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The effects of TDZ ( N -phenyl- N ′-1,2,3-thidiazol-5-ylurea) and CPPU ( N -(2-chloro-4-pyridil)- N ′-phenylurea) on bud initiation of Amaranthus edulis thin cell layers (tTCLs) explants were investigated. tTCLs (0.2–0.4 mm thick) excised from 1 week old seedlings were cultured on Murashige and Skoog (MS) basal salts medium supplemented with varying concentrations of TDZ or CPPU ranging from 1 to 20 μ M and 1 week after culture, buds were initiated. The optimal concentrations were 2 μ M for TDZ and 10 μ M for CPPU. Buds developed into normal plants after transfer onto a MS basal salts medium containing BA to obtain normal internode elongation and onto a MS basal salts medium containing IBA or TDZ for rooting.

Expansion in apple fruit (Malus domestica Borkh. cv. Braeburn) during early development was induced by injecting 2,4-dichlorophenoxyacetic acid (2,4-D) through the calyx of the fruit and by dipping the apples in N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU). Cell wall composition was analysed, focusing on the hemicellulose fraction containing xyloglucan, a polysaccharide believed to play an important role in cell expansion. Changes were observed in the yields of the cell wall fractions of the fruit treated with either 2,4-D or CPPU, although the monosaccharide composition of the fractions exhibited few differences. There was no decrease in the molecular weight of the xyloglucan from treated fruit. These results are discussed in terms of current cell wall expansion mechanisms.

The paper presents a novel approach for the determination of three cytokinin compounds, thidiazuron (TDZ), 1,3-diphenylurea (1,3-DPU) and forchlorfenuron (CPPU), in fruit and vegetables samples using liquid chromatography with electrospray ionization and time-of-flight mass spectrometry (LC—ESI—TOFMS). Analytes were extracted from the sample matrix with ethanol, and the extract, after dilution with water, was submitted to dispersive liquid—liquid microextraction (DLLME). Once acetonitrile and 1,2-dichloroethane had been selected as extraction and disperser solvents, respectively, the influence of the following experimental parameters was studied using a Plackett—Burman design: volume of extraction and disperser solvents, sample mass and time and speed of centrifugation. The best analytical conditions were 250 µL 1,2-dichloroethane, 1.5 mL acetonitrile, 5 g sample mass, and centrifugation at 3000 rpm for 3 min. The optimized method provided DLs in the range 0.02—0.05 ng g−1, depending on the compound. Satisfactory recovery values between 89 and 106% were obtained for spiked samples (kiwifruit, watermelon, grape and tomato) in the 0.2—1.0 ng g−1 concentration range, depending on the compound. None of the target analytes was detected in any of the samples analyzed.

Rapid analytical methods enabling the determination of diverse targets are essential in a number of research areas, from clinical diagnostics to feed and food quality and safety. Herein, the development of a quantitative immunochromatographic assay for the detection of the synthetic phytoregulator forchlorfenuron (CPPU) is described. The competitive lateral flow immunoassay (LFIA) was based on the immobilization onto a nitrocellulose membrane of an ovalbumin-CPPU conjugate (test line) and on the use of an immunodetection ligand consisting of carbon nanoparticles labeled with an anti-CPPU monoclonal antibody through interaction with a secondary antibody. The presence of CPPU in horticultural samples was visually interpreted by the decrease in the black signal intensity of the test line, according to the competitive character of the format. The quantitative determination of the analyte was easily performed by a two-step procedure consisting of flatbed scanning of the strips followed by computer-based image analysis of the pixel gray volumes of the test lines. Under optimized conditions, the immunochromatographic test afforded a limit of quantification in buffer of 89. ng/L. The accuracy of the strip test was assessed by the analysis of fruit samples with incurred residues, and the obtained results were compared with those derived from two reference methods, ELISA and HPLC. The LOQ of the CPPU-specific LFIA in kiwifruits and grapes was established at 33.4 οg/kg. The excellent analytical performance of the developed strip test demonstrates the potential of immunochromatographic assays for the quantitative monitoring of small organic molecules in complex matrices. © 2012 Elsevier B.V., This work was supported by the Ministerio de Educación y Ciencia (AGL2009-12940-C02/01/02/ALI), and cofinanced by FEDER Funds. Celia Suárez Pantaleón was hired by the CSIC under a predoctoral I3P contract.

The significance of changes in cytokinin content during early fruit growth was examined in the kiwifruit (Actinidia deliciosa var. deliciosa cv. Hayward). Fruit growth was modified by the reduction of seed number or by the application of the synthetic phenylurea cytokinin N-(2-chloro-4-pyridyl)-N'-phenylurea (CPPU). The influence of these treatments on cell division was monitored by flow cytometry and changes in the endogenous cytokinins were measured at days 10 and 20 after anthesis, using high-performance liquid chromatography and radioimmunoassay. Total cytokinin levels appeared not to be limiting growth since the highest total cytokinin concentration was detected in unpollinated fruit, which abscised by day 25 after anthesis. However, compared with control fruit which had the highest concentration of zeatin (Z) 10 days post anthesis, Z levels were low in unpollinated fruit. It is hypothesised that an increase in Z is the critical change in cytokinin metabolism required for the initiation of cell division and fruit growth. The synthetic cytokinin CPPU promoted fruit development, but there was a decrease in the endogenous cytokinin concentration. Zeatin was not detected in CPPU-treated fruit. Cell division was reduced in unpollinated fruitlets but there was no significant difference (P > 0.05) between the other treatments. Differences in final fruit size appeared to be due to cell expansion.

Field trials were carried out in three provinces of China to study the dissipation and residue of forchlorfenuron in citrus fruits. The results had shown that the degradation rate of forchlorfenuron in citrus fruits followed the first-order kinetics equation C = A∙eBt. The half-lives of forchlorfenuron were 15.8-23.0 days, the final residues of forchlorfenuron in pulp were all ≤0.002 mg/kg, and most of the residues were concentrated in the peel. The risk assessment revealed that no significant potential health risk would be induced by forchlorfenuron in citrus fruits. Therefore, it could be safe to apply forchlorfenuron in citrus fruits, and the results of this study could also be regarded as a reference to the setting of maximum residue limit for forchlorfenuron in citrus fruits in China.

