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Query: EC:3.4.25.1 (proteasome)
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By screening for ethylene response mutants in Arabidopsis, a novel mutant, eer2, was isolated which displays enhanced ethylene responses. On a low nutrient medium (LNM) light-grown eer2 seedlings showed a significant hypocotyl elongation in response to low levels of 1-amino-cyclopropane-1-carboxylate (ACC), the precursor of ethylene, compared with the wild type, indicating that eer2 is hypersensitive to ethylene. Treatment with 1-MCP (1-methylcyclopropene), a competitive inhibitor of ethylene signalling, suppressed this hypersensitive response, demonstrating that it is a bona fide ethylene effect. By contrast, roots of eer2 were less sensitive than the wild type to low concentrations of ACC. The ethylene levels in eer2 did not differ from the wild type, indicating that ethylene overproduction is not the primary cause of the eer2 phenotype. In addition to its enhanced ethylene response of hypocotyls, eer2 is also affected in the pattern of senescence and its phenotype depends on the nutritional status of the growth medium. Furthermore, linkage analysis of eer2 suggests that this mutant defines a new locus in ethylene signalling.
J Exp Bot 2005 Sep
PMID:The Arabidopsis mutant eer2 has enhanced ethylene responses in the light. 1604 54

Arabidopsis plants possess a family of nine AtAtg8 gene homologues of the yeast autophagy-associated Apg8/Aut7 gene. To gain insight into how these genes function in plants, first, the expression patterns of five AtAtg8 homologues were analysed in young Arabidopsis plants grown under favourable growth conditions or following exposure to prolonged darkness or sugar starvation. Promoters, plus the entire coding regions (exons and introns) of the AtAtg8 genes, were fused to the beta-glucuronidase reporter gene and transformed into Arabidopsis plants. In all plants, grown under favourable growth conditions, beta-glucuronidase staining was much more significant in roots than in shoots. Different genes showed distinct spatial and temporal expression patterns in roots. In some transgenic plants, beta-glucuronidase staining in leaves was induced by prolonged darkness or sugar starvation. Next, Arabidopsis plants were transformed with chimeric gene-encoding Atg8f protein fused to N-terminal green fluorescent protein and C-terminal haemagglutinin epitope tags. Analysis of these plants showed that, under favourable growth conditions, the Atg8f protein is efficiently processed and is localized to autophagosome-resembling structures, both in the cytosol and in the central vacuole, in a similar manner to its processing and localization under starvation stresses. Moreover, treatment with a cocktail of proteasome inhibitors did not prevent the turnover of this protein, implying that its turnover takes place in the vacuoles, as occurs in yeasts. The results suggest that, in plants, the cellular processes involving the Atg8 genes function efficiently in young, non-senescing tissues, both under favourable growth conditions and under starvation stresses.
J Exp Bot 2005 Nov
PMID:The autophagy-associated Atg8 gene family operates both under favourable growth conditions and under starvation stresses in Arabidopsis plants. 1615 55

In this work, the effect of ethylene on flower opening of cut rose (Rosa hybrida) cv. Samantha was studied. However, although ethylene hastened the process of flower opening, 1-MCP (1-methylcyclopropene), an ethylene action inhibitor, impeded it. Ethylene promoted ethylene production in petals, but 1-MCP did not inhibit this process. Of the four ethylene biosynthetic genes tested, Rh-ACS1 and Rh-ACS2 were undetectable; Rh-ACS3 and Rh-ACO1 expression was enhanced by ethylene slightly and greatly, respectively. However, their mRNA amounts were not inhibited by 1-MCP compared with controls. Expression of seven signalling component genes was also studied, including three ethylene receptors (Rh-ETR1, Rh-ETR3, and Rh-ETR5), two CTRs (Rh-CTR1 and Rh-CTR2), and two transcription factors (Rh-EIN3-1 and Rh-EIN3-2). Transcripts of Rh-ETR5, Rh-EIN3-1, and Rh-EIN3-2 were accumulated in a constitutive manner and had no or little response to ethylene or 1-MCP, while transcript levels of Rh-ETR1 and Rh-CTR1 were substantially elevated by ethylene, and those of Rh-ETR3 and Rh-CTR2 were greatly enhanced by ethylene; 1-MCP reduced all the four genes to levels much less than those in control flowers. These results show that ethylene triggers physiological responses related to flower opening in cut rose cv. Samantha, and that continued ethylene perception results in flower opening. Ethylene may regulate flower opening mainly through expression of two ethylene receptor genes (Rh-ETR1 and Rh-ETR3) and two CTR (Rh-CTR1 and Rh-CTR2) genes.
J Exp Bot 2006
PMID:Transcriptional regulation of ethylene receptor and CTR genes involved in ethylene-induced flower opening in cut rose (Rosa hybrida) cv. Samantha. 1684 35

