine the abscission zone (AZ) corresponding to the point of mature fruit detachment and to determine whether fruit detachment occurs as a result of abscission or physical separation in rabbiteye blueberry (Vaccinium ashei). Anatomical studies indicated the presence of an AZ at the PPJ. Greater than 92% of the natural detachment of mature fruit occurred at the PPJ. The morphology of the fracture plane at the PPJ in naturally detached fruit was even and uniform, consistent with fruit detachment through abscission at this location. Abscission agents such as methyl jasmonate (20 mm) and ethephon (1000 mg·L−1) enhanced the extent of fruit detachment at the PPJ, further indicating that mature fruit detachment through abscission occurred primarily at this location. Additionally, the fracture plane at the PPJ during fruit detachment in response to abscission agent applications was flattened and even, further supporting the conclusion that fruit detachment at this location occurred through abscission. In contrast, the majority of the fruit detachment in response to mechanical shaking occurred at the FPJ. Analysis of the morphology of the fracture plane at the FPJ during detachment in response to mechanical shaking indicated that fruit detachment at this location was associated with extensive tearing and mechanical disruption of cells, consistent with physical separation. Together, data from this study indicate that mature fruit detachment resulting from abscission occurs primarily at the PPJ, whereas fruit detachment during mechanical shaking occurs primarily at the FPJ as a result of physical breakage at this weak junction.

A portable, handheld, mechanical shaking device was developed and its effects on fruit detachment in rabbiteye bluebbery (Vaccinium ashei Reade) and southern highbush blueberry (hybrids of mostly Vaccinium corymbosum L. and Vaccinium darrowi Camp.) were evaluated. The instrument facilitated effective detachment of fruit within a branch, especially in rabbiteye blueberry (greater than 75%). Approximately 3 to 4 seconds of shaking was sufficient to detach the majority of the fruit. Differences in the extent of fruit detachment were observed across the genotypes, especially among southern highbush blueberry genotypes. The majority of fruit detachment in rabbiteye blueberry and most of the southern highbush blueberry genotypes occurred at the point of attachment of the pedicel to the berry, although a significant portion detached with the stem intact, resulting in stemmy fruit. Although only a small proportion of the detached fruit was immature in the rabbiteye blueberry genotypes, up to 23% of the detached fruit was immature in the southern highbush blueberry genotypes. Application of the abscission agents methyl jasmonate (MeJa; 20 mm) and ethephon (1000 mg·L−1) reduced the time required for fruit detachment on mechanical shaking by up to 5-fold. Together, these data indicate that the mechanical shaking device developed here is an effective tool for studying fruit detachment in blueberry. This instrument has potential applications in blueberry research programs evaluating fruit production. It can be used in breeding programs to aid in the selection of genotypes with fruit detachment characteristics that are potentially better suited for mechanical harvesting, and also in programs involving the screening and evaluation of abscission agents in blueberry.

Two abscission agents, ethephon and methyl jasmonate, were investigated in five studies to determine their potential for increasing fruit detachment during harvest in rabbiteye (Vaccinium ashei Reade) and southern highbush (hybrids based largely on Vaccinium corymbosum L. and Vaccinium darrowi Camp.) blueberry. In the first study with a rabbiteye blueberry genotype, T-451, ethephon applications up to 1000 mg·L−1 did not affect fruit drop but reduced fruit detachment force (FDF) by up to 21%. In the second study with two southern highbush blueberry genotypes, ethephon (up to 1500 mg·L−1) and methyl jasmonate (MeJa; up to 10 mm) applications resulted in significant fruit drop in ‘Star’ but neither of the growth regulators affected the fruit detachment characteristics of ‘Farthing’. In a third study with rabbiteye blueberry genotypes, MeJa applications of 10, 20, and 30 mm displayed an increasing linear trend in fruit drop in ‘Climax’ and linear and quadratic trends in fruit drop in ‘Powderblue’. In a fourth study with ‘Powderblue’, MeJa (20 mm) and ethephon (1000 mg·L−1) applications resulted in rapid and significant fruit drop. The fruit drop induced by MeJa in this study was attenuated by the coapplication of aminoethoxyvinylglycine (AVG), an ethylene biosynthesis inhibitor, suggesting that MeJa induced fruit detachment partly through its effects on ethylene biosynthesis. In another study with the southern highbush blueberry genotype, O’Neal, MeJa applications (20 mm) induced significant fruit drop but ethephon (1000 mg·L−1) applications did not affect fruit detachment. Overall, MeJa applications (20 mm or greater) generally induced rapid and extensive fruit abscission, often within 1 day after treatment. Applications of MeJa resulted in leaf yellowing and necrosis of leaf tips and margins, especially at high rates of application (20 mm or greater). Ethephon applications resulted in the abscission of mature and immature berries. Both ethephon and MeJa applications resulted in the detachment of the pedicel along with the fruit. Together, these data suggest that although ethephon and MeJa have the potential to be used as harvest aids in blueberry, the rates of application require further optimization to minimize potential phytotoxicity. Additionally, effective de-stemming of the berries may be essential if these compounds are to be used as harvest aids.