A selective and sensitive LC-MS/MS method is presented for simultaneous determination of 12 plant growth regulators, viz., indol-3-acetic acid, indol-3-butyric acid, kinetin, zeatin, 6-benzyl aminopurine, gibberellic acid, abscisic acid, chlormequat chloride, forchlorfenuron, paclobutrazole, daminozide, and 2,4-dichlorophenoxy acetic acid, in bud sprouts and grape berries. The sample preparation method involved extraction of homogenized sample (5 g) with 40 mL methanol (80%), and final determination was by LC-MS/MS in the multiple reaction monitoring (MRM) mode with time segmentation for quantification supported by complementary analysis by quadrupole-time of flight (Q-TOF) MS with targeted high-resolution MS/MS scanning for confirmatory identification based on accurate mass measurements. The recovery of the test compounds ranged within 90–107% with precision RSD less than 5% (n = 6). The method could be successfully applied in analyzing incurred residue samples, and the strength of accurate mass analysis could be utilized in identifying the compounds in cases where the qualifier MRM ions were absent or at an S/N less than 3:1 due to low concentrations.

Forchlorfenuron is a synthetic phytohormone with cytokinin-like activity used worldwide as a plant growth regulator to increase fruit size in a number of crops, mostly in kiwifruit and grape vines. A monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) for the determination of forchlorfenuron has been characterized and optimized. The selected immunoreagents afforded a highly selective assay with a limit of detection of 10 ng L−1 in buffer. This direct competitive ELISA was validated in terms of trueness, precision, and robustness using both commercial juice and whole fruit samples. Recoveries from fortified kiwifruit juices and white and red musts were between 97 % and 131 %, with relative standard deviations below 16 %. When homogenized whole fruits were analysed after acetonitrile extraction, recoveries between 96 % and 113 % were found, with a limit of quantification of 5 μg kg−1. The proposed immunoassay was validated by comparison with a reference chromatographic method using fruits from in-field treated grape and kiwifruit vines. Linear regression analysis of ELISA and HPLC–UV determinations showed an excellent correlation (r 2 = 0.998), whereas analysis of the slope (0.99 ± 0.01) and of the intercept (−1 ± 3) clearly proved that the developed competitive immunoassay provided results that were statistically comparable to those obtained by the instrumental method for the analysis of forchlorfenuron in fruits at trace levels., This work was supported by Ministerio de Educación y Ciencia (PET2006_0423_00 and AGL2009-12940-C02/01/02/ALI), Ecología y Protección Agricola S.L., and FEDER Funds. C.S.-P., F.A.E.-T., and J.V.M. were hired by the CSIC under a predoctoral I3P contract, a postdoctoral JAE contract, and a Ramón y Cajal contract, respectively. All contracts were cofinanced by Ministerio de Ciencia e Innovación and the European Social Fund.

The induction of parthenocarpic fruit set was investigated using the apple cvs. Golden Delicious and Jonagold. The gibberellins GA3, GA4, GA5 and GA7 and the synthetic phenylurea-type cytokinin CPPU (N-(2-chloro-4-pyridyl)-N-phenylurea), were applied alone and in combination to unpollinated flowers at the end of petal fall. Gibberellins induced only a marginal final set of parthenocarpic fruits. CPPU sprays were more effective, particularly in the first year. When applied in combination, CPPU and gibberellins had a positive synergistic effect on parthenocarpic fruit set and fruit size, but a negative effect on flower induction the next year. After CPPU + GA sprays, percent fruit set was similar, or greater, compared to natural pollinated trees. The parthenocarpic fruits induced by CPPU + GA had an increased length to diameter ratio. CPPU stimulated, and GA4 and GA7 reduced, the russeting of the parthenocarpic fruits. The internal quality of the fruits was hardly affected, but Ca-deficiency symptoms occurred more frequently in parthenocarpic fruits.

Synthetic auxin treatment for promoting the fruit growth of tomato without pollination sometimes causes puffy fruits. The newly developed cytokinin-like substance, forchlorfenuron (1- (2-chloro-4-pyridyl) -3-phenylurea, CPPU). when applied at anthesis, reduces tomato puffiness of auxin-induced parthenocarpic fruits. Addition of 50300 mg•liter-1 to an auxin solution did not decrease fruit weight but thickened the peduncle, enlarged columella, and elongated the calyxes beyond those of the control fruits.

An effective method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed and optimized to obtain a complete separation of five representative plant growth regulators (PGRs) [gibberellic acid, 2,4-dichlorophenoxyacetic acid (2,4-D), thidiazuron, forchlorfenuron, and paclobutrazol] in fruits. Extraction was performed with acetonitrile containing 0.1% (v/v) acetic acid, applying modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) methodology. LC-MS/MS conditions including composition of mobile phases and mass spectrometry (MS) conditions were evaluated to achieve the highest sensitivity in MS detection. All of the data acquisition was employed in the segmented multiple-reaction monitoring mode for the selected negative and positive transition ions. The octadecylsilyl (C18) dispersive solid-phase extraction (SPE) sorbent was found to provide the more satisfied recoveries than primary secondary amine (PSA) and graphitized carbon black (GCB) for five target PGRs. The optimized method allowed for recoveries of 76-112% for the five PGRs from fruit samples with relative standard deviation (RSD) values less than 10%. Limits of quantification (0.5-16.5 μg/kg) were lower than the maximum limit of residues established for PGRs. The results demonstrated that the developed LC-MS/MS and QuEChERS extraction method is highly effective for analyzing trace amounts of target PGRs in fruit samples. Finally, the method was successfully used to detect residual PGRs in Beijing, China, in 2010. The concentrations of 2,4-D (5.1-1503 μg/kg) and paclobutrazol (1-1381 μg/kg) found in orange and peach, respectively, suggesting that the use of these PGRs in these fruits should be regulated in China in the future.