Ethylene initiates the ripening and senescence of climacteric fruit, whereas polyamines have been considered as senescence inhibitors. Ethylene and polyamine biosynthetic pathways share S-adenosylmethionine as a common intermediate. The effects of 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception, on ethylene and polyamine metabolism and associated gene expression was investigated during ripening of the model climacteric fruit, tomato (Solanum lycopersicum L.), to determine whether its effect could be via polyamines as well as through a direct effect on ethylene. 1-MCP delayed ripening for 8 d compared with control fruit, similarly delaying ethylene production and the expression of 1-aminocyclopropane-1-carboxylic acid (ACC)-synthase and some ethylene receptor genes, but not that of ACC oxidase. The expression of ethylene receptor genes returned as ripening was reinitiated. Free putrescine contents remained low while ripening was inhibited by 1-MCP, but increased when the fruit started to ripen; bound putrescine contents were lower. The activity of the putrescine biosynthetic enzyme, arginine decarboxylase, was higher in 1-MCP-treated fruit. Activity of S-adenosylmethionine-decarboxylase peaked at the same time as putrescine levels in control and treated fruit. Gene expression for arginine decarboxylase peaked early in non-treated fruit and coincident with the delayed peak in putrescine in treated fruit. A coincident peak in the gene expression for arginase, S-adenosylmethionine-decarboxylase, and spermidine and spermine synthases was also seen in treated fruit. No effect of treatment on ornithine decarboxylase activity was detected. Polyamines are thus not directly associated with a delay in tomato fruit ripening, but may prolong the fully-ripe stage before the fruit tissues undergo senescence.
J Exp Bot 2006
PMID:Inhibition of the ethylene response by 1-MCP in tomato suggests that polyamines are not involved in delaying ripening, but may moderate the rate of ripening or over-ripening. 1692 Jul 66

The feedback regulation of ethylene biosynthesis in banana [Musa sp. (AAA group, Cavendish subgroup) cv. Grand Nain] fruit was investigated in an attempt to clarify the opposite effect of 1-methylcyclopropene (1-MCP), an ethylene action inhibitor, before and after the onset of ripening. 1-MCP pre-treatment completely prevented the ripening-induced effect of propylene in pre-climacteric banana fruit, whereas treatment after the onset of ripening stimulated ethylene production. In pre-climacteric fruit, higher concentrations of propylene suppressed ethylene production more strongly, despite their earlier ethylene-inducing effect. Exposure of the fruit ripened by propylene to 1-MCP increased ethylene production concomitantly with an increase in 1-aminocyclopropane-1-carboxylate (ACC) synthase activity and ACC content, and prevented a transient decrease in MA-ACS1 transcripts in the pulp tissues. In contrast, in the peel of ripening fruit, 1-MCP prevented the increase in ethylene production and subsequently the ripening process by reduction of the increase in MA-ACS1 and MA-ACO1 transcripts and of ACC synthase and ACC oxidase activities. These results suggest that ethylene biosynthesis in ripening banana fruit may be controlled negatively in the pulp tissue and positively in the peel tissue. This differential regulation by ethylene in pulp and peel tissues was also observed for MA-PL, MA-Exp, and MA-MADS genes.
J Exp Bot 2007
PMID:Differential feedback regulation of ethylene biosynthesis in pulp and peel tissues of banana fruit. 1718 40

Cell wall disassembly in ripening fruit is highly complex, involving the dismantling of multiple polysaccharide networks by diverse families of wall-modifying proteins. While it has been reported in several species that multiple members of each such family are expressed in the same fruit tissue, it is not clear whether this reflects functional redundancy, with protein isozymes from a single enzyme class performing similar roles and contributing equally to wall degradation, or whether they have discrete functions, with some isoforms playing a predominant role. Experiments reported here sought to distinguish between cell wall-related processes in ripening melon that were softening-associated and softening-independent. Cell wall polysaccharide depolymerization and the expression of wall metabolism-related genes were examined in transgenic melon (Cucumis melo var. cantalupensis Naud.) fruit with suppressed expression of the 1-aminocyclopropane-1-carboxylate oxidase (ACO) gene and fruits treated with ethylene and 1-methylcyclopropene (1-MCP). Softening was completely inhibited in the transgenic fruit but was restored by treatment with exogenous ethylene. Moreover, post-harvest application of 1-MCP after the onset of ripening completely halted subsequent softening, suggesting that melon fruit softening is ethylene-dependent. Size exclusion chromatography of cell wall polysaccharides, from the transgenic fruits, with or without exogenous ethylene, indicated that the depolymerization of both pectins and xyloglucans was also ethylene dependent. However, northern analyses of a diverse range of cell wall-related genes, including those for polygalacturonases, xyloglucan endotransglucosylase/hydrolases, expansin, and beta-galactosidases, identified specific genes within single families that could be categorized as ethylene-dependent, ethylene-independent, or partially ethylene-dependent. These results support the hypothesis that while individual cell wall-modifying proteins from each family contribute to cell wall disassembly that accompanies fruit softening, other closely related family members are regulated in an ethylene-independent manner and apparently do not directly participate in fruit softening.
J Exp Bot 2007
PMID:Ethylene regulation of fruit softening and cell wall disassembly in Charentais melon. 1730 29