Plant responses to drought stress vary depending on the severity of stress and the stage of drought progression. To improve the understanding of such responses, the leaf physiology, abscisic acid (ABA) concentration, and expression of genes associated with ABA metabolism and signalling were investigated in Petunia × hybrida. Plants were exposed to different specific substrate water contents (θ = 0.10, 0.20, 0.30, or 0.40 m(3)·m(-3)) to induce varying levels of drought stress. Plant responses were investigated both during the drying period (θ decreased to the θ thresholds) and while those threshold θ were maintained. Stomatal conductance (g(s)) and net photosynthesis (A) decreased with decreasing midday leaf water potential (Ψ(leaf)). Leaf ABA concentration increased with decreasing midday Ψ(leaf) and was negatively correlated with g(s) (r = -0.92). Despite the increase in leaf ABA concentration under drought, no significant effects on the expression of ABA biosynthesis genes were observed. However, the ABA catabolism-related gene CYP707A2 was downregulated, primarily in plants under severe drought (θ = 0.10 m(3)•m(-3)), suggesting a decrease in ABA catabolism under severe drought. Expression of phospholipase Dα (PLDα), involved in regulating stomatal responses to ABA, was enhanced under drought during the drying phase, but there was no relationship between PLDα expression and midday Ψ(leaf) after the θ thresholds had been reached. The results show that drought response of plants depends on the severity of drought stress and the phase of drought progression.

Shading during early fruit development reduces fruit growth and initiates fruit abscission in apple (Malus ×domestica). The mechanisms mediating the decline in fruit growth in response to shading are not well understood. In this study, the effects of shading during early fruit development on cell production and expansion were investigated. Additionally, the effects of shading on the expression of genes associated with carbohydrate metabolism, fruit growth, and cell production and expansion were investigated to develop a better understanding of the molecular mechanisms and to identify genes that mediate the reduction in fruit growth. Shading of isolated branches or entire trees ≈15 to 18 days after full bloom resulted in a sharp decline in fruit growth by 3 days after treatment. Reduction in fruit growth was consistently mediated by a decline in cell production within 3 days after treatment. Reduced fruit growth was also associated with lower cell size by 3 to 7 days after shading in two different years. These data indicate that the reduction in fruit growth as a result of shading is mediated by a reduction in cell production and expansion. The expression of two sorbitol dehydrogenase (SDH) genes, MdSDH1 and MdSDH2, was higher in the shaded fruit by up to 10-fold, suggesting an increase in SDH activity to meet the immediate respiratory demands of the developing fruit. The auxin response factor (ARF), MdARF106, displayed ≈3-fold higher expression in the shaded fruit, suggesting its involvement in regulating mechanisms that mediate the reduction in fruit growth. Two A2-type cyclins, MdCYCA2;2 and MdCYCA2;3, which are positively associated with cell production, displayed lower expression in the shaded fruit by up to 4.6-fold. Conversely, MdKRP4 and MdKRP5, cell cycle genes negatively associated with cell production, displayed 3.9- and 5.3-fold higher expression in the shaded fruit, respectively. Additionally, two genes associated with cell expansion, MdCOB1 (cobra1) and MdEXPA10;1 (expansin), displayed lower expression in the shaded fruit. Together, these data indicate that shading results in coordinated changes in the expression of carbohydrate metabolism-related genes, key transcription factors related to fruit growth, and genes associated with cell production and expansion. These changes may subsequently decrease the progression of the primary processes that mediate fruit growth.

