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Forchlorfenuron (CPPU) is often applied during the cultivation of kiwifruit to produce larger fruit. To address degradation patterns of CPPU during simulated cold chain logistics and simulated shelf life of the fruit after harvest, appropriate storage methods and safe consumption behavior can be investigated. In this study, an ultra-high-performance liquid chromatography-tandem mass spectrometry method was adopted to detect CPPU residues under different conditions. CPPU in kiwifruit stored at 6 °C had a half-life of 40.8–77.0 days. However, when kiwifruit was stored at 0 °C under simulated cold chain storage conditions, the half-life of CPPU was 63.0–115.5 days, implying that lower storage temperatures can reduce the degradation rate of CPPU. The residues of CPPU in kiwifruit pulp declined with time, and the reduction followed the first-order kinetics equation. More CPPU residues were present in the pulp of postharvest kiwifruit treated with exogenous ethylene than in the pulp of untreated kiwifruit. Thus, using exogenous ethylene for artificial ripening after harvest is not recommended. We determined that the appropriate cold chain storage temperature is 6 °C. It is recommended that the public select kiwifruit stored for at least 2 weeks. The estimated chronic and acute dietary risk quotients of CPPU are ≤ 0.79% and ≤ 0.11%, respectively. Therefore, it is highly unlikely that consumers will be poisoned by CPPU due to kiwifruit consumption. Our results provide scientific evidence regarding the adoption of appropriate kiwifruit storage methods and consumption behavior to enhance consumption safety.

An imidazolium ionic-liquid-modified phenolic resin (ILPR) was synthesized using 3-aminophenol as a functional monomer, glyoxylic acid as a green cross-linker, and polyethylene glycol 6000 as a porogen. The obtained ILPR showed better extraction of benzoylurea plant hormones thidiazuron and forchlorfenuron than the unmodified phenolic resin because the imidazolium IL provides more interaction modes with the analytes. ILPR, as a tailored adsorbent for solid-phase extraction, was coupled with high-performance liquid chromatography (ILPR‒SPE‒HPLC) for the simultaneous determination of thidiazuron and forchlorfenuron in cucumbers. Good linearity of the ILPR‒SPE‒HPLC method was obtained, ranging from 0.0100 to 5.00 μg g−1 with a correlation coefficient (r) ≥ 0.9999. The recoveries of spiked samples ranged from 91.4% to 100.7% with a relative standard deviation of ≤ 6.0%.

MSc (Environmental Sciences), North-West University, Potchefstroom Campus Soybean is an important protein and oilseed crop throughout the world due to the commercial use of soybean meal, oil and sub-products. South Africa is the largest soybean producer in Africa and produces on average 1.8 t ha-1. Although soybean productivity in South Africa has increased over the past decades, yield gaps can be narrowed by adopting improved agricultural practices that increase yield and decrease yield loss due to genetic barriers and environmental stresses. Available biotechnologies, in particular plant growth regulators (PGRs), have been reported to increase the tolerance of crops to environmental stresses, increase harvestable yield and to enhance growth and yield components. The aims of this study were to evaluate the efficacy of PGRs at different application times and rates to increase soybean yield in South Africa. To determine the optimal time of application, five field trials were conducted at three localities, viz. Bethlehem and Kroonstad in the Free State province and Potchefstroom in the North-West province, South Africa. Two field trials and one pot trial were conducted to investigate application rates at Potchefstroom. The cultivar PAN 1521 R was planted in the field trials while LS 6161 R was used in the pot trial. Foliar treatments consisted of a control (untreated) and three PGRs, to which a specific identification code was assigned, viz. A2019 forchlorfenuron (CPPU), B2019 naphthalene acetic acid (NAA) and C2019 gibberellic acid + abscisic acid (GA3 + S-ABA). Plant growth regulators were applied at two different growth stages (R1 and R3). To determine the optimal application time, applications were done at R1 and R3, as well as at both R1 + R3 (double application). Three concentrations of each PGR were applied at growth stages R1 + R3 (double application) to determine the optimal rate of application. No optimum application time or rate could be determined for any of the PGRs evaluated on soybean in this study. None of the three PGRs evaluated at different application times significantly increased the yield of soybean. Application of PGRs to soybean planted in Bethlehem (cold zone) were more effective in terms of nodes plant-1, pods node-1 and pods plant-1 than PGR applications at Potchefstroom and Kroonstad. Plant growth regulator treatments increased growth parameters of soybean, however these effects varied between Potchefstroom, Bethlehem and Kroonstad and were not consistent. None of the application rates of the respective PGRs, increased any of the growth and yield components in the field trials or the pot trial at Potchefstroom. It could possibly be ascribed to the low application rates used and the warm and dry climatic conditions experienced during this study. Application of PGRs at too low concentrations will have no effect on growth and yield parameters. Although increases in growth and yield parameters were reported for other crops, similar results were not found during this study. More research on the PGR, rate and application time for soybean is therefore needed in South Africa. Masters

