GFP in plants

Haseloff, Jim; Amos, Brad

doi:10.1016/0168-9525(95)90186-8

Cited by 240 publications

(147 citation statements)

References 5 publications

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“…In the onion epidermal cells, GFP alone was detected in the nucleus and cytoplasm ( Figure 4G). This pattern is consistent with previous observations of the GFP localization in plant cells (Haseloff and Amos 1995;Chiu et al 1996). Fluorescence of fused proteins, i.e.…”

Section: Localization Of Rsbamy1 and Starch Granules In The Taprootsupporting

confidence: 81%

Overexpression of an extraplastidic β-amylase which accumulates in the radish taproot influences the starch content of Arabidopsis thaliana

Takahashi

Kuboi

Fujiwara

et al. 2012

Plant Biotechnology

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Recent studies have revealed that β-amylases (EC 3.2.1.2) located in plastids participate in endogenous starch degradation. On the other hand, many plants possess a respectable amount of extraplastidic β-amylases. Because of their existence outside of the plastids, it has been believed that extraplastidic β-amylases do not contribute to starch degradation in plants. Here, we investigated the role of the major extraplastidic β-amylase of the radish (Raphanus sativus) (RsBAMY1) in starch metabolism. Amylase activity was elevated in the growing taproot of the radish. During the elevation of amylase activity, the starch content decreased, but the contents of soluble sugars such as maltose, glucose, fructose and sucrose increased. In the radish taproot, RsBAMY1 protein accumulated in the primary cambium and anomalous cambium of the storage parenchyma. The starch granules, however, were found just inside the cambium and adjacent to the anomalous cambium. When the RsBAMY1 gene was overexpressed in Arabidopsis, the plants contained lower starch contents than the wild-type plant. These results suggest that the extraplastidic β-amylase may affect the starch metabolism in some plants.Key words: Cambium, maltose, Raphanus sativus, storage organ.β-Amylase (EC 3.2.1.2) is an amylolytic enzyme which hydrolyses α-1,4 glycosidic linkages at the non-reducing end to β-maltose. Recent molecular genetic studies have provided convincing evidence that plastidic β-amylases are involved in endogenous starch degradation in Arabidopsis leaves (Smith et al. 2005;Fettke et al. 2009;Zeeman et al. 2010). β-Maltose is a primary product of starch degradation in Arabidopsis leaves (Weise et al. 2005). The Arabidopsis genome has nine β-amylase genes. Among them, the four β-amylase proteins (BAMs 1-4) are localized in the plastids, and BAMs 1, 3, and 4 were shown to contribute to starch degradation (Kaplan and Guy 2005;Edner et al. 2007;Fulton et al. 2008;Valerio et al. 2011). The bam3 or bam4 mutant showed a strong starch excess phenotype (Kaplan and Guy 2005;Fulton et al. 2008). The bam1 mutant showed no starch excess phenotype (Kaplan and Guy 2005;Fulton et al. 2008), but a multiple mutant which was produced by crossing the bam1 mutant with the bam3 or bam4 mutant showed a higher starch content compared to bam3 or bam4 (Fulton et al. 2008). The isoforms BAM1 and BAM3 had amylolytic activities, but BAM4 did not (Li et al. 2009), suggesting that BAM4 may modulate the rate of starch degradation as a maltose sensor (Fulton et al. 2008). These results indicate that the plastidic β-amylases directly or indirectly contribute to starch degradation in plants.On the other hand, it is known that a large amount of β-amylase activities, most of which are extraplastidic, are found in many vegetative organs (Beck and Ziegler 1989;Ziegler 1999), such as pea leaves (Ziegler and Beck 1986) and hedge bindweed rhizomes (Van Damme et al. 2001). Despite their abundance, the extraplastidic β-amylases have not been considered to degrade endogenous starch beca...

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Section: Localization Of Rsbamy1 and Starch Granules In The Taprootsupporting

confidence: 81%

Overexpression of an extraplastidic β-amylase which accumulates in the radish taproot influences the starch content of Arabidopsis thaliana

Takahashi

Kuboi

Fujiwara

et al. 2012

Plant Biotechnology

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“…This pattern is consistent with previous observations of GFP localization (Haseloff and Amos, 1995; Chiu et al, 1996). To determine whether the increased nuclear fluorescence observed in SAR-induced seedlings reflects nuclear accumulation of NPR1-GFP, we analyzed localization of the fusion protein by subcellular fractionation and protein gel blot analysis.…”