A videira 'Itália' (Vitis vinifera L.) é a cultivar de uva fina para mesa mais consumida no Brasil. A qualidade dos cachos é uma característica fundamental, sendo o tamanho das bagas o componente mais valorizado pelos consumidores. Uma das alternativas para incrementar a qualidade das bagas é o uso de biorreguladores. Avaliaram-se, em três ciclos de produção, os efeitos de doses de ácido giberélico (AG3) isolado e associado com forchlorfenuron (CPPU), na qualidade dos cachos de uva Itália produzida em Porto Feliz-SP. A aplicação dos biorreguladores foi realizada aos 25 dias após o florescimento, em delineamento experimental inteiramente casualizado, em fatorial 4X4 (zero, 10; 20 e 30 mg L-1 AG3 X zero, 5; 10 e 15 mg L-1 CPPU), com oito repetições para o primeiro ciclo, e fatorial 3X3 (zero, 20 e 30 mg L-1 AG3 X zero, 10 e 20 mg L-1 CPPU) com dez repetições para o segundo e terceiro ciclos. A mistura de 20 mg L-1 de AG3 com 10 mg L-1 de CPPU promoveu o incremento do comprimento e do diâmetro das bagas sem prejuízo da massa dos cachos, melhorando sua qualidade, em Porto Feliz-SP. O uso de CPPU isoladamente acarretou em redução do teor de sólidos solúveis das bagas. 'Itália' grape (Vitis vinifera L.) is one of the main varieties for fresh market in Brazil. Cluster quality is determinant for commercialization and the berry size is highly valued by consumers. An alternative for improving berry size is the use of growth regulators. The effect of increasing concentrations of gibberellic acid (GA3) and forchlorfenuron (CPPU) applied alone or in mixtures was evaluated during three production cycles. Growth regulators were applied 25 days after flowering in the "pea berry size" phenologic stage. Experimental design was completely randomized 4x4 factorial (zero, 10, 20 and 30 mg L-1 GA3 X zero, 5, 10 and 15 mg L-1 CPPU) with eight replications for the first cycle, and 3X3 factorial (zero, 20 and 30 mg L-1 GA3 X zero, 10 and 20 mg L-1 CPPU) with ten replications for the others cycles. The application of GA3 at 20 mg L-1 associated with CPPU at 10 mg L-1 improved berry size without affecting cluster mass, leading to higher grape quality in Porto Feliz, SP. CPPU single application reduced total soluble solids content of the berries.

A videira 'Itália' (Vitis vinifera L.) é a cultivar de uva fina para mesa mais consumida no Brasil. A qualidade dos cachos é uma característica fundamental, sendo o tamanho das bagas o componente mais valorizado pelos consumidores. Uma das alternativas para incrementar a qualidade das bagas é o uso de biorreguladores. Avaliaram-se, em três ciclos de produção, os efeitos de doses de ácido giberélico (AG3) isolado e associado com forchlorfenuron (CPPU), na qualidade dos cachos de uva Itália produzida em Porto Feliz-SP. A aplicação dos biorreguladores foi realizada aos 25 dias após o florescimento, em delineamento experimental inteiramente casualizado, em fatorial 4X4 (zero, 10; 20 e 30 mg L-1 AG3 X zero, 5; 10 e 15 mg L-1 CPPU), com oito repetições para o primeiro ciclo, e fatorial 3X3 (zero, 20 e 30 mg L-1 AG3 X zero, 10 e 20 mg L-1 CPPU) com dez repetições para o segundo e terceiro ciclos. A mistura de 20 mg L-1 de AG3 com 10 mg L-1 de CPPU promoveu o incremento do comprimento e do diâmetro das bagas sem prejuízo da massa dos cachos, melhorando sua qualidade, em Porto Feliz-SP. O uso de CPPU isoladamente acarretou em redução do teor de sólidos solúveis das bagas. Também disponível em: .

To obtain highly-specific and selective forchlorfenuron binders, a collection of functionalized derivatives with different spacer arm locations and lengths was prepared. By immunization with target-mimicking haptens, a large battery of monoclonal and polyclonal antibodies against this synthetic cell regulator was produced and exhaustively characterized in two immunoassay formats using homologous and heterologous conjugates. Antibodies with IC50 values lower than 0.3 nM were successfully raised from the prepared immunogens, thus evidencing the efficacy of the explored strategies. In order to identify significant epitopes in the antibody-antigen interaction, a series of new chemical forchlorfenuron analogues, with slight modifications at both rings of the target molecule, were synthesized and evaluated in competitive assays. As a novel approach in hapten recognition studies, data processing was performed by computational classification methods based on hierarchical clustering. This strategy was shown to be highly valuable for a straightforward profiling of antibodies according to analogue recognition patterns. A relationship could be established between the antigen binding properties of antibodies and the structure of the immunogen. Whereas antibodies with equivalent affinities had been obtained from all of the derivatives, their specificity was found to be largely influenced by the differential exposition of the molecule to the immune system., This work was supported by Ministerio de Educación y Ciencia (AGL2009-12940-C02/01/02/ALI), and cofinanced by FEDER Funds. C.S.P. and J.V.M. were hired by the CSIC, the former under a predoctoral I3P contract and the latter under a Ramón y Cajal postdoctoral contract.

Effects of the synthetic cytokinin CPPU (N‐(2‐chloro‐4‐pyridyl)‐N'‐phenylurea; common name forchlorfenuron) on kiwifruit (Actinidia deliciosa) cv. Hayward fruit growth, maturity, and storage quality were examined. CPPU was applied to fruitlets 21 days after flowering, either as a 5 mg/ litre dip or as a 5 mg/litre spray. Dipping increased mean fresh weight of fruit at harvest by 43% and spraying by 33%. Most of the increase in fruit size was because of an increase in the volume of “small” isodiametric parenchyma cells in the outer pericarp. The volume of “large” ovoid parenchyma cells in the outer pericarp was not affected by CPPU treatment. CPPU application increased the rate of accumulation of soluble solids in fruit prior to harvest but did not increase the proportion of total dry matter. CPPU also caused increased growth of the outer pericarp at the stylar end of the fruit, resulting in small protuberances. Morphological attributes of fruit including the ratio of length to diameter, and flatn...

CPPU, a cytokinin-like substance derived from phenylurea, was applied to shoots, fruits, and to shoots + fruits of mature trees of apple cultivars ‘Golden Delicious’ clone B and ‘Red Delicious’ clone Clear Red 25 days after full bloom. While no influence on fruit morphogenesis was detected for the shoot only treatment, the fruit only and shoot + fruit applications enhanced fruit size and weight, though often inducing irregular elongation, a slight delay in colouring and a lower sugar content. No differences were recorded in the contents and degradation of pectic substances.