Plant growth regulators (PGRs) are commonly used to regulate maturation in strawberry. Despite this, comprehensive assessments of the metabolomic effects of PGRs on strawberry maturation are lacking. In this study, a nuclear magnetic resonance-based approach, combined with multivariate and pathway analysis, was used to evaluate the regulatory effects of gibberellin, forchlorfenuron, and brassinolide, applied at two different maturation stages, on the expression of metabolites in strawberry. The results demonstrated that the PGRs differentially influenced metabolism, whether applied at the same or different maturation stages. Additionally, we also discovered that these different PGRs exhibited some similar metabolic trends when applied at the same growth period. Our findings validate the use of NMR-based metabolomics for identifying subtle changes in the expression of metabolites associated with PGR application.

Seaweed tissue culture is one of the potential technologies that can be used to increase seaweed production to cope with increasing demand. Seaweed tissue culture can be divided into two methods, namely, direct regeneration and callus induction (indirect regeneration). These methods provide several advantages, including generating seaweed cultures that are disease-free and able to mature faster to support mass production. The present study aims to develop a suitable protocol for tissue culture of Sargassum polycystum C. Agardh. We investigated different methods of sterilizing the stipe, stolon and leaf explant surfaces to allow for axenic tissue growth. In addition, we also investigated the effects of different plant growth regulators (PGRs) on seaweeds that are grown via direct regeneration and callus induction method. Our results showed that a combination of physical (brushing using soft brush under microscope) and chemical treatments (detergent, povidone iodine (PI) and antibiotic solution of streptomycin sulphate) was most suitable for growing axenic stipe of S. polycystum. For obtaining axenic leaf and stolon explants, the best treatment option was using germanium oxide (GeO2) as a pre-treatment, followed by treatments with detergent, PI and streptomycin sulphate antibiotic solution. We tested the effects of nine different plant growth regulators on the growth of tissue explants, which include indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 1-napthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), picloram, forchlorfenuron (CPPU), uniconazole, 6-benzylaminopurine (BAP) and kinetin. Results showed that only bud explant was able to induce callus growth when treated with kinetin (4.65 μM and 13.94 μM) and uniconazole (3.00 μM, 7.00 μM and 10.00 μM). In addition, only stipe explant showed regeneration of adventitious bud when grown in the absence of plant growth regulators.

The current study prepared solid dispersions of forchlorfenuron (CPPU) and γ-cyclodextrin (γCD) or CPPU and 2-hydroxypropyl-γ-cyclodextrin (HPγCD) via cogrinding and coprecipitation to assess their physicochemical properties and their effect on plant growth. According to phase solubility diagrams, both CPPU/γCD and CPPU/HPγCD formed an inclusion complex at a molar ratio of 1/1. According to differential scanning calorimetry and powder X-ray diffraction, a ground mixture (GM) of CPPU and γCD (molar ratio = 1/1), a GM of CPPU and HPγCD (molar ratio = 1/1), and a coprecipitate (CP) of CPPU and γCD (molar ratio = 1/1) formed an inclusion complex. According to 1H-1H nuclear Overhauser effect spectroscopy NMR spectroscopy of the GMs and CP, the aromatic rings of the CPPU molecule are presumably included in CD from the wider to the narrower rim of its ring. Cultivation of broccoli sprouts with the GMs and CP resulted in no differences in the length of sprouts in comparison to a commercial preparation (Fulmet)., This is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes.

Krymsk® 5 (VSL-2) is a semi-dwarf cherry rootstock adaptable to a range of climates. The present study aimed to establish the first efficient in vitro propagation protocol for this rootstock. Therefore, six cytokinines, four adenine type (6-benzyladenine, 2-isopentenyladenine, kinetin and meta-topolin) and two phenylureas (thidiazuron and forchlorfenuron) at three (2.4 μΜ, 4.8 μΜ and 9.6 μΜ) concentrations plus three (0.24 μΜ, 0.48 μΜ, 0.96 μΜ) for thidiazuron only were tested during the multiplication stage. 6-Benzyladenine was the most efficient cytokinin, based on the number of shoots produced (3.5 shoots at 9.6 μΜ) and the number of nodes per explant (10 nodes at 9.6 μΜ) whereas the other aromatic adenine tested, i.e. meta-topolin, presented the highest number of nodes per cm and node per shoot. During the rooting stage the synthetic auxins 1-naphthaleneacetic acid (1-NAA) and indolebutyric acid (IBA) were tested at concentrations of 0, 2.5, 5, 10 and 20 μΜ both separately and in all possible combinations. The percentage of successfully rooted explants reached 95% under the combination of 20 μΜ IBA plus 5 μΜ 1-NAA, whereas the highest number of roots recorded was 8.5 roots for the treatment 20 μΜ ΙΒΑ plus 2.5 μΜ 1-NAA. Furthermore, two different carbon sources were compared, the widely used sucrose and the endogenous sugar ratio of mother plant softwood shoot, sampled during late of May. Endogenous sugar ratio proved to be the preferable carbon source, since it increased the number of shoots produced and almost doubled the number of produced nodes per explant.

