Identification of superior reference genes for data normalisation of expression studies via quantitative PCR in hybrid roses (Rosa hybrida)

Background: Gene expression studies are a prerequisite for understanding the biological function of genes. Because of its high sensitivity and easy use, quantitative PCR (qPCR) has become the gold standard for gene expression quantification. To normalise qPCR measurements between samples, the most prominent technique is the use of stably expressed endogenous control genes, the so called reference genes. However, recent studies show there is no universal reference gene for all biological questions. Roses are important ornamental plants for which there has been no evaluation of useful reference genes for gene expression studies. Results: We used three different algorithms (BestKeeper, geNorm and NormFinder) to validate the expression stability of nine candidate reference genes in different rose tissues from three different genotypes of Rosa hybrida and in leaves treated with various stress factors. The candidate genes comprised the classical "housekeeping genes" (Actin, EF-1α, GAPDH, Tubulin and Ubiquitin), and genes showing stable expression in studies in Arabidopsis (PP2A, SAND, TIP and UBC). The programs identified no single gene that showed stable expression under all of the conditions tested, and the individual rankings of the genes differed between the algorithms. Nevertheless the new candidate genes, specifically, PP2A and UBC, were ranked higher as compared to the other traditional reference genes. In general, Tubulin showed the most variable expression and should be avoided as a reference gene. Conclusions: Reference genes evaluated as suitable in experiments with Arabidopsis thaliana were stably expressed in roses under various experimental conditions. In most cases, these genes outperformed conventional reference genes, such as EF1-α and Tubulin. We identified PP2A, SAND and UBC as suitable reference genes, which in different combinations may be used for normalisation in expression analyses via qPCR for different rose tissues and stress treatments. However, the vast genetic variation found within the genus Rosa, including differences in ploidy levels, might also influence expression stability of reference genes, so that future research should also consider different genotypes and ploidy levels.

Efficacy of nano-silver in alleviating bacteria-related blockage in cut rose cv. Movie Star stems

Postharvest treatments with nano-silver (NS) significantly improve water relations and therefore prolong the vase life of several cut flowers, including rose (Rosa hybrida cv. Movie Star). The efficacy of NS in alleviating bacterial related blockage in the stem-ends of cut cv. Movie Star was further investigated. Four dominant bacteria strains Pseudomonas fluorescens, Aeromonas sp., Comamonas acidovorans and Chryseomonas luteola were isolated from the stem-ends of cut roses. High numbers of the isolated bacteria at 10 8colony forming unitsmL -1 vase solution led to a sharp reduction in vase life, flower fresh weight, and water uptake. In vitro assessments of the antibacterial activity of NS against the four bacterial strains was >80% at 5mgL -1 and nearly 100% at 50mgL -1. Bacterial blockage in the stem-ends of cut cv. Movie Star roses with and without NS pulse treatments was assessed during the vase period using scanning electron microscopy. Following a 50mgL -1 NS pulse treatment, there were few bacterial cells on the cut surface of the stems even on day 7. Moreover, no obvious bacterial blockage was observed inside the xylem vessels. In contrast, the cut surface of control stems was covered with bacteria and associated amorphous substances, and numerous bacteria were found in the xylem vessels. © 2012 Elsevier B.V.

Diagnóstico de la fertilidad en rosas para corte mediante análisis foliares

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Gibberellic acid, ozone and 1-methylcyclopropene on the gray mold control in avant garde rose

Gray mold caused by Botrytis cinerea is considered the major disease of greenhouse grown flowers. The goal of this study was to evaluate the effects of gibberellic acid (GA3), ozone, and 1-MCP, applied on postharvest, on the gray mold control in 'Avant Garde' rose. Rose flowers were artificially inoculated with B. cinerea (104 conidia ml-1) and non-inoculated. After treatments, roses were stored under room conditions (20±2°C/80±5% RH) and checked for gray mold incidence and severity. Spraying of GA3 at 25, 50, and 75 mg L-1 on non-inoculated roses reduced the area under the disease progress curve (AUDPC) of gray mold incidence in 41, 40 and 54%, respectively. Continuous application of ozone at 2.7 ppm reduced 14-folds B. cinerea sporulation. On the other hand, 1-MCP did not control gray mould in rose. These results showed that GA3 sprays and ozone contribute to postharvest control of gray mold in cut rose and can be utilized on integrated disease management.

Effects of vase solution pH and abscisic acid on the longevity of cut 'Baccara' roses

Abscisic acid (ABA) supplied in the vase solution can induce stomatal closure in the leaves of cut flowers, including roses (Rosa hybrida L.). This effect may be beneficial in reducing water deficit stress. Extracellular pH can affect active ABA concentrations in the apoplast of guard cells, with sap alkalisation enhancing the physiological activity of ABA. Accordingly, it was hypothesized that vase solution pH may affect ABA-mediated stomatal closure of cut roses. Two experiments were conducted to study the interaction of vase solution pH and ABA. In the first, cut 'Baccara' roses were held in vase solutions with +/- 10(-5) M ABA at pH 6, pH 7 and pH 8. In the second experiment, roses were held with +/- 10(-5) M ABA at pH 6 and pH 8 in the presence and absence of 1 mg l(-1) AgNO3 as a bactericide. Supply of ABA increased vase life and reduced vase solution usage of flowers held in low pH 6 solutions, indicating induction of stomatal closure. Conversely, ABA supplied at pH 8 was associated with reduced vase life. This negative result was associated with enhanced development of vase solution microbes at high pH, which overrode any potential pH-mediated ABA efficacy effects.