A method using solid-phase extraction for cleanup, followed by high-performance liquid chromatography with ultraviolet detection (HPLC/UV), was developed for the determination of forchlorfenuron residues in Fruit and vegetable. The forchlorfenuron is extracted from the sample (15 g) with ethyl acetate (30 mL), and the extract is loaded onto an ENVI-18 column. The forchlorfenuron is eluted with methanol-water as the mobile phase (65:35) and determined by HPLC/UV (260 nm). The ENVI-18 column was found to provide effective cleanup, removing the greatest number of sample matrix interferences. In this method, the correlation coefficient (R2) was 1.0000 with the linear range of forchlorfenuron from 0.1 µg mL-1 to 10.0 µg mL-1. At the low, medium and high three fortification levels of 0.03-5.33 mg kg−1, forchlorfenuron average recoveries range from 84.7 % to 103.0 % and relative standard deviations (RSD) were in the range of 0.6-4.9 % for all analytes. The limits of detection (LOD) of forchlorfenuron (minimum amount injected that resulted in a peak about three times the sample baseline) was 0.02 mg kg-1. This method was practicable for routine residue analysis of forchlorfenuron in kiwi, grape, tomato and cucumber matrices.

This paper describes the application of liquid chromatography time-of-flight mass spectrometry (LC/TOF-MS), with electrospray ionization, for residue analysis of forchlorfenuron in tomato, zucchini and watermelon. The assessed method includes a sample preparation step based on the buffered QuEChERS approach. The TOF-MS fragmentation pattern of forchlorfenuron was studied at different fragmention voltages in the range of 120-270 V. Analyses were carried out under full-scan conditions by using the extracted ion chromatogram (XIC) of the m/z 248 ion with a 0.2 Da window. The linearity of the analytical response across the studied range of concentrations (10-500 microg/kg) was excellent, obtaining correlation coefficients higher than 0.999, and relative standard deviations of the response factors lower than 14%, for the 15 linear calibration curves of forchlorfenuron evaluated along the complete validation study. No significant matrix effects were observed. The signal-to-noise ratios obtained for the 10 microg/kg forchlorfenuron in matrix matched standards were70 for all three matrices. Recovery studies were carried out on spiked tomato, zucchini and watermelon blank samples, at three concentration levels (10, 50, and 200 microg/kg) performing five replicates at each level. Forchlorfenuron mean recoveries ranged between 80% and 87% in watermelon and zucchini, and between 65% and 71% in tomato, obtaining in all cases relative standard deviation values lower than 10%. The method readily achieved a lowest validated level of 10 microg/kg, which was fit-for-purpose in residue monitoring applications.

Dry seeds from two lines of common bean (Phaseolus vulgaris L.) and one cultivar of faba bean (Vicia faba L.) were germinated on Murashige and Skoog (MS) medium containing B5 vitamins, 30 g sucrose/liter, and either 2.5, 5.0, or 7.5 µM benzyladenine (BA). Axenic seed cultures were grown at 22 to 24C in darkness and under continuous light from cool-white fluorescent tubes (40 µmol·m -2 ·s -1 ). Explant tissues were prepared from cotyledonary nodes (CN) and primary nodes (PN) of 14-day-old seedlings. Explants were cultured on corresponding seedling growth medium and maintained under continuous cool-white light (40 µmol·m -2 ·s -1 ). The percentages of CN and PN (in one line of common bean) explants that regenerated shoots and the number of shoots per explant (in all germplasm) were highest when nodal tissues were prepared from seedlings germinated in darkness. These responses were optimal on medium containing 5 µM BA during seedling growth and subsequent culture of explants. The number of shoots per explant was two to five times higher on explants cultured on medium with 0.25 to 1.0 µM forchlorfenuron (CPPU) or thidiazuron (TDZ) than on medium with 5 µM BA. Higher (2.5 and 5 µM) CPPU and TDZ concentrations inhibited shoot elongation and stimulated callus production. Histological analyses indicated that adventitious meristems formed 6 to 8 days after explant culture. Progenies from regenerated plants appeared similar to plants raised from the original seed stocks. Chemical names used: N- (phenylmethyl) -1 H- purin-6-amine (benzyladenine, BA); N- (2-chloro-4-pyridyl)- N'- phenylurea (forchlorfenuron, CPPU); N- phenyl -N' -1,2,3-thiadiazol-5-ylurea (thidiazuron, TDZ). Tissue culture of grain legumes using cotyledonary node ex- plants from in vitro-grown seedlings has been documented for organogenic competence (Jackson and Hobbs, 1990; McClean and Grafton, 19889; Wright et al., 1986). BA supplements (5 to 10 µM) in the medium during germination and subsequent explant culture were one of the most critical factors for organ- ogenic differentiation and shoot multiplication in this regener- ation system. N- (2-furanylmethyl)-1 H- purin-6-amine (kinetin) was ineffective (McClean and Grafton, 1989) for inducing mul- tiple-shoot organogenesis in common bean or in pea (Pisum sativum L.) (Jackson and Hobbs, 1990), and neither (E)-2-methyl- 4-(1H-purin-6-ylamino)-2-buten-1-ol (zeatin) nor N- (3-methyl- 2-butenyl)-1 H- purin-6-amine (2iP) could be substituted for BA in soybean (Glycine max (L.) Merr.) shoot regeneration studies

Solutions of CPPU and BA were applied to ovaries of melon (Cucumis melo) flowers with or without pollination, and the effects on fruit set, growth, and sugar content were investigated. Treatment with CPPU increased fruit set in both seeded and seedless melons. Even at low concentrations, CPPU had a strong effect on fruit set in the seeded melons. In seedless melons, CPPU induced 100% parthenocarpic fruit set when applied with 10 mg·L–1; lower concentrations were much less effective. Treatment with BA increased fruit set in seeded melons, but was not particularly effective in the absence of pollination. During the first 10 days after anthesis, CPPU promoted fruit growth, but between 8 and 13 days after anthesis, the growth rate was lower than in the controls. Treatment with CPPU had little effect upon soluble solids (SS) levels in seeded fruit. SS content was significantly lower in seedless than in seeded fruit; this difference was larger in the placenta than in the mesocarp. Sucrose levels of both seeded and seedless fruits were consistently higher than glucose and fructose levels. High concentrations of CPPU reduced sucrose levels in the placenta of seedless fruit. These results indicate that seeds play an important role in sugar accumulation and melon fruit growth during later stages of development. Chemical names used: [1-(2-chloro-4-pyridyl)-3-phenylurea] (CPPU); 6-benzylaminopurine (BA).

