ABSTRACT
KEY MESSAGE: Analyses of the function of Arabidopsis Calmodulin7 (CAM7) in concert with multiple regulatory proteins involved in various signal transduction processes. Calmodulin (CaM) plays various regulatory roles in multiple signaling pathways in eukaryotes. Arabidopsis CALMODULIN 7 (CAM7) is a unique member of the CAM family that works as a transcription factor in light signaling pathways. CAM7 works in concert with CONSTITUTIVE PHOTOMORPHOGENIC 1 and ELONGATED HYPOCOTYL 5, and plays an important role in seedling development. Further, it is involved in the regulation of the activity of various Ca2+-gated channels such as cyclic nucleotide gated channel 6 (CNGC6), CNGC14 and auto-inhibited Ca2+ ATPase 8. Recent studies further indicate that CAM7 is also an integral part of multiple signaling pathways including hormone, immunity and stress. Here, we review the recent advances in understanding the multifaceted role of CAM7. We highlight the open-ended questions, and also discuss the diverse aspects of CAM7 characterization that need to be addressed for comprehensive understanding of its cellular functions.
Subject(s)
Calmodulin/metabolism , Plant Development , Stress, Physiological , Amino Acid Sequence , Apoptosis , Calmodulin/chemistry , Morphogenesis , Plant Growth Regulators/metabolismABSTRACT
Arabidopsis MYC2 bHLH transcription factor plays a negative regulatory role in blue light (BL)-mediated seedling development. HY5 bZIP protein works as a positive regulator of multiple wavelengths of light and promotes photomorphogenesis. Both MYC2 and HY5, belonging to two different classes of transcription factors, are the integrators of multiple signaling pathways. However, the functional interrelations of these two transcription factors in seedling development remain unknown. Additionally, whereas HY5-mediated regulation of gene expression has been investigated in detail, the transcriptional regulation of HY5 itself is yet to be understood. Here, we show that HY5 and MYC2 work in an antagonistic manner in Arabidopsis seedling development. Our results reveal that HY5 expression is negatively regulated by MYC2 predominantly in BL, and at various stages of development. On the other hand, HY5 negatively regulates the expression of MYC2 at various wavelengths of light. In vitro and in vivo DNA-protein interaction studies suggest that MYC2 binds to the E-box cis-acting element of HY5 promoter. Collectively, this study demonstrates a coordinated regulation of MYC2 and HY5 in blue-light-mediated Arabidopsis seedling development.
Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Arabidopsis/genetics , Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics , Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/metabolism , Basic-Leucine Zipper Transcription Factors/genetics , Basic-Leucine Zipper Transcription Factors/metabolism , Seedlings/genetics , Anthocyanins/metabolism , Arabidopsis/growth & development , Arabidopsis/metabolism , Arabidopsis/radiation effects , Chlorophyll/metabolism , Gene Expression Regulation, Plant/genetics , Gene Expression Regulation, Plant/radiation effects , Gravitropism/genetics , Light , Microscopy, Fluorescence , Mutation , Phenotype , Plant Roots/genetics , Plant Roots/metabolism , Promoter Regions, Genetic , Protein Binding , Seedlings/growth & development , Seedlings/metabolism , Signal Transduction/genetics , Signal Transduction/radiation effects , Two-Hybrid System TechniquesABSTRACT
PURPOSE: Environmental changes generate free radicals and reactive oxygen species (ROS) resulting in abiotic stress in plants. This causes alterations in germination, morphology, growth and development ultimately leading to yield loss. Gamma irradiation was used to experimentally induce oxidative damage in an important pulse crop Vigna radiata (L.) Wilczek or mung bean. Our research was aimed towards augmentation of oxidative stress tolerance through treatment with a group of aliphatic amines known as polyamines. MATERIALS AND METHODS: We used sub-lethal doses of gamma irradiation to generate oxidative damage which was evaluated using Nitro blue tetrazolium (NBT) staining, total antioxidant activity, 1, 1-Diphenyl-2-picryl hydrazyl (DPPH) radical scavenging assay, proline content and lipid peroxidation. Changes in internal free polyamines and messenger ribonucleic acid (mRNA) expression of key rate-limiting S-adenosylmethionine decarboxylase (SAMDC) enzyme in polyamine biosynthetic pathway was studied using real-time polymerase chain reaction (PCR). RESULTS: We observed increased oxidative damage with higher irradiation dose which was partially alleviated by putrescine treatment. Internal levels of putrescine and spermidine increased with 1 mM (50 and 100 Gy) and 2 mM putrescine treatment. Expression of SAMDC also increased with putrescine treatment. CONCLUSION: This study shows that treatment with putrescine can partially alleviate oxidative damage caused by gamma rays.