BACKGROUND Gray mold is a ubiquitous destructive plant disease found worldwide. To avoid the shortcomings of conventional spraying systems for controlling this disease, such as high selection pressure on Botrytis cinerea for resistance and fungicide waste resulting from spray drift, a precisely targeted application strategy of dipping young cucumber fruit in a mixture of fungicide and forchlorfenuron (plant growth regulator, PGR) during the bloom period to control cucumber gray mold was developed in the current study. RESULTS Without leaving above-limit residues in cucumber fruits, dipping in fludioxonil at 30 mg L-1 provided a greater efficacy (85.4%) against cucumber gray mold than did spraying at 100 mg L-1 (76.4%). Importantly, fludioxonil mixed with forchlorfenuron from 25 to 35 mg L-1 increased the yield of cucumbers by 26.2-36.7% compared to dipping fruit only in forchlorfenuron. The increased yield may be a benefit of controlling gray mold. CONCLUSION Dipping fruit in fungicides and PGRs seems to be a potential precisely targeted application strategy to not only control cucumber gray mold effectively but also, through the action of PGRs, to increase the cucumber yield. This novel application method is believed to have a bright prospect in cucumber production in Chinese solar glasshouses. © 2018 Society of Chemical Industry.

The advantages of using smart materials as immunosorbents in the analysis of complex matrices by ion mobility spectrometry (IMS) have been highlighted in this study. A novel analytical method has been proposed for the sensitive, selective, and fast determination of residues of the plant growth regulator forchlorfenuron in fruit juices. Three different monoclonal antibodies (s3#22, p2#21, and p6#41) were employed for the production of immunosorbents, based on Sepharose gel beads, which were characterized in terms of loading capacity, solvent resistance, and repeatability for its use in solid-phase extraction (SPE). Immunosorbents that were prepared with antibody p6#44 provided the best performance, with a loading capacity of 0.97 μg, a 10% (v/v) 2-propanol tolerance, and a reusability of at least eight uses. The SPE procedure involved the use of a column with 0.15 g Sepharose beads, containing 0.5 mg antibody, which was loaded to 20 mL of the sample, washed with 2 mL of water plus 2 mL of 10% (v/v) 2-propanol, and eluted with 2 mL of 2-propanol. The cleaned extract was directly analyzed by IMS, giving a limit of detection of 2 μg L−1 with a relative standard deviation of 7.6%. Trueness was assessed by the analysis of blank grape and kiwifruit juice samples spiked with forchlorfenuron concentrations from 10 to 400 μg L−1, with recoveries from 80 to 115%. The analytical performance of the proposed immunosorbent was compared with conventional extraction and cleanup methods, such as QuEChERS and C18-based SPE, giving the cleanest extracts for accurate determinations of forchlorfenuron by IMS.

Physiological (metabolite analysis) and molecular (gene expression) approaches were used to understand the mechanism underlying russet formation in response to the application of GA3 and CPPU (Forchlorfenuron) in a Japanese table grape cultivar ‘Shine Muscat’. Several different concentrations of GA3 and GA3 + CPPU [25 mg L−1 GA3 (A), 25 mg L−1 GA3 + 5 mg L−1 CPPU (B), 25 mg L−1 GA3 + 10 mg L−1 CPPU (C), and 25 mg L−1 GA3 + 15 mg L−1 CPPU (D)] were applied to grape berry clusters at two weeks after flowering (WAF). No russet was observed on the berries treated with the ‘C’ combination. Lower levels of phenylalanine ammonia-lyase (PAL) activity was observed in the treated samples, relative to the untreated material. Reduced peroxide (POD) activity was also observed in response to different treatments, while the expression of Peroxidase 17 and Phenylalanine ammonia-lyase G1 genes mirrored lignin content. Increased activity of 4-coenzyme A ligase (4CL) may contribute to decreasing the level of russet and help to improve grape berry quality.