Unfertilized ovules were excised 19-21 days before anthesis and cultured in a liquid medium to induce embryogenic callus from 'Kyoho' grape (Vitis×labruscana). The culture medium consisted of half strength MS supplemented with 1.0μM 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 0.2-5.0μM N-(1, 2, 3-thiadiazol-5-yl)-N'-phenylurea (TDZ, thidiazuron), or 1.0μM 2, 4-D and 0.2μM N-(2-chloro-4-pyridyl)-N'-phenylurea (CPPU). After 3-4 months of culture, the embryogenic calli produced from the ovules were successfully subcultured to maintain a high embryogenic potential for over one year. When these adventitious embryos were transferred to a modified 1/2MS hormone-free medium, plantlets were regenerated 1-2.5 months later. This is the first embryogenic callus induction and plant regeneration from 'Kyoho' grape.

The development of immunoassays for the detection of the plant growth regulator forchlorfenuron (CPPU) is described. To achieve that purpose, a set of CPPU derivatives has been obtained by the previous synthesis of the adequate p-aminophenyl alkanoic acid. Protein conjugates of these compounds have been used as immunogens to produce rabbit polyclonal antibodies and a collection of mouse monoclonal antibodies. Additionally, a battery of structural analogues of the target analyte has been synthesized and used for the characterization of antibody binding. This strategy has demonstrated that most antibodies followed Landsteiner's principle, although some monoclonal antibodies showing important deviations from this behavior have also been found. Finally, different assay formats have been developed with a variety of antibodies and conjugates, and a rapid procedure has been optimized for the indirect ELISA format using monoclonal and polyclonal antibodies. In the indirect competitive ELISA, assay IC50 values for CPPU below 0.5 nM were found with LODs as low as 0.013 nM.

The rate of degradation of forchlorfenuron, a cytokinin-based plant growth regulator (PGR) was explored in typical grapevine soils of India with simultaneous evaluation of its effect on biochemical attributes of the test soils in terms of the activities of specific soil microbial enzymes. In all the test soils, namely clay, sandy-loam and silty-clay, the dissipation rate was faster at the beginning, which slowed down with time, indicating a non-linear pattern of degradation. Degradation in soils could best be explained by two-compartment 1st+1st order kinetics with half-life ranging between 4-10 days. The results suggest that organic matter might be playing a major role in influencing the rate of degradation of forchlorfenuron in soil. The rate of degradation in sandy-loam soil was fastest followed by clay and silty-clay soils, respectively. Comparison of the rate of degradation in natural against sterilized soils suggests that microbial degradation might be the major pathway of residue dissipation. Changes in soil enzyme activities as a consequence of forchlorfenuron treatment were studied for extra-cellular enzymes namely acid phosphatase, alkaline phosphatase and beta -glucosidase and intracellular enzyme-dehydrogenase. Although small changes in enzyme activities were observed, forchlorfenuron did not have any significant deleterious effect on the enzymatic activity of the test soils. Simple correlation studies between degradation percentage and individual enzyme activities did not establish any significant relationships. The pattern and change of enzyme activity was primarily the effect of the incubation period rather than the effect of forchlorfenuron itself.

The kiwi, as a dioecious plant, produces male and female flowers in different individuals which makes the pollination difficult - a necessary condition for good yields. A trial was carried out in an orchard in the region of Guarapuava, state of Paraná, Brazil, aiming at studying the effect of CPPU on the fruit development of kiwi cv. Bruno. The treatment consisted of spraying the flowers when 50% of them were opened with an aqueous solution of CPPU at 0, 2.5, 5.0, 7.5, 10.0 and 12.5 mg L–1, and also a treatment with hand-pollination was done. The treatment with CPPU and hand-pollination increased the fruit set, without changing the size and the weight of the fruits. The treatment with CPPU at 5 mg L–1 increased the percentage of fruit set from 54.6% on the control to 87.5%. Nevertheless, the treatment with CPPU at 12.5 mg L–1 decreased the total soluble solids content of the mosto. O kiwi, como planta dióica, produz flores masculinas e femininas em indivíduos diferentes, o que dificulta a sua polinização, condição necessária para bons rendimentos de produção. Nesse contexto, um experimento foi conduzido em pomar na região de Guarapuava, Estado do Paraná, com o objetivo de estudar o efeito do CPPU na frutificação do kiwi cv. Bruno. Os tratamentos consistiram de pulverizações dirigidas às flores, quando 50% destas se encontravam abertas, de soluções aquosas de CPPU a 0; 2,5; 5,0; 7,5; 10,0 e 12,5 mg L–1 , além de um tratamento em que se realizou polinização manual. Os tratamentos com CPPU e com polinização manual aumentaram significativamente a porcentagem de pegamento dos frutos sem interferir em seu tamanho e peso. O tratamento com CPPU a 5,0 mg L–1 aumentou a porcentagem de pegamento dos frutos de 54,6% na testemunha para 87,5%. No entanto, o tratamento com CPPU a 12,5 mg L–1 reduziu o teor de sólidos solúveis totais do mosto.

In order to study the effect of KT-30 (N-(2-chloro-4-pyridyl)-N''-phenylurea) treatment on the fruit growth of ‘Monty’ kiwifruit, the changes in some chemical components of the fruit throughout the growing season were investigated, and cell size and starch distribution were observed histochemically. The treatment was done by dipping the flowers into 10ppm KT-30 solution on the 7th d after pollination.1. The fruit weight increase of the kiwifruit was accelerated with KT-30 treatment by 35-42 d. Weight of the treated fruit was about 1.4 times that of the control on the 170th d after pollination. Some deformity in shape attributable to KT-30 treatment was observed.2. In the control and treated fruits, starch granules were observed from the 63rd d after pollination in the pericarp, and from the 105th d in the central core by light microscopy. There was no difference in the number of starch granules between the control and treated fruits. There was no change in cell size with KT-30 treatment. It suggested that fruit enlargement with KT-30 treatment was caused by increase of cell number.3. The analysis of sugar by high-performance liquid chromatography (HPLC) revealed the presence of fructose, glucose and sucrose, and total sugar content in the fruit increased about 1.2 times by KT-30 treatment compared with the control on the 170th d after pollination. This suggested that KT-30 treatment promoted the growth of kiwifruit.4. As for free organic acids, more citric and quinic acids were found in the treated fruit than in the control, but almost the same pattern was observed in the changes throughout the growing season.5. Changes in total pectin contents throughout the growing season were similar in both the control and treated fruits, whereas the content was slightly higher in the control. Total pectin was largely composed of hydrochloric acid soluble pectin (HSP) in both control and treated fruits.6. The pericarp of the control fruit became deep green throughout the growing season, while that of the treated fruit became pale. Chlorophyll content tended to show a higher value in the treated fruit than in the control up to 35 d after pollination, but thereafter it decreased steadily. In the control fruit, however, chlorophyll content tended to increase throughout the growing period. It was supposed that chlorophyll content in kiwifruit was affected by KT-30 treatment.

