Alfalfa (Medicago sativa) is a high-quality legume forage crop worldwide, and alfalfa production is often threatened by abiotic environmental stresses. GRAS proteins are important transcription factors that play a vital role in plant development, as well as in response to environmental stress. In this study, the availability of alfalfa genome “Zhongmu No.1” allowed us to identify 51 GRAS family members, i.e., MsGRAS. MsGRAS proteins could be classified into nine subgroups with distinct conserved domains, and tandem and segmental duplications were observed as an expansion strategy of this gene family. In RNA-Seq analysis, 14 MsGRAS genes were not expressed in the leaf or root, 6 GRAS genes in 3 differentially expressed gene clusters were involved in the salinity stress response in the leaf. Moreover, qRT-PCR results confirmed that MsGRAS51 expression was induced under drought stress and hormone treatments (ABA, GA and IAA) but down-regulated in salinity stress. Collectively, our genome-wide characterization, evolutionary, and expression analysis suggested that the MsGRAS proteins might play crucial roles in response to abiotic stresses and hormonal cues in alfalfa. For the breeding of alfalfa, it provided important information on stress resistance and functional studies on MsGRAS and hormone signaling.
Seed aging is a major challenge for food security, agronomic production, and germplasm conservation, and reactive oxygen species (ROS) and methylglyoxal (MG) are highly involved in the aging process. However, the regulatory mechanisms controlling the abundance of ROS and MG are not well characterized. To characterize dynamic response of antioxidant and glyoxalase systems during seed aging, oat (Avena sativa L.) aged seeds with a range of germination percentages were used to explore physiological parameters, biochemical parameters and relevant gene expression. A reference transcriptome based on PacBio sequencing generated 67,184 non-redundant full-length transcripts, with 59,050 annotated. Subsequently, eleven seed samples were used to investigate the dynamic response of respiration, ROS and MG accumulation, antioxidant enzymes and glyoxalase activity, and associated genes expression. The 48 indicators with high correlation coefficients were divided into six major response patterns, and were used for placing eleven seed samples into four groups, i.e., non-aged (Group N), higher vigor (Group H), medium vigor (Group M), and lower vigor (Group L). Finally, we proposed a putative model for aging response and self-detoxification mechanisms based on the four groups representing different aging levels. In addition, the outcomes of the study suggested the dysfunction of antioxidant and glyoxalase system, and the accumulation of ROS and MG definitely contribute to oat seed aging.
Abiotic stress disturbs plant cellular redox homeostasis, inhibiting seed germination and plant growth. This is a crucial limitation to crop yield. Glutathione reductase (GR) is an important component of the ascorbate-glutathione (AsA-GSH) cycle which is involved in multiple plant metabolic processes. In the present study, GRs in A. sativa (AsGRs) were selected to explore their molecular characterization, phylogenetic relationship, and RNA expression changes during seed imbibition under abiotic stress. Seven AsGR genes were identified and mapped on six chromosomes of A, C, and D subgenomes. Phylogenetic analysis and subcellular localization of AsGR proteins divided them into two sub-families, AsGR1 and AsGR2, which were predicted to be mainly located in cytoplasm, mitochondrion, and chloroplast. Cis-elements relevant to stress and hormone responses are distributed in promoter regions of AsGRs. Tissue-specific expression profiling showed that AsGR1 genes were highly expressed in roots, leaves, and seeds, while AsGR2 genes were highly expressed in leaves and seeds. Both AsGR1 and AsGR2 genes showed a decreasing-increasing expression trend during seed germination under non-stress conditions. In addition, their responses to drought, salt, cold, copper, H2O2, and ageing treatments were quite different during seed imbibition. Among the seven AsGR genes, AsGR1-A, AsGR1-C, AsGR2-A, and AsGR2-D responded more significantly, especially under drought, ageing, and H2O2 stress. This study has laid the ground for the functional characterization of GR and the improvement of oat stress tolerance and seed vigor.
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