Subject(s)
Oxidative Stress/drug effects , Oxidative Stress/radiation effects , Polyamines/pharmacology , Vigna/metabolism , Antioxidants/metabolism , Biphenyl Compounds/metabolism , Gene Expression Regulation, Plant/drug effects , Gene Expression Regulation, Plant/radiation effects , Lipid Peroxidation/drug effects , Lipid Peroxidation/radiation effects , Picrates/metabolism , Plant Leaves/drug effects , Plant Leaves/metabolism , Plant Leaves/radiation effects , Plant Proteins/genetics , Proline/metabolism , Vigna/drug effects , Vigna/genetics , Vigna/radiation effectsABSTRACT
PURPOSE: To investigate the effects of gamma pre-exposure on cadmium accumulation in Plantago ovata seedlings. Metallothionein (MT) localization was also studied following Cadmium (Cd) treatment in P. ovata. MATERIALS AND METHODS: DNA damage was determined by alkaline comet assay. MT gene and protein expression were studied by real-time polymerase chain reaction and flow cytometry, respectively, in root and shoot tissues. Metal accumulation (Cd, zinc [Zn], iron [Fe]) was evaluated by Atomic Absorption Spectroscopy. RESULTS: Cd treatment decreased seed germination rate, biomass and free radical scavenging activity and increased DNA damage in a dose-dependent manner. When P. ovata seeds were pre- exposed to 5 Gy gamma dose (prior to Cd treatment) seed germination rate, biomass and free radical scavenging activity increased significantly. MT genes (PoMT1, PoMT2 and PoMT3) and MT protein expression enhanced when 5 Gy gamma-irradiated seeds were grown in Cd containing medium and Cd accumulation also increased in a dose-dependent manner. CONCLUSIONS: Higher Cd accumulation in P. ovata seedlings may be attributed to the upregulation of PoMT genes in gamma pretreated seedlings. Localization of metallothionein in cytosol and nucleus indicated its positive role against Cd-mediated cytotoxic and genotoxic effects.
Subject(s)
Adaptation, Physiological/physiology , Cadmium/pharmacokinetics , Cadmium/toxicity , Metallothionein/metabolism , Plantago/physiology , Stress, Physiological/physiology , Adaptation, Physiological/drug effects , Adaptation, Physiological/radiation effects , Dose-Response Relationship, Drug , Gamma Rays , Metals/metabolism , Plantago/drug effects , Plantago/radiation effects , Radiation Dosage , Seedlings/drug effects , Seedlings/physiology , Seedlings/radiation effects , Stress, Physiological/drug effects , Stress, Physiological/radiation effectsABSTRACT
Effects of gamma rays on the free polyamine (PA) levels were studied in Vigna radiata (L.) Wilczek. Seeds exposed to different doses of gamma rays were checked for damage on phenotype, germination frequency and alteration in photosynthetic pigments. Free polyamine levels were estimated from seeds irradiated in dry and water imbibed conditions. Polyamine levels of seedlings grown from irradiated seeds, and irradiated seedlings from unexposed seeds were also measured. Damage caused by gamma irradiation resulted in decrease in final germination percentage and seedling height. Photosynthetic pigments decreased in a dose dependent manner as marker of stress. Polyamines decreased in irradiated dry seeds and in seedlings grown from irradiated seeds. Radiation stress induced increase in free polyamines was seen in irradiated imbibed seeds and irradiated seedlings. Response of polyamines towards gamma rays is dependent on the stage of the life cycle of the plant.