A simplified method for the determination of forchlorfenuron in agricultural products by HPLC with UV detection was investigated. A chopped sample homogenate from agricultural products was extracted with acetone. The extract was filtrated and concentrated. The residues was loaded onto a Chem Elut column and extracted with ethyl acetate. The crude extract was purified on Oasis HLB and Bond Elut PSA mini-columns using a mixture of methanol and ethyl acetate. Forchlorfenuron was analyzed by HPLC with UV detection (263 nm). HPLC separation was performed on an ODS column with methanol-water as the mobile phase. Recoveries of forchlorfenuron from several agricultural products fortified at the level of 0.1 microg/g were in the range of 87.6-99.5%. The limit of detection (S/N=3) was 0.005 microg/g in the sample.

This paper describes the application of liquid chromatography-tandem mass spectrometry (LC/MS-MS) for analysis of residues of forchlorfenuron (CPPU), a new plant growth regulator, in watermelons, after a sample preparation step based on the buffered Quick, Easy, Cheap, Effective, Rugged and Safe extraction method. Analytical determinations were carried out in a triple quadrupole system fitted with an electrospray interphase operating in the positive ionisation mode (ESI+). Three simultaneous MS-MS transitions of the quasi-molecular ion m/z 248 (precursor ion) were monitored for data acquisition (248129, 248155, and 248248), using the transition 248129 for quantitation. Recovery studies on watermelons at levels of 1-200 microg/kg, performing five replicates at each level and using bracketing single-level calibration with matrix-matched standards for quantitation, gave forchlorfenuron mean recoveries ranging from 82 to 106% with relative standard deviations (RSD) lower than 18%. The limit of determination was established at 1 microg/kg. The method was applied to evaluate the persistence of forchlorfenuron residues in watermelons grown in plastic greenhouses, after applying an individual spray treatment to the flower ovary at the anthesis stage (45 mu g/flower and 60 mu g/flower for cv "Extazy" and cv "reina de corazones" watermelons, respectively). One month after treatment, 20 "Extazy" watermelon units (1.5-2 kg/unit) and 20 "Reina de corazones" watermelon units (4-5 kg/unit) were collected and analyzed individually. None of the samples contained forchlorfenuron residues higher than 1 microg/kg.

O presente trabalho teve por objetivo verificar o efeito da aplicação de diferentes concentrações do ácido giberélico (AG3), do thidiazuron (TDZ) e do forchlorfenuron (CPPU) no aumento do tamanho dos cachos e das bagas e no teor de sólidos solúveis totais de uva sem semente, cv. BRS Clara. As plantas foram conduzidas no sistema de latada, sobre o porta-enxerto IAC 572, no espaçamento 2,5 x 2,0m e com irrigação por microaspersão. Os experimentos foram conduzidos na Estação Experimental de Viticultura Tropical, da Embrapa Uva e Vinho, em Jales-SP. Os reguladores de crescimento foram aplicados via pulverização localizada no cacho, utilizando-se de concentrações de 0 a 4mg.L-1 de CPPU; 0 a 10mg.L-1 de TDZ, e de 0 a 90mg.L-1 de AG3, isolados ou em conjunto. De modo geral, a aplicação dos reguladores de crescimento promove a melhoria da qualidade dos cachos da cv. BRS Clara; o uso do TDZ e do CPPU em conjunto com o AG3 produz um efeito sinérgico, proporcionando melhor resposta do que o uso isolado do AG3; os tratamentos com 60mg.L-1 de AG3; 20mg.L-1 de AG3 + 4mg.L-1 de CPPU, e 10mg.L-1 de AG3 + 5mg.L-1 de TDZ proporcionam os melhores resultados para o aumento do diâmetro das bagas; a aplicação de AG3 antes da floração da cv. BRS Clara provoca abortamento excessivo, reduzindo a qualidade comercial dos cachos; a utilização de concentrações elevadas dos reguladores reduz o teor de sólidos solúveis totais. The objective of this study was to evaluate the effect of different concentrations of gibberellic acid (AG3), thidiazuron (TDZ) and forchlorfenuron (CPPU) on the size of bunches and berries and on the total soluble solids content, when applied in different concentrations on the seedless grapes, cv. BRS Clara. The grapevines were grafted on the rootstock IAC 572, trained in the pergola system and, spaced 2.5 x 2.0m and irrigated by micro sprinklers. The experiments were carried out at the Embrapa Grape and Wine Experimental Station for Tropical Viticulture in Jales, SP. The bunches were directly sprayed with the plant growth regulators, either individually or combined, in concentrations from 0 to 4mg.L-1 of CPPU; 0 to 10mg.L-1of TDZ and 0 to 90mg.L-1 of AG3. Generally, the application of growth regulators improved the quality of the bunches of the cv. BRS Clara; the use of TDZ and CPPU, combined with AG3, produce a synergistic effect, causing a better response than the isolated use of AG3; the treatments with 60mg.L-1 of AG3, 20mg.L-1 of AG3 + 4mg.L-1 of CPPU and 10mg.L-1 of AG3 + 5mg.L-1 of TDZ provide the best results for increase in the berries diameter; the application of AG3 before flowering cause excessive abortion which reduced the commercial value of the bunches. High concentrations of plant growth regulators decrease the content of total soluble solids.