The objective of this study was to investigate the effect of the preharvest application of forchlorfenuron (CPPU) and perforated polyethylene bag packaging (PPE) on maintaining the postharvest quality of red-fleshed cv. ‘Da-Hong’ pitaya (Hylocereus polyrhizus sp.) fruit. On the flowering day, 100 mg·L−1 CPPU was sprayed on the bracts and water was used as the control. After harvest, all fruits were divided into three package treatments, which were packed without bags, packed with and without PPE bags, and stored at 5 ± 0.5 °C and 90 ± 5% relative humidity for 21 days, followed by 7 days at 20 °C and 75 ± 5% relative humidity without bags for quality evaluation. Significantly higher bract thickness (2.26 vs. 1.44 mm), longer fruit length (120.5 vs. 109.04 mm), and greater firmness (1.56 vs. 1.04 kg·cm−2) were recorded for the CPPU treated fruit at harvest. Preharvest application of CPPU with perforated packaging resulted in significantly greener bracts, a lower yellow index, fewer chilling incidences, and a lower decay ratio, but there was a slight decrease in respiration rate during cold storage at 5 °C for 21 days. However, all criteria reached the threshold when fruits were transferred to 20 °C for 7 days. In conclusion, preharvest CPPU application plus perforated packaging is the best combination for the long-term storage of red-fleshed pitaya fruit at 5 °C.

Grapevine (Vitis vinifera L.) is considered to be one of the most popular and widespread fruit crops in the world. Numerous value added products are prepared from grape fruit and investments are being made to establish new viticulture region (Hoff et al., 2017; Imran et al., 2017). CPPU [forchlorfenuron N-(2-chloro-4-pyridyl)-N-phenylurea] is a synthetic cytokine-like plant regulator which promotes grape berry set and development. The influence of CPPU [forchlorfenuron N-(2-chloro-4-pyridyl)-N-phenylurea] on berry development of 'Shine Muscat' (Vitis labruscana Bailey×V vinifera L.) grapes was evaluated under field conditions. A concentration response was observed over a range of 0, 5, and 10 mgL-1 CPPU that was applied to fruitlets (mean diameter 6mm) at 2 weeks after full bloom. Gas-chromatography mass-spectrometry (GC-MS) revealed that volatile compounds such as terpenoids and aromatics; especially linalool, geraniol and benzyl alcohols, were greatly reduced in CPPU-treated grapes. In contrast, aliphatics, such as hexanol, were increased in CPPU-treated berries. RNA sequencing (RNA-Seq) was conducted to identify differentially expressed genes (DEGs) that were induced by CPPU, especially those related to volatile biosynthesis. A total of 494, 1237, and 1085 DEGs were detected in CPPU0-vs-CPPU5, CPPU0-vs-CPPU10, and CPPU5-vs-CPPU10 treatments, respectively. The results were compared against two databases (Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG)) to annotate gene descriptions and assign a pathway to each gene. GO covers three domains: biological processes, molecular functions and cellular components. Pathway enrichment annotation demonstrated that highly ranked genes were associated with the fatty acid degradation and biosynthesis, phenylpropanoid metabolism and biosynthesis, carotenoid biosynthesis, and plant hormone signal transduction. Analysis with qRT-PCR of twelve selected transcripts validated the data obtained by RNA-seq. Additionally, we also found that genes such as CCDs (carotenoid cleavage dioxygenase), LOX (lipoxygenase), GGDP reductase (geranylgeranyl diphosphate reductase), PAL (phenylalanine ammonia-lyase) and some hormones related genes, were closely involved in the formation of volatiles compounds in CPPU treated berries. In summary, our results provide the first sequential transcriptomic atlas of CPPU treated grape berries which significantly increases our understanding of volatile metabolites and biosynthesis pathways in grape affected by CPPU.

In this study, we have successfully synthesized a forchlorfenuron (CPPU)-imprinted polymer in dimethyl sulfoxide by molecular imprinting technique. In the molecular imprinting, β-cyclodextrin was used as the monomer with 1,6-hexamethylene diisocyanate (HMDI) as the cross-linking agent that formed a complex with forchlorfenuron by inclusion interactions. The adsorption equilibrium was attained in approximately 30 min and the maximum binding capacity was 26.79 mg/g, which indicated that the adsorption kinetics was relatively fast. The results of adsorption and selectivity experiments have shown that the imprinted polymer was able to bind forchlorfenuron specifically and reversibly. The specific recognition of the imprinted polymer for forchlorfenuron resulted from the cooperative effects of inclusion interaction and hydrophobic interaction. This imprinted polymer was also used to accurately determine forchlorfenuron residue in real fruit samples. In the standard spiked strawberry sample, the adsorption recoveries for forchlorfenuron were as high as 90.3, 84.5, and 90.8%, respectively. The binding specificity of the imprinted polymer for forchlorfenuron resulted from the imprinting effect. Therefore, as a reusable material possessing high affinity and selectivity, the imprinted polymer is promising in application to detect pesticide residues in fruit samples. In addition, because β-cyclodextrin is water soluble and can include neutral compounds, this molecularly imprinted polymer can be used to recognize pesticides in aqueous solutions. Graphical abstract Schematic demonstration of molecular imprinting and re-binding of forchlorfenuron.

