The F-box protein RhSAF destabilizes the gibberellic acid receptor RhGID1 to mediate ethylene-induced petal senescence in rose.

Roses are among the most popular ornamental plants cultivated worldwide for their great economic, symbolic, and cultural importance. Nevertheless, rapid petal senescence markedly reduces rose (Rosa hybrida) flower quality and value. Petal senescence is a developmental process tightly regulated by various phytohormones. Ethylene accelerates petal senescence, while gibberellic acid (GA) delays this process. However, the molecular mechanisms underlying the crosstalk between these phytohormones in the regulation of petal senescence remain largely unclear. Here, we identified SENESCENCE-ASSOCIATED F-BOX (RhSAF), an ethylene-induced F-box protein gene encoding a recognition subunit of the SCF-type E3 ligase. We demonstrated that RhSAF promotes degradation of the GA receptor GIBBERELLIN INSENSITIVE DWARF1 (RhGID1) to accelerate petal senescence. Silencing RhSAF expression delays petal senescence, while suppressing RhGID1 expression accelerates petal senescence. RhSAF physically interacts with RhGID1s and targets them for ubiquitin/26S proteasome-mediated degradation. Accordingly, ethylene-induced RhGID1C degradation and RhDELLA3 accumulation are compromised in RhSAF-RNAi lines. Our results demonstrate that ethylene antagonizes GA activity through RhGID1 degradation mediated by the E3 ligase RhSAF. These findings enhance our understanding of the phytohormone crosstalk regulating petal senescence and provide insights for improving flower longevity.

Phytosulfokine α (PSKα) delays senescence and reinforces SUMO1/SUMO E3 ligase SIZ1 signaling pathway in cut rose flowers (Rosa hybrida cv. Angelina).

Roses are widely used as cut flowers worldwide. Petal senescence confines the decorative quality of cut rose flowers, an impressively considerable economic loss. Herein, we investigated the SUMO1/SUMO E3 ligase SIZ1 signaling pathway during bud opening, and petal senescence of cut rose flowers. Our results exhibited that the higher expression of SUMO1 and SUMO E3 ligase SIZ1 during bud opening was accompanied by lower endogenous H2O2 accumulation arising from higher expression and activities of SOD, CAT, APX, and GR, promoting proline accumulation by increasing P5CS expression and activity and enhancing GABA accumulation by increasing GAD expression and activity. In harvested flowers, lower expressions of SUMO1 and SUMO E3 ligase SIZ1 during petal senescence were associated with higher endogenous H2O2 accumulation due to lower expression and activities of SOD, CAT, APX, and GR. Therefore, promoting the activity of the GABA shunt pathway as realized by higher expression and activities of GABA-T and SSADH accompanied by increasing OAT expression and activity for sufficiently supply proline in rose flowers during petal senescence might serve as an endogenous antisenescence mechanism for slowing down petals senescence by avoiding endogenous H2O2 accumulation. Following phytosulfokine α (PSKα) application, postponing petal senescence in cut rose flowers could be ascribed to higher expression of SUMO1 and SUMO E3 ligase SIZ1 accompanied by higher expression and activities of SOD, CAT, APX, and GR, higher activity of GABA shunt pathway as realized by higher expression and activities of GAD, GABA-T, and SSADH, higher expression and activities of P5CS and OAT for supplying proline and higher expression of HSP70 and HSP90. Therefore, our results highlight the potential of the PSKα as a promising antisenescence signaling peptide in the floriculture industry for postponing senescence and extending the vase life of cut rose flowers.

Ethylene-regulated asymmetric growth of the petal base promotes flower opening in rose (Rosa hybrida).