As cultivares de uvas apirênicas (Vitis vinifera L.), via de regra, possuem bagas de tamanhos reduzidos, necessitando de ajustes no manejo para a melhoria da qualidade dos cachos, sendo a aplicação de reguladores de crescimento um dos tratamentos mais eficazes. Objetivou-se com o presente trabalho avaliar os efeitos do forchlorfenuron (CPPU: 0 e 10 mgL-1) combinado ao ácido giberélico (GA3: 0, 25, 50, 75 e 100 mgL-1) durante o primeiro ciclo de produção das cultivares Centennial Seedless, Flame Seedless e Thompson Seedless. As características avaliadas foram o comprimento, diâmetro e peso de bagas. Para a 'Centennial Seedless', maiores comprimentos, diâmetro e peso de bagas foram obtidos com 100 mgL-1 de GA3; e a concentração de 100 mgL-1 de GA3 adicionada a 10 mgL-1 de CPPU foi a mais responsiva para a 'Flame Seedless' e 'Thompson Seedless', observando-se atrasos na maturação, em termos de sólidos solúveis totais, para todas as cultivares. The Seedless grapes (Vitis vinifera L.), have always decrease in berry size, requiring managements to increase cluster quality. The spray with growth regulators is one of most effective the treatment to enlarge berry size. The objective of this work was to determine the effects of Forchlorfenuron (CPPU: 0 e 10mgL-1) associated with Giberelic acid (GA3: 0, 25, 50, 75 e 100 mgL-1) during the first cycle of production of the cultivars Centennial Seedless, Flame Seedless and Thompson Seedless. The appraised characteristics were length, diameter and weight of berries. For Centennial Seedless, larger lengths, diameter and weight of berries were obtained with 100 mL-1 of GA3; and the concentration of 100 mL-1 of GA3 added to 10 mL-1 of CPPU was the most responsive for Flame and Thompson Seedless, being observed delays in maturation, in terms of total soluble solids, for all cultivars.

Persistence of forchlorfenuron residues in grape berries at harvest following its dip application as single or split doses to grape berry clusters and periodic dissipation of forchlorfenuron residues in grape berries following foliar spray application were studied. Periodic dissipation of forchlorfenuron residues following its fortification in soil and water were also studied. Splitting the dip application concentration of forchlorfenuron to grape berries reduced its residues in the berries at harvest, which persisted for more than 65 days from all treatments. In case of foliar application, however, the residues of forchlorfenuron in/on the grape berries persisted for 15-20 days only from three treatment concentrations of 2, 3 and 4 ml/l and dissipated with half-lives of 3.4-4.5 days. The residues of forchlorfenuron dissipated faster in soils maintained at field capacity moisture condition than in air dry soils. There was wide variation in its residue persistence in soil (DT50 = 15.1-121.3 days) depending on soil type and moisture condition. Forchlorfenuron residues persisted for more than 30 days in water and its dissipation was fastest at a water salinity level of 3.85 mmho/ cm although the rate of dissipation was not significantly affected by the change in salinity level from

Clematis spp. L. is a twining vine covered in showy blooms. Typical growth of hybrids is from the main leader, producing a thin, unbranched plant with one cyme. Apical dominance is released by cutting back the vine during production. Cutting back, or pinching, of a plant is labor intensive and compromises bloom for vegetative growth at time of sales. The purpose of this project was to eliminate manual pinching by treating young plants with chemical plant growth regulators (PGRs) that enhance branching without removal of the apical meristem. The first project evaluated the use of Atrimmec (dikegulac sodium), Fascination (BA+GA4+7), Florel (ethephon), and Dropp 50 (thidiazuron) on Clematis cultivars Ernest Markham, and Hagley Hybrid, and Clematis viticella 'Polish Spirit.' Plants treated with 800 mg·L-1 Atrimmec, or 800 or 1200 mg·L-1 Fascination experienced an increase in branch numbers. The second experiment manipulated the ratio of the components of Fascination, 6-BA and GA4+7, to reduce phytotoxicity experienced in the first experiment. The optimal ratio to enhance branching was 1:1, which is the stock solution for Fascination. All ratios produced phytotoxic symptoms. A third experiment tested lower rates of thidiazuron and added CPPU (forchlorfenuron) to the list of PGRs to test. The last experiment took the most effective PGR treatments, Atrimmec at 800 mg·L-1, and Fascination at 800 or 1200 mg·L-1, and compared them to the current production practices of pinching. Large flowering cultivars of clematis were used, including 'Comotesse de Bouchard,' 'Ernest Markham,' and 'Hagley Hybrid.' Atrimmec increased branch numbers and suppressed leader lengths without a mechanical pinch. Results from Fascination varied by cultivar. Master of Science

A novel method based on the monolithic column solid-phase extraction( SPE) coupled w ith high performance liquid chromatography in online mode w as developed for the determination of trace forchlorfenuron in fruits. T he enrichment factor of SPE for forchlorfenuron w as 214. T he method w as fully validated. It show ed a linear range of 0. 01- 50. 00 μg / L and a limit of detection of 25 ng / L,w ith the relative standard deviation( R SD) of 3. 9%. T he proposed method w as successfully applied for the determination of forchlorfenuron in grape,kiw ifruit and w atermelon samples,w ith the recoveries of 87. 0%-120. 7% and the R SDs of 0. 6%- 9. 2%. T his online method w as proved to be selective,sensitive and convenient for the determination of trace forchlorfenuron in complex samples.

Flame Seedless grapes were sprayed with N-(2-chloro-4-pyridyl)-N-phenylurea (CPPU) at 0, 2.5, and 5.0 ppm to develop rachis resistant to browning and dehydration. Rachis polyphenol oxidase (PPO) activity was determined during cluster development. Cluster components were weighed at commercial (CM), and physiological maturity (PM). PPO activity, rachis color changes (L* and a*), and cluster weight loss were evaluated at 0 °C for 8, 16, 32, and 56 days. CPPU-treated rachis had a decrease of 36% in PPO activity and a week delay in peak activity. At PM, dry weight of CPPU-treated rachis increased by 3 g. Postharvest rachis PPO activity declined with CPPU application, and color changes followed the same pattern for CM and PM. After 32 days of storage, L* and a* in lateral branches were significantly superior in CPPU treatments. Weight losses below 2.1% were significantly lowest in CPPU-treated clusters for 16 days of storage regardless of cluster maturity. Keywords: Polyphenol oxidase; table grapes; rachis, dar...

A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed for the determination of the plant growth regulator (PGR) (paclobutrazol, forchlorfenuron, isopentennyladenine and 6-benzylaminopurine) residues in amphisarcas. The sample was extracted with acetonitrile, then cleaned up by MCX solid phase extraction. The HPLC separation was performed on an Agilent XDB-C18 column with 5 mmol/L ammonium acetate solution and acetonitrile containing 0.1% (volume percentage) formic acid as the mobile phases in a gradient elution mode. The PGRs were determined by MS/MS in positive electrospray ionization mode, and quantified by matrix-matched external standard method. The calibration curves showed good linearity in each range with correlation coefficients greater than 0.999. The limits of quantification (LOQs, S/N > 10) were 0.04-1.35 g/kg and the limits of detection (LODs, S/N > 3) were 0. 01-0 41 microg/kg for the four PGRs spiked in cucumber and apple. The recoveries of the four PGRs spiked at three levels ranged from 81.0% to 93.%, with the relative standard deviations (RSDs) of 3.5%-9.5%. The sensitivity, accuracy and precision of the method meet the technical standards of the pesticide determination. Therefore the method can be applied to the determination of the four PGRs in amphisarcas.