Effect of seasonal variation and storage temperature on leaf chlorophyll fluorescence and vase life of cut roses

Low temperature injury (LTI) of roses (Rosa hybrida L.) is difficult to assess by visual observation. Relative chlorophyll fluorescence (CF; F-v/F-m) is a non-invasive technique that provides an index of stress effects on photosystem 11 (PS 11) activity. This instrumental technique allows determination of the photosynthetic efficiency of plant tissues containing chloroplasts, such as rose leaves. In the present study, pre- and Post-Storage measurements of F-v/F-m were carried out to assess LTI in 'First Red' and 'Akito' roses harvested year round. Relationships between the pre-harvest environment conditions of temperature, relative humidity and photon flux density (PFD), F-v/F-m, and, vase life duration after storage are reported. After harvest, roses were stored at 1, 5 and 10 degrees C for 10 days. Non-stored roses were the control treatment. F-v/F-m ratios were reduced following storage, suggesting LTI of roses. However, reductions in F-v/F-m were not closely correlated with reduced vase life duration and were seasonally dependent. Only during winter experiments was F-v/F-m of roses stored at 1 degrees C significantly (P <= 0.001) lower compared to F-v/F-m of non-stored control roses and roses stored at 5 and 10 degrees C. Thus, the fall of F-v/F-m was due to an interaction of growing season and storage at 1 degrees C. Vase lives of roses grown during winter were significantly (P <= 0.001) shorter compared to roses grown during summer. Length of vase life was intermediate for roses grown during autumn and spring. Because of the lack of correlation between F-v/F-m and post-storage vase life it is concluded that the CF parameter F-v/F-m is nota practical index for assessing LTI in cold-stored roses. Higher PFD and temperature in summer were positively and significantly correlated with maintenance of post-storage FvIF ratios and longer vase life. It is suggested that shorter vase lives and lower post-storage F-v/F-m values after storage at 1 degrees C are consequences of reduced photosynthesis and smaller carbohydrate pools in winter-harvested roses. (c) 2004 Elsevier B.V All rights reserved.

Fotocontrol de la productividad y elongación de tallos de tres cultivares de Rosa x hybrida L. bajo cubiertas de polietileno fotoselectivas

Se evaluaron los cambios en la radiación transmitida a través de films fotoselectivos (FS) fluorescentes y el impacto de estos cambios sobre la producción de tres cultivares de rosa para corte Fuego Negro, Maroussia y Anna. Se observó que los FS nuevos o expuestos a la radiación solar vs. noFS disminuyen la transmisión de radiación azul (A) (-28,4 a -32,9%, respectivamente), incrementan el R produciendo una relación R:RL mayor (+3,6%), si bien transmiten algo menos de radiación fotosintéticamente activa que los noFS nuevos. El número de rosas producidas fue significativamente mayor bajo el FS vs. noFS en los tres cv (+24, +32 y +36% en Anna, Fuego Negro y Maroussia, respectivamente), con un peso fresco y seco (PF y PS) significativamente mayor y tallos florales más largos en Anna y Maroussia (50,69 y 43,91 cm vs. 38,91 y 40,04 cm en invierno y primavera, respectivamente), y pimpollos significativamente más largos y de mayor PF y PS en los tres cv. Mayor relación R:RL y menor UV-A y A en la radiación transmitida por films FS aumentaron la cantidad y calidad de determinados cultivares de rosas mostrando una alternativa a los reguladores químicos de crecimiento.

Sodium hypochlorite sterilization of culture medium in micropropagation of Gerbera hybrida cv. Essandre.

Micropropagation requires controlling contamination that might compromise the success of the process. Thermal sterilization is traditionally used; however, costs deriving from equipment acquisition and maintenance render this technique costly. With the purpose of finding an alternative to thermal sterilization, this research aimed at assessing the efficiency and ideal concentration of sodium hypochlorite for sterilization of culture media and glassware used during rooting of micropropagated Gerbera hybrida cv. Essandre. Two experiments were carried out. In the first one, treatments consisted of control I (no sterilization), control II (thermal sterilization), and total active chlorine concentrations of 0.0005, 0.001, 0.002 and 0.003%. In the second experiment, based on the results observed in the first experiment, treatments consisted of control I (thermal sterilization) and II (chemical sterilization), and total active chlorine concentrations of 0.002, 0.0025 and 0.003%. Plant behavior was assessed based on the length of aerial part and roots, number of roots, and dry biomass of plants. Results showed that the addition of an active chlorine concentration of 0.003% to culture media provided total control of contaminants, and there were no significant differences regarding the variables analyzed between plants obtained with thermal sterilization and with sodium hypochlorite sterilization. Thus, chemical sterilization can be used as a replacement for thermal sterilization of nutrition media for rooting of gerbera in vitro.

Family portrait of siblings Wladimir, Rosa, Lena and Georg Eliasberg; New York Portraits Family

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Family portrait of siblings Wladimir, Rosa, Lena and Georg Eliasberg; New York Portraits Family

Digital Image