Fruit size is a valuable commercial trait in blueberry. The cellular basis of variation in fruit size among rabbiteye blueberry (Vaccinium ashei) genotypes was investigated. Twenty genotypes, including cultivars and advanced selections from the University of Georgia blueberry breeding program, were analyzed. Among the 20 genotypes, fruit weight and fruit diameter varied by over threefold and 1.6-fold, respectively. Regression analysis indicated a linear relationship between fruit weight and fruit diameter (R2 = 0.97, P < 0.001), suggesting that fruit diameter is a good predictor of fruit weight. Among the 20 genotypes, mesocarp cell number and cell area varied by almost 2.5-fold and 1.5-fold, respectively. Although fruit diameter and cell number were significantly related (R2 = 0.79, P < 0.001), no relationship could be established between fruit diameter and cell area. These data indicate that variation in fruit size among rabbiteye blueberry genotypes is primarily facilitated by variation in cell number. Two small and two large fruit size genotypes were further analyzed. Differences in cell number among some of these genotypes were apparent at bloom suggesting that cell production before bloom is an important mechanism contributing to variation in final cell number. Differences in final cell number among other genotypes were manifested during fruit development, indicating that cell production during fruit development was also instrumental in determining variation in final cell number. This study suggests that fruit size variation in rabbiteye blueberry genotypes is determined by mechanisms that regulate cell production before bloom and during fruit development.

Ripening in blueberry fruit is irregular and occurs over an extended period requiring multiple harvests, thereby increasing the cost of production. Several phytohormones contribute to the regulation of fruit ripening. Certain plant growth regulators (PGRs) can alter the content, perception, or action of these phytohormones, potentially accelerating fruit ripening and concentrating the ripening period. The effects of three such PGRs&mdash, ethephon, abscisic acid, and methyl jasmonate&mdash, on fruit ripening were evaluated in the rabbiteye blueberry (Vaccinium virgatum) cultivars &lsquo, Premier&rsquo, and &lsquo, Powderblue&rsquo, Application of ethephon, an ethylene-releasing PGR, at 250 mg L&minus, 1 when 30&ndash, 40% of fruit on the plant were ripe, accelerated ripening by increasing the proportion of blue (ripe) fruit by 1.5&ndash, 1.8-fold within 4 to 7 days after treatment in both cultivars. Ethephon applications did not generally alter fruit quality characteristics at harvest or during postharvest storage, except for a slight decrease in juice pH at 1 day of postharvest storage and an increase in fruit firmness and titratable acidity after 15 days of postharvest storage in Powderblue. In Premier, ethephon applications decreased the proportion of defective fruit at 29 days of postharvest storage. Abscisic acid (600&ndash, 1000 mg L&minus, 1) and methyl jasmonate (0.5&ndash, 1 mM) applications did not alter the proportion of ripe fruit in either cultivar. These applications also had little effect on fruit quality characteristics at harvest and during postharvest storage. None of the above PGR applications affected the development of naturally occurring postharvest pathogens during storage. Together, data from this study indicated that ethephon has the potential to accelerate ripening in rabbiteye blueberry fruit, allowing for a potential decrease in the number of fruit harvests.