Flowers are the core reproductive structures and key distinguishing features of angiosperms. Flower opening to expose stamens and gynoecia is important in cases where pollinators much be attracted to promote cross-pollination, which can enhance reproductive success and species preservation. The floral opening process is accompanied by the coordinated movement of various floral organs, particularly petals. However, the mechanisms underlying petal movement and flower opening are not well understood. Here, we integrated anatomical, physiological, and molecular approaches to determine the petal movement regulatory network using rose (Rosa hybrida) as a model. We found that PETAL MOVEMENT-RELATED PROTEIN1 (RhPMP1), a homeodomain transcription factor (TF) gene, is a direct target of ETHYLENE INSENSITIVE3, a TF that functions downstream of ethylene signaling. RhPMP1 expression was upregulated by ethylene and specifically activated endoreduplication of parenchyma cells on the adaxial side of the petal (ADSP) base by inducing the expression of RhAPC3b, a gene encoding the core subunit of the Anaphase-Promoting Complex. Cell expansion of the parenchyma on the ADSP base was subsequently enhanced, thus resulting in asymmetric growth of the petal base, leading to the typical epinastic movement of petals and flower opening. These findings provide insights into the pathway regulating petal movement and associated flower-opening mechanisms.�.

Auxin Regulates Sucrose Transport to Repress Petal Abscission in Rose (Rosa hybrida).

Developmental transitions in plants require adequate carbon resources, and organ abscission often occurs due to competition for carbohydrates/assimilates. Physiological studies have indicated that organ abscission may be activated by Suc deprivation; however, an underlying regulatory mechanism that links Suc transport to organ shedding has yet to be identified. Here, we report that transport of Suc and the phytohormone auxin to petals through the phloem of the abscission zone (AZ) decreases during petal abscission in rose (Rosa hybrida), and that auxin regulates Suc transport into the petals. Expression of the Suc transporter RhSUC2 decreased in the AZ during rose petal abscission. Similarly, silencing of RhSUC2 reduced the Suc content in the petals and promotes petal abscission. We established that the auxin signaling protein RhARF7 binds to the promoter of RhSUC2, and that silencing of RhARF7 reduces petal Suc contents and promotes petal abscission. Overexpression of RhSUC2 in the petal AZ restored accelerated petal abscission caused by RhARF7 silencing. Moreover, treatment of rose petals with auxin and Suc delayed ethylene-induced abscission, whereas silencing of RhARF7 and RhSUC2 accelerated ethylene-induced petal abscission. Our results demonstrate that auxin modulates Suc transport during petal abscission, and that this process is regulated by a RhARF7-RhSUC2 module in the AZ.

Sugar availability suppresses the auxin-induced strigolactone pathway to promote bud outgrowth.

Apical dominance occurs when the growing shoot tip inhibits the outgrowth of axillary buds. Apically-derived auxin in the nodal stem indirectly inhibits bud outgrowth via cytokinins and strigolactones. Recently, sugar deprivation was found to contribute to this phenomenon. Using rose and pea, we investigated whether sugar availability interacts with auxin in bud outgrowth control, and the role of cytokinins and strigolactones, in vitro and in planta. We show that sucrose antagonises auxin's effect on bud outgrowth, in a dose-dependent and coupled manner. Sucrose also suppresses strigolactone inhibition of outgrowth and the rms3 strigolactone-perception mutant is less affected by reducing sucrose supply. However, sucrose does not interfere with the regulation of cytokinin levels by auxin and stimulates outgrowth even with optimal cytokinin supply. These observations were assembled into a computational model in which sucrose represses bud response to strigolactones, largely independently of cytokinin levels. It quantitatively captures our observed dose-dependent sucrose-hormones effects on bud outgrowth and allows us to express outgrowth response to various combinations of auxin and sucrose levels as a simple quantitative law. This study places sugars in the bud outgrowth regulatory network and paves the way for a better understanding of branching plasticity in response to environmental and genotypic factors.

Hydrogen gas alleviates postharvest senescence of cut rose 'Movie star' by antagonizing ethylene.