Kiwifruit (Actinidia chinensis) is a rich nutritious fruit crop owing to a markedly higher content of vitamin C and minerals. To promote fruit set and to increase the yield of kiwifruit, forchlorfenuron (CPPU) has been widely applied. However, the molecular details regarding CPPU controlling kiwifruit development, especially at the fastest fruit growth stage, remain unknown. In the present study, we measured the effect of CPPU on developmental regulation in red-fleshed kiwifruit (Actinidia chinensis 'Hongyang'). Additionally, a cytological analysis was performed to clarify the precise changes in the cell structure of the CPPU-treated kiwifruits. Moreover, the concentration of endogenous phytohormones, including indoleacetic acid (IAA), zeatin (ZT), gibberellic acid 3 (GA3), and abscisic acid (ABA), were measured by Enzyme-linked Immunosorbent Assay (ELISA). Furthermore, RNA-Seq was performed to dissect the complicated molecular mechanisms, with a focus on biosynthesis, metabolism, and signaling compounds, such as endogenous hormones, sugars, and L-ascorbic acid. Our results demonstrated that CPPU treatment not only regulates the size and weight of a single fruit but also improves the quality in 'Hongyang' kiwifruit through the accumulation of both soluble sugar and vitamin C. It was also seen that CPPU regulates kiwifruit development by enhancing cell expansion of epidermal cells and parenchyma cells, while, promoting cell division of subepidermal cells. Additionally, CPPU significantly increased the gibberellin and cytokinin biosynthetic pathway and signaling, while repressing auxin and ABA biosynthetic pathway; thus, signaling plays an essential role in CPPU controlling kiwifruit development. Notably, transcriptomic analysis revealed that a total of 2244 genes, including 352 unannotated genes, were differentially expressed in kiwifruits because of CPPU treatment, including 127 transcription factors. These genes are mainly enriched in plant hormone signal transduction, photosynthesis, MAPK signaling pathway, starch and sucrose metabolism, and phenylpropanoid biosynthesis. Overall, our results highlight that CPPU regulation of kiwifruit development is mainly associated with an antagonistic and/or synergistic regulatory role of endogenous phytohormones, and enhancing the energy supply. This provides new insights into the molecular details of CPPU controlling kiwifruit development at the fastest fruit growth stage, which is of agricultural importance for kiwifruit breeding and crop improvement.

Resumen Introducción. La producción de vainilla en México ha disminuido en un 70 % en los últimos quince años, debido a la caída de sus frutos que ocurre en condiciones climáticas extremas, cuando la temperatura es mayor a 32 °C y la humedad relativa es menor a 80 %. Objetivo. El objetivo fue determinar el efecto de algunos reguladores de crecimiento (RC) comerciales sobre la retención de frutos de vainilla (Vanilla planifolia Jacks ex Andrews) cultivar Mansa. Materiales y métodos. El estudio se realizó en el 2016 en el municipio de Tecolutla, Veracruz, México. Los tratamientos evaluados fueron: clorhidrato de aminoetoxivinilglicina (AVG; 250 ppm), 1-ácido naftalenacético (ANA; 27 ppm), Benciladenina (BA; 150 ppm), Forclorfenurón (CPPU; 5 ml l-1 de agua), Ácido 2,4-diclorofenoxiacético (2,4-D; 32 ppm), 2,4-D (32 ppm) + CPPU (5 ml l-1 de agua) y testigo, que consistió en aplicar solamente un acidificante (8 ml en 10 l de agua). Los tratamientos se aplicaron en tres ocasiones en los mismos frutos a los 6, 20 y 31 días después de las últimas flores polinizadas por racimo. Resultados. Todos los RC retuvieron más frutos etiquetados que el testigo. Los tratamientos 2,4-D y 2,4-D + CPPU retuvieron más frutos por racimo y produjeron frutos significativamente más largos que el testigo. Además, la mezcla 2,4-D + CPPU produjo frutos con mayor diámetro que el testigo. Los reguladores de crecimiento AVG, ANA BA y CPPU no difirieron en el número de frutos etiquetados, frutos por racimo, longitud del fruto y diámetro del fruto. Conclusión. Los RC tuvieron un efecto positivo en la retención de frutos de vainilla, principalmente la mezcla 2,4-D + CPPU, con la cual se obtuvieron más frutos etiquetados y por racimo, con respecto al testigo. El uso de RC fue efectivo para reducir el aborto en vainilla. Abstract Introduction. The vanilla production in Mexico has decreased by 70 % in the last fifteen years, due to the fall of its fruits. This occurs in extreme climatic conditions when the temperature is higher than 32 °C and the relative humidity is less than 80 %. Objective. The objective was to determine the effect of some commercial growth regulators (GR) on the retention of vanilla (Vanilla planifolia Jacks ex Andrews) fruits Mansa cultivar. Materials and methods. The study was carried out in 2016 in the Tecolutla Municipality, Veracruz, Mexico. The treatments evaluated were: aminoethoxyvinylglycine hydrochloride (AVG; 250 ppm), 1-naphthaleneacetic acid (ANA; 27 ppm), benzyl-adenine (BA; 150 ppm), forchlorfenuron (CPPU; 5 ml l-1 of water), 2,4-Dichlorophenoxyacetic acid (2,4-D; 32 ppm), 2,4-D (32 ppm) + CPPU (5 ml l-1 of water) and control, which consisted of applying only one acidifier (8 ml in 10 l of water). Treatments were applied three times to the same fruits, at 6, 20 and 31 days after the last flowers pollinated by bunch. Results. All GR retained more labelled fruits than the control. Treatments 2,4-D and 2,4-D + CPPU retained more fruits per raceme and produced significantly longer fruits than the control. In addition, the 2,4-D + CPPU mixture produced fruits with larger diameter than the control. The growth regulators AVG, ANA BA and CPPU did not differ in the number of labelled fruits, fruits per raceme, fruit length, and fruit diameter. Conclusion. GR had a positive effect on vanilla fruit retention, mainly the 2,4-D + CPPU mixture with which more labelled fruits and per bunch were obtained compared to the control. The use of GR was effective in reducing vanilla abortion.