Polyamines (PAs) are ubiquitous, polycationic biogenic amines that are implicated in many biological processes, including plant growth and development, but their precise roles remain to be determined. Most of the previous studies have involved three biogenic amines: putrescine (Put), spermidine (Spd) and spermine (Spm), and their derivatives. We have expressed a yeast spermidine synthase (ySpdSyn) gene under constitutive (CaMV35S) and fruit-ripening specific (E8) promoters in Solanum lycopersicum (tomato), and determined alterations in tomato vegetative and fruit physiology in transformed lines compared with the control. Constitutive expression of ySpdSyn enhanced intracellular levels of Spd in the leaf, and transiently during fruit development, whereas E8-ySpdSyn expression led to Spd accumulation early and transiently during fruit ripening. The ySpdSyn transgenic fruits had a longer shelf life, reduced shriveling and delayed decay symptom development in comparison with the wild-type (WT) fruits. An increase in shelf life of ySpdSyn transgenic fruits was not facilitated by changes in the rate of water loss or ethylene evolution. Additionally, the expression of several cell wall and membrane degradation-related genes in ySpdSyn transgenic fruits was not correlated with an extension of shelf life, indicating that the Spd-mediated increase in fruit shelf life is independent of the above factors. Crop maturity, indicated by the percentage of ripening fruits on the vine, was delayed in a CaMV35S-ySpdSyn genotype, with fruits accumulating higher levels of the antioxidant lycopene. Notably, whole-plant senescence in the transgenic plants was also delayed compared with WT plants. Together, these results provide evidence for a role of PAs, particularly Spd, in increasing fruit shelf life, probably by reducing post-harvest senescence and decay.

Fruit size regulation was studied in the apple cultivar 'Gala' and a large fruit size spontaneous mutant of 'Gala', 'Grand Gala' (GG). GG fruits were 15% larger in diameter and 38% heavier than 'Gala' fruits, largely due to an increase in size of the fruit cortex. The mutation in GG altered growth prior to fruit set and during fruit development. Prior to fruit set, the carpel/floral-tube size was enhanced in GG and was associated with higher cell number, larger cell size, and increased ploidy through endoreduplication, an altered form of the cell cycle normally absent in apple. The data suggest that the mutation in GG promotes either cell production or endoreduplication in the carpel/floral-tube cells depending on their competence for division. Ploidy was not altered in GG leaves. During fruit growth, GG fruit cells exited cell production earlier, and with a DNA content of 4C suggesting G2 arrest. Cell size was higher in GG fruits during exit from cell production and at later stages of fruit growth. Final cell diameter in GG fruit cortex cells was 15% higher than that in 'Gala' indicating that enhanced fruit size in GG was facilitated by increased cell size. The normal progression of cell expansion in cells arrested in G2 may account for the increase in cell size. Quantitative RT-PCR analysis indicated higher MdCDKA1 expression and reduced MdCYCA2 expression during early fruit development in GG fruits. Together, the data indicate an important role for cell expansion in regulating apple fruit size.

Understanding leaf and fruit abscission is essential in order to develop strategies for controlling the process in fruit crops. Mechanisms involved in signalling leaf and fruit abscission upon induction by abscission agents were investigated in Citrus sinensis cv. 'Valencia'. Previous studies have suggested a role for phospholipid signalling; hence, two phospholipase D cDNA sequences, CsPLDalpha1 and CsPLDgamma1, were isolated and their role was examined. CsPLDalpha1 expression was reduced in leaves but unaltered in fruit peel tissue treated with an ethylene-releasing compound (ethephon), or a fruit-specific abscission agent, 5-chloro-3-methyl-4-nitro-1H-pyrazole (CMNP). By contrast, CsPLDgamma1 expression was up-regulated within 6 h (leaves) and 24 h (fruit peel) after treatment with ethephon or CMNP, respectively. CsPLDalpha1 expression was diurnally regulated in leaf blade but not fruit peel. CsPLDgamma1 exhibited strong diurnal oscillation in expression in leaves and fruit peel with peak expression around midday. While diurnal fluctuation in CsPLDalpha1 expression appeared to be light-entrained in leaves, CsPLDgamma1 expression was regulated by light and the circadian clock. The diurnal expression of both genes was modulated by ethylene-signalling. The ethephon-induced leaf abscission and the ethephon- and CMNP-induced decrease in fruit detachment force were enhanced by application during rising diurnal expression of CsPLDgamma1. The results indicate differential regulation of CsPLDalpha1 and CsPLDgamma1 in leaves and fruit, and suggest possible roles for PLD-dependent signalling in regulating abscission responses in citrus.