KEY MESSAGE: H2 prolonged the vase life and improved the vase quality of cut roses through repressing endogenous ethylene production and alleviating ethylene signal transduction during the entire senescing period. Recently, the application of hydrogen gas (H2) was shown to improve postharvest quality and longevity in perishable horticultural products, but the specific regulation mechanism remains obscure. Here, endogenous ethylene production and the expression of genes in ethylene biosynthesis and signalling pathway were investigated to explore the crosstalk between H2 and ethylene during the senescence of cut roses. Our results revealed that addition of exogenous ethylene by ethephon accelerated the senescence of cut roses, in which 100 mg L-1 ethephon displayed the most obvious senescent phenotype. While the applied different concentrations (1%, 10%, 50% and 100%) of hydrogen-rich water (HRW) conducted different affects in alleviating the senescence of cut roses, and 1% HRW displayed the best ornamental quality and the longest vase life by reducing ethylene production, supported by the decrease of 1-aminocyclopropene-1-carboxylate (ACC) accumulation, ACC synthase (ACS) and ACC oxidase (ACO) activities, and Rh-ACS3 and Rh-ACO1 expressions in ethylene biosynthesis. In addition, HRW increased the transcripts of ethylene receptor genes Rh-ETR1 at blooming period from day 4 to day 6 and suppressed Rh-ETR3 at senescence phase at day 8 after harvest. Furthermore, the relevant affection of HRW on Rh-ETR1 and Rh-ETR3 expressions still existed when the ethylene production was compromised by adequate addition of exogenous ethylene in HRW-treated cut rose petals, and HRW directly repressed the protein level of Rh-ETR3 in a transient expression assay. Overall, the results suggested that H2 is involved in neutralizing ethylene-mediated postharvest in cut flowers.

Response of Roses (Rosa hybrida L. 'Herbert Stevens') to Foliar Application of Polyamines on Root Development, Flowering, Photosynthetic Pigments, Antioxidant Enzymes Activity and NPK.

The effect of foliar application of polyamines on roses (Rosa hybrida cv. 'Herbert Stevens') was investigated in a factorial experiment based on a completely randomized design with three replications in a greenhouse. Two factors were applied including polyamine type (putrescine, spermidine, and spermine) and polyamine concentration (0, 1, 2 and 4 mM). The recorded traits included root fresh and dry weight, root length, number of flowers, flower longevity, chlorophyll content, carotenoids, antioxidant enzymes activity (catalase, ascorbate peroxidase and guaiacol peroxidase) and some macronutrients such as nitrogen, phosphorus and potassium. The results showed that among polyamines, putrescine had the greatest effect on root dry weight; spermidine showed the greatest effect on root length, chlorophyll content, plant phosphorus and spermine affected root fresh weight and flower longevity most strongly. Polyamine concentration of 1 mM had the strongest effect on flower longevity, carotenoids, nitrogen and phosphorus content. The highest potassium rate was observed in treatments with the concentration of 4 mM. Polyamine treatments had no significant effect on the number of flowers per plant and antioxidant enzymes.

RhERF113 Functions in Ethylene-Induced Petal Senescence by Modulating Cytokinin Content in Rose.

In rose (Rosa hybrida), flower senescence is accelerated by ethylene and delayed by cytokinins (CTKs). However, the effectors that regulate these processes are not currently understood. In this study, we identified an APETALA2/ethylene-responsive factor (AP2/ERF) gene, RhERF113, which was induced by ethylene and up-regulated during flower senescence in most floral organs, including sepal, petal, stamen and pistil. The virus-induced gene silencing (VIGS) of RhERF113 expression accelerated rose flower senescence, which was accompanied by a lower CTK content in the flowers. This accelerated senescence could be restored by exogenous CTK treatment. Moreover, the expression levels of genes related to CTK biosynthesis and signaling, including ISOPENTENYL TRANSFERASE 5 (RhIPT5), RhIPT8, HISTIDINE KINASE 2 (RhHK2), RhHK3, CYTOKININ RESPONSE REGULATOR 3 (RhCRR3), RhCRR5, RhCRR8, HOMEOBOX PROTEIN 6 (RhHB6) and PATHOGENESIS-RELATED 10.1 (RhPR10.1), were decreased in the RhERF113-silenced rose flowers. Taken together, our results demonstrate that RhERF113 delays ethylene-induced flower senescence by increasing the CTK content of the floral tissues.

