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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.

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.

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.

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 formation and inhibit tuberization in potato. Outgrowth of axillary buds following the decapitation of the apical meristem 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.

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.

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.

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.

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.

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.

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.