Additional index words. apical dominance, epinasty, photoperiod, root anaerobiosis, soil moisture status, stomatal conductance, tuberization Abstract. Plant growth regulators (PGRs) play important roles in the way plants grow and develop. Myriad processes important to horticultural crops are regulated by PGRs. Changes in the presence, balance, and distribution of PGRs communicate developmental, stress-related, or environmental cues that alter growth. Short-distance communication involves changes in biosynthesis or metabolic conversion, whereas longer-distance communication may also require export and translocation of PGRs, their precursors or metabolites. Examples are presented that demonstrate PGR communication between roots and shoots in horticultural commodities. For example, increased duration and intensity of flooding stress can result in synthesis of 1-aminocyclopropane-1-carboxylic acid (ACC), precursor of the PGR ethylene, in roots. ACC transported to the shoot through the transpiration stream is converted to ethylene and causes leaf epinasty. Roots sense the onset of water stress and can communicate the need to close leaf stomata by altering abscisic acid (ABA) levels in the shoot. Daylength and temperature regulate synthesis and transport of gibberellins, which promote stem elongation and stolon formationandinhibit tuberization inpotato.Outgrowth ofaxillarybuds followingthedecapitation ofthe apicalmeristem is dependent on synthesis and transport of cytokinin from root to the axillary buds as well as the balance of indole-3-acetic acid (IAA) cytokinin, and additional messengers. Current research in the field of long-distance communication within plants is uncovering novel messengers and altering our view of the central roles for PGRs in such signaling.

Background Fruit growth in apple (Malus × domestica Borkh.) is mediated by cell production and expansion. Genes involved in regulating these processes and thereby fruit growth, are not well characterized. We hypothesized that the apple homolog(s) of AINTEGUMENTA (ANT), an APETALA2–repeat containing transcription factor, regulates cell production during fruit growth in apple. Results Two ANT genes, MdANT1 and MdANT2, were isolated from apple and their expression was studied during multiple stages of fruit development. MdANT1 and MdANT2 expression was high during early fruit growth coincident with the period of cell production, rapidly declined during exit from cell production, and remained low during the rest of fruit development. The effects of increase in carbohydrate availability during fruit growth were characterized. Increase in carbohydrate availability enhanced fruit growth largely through an increase in cell production. Expression of MdANT1 and MdANT2 increased sharply by up to around 5-fold in response to an increase in carbohydrate availability. Expression of the ANT genes was compared across two apple genotypes, ‘Gala’ and ‘Golden Delicious Smoothee’ (GS), which differ in the extent of fruit growth, largely due to differences in cell production. In comparison to ‘Gala’, the larger fruit-size genotype, GS, displayed higher levels and a longer duration of MdANT1 and MdANT2 expression. Expression of the ANTs and cell cycle genes in the fruit core and cortex tissues isolated using laser capture microdissection was studied. During early fruit growth, expression of the MdANTs was higher within the cortex, the tissue that constitutes the majority of the fruit. Additionally, MdANT1 and MdANT2 expression was positively correlated with that of A- and B-type CYCLINS, B-type CYCLIN-DEPENDENT-KINASES (CDKBs) and MdDEL1. Conclusions Multiple lines of evidence from this study suggest that MdANT1 and MdANT2 regulate cell production during fruit growth in apple. ANTs may coordinate the expression of cell proliferation genes and thereby affect the competence of cells for cell production during fruit growth. Together, data from this study implicate MdANT1 and MdANT2 in the regulation of fruit growth in apple.