Genome-wide transcriptome analysis of the salt stress tolerance mechanism in Rosa chinensis.

Plants regulate responses to salt stress using biological pathways, such as signal perception and transduction, photosynthesis, and energy metabolism. Little is known about the genetics of salt tolerance in Rosa chinensis. Tineke and Hiogi are salt-tolerant and salt-sensitive varieties of R. chinensis, respectively, and are good choices for studying salt-tolerance genes. We studied leaf and root tissues from 1-year-old Hiogi and Tineke plants simultaneously grown under the same conditions. A 0.4%-mmol/L salt ion mixture was added to the basic growth medium. Illumina sequencing was used to identify differentially expressed transcripts. GO and KEGG pathway enrichment analyses were performed to identify differentially expressed genes. We identified many differentially expressed genes associated with salt tolerance. The abscisic acid-dependent signaling pathway was the main pathway that mediated the salt stress response in R. chinensis. Two pathways (plant hormone signal transduction and glutathione metabolism) were also active in salt stress responses in R. chinensis. The difference in salt tolerance in the cultivars was due to different gene sensitivity to salt in these two pathways. Roots also play a role in salt stress response. The effects of salt stress in the roots are eventually manifested in the leaves, causing changes in processes such as photosynthesis, which eventually result in leaf wilting. In Tineke, Snrk2, ABF, HSP, GSTs, and GSH1 showed high activity during salt stress, indicating that these genes are markers of salt tolerance.

Heavy metal accumulation imparts structural differences in fragrant Rosa species irrigated with marginal quality water.

Wastewater is an alternative to traditional sources of renewable irrigation water in agriculture, particularly in water-scarce regions. However, the possible risks due to heavy metals accumulation in plant tissues are often overlooked by producers. The present study aimed to identify heavy metals-induced structural modifications to roots of scented Rosa species that were irrigated with water of marginal quality. The chemical and mineral contents from the experimental irrigation canal water (control) and treated wastewater were below the limits recommended by the Pakistan Environmental Protection Agency (Pak-EPA) for medicinal plants. The experimentally untreated wastewater contained electrical conductivity (EC), chemical oxygen demand (COD), biological oxygen demand (BOD), and heavy metals (Co, Cu, Cd, Pb) that were above the recommended limits. The responses by wastewater-treated Rosa species (Rosa damascena, R. bourboniana, R. Gruss-an-Teplitz, and R. centifolia) were evaluated. The experimental data revealed that treated wastewater significantly increased the thickness of collenchyma (cortex and pith) and parenchyma tissues (vascular bundle, xylem, and phloem) of R. Gruss-an-Teplitz. Root dermal tissues (epidermis) of R. bourboniana also responded to treated wastewater. R. damascena and R. centifolia were the least affected species, under the experimental irrigation conditions. Collenchyma and dermal tissues were thicker in R. damascena and R. Gruss-an-Teplitz under untreated wastewater conditions. In parenchyma tissues, vascular bundles were thicker in R. damascena in untreated wastewater conditions, while the xylem and phloem of R. Gruss-an-Teplitz were thicker where treated wastewater was applied. In tissues other than the vascular bundle, the differences in anatomical metrics due to the experimental irrigation treatments were greater during the second year of the experiment than in the first year. The contents of metals other than chromium in the roots and stems of roses were below the WHO limits, under all of the experimental irrigation conditions. Rosa centifolia contained higher heavy metals content than the other experimental species, and heavy metals content was associated with anatomical changes due to the treatments. We conclude that, under conditions of wastewater irrigation, R. Gruss-an-Teplitz was highly resistant; R. damascena was moderately resistant while R. bourboniana and R. centifolia were the most susceptible to irrigation with marginal quality water. This is the first report of plant tissue responses to wastewater irrigation by the experimental species. Regarding the accumulation of heavy metals in rose plant tissues, the results confirm that untreated wastewater must be treated to grow Rosa species where water is scarce.

Hibiscus rosa sinensis mediate anxiolytic effect via modulation of ionotropic GABA-A receptors: possible mechanism of action.