The ability of forchlorfenuron (CPPU), a substituted phenylurea compound, for inducing somatic embryogenesis in peanut (Arachis hypogaea L.) seedlings has been demonstrated. CPPU promoted somatic embryogenesis at a range of concentrations in all three peanut cultivars tested. Embryogenic response was dependent on applied CPPU concentrations. Exposure of seedlings for only two days to CPPU induced somatic embryogenesis, but the most effective treatment was to induce seed germination on media supplemented with either 2.5 or 4.0 μM CPPU and to maintain the seedlings on the same medium. Number of somatic embryos and the frequency of embryogenesis was higher for younger seedlings (up to 9 days), regardless of the CPPU concentrations and seedlings older than 21 days failed to produce somatic embryos. Removal of cotyledons from the seeds drastically reduced the embryogenic potential of the seedlings. Somatic embryos developed into whole plants following their separation and subculture on a medium lacking growth regulators. The induction of somatic embryos using CPPU as a sole growth regulator may provide a useful system to study the role of this compound in plant morphogenesis.

We have found that a synthetic urea derivative, N -(2-chloro-4-pyridyl)- N ′-phenylurea (4PU30, forchlorfenuron), has even higher cytokinin activity than N 6 -benzyladenine (BA). Using [ 3 H]PU30 (2.83 TBq/mmol) as a probe, we confirmed chromatographically the presence of a high-affinity cytokinin-specific binding protein (CSBP) in whole-cell extract from etiolated mung bean seedlings. The apparent molecular weight of CSBP was estimated to be 21 kDa. The association constants (K a ′s) of CSBP for 4PU30 and BA were calculated to be 4 × 10 10 ) M −1 and 3 × 10 9 M −1 , respectively. Various active cytokinins showed mutually competitive binding to CSBP, and their affinities for CSBP corresponded well to their cytokinin activities at the tissue level.

Certain phenylurea derivatives: N1-(2-chloro-4-pyridyl)-NZ phenylurea (4-PU30, Fulmet, Forchlorfenuron) (Takahashi et al. 1978), N1-(1,2,3-thiadiazol-5y1)-N521-phenylurea (Thidiazuron, Dropp-50) (Baskakov et al., 1981; Kulaeva, 1982; Mok et al., 1982), N1-(pyrrolidino-3-sulfonyl-phenyl)-N521-phenylurea (PSPPU) (Karanov et al., 1985) and N1-(4-methoxy-6-methyl-sim.triazine-2yl)-N5- (2-chlorophenyl-sulfonyl) urea (Glean-75, DPX-4189, Chlorsulfuron) (Levit, 1983) exhibit cytokinin activity exceeding that of adenine derivatives (e.g. kinetin, BA, zeatin).

A study was conducted to determine the effects of post-bloom applications of forchlorfenuron (CPPU) at 5 and 10 ppm, and the practice of cane girdling on fruit quality of two table grape cultivars (cvs.) « Perlette » (seedless) and « Italia » (seeded). For « Perlette » cv, CPPU treatments increased significantly berry weight and crop per vine, relatively to the control. However, soluble solids decreased while the acidity increased, reflecting a late maturation of berries from the treated vines. The delay of fruit maturity was also observed in « Italia » grapes, measured by a significant decrease in fruit soluble solids content. Girdling had a slight and non significant effect on fruit quality for both cvs.

In Spain, the adoption of the triploide ‘Queen of Hearts’ (Citrullus lanatus Thunb.) watermelon cultivar has brought important changes in the production of seedless watermelon thanks to its magnificent acceptance by the market. The experiments on triploid watermelon presented here examined innovative production techniques that would guarantee the productivity and quality of this cultivar in plastic greenhouses and improve cost-effectiveness, serving growers. Crop intensification was tested under a “temporary trellis” management system, increasing plant density from 2500 plants/ha in the traditional or creeping crop system to 10,000 plants/ha. The “temporary trellis” system was combined with fruit set through the localized application of a 150- to 200-mg·L−1 solution of 1-(2-chloro-4-pyridyl)-3-phenylurea (CPPU). The increase of grafted watermelon plant density to 1 plant/m2 gave rise to an increase in early harvest measured as the number of fruits per surface area compared with the traditional system with a density of 0.25 plant/m2. In addition, a process of temporary trellising facilitates choosing female flowers for applying CPPU. The °Brix and pulp firmness quality parameters did not show significant differences between “temporary trellised” and creeping crops.

Pollination is one problem with intensive seedless watermelon (Citrullus lanatus Thunb.) production under unfavorable environmental conditions (low solar radiation and temperature) due to the lowered activity of pollinating insects, such as the bee (Apis mellifera L.). An alternative to overcome these problems is the use of plant growth regulators. For this reason, experiments were conducted for 2 years in plastic greenhouses in the fields of Almeria, southwestern Spain, to evaluate the production and quality of ‘Reina de corazones’ triploid watermelon grafted onto RS841 rootstock (Cucurbita maxima × Cucurbita moschata). Two phytoregulators were used for the development of the ovary: 1-(2-chloro-4-pyridyl)-3-phenylurea (CPPU) and 2,4-dichlorophenoxyacetic acid (2,4-D). Concentrations of CPPU evaluated were 50, 100, 150, and 200 mg·L−1. An application of 0.6 mL was applied to each ovary in addition to 4, 6, 8, or 12 mg·L−1 of 2,4-D to the foliar mass at a proportion of 1000 L·ha−1. Results showed that the production and number of fruit obtained with CPPU treatments were similar to what is obtained by using bees for fruit pollination. Maximum production was reached at concentrations of 100–200 mg·L−1. Average production was 32% to 83% higher than results from 2,4-D at 8 mg·L−1. The number of fruit per plant was 33% to 35% higher as well. In the first assay, a positive correlation was also observed between production and CPPU concentration. CPPU treatments had a lower accumulation of sugars than those with 2,4-D; nevertheless, both treatments showed values of commercially acceptable soluble solids.