In this study, a modified quick, easy, cheap, effective, rugged, and safe method combined with ultra-high performance liquid chromatography and tandem mass spectrometry was developed for the multiclass determination of 28 plant growth regulators in various fruits. Different extraction solvents and adsorbents, including primary secondary amine, octadecylsilyl, graphitized carbon black, and zirconia-based sorbent, were investigated. Internal calibration and isotope internal standards, chlormequat chloride-d4 , mepiquat chloride-d6 , indole-3-acetic acid-d2 , and forchlorfenuron-d5 were used to improve accuracy. For method validation, good linearity, low limits of detection and quantification were obtained. At three spiked concentrations (10, 50, and 100 µg/kg), satisfactory recoveries with relative standard deviations of 2.4-17.5% were obtained for strawberries (75.2-119.8%), grapes (70.5-114.0%), tangerines (71.7-115.4%), apples (72.7-115.4%), and kiwi fruits (71.7-119.2%). Samples analysis revealed that 15.6% of the samples (n = 96) were contaminated with one or two kinds of plant growth regulators, including chlormequat chloride, forchlorfenuron, paclobutrazol, 2,4-dichlorophenoxyacetic acid, 2-diethylaminoethyl hexanoate, and mepiquat chloride. Similar results were obtained by ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry, indicating the robustness, effectiveness, and suitability of the developed method for routine monitoring of plant growth regulator residues in fruits.

Malignant mesothelioma (MM) is one of the most aggressive cancer types with a patient’s life expectancy of typically less than one year upon diagnosis. The urgency of finding novel therapeutic approaches to treat mesothelioma is evident. Here we tested the effect of the plant-growth regulator forchlorfenuron (FCF), an inhibitor of septin function(s) in mammalian cells, on the viability and proliferation of MM cell lines, as well as other tumor cell lines derived from lung, prostate, colon, ovary, cervix and breast. Exposure to FCF strongly inhibited proliferation of human and mouse (most efficiently epithelioid) MM cells and all other tumor cells in a concentration-dependent manner and led to cell cycle arrest and cell death. The role of septin 7 (SEPT7), a presumably essential target of FCF in MM cells was confirmed by an shRNA strategy. FCF was robustly inhibiting tumor cell growth in vitro at low micromolar (IC50: ≈20-60µM) concentrations and more promisingly also in vivo. Initial experiments with FCF analogous revealed the importance of FCF’s chloride group for efficient cell growth inhibition. FCF’s rather low systemic toxicity might warrant for an extended search for other related and possibly more potent FCF analogues to target MM and putatively other septin-dependent tumors.

