Light-hormone interaction in the red-light-induced suppression of photomorphogenesis in rice seedlings.

Red light perceived by the shoot bottom suppresses photomorphogenesis in rice seedlings mediated by phytochrome A. Shoots of these seedlings grown in red light having their shoot bottom exposed were deficient in chlorophyll and accumulated high concentration of trans-zeatin riboside. However, reduced presence of isopentynyl adenosine, dihydrozeatin riboside was observed in shoots of red-light-grown non-green seedlings in comparison to green seedling. The message abundance of cytokinin receptor (OsHK5), transporters (OsENT1, OsENT2), and response regulators (OsRR4, OsRR10) was downregulated in these red-light-grown non-green seedlings. Attenuation of greening process was reversed by application of exogenous cytokinin analogue, benzyladenine, or supplementing red light with blue light. In the same vein, the suppression of gene expression of cytokinin receptor, transporters, and type-A response regulators was reversed in red-light-grown seedlings treated with benzyladenine suggesting that the disarrayed cytokinin (CK) signaling cascade is responsible for non-greening of seedlings grown in red light. The reversal of red-light-induced suppression of photomorphogenesis by blue light and benzyladenine demonstrates the interaction of light and cytokinin signaling cascades in the regulation of photomorphogenesis. Partial reversal of greening process by exogenous application of benzyladenine suggests, apart from CKs perception, transportation and responsiveness, other factors are also involved in modulation of suppression of photomorphogenesis by red light.

Expression of chickpea CIPK25 enhances root growth and tolerance to dehydration and salt stress in transgenic tobacco.

Calcium signaling plays an important role in adaptation and developmental processes in plants and animals. A class of calcium sensors, known as Calcineurin B-like (CBL) proteins sense specific temporal changes in cytosolic Ca(2+) concentration and regulate activities of a group of ser/thr protein kinases called CBL-interacting protein kinases (CIPKs). Although a number of CIPKs have been shown to play crucial roles in the regulation of stress signaling, no study on the function of CIPK25 or its orthologs has been reported so far. In the present study, an ortholog of Arabidopsis CIPK25 was cloned from chickpea (Cicer arietinum). CaCIPK25 gene expression in chickpea increased upon salt, dehydration, and different hormonal treatments. CaCIPK25 gene showed differential tissue-specific expression. 5'-upstream activation sequence (5'-UAS) of the gene and its different truncated versions were fused to a reporter gene and studied in Arabidopsis to identify promoter regions directing its tissue-specific expression. Replacement of a conserved threonine residue with an aspartic acid at its catalytic site increased the kinase activity of CaCIPK25 by 2.5-fold. Transgenic tobacco plants overexpressing full-length and the high active versions of CaCIPK25 displayed a differential germination period and longer root length in comparison to the control plants. Expression of CaCIPK25 and its high active form differentially increased salt and water-deficit tolerance demonstrated by improved growth and reduced leaf chlorosis suggesting that the kinase activity of CaCIPK25 was required for these functions. Expressions of the abiotic stress marker genes were enhanced in the CaCIPK25-expressing tobacco plants. Our results suggested that CaCIPK25 functions in root development and abiotic stress tolerance.

Comparative proteomics analysis by DIGE and iTRAQ provides insight into the regulation of phenylpropanoids in maize.

The maize pericarp color1 (p1) gene encodes a Myb transcription factor that regulates the accumulation of 3-deoxyflavonoid pigments called phlobaphenes. The Unstable factor for orange1 (Ufo1) is a dominant epigenetic modifier of the p1 that results in ectopic pigmentation in pericarp. Presence of Ufo1-1 correlates with pleiotropic growth and developmental defects. To investigate the Ufo1-1-induced changes in the proteome, we conducted comparative proteomics analysis of P1-wr; Ufo1-1 pericarps using the 2-D DIGE and iTRAQ techniques. Most of the identified proteins were found to be involved in glycolysis, protein synthesis and modification, flavonoid and lignin biosynthesis and defense responses. Further, immunoblot analysis of internode protein extracts demonstrated that caffeoyl CoA O-methyltransferase (COMT) is post-transcriptionally down regulated in P1-wr; Ufo1-1 plants. Consistent with the down regulation of COMT, the concentrations of p-coumaric acid, syringaldehydes, and lignin are reduced in P1-wr; Ufo1-1 internodes. The reductions in these phenylpropanoids correlate with the bent stalk and stunted growth of P1-wr; Ufo1-1 plants. Finally, over-expression of the p1 in transgenic plants is also correlated with a lodging phenotype and reduced COMT expression. We conclude that ectopic expression of p1 can result in developmental defects that are correlated with altered regulation and synthesis of phenylpropanoid compounds including lignin. BIOLOGICAL SIGNIFICANCE: Transcription factors have specific expression patterns that ensure that the biochemical pathways under their control are active in relevant tissues. Plant breeders can select for alleles of transcription factors that produce desirable expression patterns to improve a plant's growth, development, and defense against insects and pathogens. The resulting de novo accumulation of metabolites in plant tissues in significant quantities could have beneficial and/or detrimental consequences. To understand this problem we investigated how the aberrant expression of a classically-studied transcription factor pericarp color1 (p1) which regulates phenylpropanoid metabolism, affects the maize proteome in pericarp tissue. We utilized a dominant mutant Unstable factor for orange 1-1 (Ufo1-1) which reduces the epigenetic suppression of p1 in various tissues throughout the maize plant. Our proteomic analysis shows how, in the presence of Ufo1-1, key enzymes of the glycolytic and shikimic acid pathways were modulated to produce substrates required for flavonoid synthesis. The finding that the presence of Ufo1-1 affected the expression levels of various enzymes in the lignin pathway was of particular interest. We show that lignin was reduced in Ufo1-1 plants expressing p1 and was associated with the post-transcriptional down regulation of CoA O-methyltransferase (COMT) enzyme. We further correlated the down-regulation of COMT with plant bending phenotype in Ufo1-1 plants expressing p1 and to a stalk lodging phenotype of transgenic p1 plants. This study demonstrates that although there can be adverse consequences to aberrantly overexpressing transcription factors, there might also be benefits such as being able to reduce lignin content for biofuel crops. However, more research will be required to understand the genetic and epigenetic regulation of transcription factors and how their expression can be optimized to obtain desired traits in preferred tissue types. This article is part of a Special Issue entitled: Translational Plant Proteomics.