Application of transcriptomics approaches can greatly enhance our understanding of blueberry physiology. The success of transcriptomics approaches is dependent on the extraction of high-quality RNA which is complicated by the abundance of polyphenolics and polysaccharides in blueberry. Additionally, transcriptomics requires the accurate quantification of transcript abundance. Quantitative real-time polymerase chain reaction (qRT-PCR) is a robust method to determine transcript abundance. Normalization of gene expression using stably expressed reference genes is essential in qRT-PCR. An evaluation of the stability of expression of reference genes has not yet been reported in blueberry. The objectives of this study were to develop an effective procedure for extracting RNA from different organs and to evaluate potential reference genes for qRT-PCR analyses in blueberry. RNA of high quality and yield was extracted from eight and six organs of rabbiteye and southern highbush blueberry, respectively, using a modified cetyltrimethyl ammonium bromide-based method. The expression stability of 12 reference genes was evaluated. UBIQUITIN-CONJUGATING ENZYME (UBC28), RNA HELICASE-LIKE (RH8), CLATHRIN ADAPTER COMPLEXES MEDIUM SUBUNIT FAMILY PROTEIN (CACSa), and POLYUBIQUITIN (UBQ3b) were the most stably expressed genes across multiple organs in both blueberry species. Further, the expression stability of the reference genes in the branch abscission zone following treatment with fruit abscission-inducing compounds was analyzed. CACSa, RH8, and UBC28 were the most stably expressed genes in the abscission zone under abscission-inducing conditions. We suggest a preliminary evaluation of UBC28, CACSa, RH8, and UBQ3b to identify the most suitable reference genes for the experimental conditions under consideration in blueberry.

Unprecedented progress during the last three decades in our understanding of the principles of a living cell, particularly the identification of genes and signaling pathways involved in cell differentiation and organ development, has brought us a broader insight into plant biological processes. Technological advancements are revealing new and fundamental knowledge at the molecular and cellular levels, knowledge that is critical towards achieving the goal of precision-based crop improvement. Modernday genetic engineering has emerged as a promising precision-based technology for boosting up food production in the world and introducing desirable traits such as nutritional enhancement and disease and pest resistance, both important components of agricultural sustainability (Chrispeels et al. 2002; Fatima et al. 2008; Negi and Handa 2008). Achieving results that benefit the world will depend on the success of applying new knowledge to real-world field scenarios. The challenge, therefore, is also to simultaneously obtain knowledge on agroecosystem structure and function to understand how manipulation and control of specific gene expression will translate into directing processes at the ecological scale (Mattoo and Teasdale 2009). Developmental traits are coordinated at various levels in a plant and involve organ-to-organ communications via long-distance signaling processes that integrate transcription, hormonal action and environmental cues. Thus, plant architecture, root–soil–microbe interactions, flowering, fruit (and seed) development, and fruit ripening (and seed germination) are highly regulated genetic programs that are also impacted by processes such as organ abscission, organ senescence (and ripening), and programmed cell death (PCD). We note that belowground processes provide the anchor for a healthy and robust plant (Mattoo and Teasdale 2009) but in this

Plant responses to drought stress vary depending on the severity of stress and the stage of drought progression. To improve the understanding of such responses, the leaf physiology, abscisic acid (ABA) concentration, and expression of genes associated with ABA metabolism and signalling were investigated in Petunia × hybrida. Plants were exposed to different specific substrate water contents (θ = 0.10, 0.20, 0.30, or 0.40 m(3)·m(-3)) to induce varying levels of drought stress. Plant responses were investigated both during the drying period (θ decreased to the θ thresholds) and while those threshold θ were maintained. Stomatal conductance (g(s)) and net photosynthesis (A) decreased with decreasing midday leaf water potential (Ψ(leaf)). Leaf ABA concentration increased with decreasing midday Ψ(leaf) and was negatively correlated with g(s) (r = -0.92). Despite the increase in leaf ABA concentration under drought, no significant effects on the expression of ABA biosynthesis genes were observed. However, the ABA catabolism-related gene CYP707A2 was downregulated, primarily in plants under severe drought (θ = 0.10 m(3)•m(-3)), suggesting a decrease in ABA catabolism under severe drought. Expression of phospholipase Dα (PLDα), involved in regulating stomatal responses to ABA, was enhanced under drought during the drying phase, but there was no relationship between PLDα expression and midday Ψ(leaf) after the θ thresholds had been reached. The results show that drought response of plants depends on the severity of drought stress and the phase of drought progression.