The current study was designed with the aim to investigate anti-anxiety potential of Hibiscus rosa sinensis roots and its possible mechanism of action. For this purpose hole board test, elevated plus maze test and light/dark exploration test were employed. The ethanol extract of plant was administered orally at two different doses i.e. 100 and 500 mg/kg for consecutive 14 days. The results of present investigation indicate that plant extract significantly (p < 0.05) increased the number of head dips and rearings as compared to control on respective days of observation. The extract increased the time of permanence in open arms and the number of head dips in elevated plus maze. In light/dark test, our study indicate that Hibiscus rosa sinensis significantly (p < 0.05) increased the time spent in light compartment and number of entries as compared to control. In addition the anxiolytic effects of HRS at highest tested dose were blocked by flumazenil, a GABA-A receptor antagonist that indicate that Hibiscus rosa sinensis potentiated the GABAergic actions. The results propose that the ethanol extract of Hibiscus rosa sinensis has prospective anxiolytic effect in mice via inhibition of ionotropic GABA receptors, using different behavioral paradigms.

Leaf Physiological and Proteomic Analysis to Elucidate Silicon Induced Adaptive Response under Salt Stress in Rosa hybrida 'Rock Fire'.

Beneficial effects of silicon (Si) on growth and development have been witnessed in several plants. Nevertheless, studies on roses are merely reported. Therefore, the present investigation was carried out to illustrate the impact of Si on photosynthesis, antioxidant defense and leaf proteome of rose under salinity stress. In vitro-grown, acclimatized Rosa hybrida 'Rock Fire' were hydroponically treated with four treatments, such as control, Si (1.8 mM), NaCl (50 mM), and Si+NaCl. After 15 days, the consequences of salinity stress and the response of Si addition were analyzed. Scorching of leaf edges and stomatal damages occurred due to salt stress was ameliorated under Si supplementation. Similarly, reduction of gas exchange, photosynthetic pigments, higher lipid peroxidation rate, and accumulation of reactive oxygen species under salinity stress were mitigated in Si treatment. Lesser oxidative stress observed was correlated with the enhanced activity and expression of antioxidant enzymes, such as superoxide dismutase, catalase, and ascorbate peroxidase in Si+NaCl treatment. Importantly, sodium transportation was synergistically restricted with the stimulated counter-uptake of potassium in Si+NaCl treatment. Furthermore, two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) results showed that out of 40 identified proteins, on comparison with control 34 proteins were down-accumulated and six proteins were up-accumulated due to salinity stress. Meanwhile, addition of Si with NaCl treatment enhanced the abundance of 30 proteins and downregulated five proteins. Differentially-expressed proteins were functionally classified into six groups, such as photosynthesis (22%), carbohydrate/energy metabolism (20%), transcription/translation (20%), stress/redox homeostasis (12%), ion binding (13%), and ubiquitination (8%). Hence, the findings reported in this work could facilitate a deeper understanding on potential mechanism(s) adapted by rose due to the exogenous Si supplementation during the salinity stress.

In silico analysis of 3 expansin gene promoters reveals 2 hubs controlling light and cytokinins response during bud outgrowth.

Bud outgrowth is under the intricate control of environmental and endogenous factors. In a recent paper, 1 we demonstrated that light perceived by Rosa buds triggers cytokinins (CK) synthesis within 3 hours in the adjacent node followed by their transport to the bud. There, CK control expression of a set of major genes (strigolactones-, auxin-, sugar sink strength-, cells division and elongation-related genes) leading to bud outgrowth in light. Conversely, under dark condition, CK accumulation and transport to the bud are repressed and no bud outgrowth occurs. In this paper, we show that the 3 expansin genes RhEXPA1,2,3 are under the control of both light and CK during bud outgrowth. In silico analysis of promoter sequences highlights 2 regions enriched in light and CK cis-regulatory elements as well as a specific cis-element in pRhEXPA3, potentially responsible for the expression patterns observed in response to CK and light.

An Ethylene-Induced Regulatory Module Delays Flower Senescence by Regulating Cytokinin Content.