Involvement of phytochrome A in suppression of photomorphogenesis in rice seedling grown in red light.

Plants have evolved a remarkable capacity to track and respond to fluctuations of light quality and intensity that influence photomorphogenesis facilitated through several photoreceptors, which include a small family of phytochromes. Rice seedlings grown on germination paper in red light for 48 h having their shoot bottom exposed had suppressed photomorphogenesis and were deficient in chlorophyll. Seedlings grown under identical light regime having their shoot bottom covered were green and accumulated chlorophyll. Further, etiolated seedlings with their shoot bottom exposed, when grown in 4 min red/far-red cycles for 48 h, accumulated chlorophyll demonstrating the reversal of suppression of photomorphogenesis by far-red light. It implicates the involvement of phytochrome. Immunoblot analysis showed the persistence of photolabile phytochrome A protein for 48 h in seedlings grown in red light with their shoot bottom exposed, suggesting its involvement in suppression of photomorphogenesis. This was further corroborated in phyA seedlings that turned green when grown in red light having their shoot bottom exposed. Calmodulin (CaM) antagonist N-(6-aminohexyl)-5-chloro-1-napthalene sulphonamide or trifluoperazine substantially restored photomorphogenesis both in the wild type (WT) and phyA demonstrating the involvement of CaM-dependent kinases in the down-regulation of the greening process. Results demonstrate that red light-induced suppression of photomorphogenesis, perceived in the shoot bottom, is a red high irradiance response of PhyA.

Signaling events leading to red-light-induced suppression of photomorphogenesis in wheat (Triticum aestivum).

Perception of red light (400 µmol photon m²/s) by the shoot bottom turned off the greening process in wheat. To understand the signaling cascade leading to this photomorphogenic response, certain signaling components were probed in seedlings grown in different light regimes. Upon analysis the gene expression of heterotrimeric Gα and Gß were severely down-regulated in seedlings grown without vermiculite and having their shoot bottom exposed to red light (R/V-) and was similar to that of dark-grown seedlings. Supplementing the red-light-grown V- seedlings with blue light resulted in up-regulation of both Gα and Gß expression, suggesting that blue light is able to modulate G protein expression. Treatment of cytokinin analog benzyladenine to cytokinin-deficient red-light-grown R/V- seedlings resulted in up-regulation of gene expression of both Gα and Gß. To probe further, modulators of signal transduction pathway--AlF3 (G protein activator), LaCl3 (Ca(2+) channel blocker), NaF (nonspecific phosphatase inhibitor), or calmodulin (CaM) antagonists trifluoperazine (TFP) and N-(6-aminohexyl)-5-chloro-1-nafthalene-sulfonamide (W-7)--were added along with Hoagland solution to the roots of 4-day-old etiolated seedlings, grown on germination paper and transferred to red light. AlF3, LaCl3, NaF failed to elicit any photomorphogenic response. However, CaM antagonists TFP and W-7 significantly reversed the red-light-induced suppression of photomorphogenesis. Phosphorylation of proteins assayed in the absence or presence of CaM antagonist TFP revealed respective up-regulation or down-regulation of phosphorylation of several plastidic proteins in R/V- seedlings. These suggest that signal transduction of red light perceived by the shoot bottom to suppress photomorphogenesis is mediated by CaM-dependent protein kinases.