Fruit development in apple cultivars varying in their ultimate fruit size was analyzed using cytology, flow cytometry (FCM), and semi-quantitative RT-PCR. Fruit size variation across cultivars was largely explained by variation in cell number. The cell division phase lasted for less than 30 days in all varieties, less than previously believed. A distinct overlap between the cell division and cell expansion phases was present. Analysis of the relative cell production rate (rCPR) showed a major peak about 10 days after full bloom (DAFB) after which it declined. Comparison of the rCPR across varieties suggested distinct patterns of cell production with `Gala' having a low but sustained rCPR, `Pixy Crunch' a short but high rCPR, and `Golden Delicious' having a high and sustained rCPR. FCM analysis also showed similar patterns with a peak in the proportion of dividing cells about 10 DAFB followed by a decline. To further understand regulation of cell number, four cell cycle related genes were cloned from `Gala'. Cyclin Dependent Kinase B (CDK B) and Cyclin B were found to be highly cell division phase specific in their expression. Analysis of gene expression by semi-quantitative RT-PCR indicated peak expression of these two genes at 5-10 DAFB, consistent with the peaks in rCPR and proportion of dividing cells. Comparison of gene expression across the varieties showed higher peak expression of the above genes in the larger-fruited `Golden Delicious' than in the smaller-fruited `Gala.' This study provides novel insight into the regulation of fruit development in apple and also suggests a role for the cell cycle genes in fruit size regulation.

Fruit size is a commercially valuable trait. Although several factors are known to affect fruit size in apple, insights into the molecular aspects of its regulation are lacking. Our research aims to understand fruit size regulation using a combination of approaches. Analysis of a large fruited mutant of `Gala', `Grand Gala' (GG), showed that it was 40% heavier than `Gala' at harvest. Increase in size of GG fruit was caused by an increase in the cell size apparent at full bloom. Flow cytometry revealed the presence of multiple levels of ploidy (up to 16C) in GG during early fruit development. Increase in ploidy of GG is hypothesized to be due to endoreduplication, a process normally absent in apple. Endoreduplication is a modification of the cell cycle where DNA replication is not followed by cell division, resulting in increased DNA content accompanied by increased cell size. To understand if the cell cycle is altered in GG, four key cell cycle regulators, MdCDKA1, MdCDKB1, MdCYCB2 and MdCYCD3 have been partially cloned from apple using RT-PCR and RACE. As cell number at the end of the cell division phase is correlated with fruit size at harvest, expression analysis of these genes can provide valuable insights into their role in the regulation of cell number and fruit size. Analysis of cell cycle gene expression in GG may provide key insights into the altered molecular regulation that leads to endoreduplication in the mutant. Parallel approaches being employed to study whether environmental and cultural factors regulate fruit size through an influence on the cell cycle will also be discussed.

Peach production is significantly reduced and severely limited by frost injury in regions frequently exposed to late spring freeze conditions. Peach flower buds become increasingly susceptible to low-temperature damage from the period of completion of rest through fruit set. Delaying dehardening and/or flower bud development is an effective way to avoid frost damage. Bio-regulator applications, affecting dormancy or bud development, can delay flowering and dehardening of the buds and can help in avoiding spring freeze injury. Spring applications of AVG and dormant oils on 8-year-old `Redhaven' peach trees were evaluated. AVG applications effectively delayed bloom by 2 to 5 days. The most effective treatment was two applications of 2000 ppm AVG, which delayed bloom by almost 5 days. Repeat applications of AVG were more effective than the single dosage treatments. The 1000 ppm, repeat application delayed bloom by 4 days. A single application of 5000 ppm AVG resulted in severe phytotoxicity. The wetting agent levels were also varied and AVG applications were most effective in combination with 0.2% `Sylgard'. AVG, apparently, delayed bloom by delaying bud development following the completion of rest. The dormant oil sprays were ineffective in achieving bloom delay. The specific leaf weight characteristics of the treated trees were not affected except for the 5000-ppm AVG application, which reduced SLW. Fruit characteristics such as maturity, weight, and soluble sugar concentration were not affected by any of the spring applications (except for the 5000-ppm AVG application, which was phytotoxic). Our studies indicate that AVG is effective in delaying bloom in peaches by up to 5 days. This has the potential to substantially increase peach yields in years with a late spring freeze.