In many plant species, including rose (Rosa hybrida), flower senescence is promoted by the gaseous hormone ethylene and inhibited by the cytokinin (CTK) class of hormones. However, the molecular mechanisms underlying these antagonistic effects are not well understood. In this study, we characterized the association between a pathogenesis-related PR-10 family gene from rose (RhPR10.1) and the hormonal regulation of flower senescence. Quantitative reverse transcription PCR analysis showed that RhPR10.1 was expressed at high levels during senescence in different floral organs, including petal, sepal, receptacle, stamen, and pistil, and that expression was induced by ethylene treatment. Silencing of RhPR10.1 expression in rose plants by virus-induced gene silencing accelerated flower senescence, which was accompanied by a higher ion leakage rate in the petals, as well as increased expression of the senescence marker gene RhSAG12 CTK content and the expression of three CTK signaling pathway genes were reduced in RhPR10.1-silenced plants, and the accelerated rate of petal senescence that was apparent in the RhPR10.1-silenced plants was restored to normal levels by CTK treatment. Finally, RhHB6, a homeodomain-Leu zipper I transcription factor, was observed to bind to the RhPR10.1 promoter, and silencing of its expression also promoted flower senescence. Our results reveal an ethylene-induced RhHB6-RhPR10.1 regulatory module that functions as a brake of ethylene-promoted senescence through increasing the CTK content.

Overexpression of Rosa rugosa anthocyanidin reductase enhances tobacco tolerance to abiotic stress through increased ROS scavenging and modulation of ABA signaling.

Anthocyanidin reductase (ANR) is a key enzyme involved in the biosynthesis of proanthocyanidins (PAs) and plays a role in the plant stress response. However, the mechanism by which ANR confers stress tolerance in plants is not understood. Here, we report the isolation of RrANR, the homologous gene from rose, and NtABF, an ABA-response related transcription factor gene from tobacco. These genes were characterized regarding their functions in stress responses through the use of transgenic, transcriptomic and physiological analyses. Over-expression of RrANR in tobacco resulted in an increased accumulation of both PAs and abscisic acid (ABA), and also enhanced stress tolerance. Transcriptomic analysis of these transgenic tobacco lines indicated that RrANR overexpression induced global transcriptomic changes, including these involved in oxidation/reduction, hormone response and secondary metabolism. Genes related to ABA biosynthesis and reactive oxygen species (ROS)-scavenging were up-regulated in RrANR transgenic lines, and these effects were phenocopied by the direct treatment of tobacco plants with PAs and ABA. Transcriptomic data from each of these treatments identified the upregulation of a putative NtABF. Furthermore, the up-regulation of NtABF in RrANR transformants or in PAs- and ABA-treated tobacco plants was associated with enhanced stress tolerance. Overexpression of NtABF in transgenic tobacco mimicked the effects of RrANR-transgenic plants with regard to the up-regulation of ROS-scavenging genes and an increase in oxidative tolerance. Taken together, our findings indicate that overexpression of RrANR results in an increase in plant tolerance to oxidative stress via increased scavenging of ROS and modulation of the ABA signaling pathway.

Phenotypic- and Genotypic-Resistance Detection for Adaptive Resistance Management in Tetranychus urticae Koch.

Rapid resistance detection is necessary for the adaptive management of acaricide-resistant populations of Tetranychus urticae. Detection of phenotypic and genotypic resistance was conducted by employing residual contact vial bioassay (RCV) and quantitative sequencing (QS) methods, respectively. RCV was useful for detecting the acaricide resistance levels of T. urticae, particularly for on-site resistance detection; however, it was only applicable for rapid-acting acaricides (12 out of 19 tested acaricides). QS was effective for determining the frequencies of resistance alleles on a population basis, which corresponded to 12 nonsynonymous point mutations associated with target-site resistance to five types of acaricides [organophosphates (monocrotophos, pirimiphos-methyl, dimethoate and chlorpyrifos), pyrethroids (fenpropathrin and bifenthrin), abamectin, bifenazate and etoxazole]. Most field-collected mites exhibited high levels of multiple resistance, as determined by RCV and QS data, suggesting the seriousness of their current acaricide resistance status in rose cultivation areas in Korea. The correlation analyses revealed moderate to high levels of positive relationships between the resistance allele frequencies and the actual resistance levels in only five of the acaricides evaluated, which limits the general application of allele frequency as a direct indicator for estimating actual resistance levels. Nevertheless, the resistance allele frequency data alone allowed for the evaluation of the genetic resistance potential and background of test mite populations. The combined use of RCV and QS provides basic information on resistance levels, which is essential for choosing appropriate acaricides for the management of resistant T. urticae.