Forchlorfenuron (CPPU), as a plant growth regulator or herbicide/pesticide, is widely used in agriculture worldwide. It is adopted by most farmers due to its high efficacy for boosting size and improving the quality of fruit. However, CPPU was implicated in, and gained notoriety due to an incident of exploding watermelon that occurred in 2011. Subsequently, the wider community became aware of the potential risks it posed to living organisms and the ecosystem. In this study, we evaluated the effects of CPPU on the survival, cardiac morphology and function, as well as hematopoietic system, of zebrafish (Danio rerio). Notably, CPPU (2.5–12.5 μg/ml) induced cardiac morphology deformation, cardiac contractile dysfunction and erythrocyte reduction in zebrafish. Consistently, the mRNA expression levels of several cardiac and hematopoietic gene markers (myl7, gata4, mef2c, amhc, vmhc and gata1) were altered by CPPU treatment. In addition, CPPU caused cytotoxicity, cytoskeleton destruction and reduced corresponding proteins (Myl7, Gata4 and Mef2c) expression in H9c2 cardiomyocytes in vitro. Taken together, this study has identified the cardiotoxicity of CPPU in different experimental models and enhanced our understanding on the mechanism underlying the toxicity of CPPU to living organisms.

The plant growth regulator forchlorfenuron is often applied to promote fruit setting and development in cucumber production. However, the effect of forchlorfenuron on the appearance and nutritional quality of cucumber is unknown. In the present study, forchlorfenuron was applied to female flowers 1 day before anthesis and at the day of anthesis. The application of forchlorfenuron successfully induced parthenocarpic fruit formation in cucumber. In addition, cytokinin-responsive genes were upregulated under forchlorfenuron treatment. Fruit treated with forchlorfenuron did not differ from pollinated fruit in shape, texture and major nutrients, such as protein, total flavonoids and vitamin C, with the exception of the lower phenolic acid content. Overall, our finding suggests that forchlorfenuron could be vital in promoting the parthenocarpic fruit set without altering the quality of cucumber.

// Nianzhu Zhang 1, * , Lu Liu 2, * , Ning Fan 2 , Qian Zhang 1 , Weijie Wang 1 , Mingnan Zheng 3 , Lingfei Ma 4 , Yan Li 2 , Lei Shi 1, 5 1 Institute of Cancer Stem Cell, Cancer Center, Dalian Medical University, Dalian, 116044, Liaoning, P.R.China 2 College of Basic Medical Sciences, Dalian Medical University, Dalian, 116044 Liaoning, P.R.China 3 Department of Gynecology and Obstetrics, Dalian Municipal Central Hospital Affiliated to Dalian Medical University, Dalian, 116033, Liaoning, P.R.China 4 The First Affiliated Hospital of Dalian Medical University, Dalian, 116011, Liaoning, P.R.China 5 State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, P.R.China * These authors contributed equally to this work Correspondence to: Lei Shi, email: powerleishi@126.com Yan Li, email: ly316@126.com Lingfei Ma, email: malingfei@hotmail.com Keywords: septin, forchlorfenuron, breast cancer, invasion, MAPK Received: April 24, 2016 Accepted: July 28, 2016 Published: August 19, 2016 ABSTRACT Septins are a novel class of GTP-binding cytoskeletal proteins evolutionarily conserved from yeast to mammals and have now been found to play a contributing role in a broad range of tumor types. However, their functional importance in breast cancer remains largely unclear. Here, we demonstrated that pharmaceutical inhibition of global septin dynamics would greatly suppress proliferation, migration and invasiveness in breast cancer cell lines. We then examined the expression and subcellular distribution of the selected septins SEPT2 and SEPT7 in breast cancer cells, revealing a rather variable localization of the two proteins with cell cycle progression. To determine the role of both septins in mediating malignant behavior of cancer cells, we used RNA silencing to specifically deplete endogenous SEPT2 or SEPT7 in highly invasive breast cancer cell line MDA-MB-231. Our findings showed that SEPT2/7 depletion had virtually identical inhibitory effects on cellular proliferation, apoptosis, migration and invasion. Moreover, the opposite performance in migration and invasion was observed after enforced expression of SEPT2/7 in the same cell line. We further demonstrated MEK/ERK activation, but not other MAPKs and AKT, was positively correlated with the protein levels of SEPT2 and SEPT7. Additionally, in SEPT2/7-overexpressing cells, the MEK specific inhibitor U0126 was able to correct the high active status of MEK/ERK while normalizing the increased invasive behaviors of these cells. Taken together, these results strongly suggest that SEPT2 and SEPT7 are involved in breast carcinogenesis and may serve as valuable therapeutic targets for breast cancer.