supporting

confidence: 82%

Nuclear Localization of NPR1 Is Required for Activation of PR Gene Expression

2000

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Systemic acquired resistance (SAR) is a broad-spectrum resistance in plants that involves the upregulation of a battery of pathogenesis-related ( PR ) genes. NPR1 is a key regulator in the signal transduction pathway that leads to SAR. Mutations in NPR1 result in a failure to induce PR genes in systemic tissues and a heightened susceptibility to pathogen infection, whereas overexpression of the NPR1 protein leads to increased induction of the PR genes and enhanced disease resistance. We analyzed the subcellular localization of NPR1 to gain insight into the mechanism by which this protein regulates SAR. An NPR1-green fluorescent protein fusion protein, which functions the same as the endogenous NPR1 protein, was shown to accumulate in the nucleus in response to activators of SAR. To control the nuclear transport of NPR1, we made a fusion of NPR1 with the glucocorticoid receptor hormone binding domain. Using this steroidinducible system, we clearly demonstrate that nuclear localization of NPR1 is essential for its activity in inducing PR genes. INTRODUCTIONPlants, like animals, are capable of mounting an immune response after a primary pathogen infection. One such response is known as systemic acquired resistance (SAR). SAR, which is often triggered by a local infection, can provide long-term resistance throughout the plant to subsequent infections by a broad range of pathogens (Ross, 1961;Kuc, 1982;Ryals et al., 1996). The activation of SAR correlates with the expression of the pathogenesis-related ( PR ) genes. Even though the functions of most PR gene products are unknown, some of these proteins have been shown to confer various degrees of pathogen resistance (Schlumbaum et al., 1986;Mauch et al., 1988; Broglie et al., 1991; Woloshuk et al., 1991;Terras et al., 1992 Terras et al., , 1995 Alexander et al., 1993;Liu et al., 1994;Ponstein et al., 1994; Zhu et al., 1994).Activation of PR gene expression and the establishment of SAR require the signal molecule salicylic acid (SA). Concentrations of SA have been shown to increase in both infected and uninfected tissues after pathogen infection (Malamy et al., 1990;Métraux et al., 1990Métraux et al., , 1991Rasmussen et al., 1991). The exogenous application of SA or its synthetic analogs, such as 2,6-dichloroisonicotinic acid (INA) and benzo(1,2,3)thiadiazole-7-carbothioic acid S -methyl ester, results in expression of the PR genes and activation of SAR (White, 1979;Ward et al., 1991; Görlach et al., 1996;Lawton et al., 1996). The essential role of SA in SAR has been demonstrated in transgenic tobacco and Arabidopsis plants that express the bacterial salicylate hydroxylase ( nahG ) gene. In these plants, SA is converted to the inactive compound catechol, and the induction of PR gene expression and SAR is inhibited (Gaffney et al., 1993; Delaney et al., 1994;Lawton et al., 1995).Transduction of the SA signal requires the function of NPR1, a protein first identified in Arabidopsis through a mutant screen (Cao et al., 1994). The npr1 (nonexpressor of PR genes) mutant f...

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“…The possibility for visualizing microtubules in fixed cells by immunofluorescence and electron microscopy is rather limited because it does not allow in vivo observation and quantification of the dynamics of microtubules. Green fluorescent protein (GFP) isolated from the jellyfish Aequorea victoria has enabled real-time in vivo visualization of cytoskeletal dynamics within a variety of eukaryotic systems, including higher plants, without causing any perceptible tissue damage (Chalfie et al, 1994;Haseloff and Amos, 1995;Haseloff et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

Microtubule Dynamics in Living Root Hairs: Transient Slowing by Lipochitin Oligosaccharide Nodulation Signals

2005

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The incorporation of a fusion of green fluorescent protein and tubulin-a 6 from Arabidopsis thaliana in root hairs of Lotus japonicus has allowed us to visualize and quantify the dynamic parameters of the cortical microtubules in living root hairs. Analysis of individual microtubule turnover in real time showed that only plus polymer ends contributed to overall microtubule dynamicity, exhibiting dynamic instability as the main type of microtubule behavior in Lotus root hairs. Comparison of the four standard parameters of in vivo dynamic instability-the growth rate, the disassembly rate, and the frequency of transitions from disassembly to growth (rescue) and from growth to disassembly (catastrophe)-revealed that microtubules in young root hairs were more dynamic than those in mature root hairs. Either inoculation with Mesorhizobium loti or purified M. loti lipochitin oligosaccharide signal molecules (Nod factors) significantly affected the growth rate and transition frequencies in emerging and growing root hairs, making microtubules less dynamic at a specific window after symbiotic inoculation. This response of root hair cells to rhizobial Nod factors is discussed in terms of the possible biological significance of microtubule dynamics in the early signaling events leading to the establishment and progression of the globally important Rhizobium/legume symbiosis.

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GFP in plants

Cited by 240 publications

References 5 publications

Overexpression of an extraplastidic β-amylase which accumulates in the radish taproot influences the starch content of Arabidopsis thaliana

Overexpression of an extraplastidic β-amylase which accumulates in the radish taproot influences the starch content of Arabidopsis thaliana

Nuclear Localization of NPR1 Is Required for Activation of PR Gene Expression

Microtubule Dynamics in Living Root Hairs: Transient Slowing by Lipochitin Oligosaccharide Nodulation Signals

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