Browsing by Subject "Tobacco"

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    Die Bedeutung von Aquaporinen und ihren Interaktionspartnern für die Zelltodregulation in Pflanzen
    (2011) Hoch, Tanja; Pfitzner, Artur J. P.
    Programmed cell death (PCD, apoptosis) is an induced cell suicide process that plays an important role during the differentiation and pathogen defense responses of plants and animals. BHRF1 (?BamHI fragment H rightward open reading frame no. 1?) is a cell-death modulating protein of the Epstein-Barr virus (EBV), a human lymphotrophic herpes virus. The expression of BHRF1 in transgenic plants led to the formation of necrotic lesions. Further experiments showed that BHRF1 associated necrotic lesions are caused due to stress, senescence and pathogen defense responses. Yeast-two-hybrid-screening of a tobacco cDNA library identified two different aquaporins as partners for interacting with BHRF1. Aquaporins were identified as water channels/carriers within red blood cells, but are also present in all other organisms. Over the last years, more information was gathered indicating that, apart from transporting water, aquaporins had other functional activities. E. g. Henzler and Steudle (2000) demonstrated that aquaporins can act as hydrogen peroxide channels in the algae Chara corallina. Furthermore, publications by Bienert et al. (2006), indicating that aquaporins in plants as well as in animals are also able to transport H2O2. Hydrogen peroxide and other reactive oxygen species (ROS) have long been recognized as important signal molecules during the pathogen defense response in plants, therefore establishing a logical connection between cell death and aquaporins for the first time. It was assumed that the aquaporins NtPIP2.2a, NtPIP2.2b und NtTIP1.1a identified during the yeast-two-hybrid-screen can act as H2O2 channels. In further experiments it could indeed be established that these aquaporins have the ability to transport H2O2 in yeast cells. Yeasts expressing aquaporins could be influenced in their H2O2 sensitivity by the expression of BHRF1. BHRF1 without transmembrane domain (BHRF1deltaTMwt) led to an enhanced H2O2 sensitivity and also to an increase in cell death. In addition, the transient expression of aquaporin could induce necrotic lesions and cell death in Nicotiana benthamiana. Deletion experiments identified a common binding domain for interacting with BHRF1 in these aquaporins. This binding domain consists of the conserved region containing the first NPA motive (?loop? B) that is also half of one pore. Further studies showed that BHRF1 interacts with all kinds of different aquaporins from plants, animals (rAQP8) and humans (hAQP1). BHRF1 most likely binds with the alpha3 helix to the highly conserved NPA region of aquaporins. A cellular protein showing sequence homology to M20 proteases and aminoacylases was isolated when looking for interaction partners of aquaporins in plants. Like BHRF1, this protein binds to the conserved NPA region of the aquaporins. Although the cellular substrate for this protein has to be found yet, an interesting observation was made. Co-expression of the isolated aminoacylase with NtTIP1.1a or NtPIP2.2b in Nicotiana benthamiana led to the inhibition of cell death induced by these aquaporins.
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    Pathogenitätsfaktoren und induzierte Resistenz in der Interaktion von Orobanche ramosa L. und Nicotiana tabacum L.
    (2005) Gonsior, Guido; Sauerborn, Joachim
    Parasitic weeds of the genus Orobanche, cause severe damage to vegetables, legumes and sunflower in Europe, the Middle East and North Africa. Recently there is evidence for increasing occurrence of branched broomrape (Orobanche ramosa L.) in rape and tobacco cultivars in France and Germany. This research concerns on biology and pathogenicity of O. ramosa with special focus on the interactions with the host plant. In addition mechanisms of induced resistance of the host plant against the parasite were considered. The results showed genetic differences between the populations of O. ramosa in Germany, which partially parasitize different hosts. As possible factors of pathogenicity the accumulation of reactive oxygen species and a high peroxidase activity together with a high antioxidative capacity caused by the phenylpropanoids Verbascosid and Oraposid were identified. The control of branched broomrape was possible by application of agents, which are described to induce disease resistance. Control of branched broomrape could be based on systemic acquired resistance (SAR) as well as on induced systemic resistance (ISR). As defense mechanisms in tobacco roots against O. ramosa the biosynthesis of the hydroxycoumarin scopoletin and a significant increase of soluble and bound phenolics was discussed. This is underlined by a high activity and an increased expression of phenylalanine ammonia-lyase (PAL) the key enzyme of the phenylpropanoid pathway. Beside phytoalexin production we recognized an increased activity of peroxidases in Orobanche ramosa infected tobacco roots. Further it could be shown that the plant hormone Ethylene as well as the regulation of the sugar metabolism have importance for the resistance of the host plant.