Effect of Mentha pulegium extract and 8-hydroxy quinoline sulphate to extend the quality and vase life of rose (Rosa hybrid) cut flower.

Rose is an ornamental plant which contains one of the world's top cut flowers. Vase life of cut rose flower is short. Extracts of Mentha pulegium and 8-hydroxy quinoline sulphate (8-HQS) were used as two preservative solutions, aiming to extend the vase life of cut rose (Rosa hybrid L.) flowers. Rose flowers were treated with a vase solution containing the extract of M. pulegium, at concentrations of 0, 10, 20 and 30%, in combination with 8-HQS at concentrations of 0, 200, 400 and 600 mg l(-1). Longevity of cut roses flowers was determined on the basis of wilting and chlorophyll retention. Cut roses flowers were kept at room temperature (20 ± 2 degrees C) under normal day light and natural ventilation. The vase life of cut flowers studied was prolonged by all 8-HQS and extract treatments. The best concentration of 8-HQS and extractwere 400 mg l(-1) and 10%, respectively. Our results indicated that the flowers treated with the extract and 8-HQS had longer vase life, higher rate of solution uptake and lower SPAD value (total chlorophyll) compared to the control. Also, cut flowers treated with the extract and 8-HQS had least bacterial colonies. The greatest longevity of vase life by 11.20 and 10.25 days was related to 400 mg I(-1) 8-HQS and 10% of extract, respectively. These treatments improved cut vase life more than the control treatment. The maximum solution uptake (1.85 ml g(-1) f.wt.) and minimum SPAD value (2.19) were calculated in 30% extract along with 200 mg l(-1) 8-HQS, and 200 mg l(-1) 8-HQS, respectively. The lowest number of bacterial colonies (55.75) was obtained in treatment of 600 mg l(-1) 8-HQS. Flower quality of specimens treated with extract and 8-HQS was better than those of the control. The experiments were repeated three times with three replicates and a completely randomized design had been used. The present study concludes that it would be possible to use preservative solutions containing extract of M. pulegium L. and 8-HQS to extend vase life of cut rose (R. hybrida L.) flowers.

Discovery of small molecule inhibitors of xyloglucan endotransglucosylase (XET) activity by high-throughput screening.

Small molecules (xenobiotics) that inhibit cell-wall-localised enzymes are valuable for elucidating the enzymes' biological roles. We applied a high-throughput fluorescent dot-blot screen to search for inhibitors of Petroselinum xyloglucan endotransglucosylase (XET) activity in vitro. Of 4216 xenobiotics tested, with cellulose-bound xyloglucan as donor-substrate, 18 inhibited XET activity and 18 promoted it (especially anthraquinones and flavonoids). No compounds promoted XET in quantitative assays with (cellulose-free) soluble xyloglucan as substrate, suggesting that promotion was dependent on enzyme-cellulose interactions. With cellulose-free xyloglucan as substrate, we found 22 XET-inhibitors - especially compounds that generate singlet oxygen ((1)O2) e.g., riboflavin (IC50 29 µM), retinoic acid, eosin (IC50 27 µM) and erythrosin (IC50 36 µM). The riboflavin effect was light-dependent, supporting (1)O2 involvement. Other inhibitors included tannins, sulphydryl reagents and triphenylmethanes. Some inhibitors (vulpinic acid and brilliant blue G) were relatively specific to XET, affecting only two or three, respectively, of nine other wall-enzyme activities tested; others [e.g. (-)-epigallocatechin gallate and riboflavin] were non-specific. In vivo, out of eight XET-inhibitors bioassayed, erythrosin (1 µM) inhibited cell expansion in Rosa and Zea cell-suspension cultures, and 40 µM mycophenolic acid and (-)-epigallocatechin gallate inhibited Zea culture growth. Our work showcases a general high-throughput strategy for discovering wall-enzyme inhibitors, some being plant growth inhibitors potentially valuable as physiological tools or herbicide leads.