Following light-induced nuclear translocation, the phytochromes induce changes in gene expression to regulate plant development. PIF3 and other PIFs (phytochrome-interacting factors), members of the bHLH (basic helix-loop-helix) family of transcriptional regulators, interact specifically with the active Pfr conformer of the phytochrome molecule, suggesting that the PIFs are key components of phytochrome signal transduction. The mechanism by which the PIFs transduce phytochrome signals is not understood. After initial studies that suggested that PIF3 was a positive regulator of phytochrome signalling, mutant studies indicated that the PIFs primarily act as negative regulators in the pathway. Furthermore, in some cases they accumulate in the dark and are degraded upon illumination by the ubiquitin-26S proteasome system. At least for PIF3, the protein degradation depends on direct interaction with the phytochrome molecule and is preceded by protein phosphorylation. In this review, the current understanding of the role of the PIFs in phytochrome-mediated photomorphogenesis will be summarized, and recent findings suggesting an unanticipated dual mechanism of action of the PIFs will be discussed.
J Exp Bot 2007
PMID:Out of the dark: how the PIFs are unmasking a dual temporal mechanism of phytochrome signalling. 1785 31

Two MdERFs (ethylene-response factors) were isolated from ripening apple (Malusxdomestica Borkh. cv. Golden Delicious) fruit. The features of their conserved motifs indicated that MdERF1 and MdERF2 belong to group VII and group IX categories in Arabidopsis, respectively. MdERF1 was expressed predominantly in ripening fruit, although a small degree of expression was also observed in non-fruit tissues, whereas MdERF2 was expressed exclusively in ripening fruit. The increased expression in ripening fruit was repressed by treatment with 1-methylcyclopropene (1-MCP: a potent antagonist of ethylene receptors), indicating that transcription is regulated positively by the ethylene signalling system. Indeed, it was a tendency for cultivars with low ethylene production to show lower MdERFs expression than those with high ethylene production. On the basis of concomitant analyses of the expression of some genes related to ripening, the functions of MdERFs and the role of ethylene in the ripening process are discussed.
J Exp Bot 2007
PMID:MdERFs, two ethylene-response factors involved in apple fruit ripening. 1805 44

A cDNA clone (OsRHC1) was obtained, which encodes a novel RING zinc finger protein sharing similar structural features (multiple transmembrane domains at the N-half; a unique RING zinc finger consensus Cys-X(2)-Cys-X(11)-Cys-X-His-X(3)-Cys-X(2)-Cys-X(6)-Cys-X(2)-Cys at the C terminus) to a group of closely related annotated proteins from both monocots and dicots. OsRHC1 was found to be localized on plasma membrane of rice cells and induced by wounding in rice lines containing Xa loci. Ecotopic expression of the OsRHC1 cDNA from rice (a monocot) in transgenic Arabidopsis thaliana (a dicot) enhanced the defence response toward Pseudomonas syringae pv. tomato DC3000, suggesting that OsRHC1 may confer broad-spectrum disease resistance. The protective effects were neutralized in the presence of MG132 or in an npr1-3 mutation background, indicating that the function of OsRHC1 is dependent on the ubiquitin-mediated protein degradation via the 26S proteasome and the presence of the key defence response regulator NPR1.
J Exp Bot 2007
PMID:Expression of a RING-HC protein from rice improves resistance to Pseudomonas syringae pv. tomato DC3000 in transgenic Arabidopsis thaliana. 1818 23

The procera (pro) mutant of tomato exhibits a well-characterized constitutive gibberellic acid (GA) response phenotype. The tomato DELLA gene LeGAI in the pro mutant background contains a point mutation that results in an amino acid change in the conserved VHVID putative DNA-binding domain in LeGAI to VHEID. This same point mutation is in four different genetic backgrounds exhibiting the pro phenotype, suggesting that this mutation co-segregates with the pro phenotype. Complementation of the mutant with a constitutively expressed wild-type LeGAI gene sequence was not conclusive due to the infertility of transgenic plants. The pro mutation alters tomato branching architecture through differential suppression of axillary bud development, indicating a role for DELLA proteins in the regulation of plant structure. Isolated gib-1 pro double mutant embryo axes, which are unable to synthesize GA, germinate faster than their wild-type counterparts, and exert greater embryo growth potential. The pro mutation is therefore regulating GA responses within the tomato embryo. Transient expression of a LeGAI-GFP (green fluorescent protein) fusion protein in onion epidermis results in its location to the nucleus, and this protein is rapidly degraded by the proteasome in the presence of GA.
J Exp Bot 2008
PMID:Procera is a putative DELLA mutant in tomato (Solanum lycopersicum): effects on the seed and vegetative plant. 1825 77


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