Jatropha curcas L. as a bio energy plant belonging to the Euphorbiaceae family is gaining progressive importance over the last years. In 2012 and 2014 field experiments were carried out to assess the effects of cytokinins 6-Benzyladenine (BA) and Forchlorfenuron (CPPU) acting as plant growth regulators (PGRs). Number of flowers per inflorescence, female-to-male ratio of flowers, fruits per infructescence, fruiting rate, number of seeds per fruit, seed size and weight as well as seed oil content were determined. It was suggested that concerning effectiveness of exogenous application of PGRs the developmental stage of flower is very important. Due to that, special interest was laid on the age of inflorescences at the time of application. Our experiments revealed a strong dependence of cytokinin effectiveness on the developmental stage of flowering. So treatment of plants with 6-Benzyladenine at the beginning of flower formation (stage 1) significantly increased the number of male and female flowers per inflorescence, whereas treatment at an advanced flowering stage (stage 2) or at the stage of fully developed flowers (stage 3) had only slight or no effects. In contrast, fruit retention was progressively increased by treatment in stage 2 and 3. Application of Forchlorfenuron significantly increased female-to-male ratio in stage 1 flowers but showed no effects on stage 2 and 3. 6-Benzyladenine as well as Forchlorfenuron showed equal effects on number of fruit inflorescences treated in stage 1. Our results show a significant decrease in seed weight due to BA- and CPPU-application while kernel weight remained stable. Concerning fruits, clusters and oil yield per tree, BA-application showed significant increasing effects. This study indicates that 6-Benzyladenine and Forchlorfenuron can improve yield affecting parameters in J. curcas when the phenological stage of flowering at time of application is precisely considered.

To investigate the feasibility of using hyperspectral imaging technology to discriminate kiwifruits treated with forchlorfenuron at different concentrations, “Hayward” kiwifruits were treated with forchlorfenuron solutions at concentrations of 2.5, 5.0, 7.5, 10.0, and 12.5 mg/L at 15 days after petal fall. Hyperspectral images of 400 samples, including 100 untreated kiwifruits and 60 treated kiwifruits at each forchlorfenuron concentration, were acquired. Mask was done for each image and mean spectrum was calculated for analysis. The original spectra were preprocessed by standard normal variate and all samples were divided into calibration set and prediction set based on Duplex algorithm according to the ratio of 2:1. The calibration set included 67 untreated kiwifruits and 200 treated kiwifruits, including 40 kiwifruits at each forchlorfenuron concentration. Other 133 kiwifruits were used as the prediction set. Twenty-nine characteristic wavelengths were selected from the full spectra from 928.19 to 1,658.19 nm using successive projection algorithm (SPA). Four discrimination models, i.e., partial least squares discriminant analysis, support vector machine, extreme learning machine, and random forests were established with the full spectra and the selected characteristic wavelengths as inputs, respectively. The results showed that the best model, with 97.7 % accuracy rate, for “Hayward” kiwifruits treated with forchlorfenuron was SVM established using selected characteristic wavelengths by SPA. This study indicates that hyperspectral imaging technology can be used to nondestructively and accurately discriminate forchlorfenuron-treated kiwifruits with different concentrations from untreated ones.

Six plant growth regulators frequently used in fruit were detected using UPLC–MS/MS including chlormequat chloride, GA3, forchlorfenuron, paclobutrazol, thidiazuron and 2,4-D. A modified QuEChERS method was used in pretreatment: each compound was extracted with acetonitrile containing 1% acetic acid and cleaned with a mixture of ostade-cylsilane (ODS) and MgSO4. The detection was performed using electrospray ionization (ESI) and multiple reaction monitoring mode (MRM) and quantified using the external standard method. Results indicated that the calibration curves of six plant growth regulators were linear within the range of 1–100 µg⋅kg-1 with correlation coefficients over 0.999. The recovery at four spiked concentrations, 1, 20, 50, and 100 µg⋅kg-1, ranged from 60.77% to 119.43% and relative standard deviation (RSD) were between 0.18% and 32.94%. The limits of detection (LOD) and limits of quantitation (LOQ) were 0.01–0.14 µg⋅kg-1 and 0.02–0.46 µg⋅kg-1, respectively, for apples, 0.01–0.18 µg⋅kg-1 and 0.03–0.59 µg⋅kg-1 for grapes, 0.01–0.08 µg⋅kg-1 and 0.04–0.28 µg⋅kg-1 for kiwis, 0.003–0.110 µg⋅kg-1 and 0.01–0.38 µg⋅kg-1 for peaches, and 0.01–0.08 µg⋅kg-1 and 0.03–0.25 µg⋅kg-1 for oranges.

Fruit drop is a major problem in mango. In order to reduce fruit drop CPPU (Forchlorfenuron) is widely used in fruit industry. To increase fruit retention trial were conducted to evaluate the effect of CPPU (Forchlorfenuron) on fruit retention and postharvest quality of 'Alphonso' mango. The present investigation was carried out in three locations in Konkan region in RBD with five treatments and 4 replications. CPPU was sprayed at a dose of 1, 2, 3, 4 ppm at the peanut and marble stage and water as control. It was observed that CPPU sprayed trees showed significantly more fruit retention and total number of fruit at harvest as compared to the control treatment. The highest yield was recorded in 3 ppm CPPU treatment at Dapoli and Roha and 4 ppm treatment at Rameshwar location. There were no significant effects on quality of fruit of CPPU trees and control fruit.