Alternaria toxin-induced resistance in rose plants against rose aphid (Macrosiphum rosivorum): effect of tenuazonic acid.

Many different types of toxins are produced by the fungus, Alternaria alternata (Fr.) Keissler. Little is known, however, regarding the influence of these toxins on insects. In this study, we investigated the toxin-induced inhibitory effects of the toxin produced by A. alternata on the rose aphid, Macrosiphum rosivorum, when the toxin was applied to leaves of the rose, Rosa chinensis. The results demonstrated that the purified crude toxin was non-harmful to rose plants and rose aphids, but had an intensive inhibitory effect on the multiplication of aphids. The inhibitory index against rose aphids reached 87.99% when rose plants were sprayed with the toxin solution at a low concentration. Further results from bioassays with aphids and high performance liquid chromatography (HPLC) analyses demonstrated that tenuazonic acid (TeA) was one of the most important resistance-related active components in the crude toxin. The content of TeA was 0.1199% in the crude toxin under the HPLC method. Similar to the crude toxin, the inhibitory index of pure TeA reached 83.60% 15 d after the rose plants were sprayed with pure TeA solution at the lower concentration of 0.060 µg/ml, while the contents of residual TeA on the surface and in the inner portion of the rose plants were only 0.04 and 0.00 ng/g fresh weight of TeA-treated rose twigs, respectively, 7 d after the treatment. Our results show that TeA, an active component in the A. alternata toxin, can induce the indirect plant-mediated responses in rose plants to intensively enhance the plant's resistances against rose aphids, and the results are very helpful to understand the plant-mediated interaction between fungi and insects on their shared host plants.

RhHB1 mediates the antagonism of gibberellins to ABA and ethylene during rose (Rosa hybrida) petal senescence.

Rose (Rosa hybrida) is one of the most important ornamental plants worldwide; however, senescence of its petals terminates the ornamental value of the flower, resulting in major economic loss. It is known that the hormones abscisic acid (ABA) and ethylene promote petal senescence, while gibberellins (GAs) delay the process. However, the molecular mechanisms underlying the antagonistic effects amongst plant hormones during petal senescence are still unclear. Here we isolated RhHB1, a homeodomain-leucine zipper I transcription factor gene, from rose flowers. Quantitative RT-PCR and GUS reporter analyses showed that RhHB1 was strongly expressed in senescing petals, and its expression was induced by ABA or ethylene in petals. ABA or ethylene treatment clearly accelerated rose petal senescence, while application of the gibberellin GA3 delayed the process. However, silencing of RhHB1 delayed the ABA- or ethylene-mediated senescence, and resulted in higher petal anthocyanin levels and lower expression of RhSAG12. Moreover, treatment with paclobutrazol, an inhibitor of GA biosynthesis, repressed these delays. In addition, silencing of RhHB1 blocked the ABA- or ethylene-induced reduction in expression of the GA20 oxidase encoded by RhGA20ox1, a gene in the GA biosynthetic pathway. Furthermore, RhHB1 directly binds to the RhGA20ox1 promoter, and silencing of RhGA20ox1 promoted petal senescence. Eight senescence-related genes showed substantial differences in expression in petals after treatment with GA3 or paclobutrazol. These results suggest that RhHB1 mediates the antagonistic effect of GAs on ABA and ethylene during rose petal senescence, and that the promotion of petal senescence by ABA or ethylene operates through an RhHB1-RhGA20ox1 regulatory checkpoint.