Greenhouse hydroponic experiments were conducted using Cd-sensitive (cv. Guiyan1) and Cd-tolerant (cv. Yunyan2) tobacco cultivars to study the ameliorative effects of exogenous glycinebetaine (GB) upon 5 μM Cd stress. The foliar spray of GB markedly reduced Cd concentrations in plants and alleviated Cd-induced soil plant analysis development (SPAD) value, plant height and root length inhibition, with the mitigation effect being more obvious in Yunyan2. External GB markedly reduced Cd-induced malondialdehyde (MDA) accumulation, induced stomatal closure, ameliorated Cd-induced damages on leaf/root ultrastructure, and increased the chlorophyll content and fluorescence parameters of Fo, Fm, and Fv/Fm in both cultivars and Pn in Yunyan2. Exogenous GB counteracted Cd-induced alterations of certain antioxidant enzymes and nutrients uptake, e.g., the depressed Cd-induced increase of superoxide dismutase (SOD) and peroxidase (POD) activities, but significantly elevated the depressed catalase (CAT) and ascorbate peroxidase (APX) activities. The results indicate that alleviated Cd toxicity by GB application is related to the reduced Cd uptake and MDA accumulation, balanced nutrients and antioxidant enzyme activities, improved PSII, and integrated ultrastructure in tobacco plants.
Tobacco easily accumulates certain heavy metals in leaves and thus poses a potential threat to human health. To systematically dissect Cr-responsive microRNAs (miRNAs) and their targets at the global level, 4 small RNA libraries were constructed from the roots of Cr-treated (Cr) and Cr-free (control) for 2 contrasting tobacco genotypes,Yunyan2 (Cr-sensitive) and Guiyan1 (Cr-tolerant). Using high-throughput-sequencing-technology, the authors identified 53 conserved and 29 novel miRNA families. Comparative genomic analysis of 41 conserved Cr-responsive miRNA families revealed that 11 miRNA families showed up-regulation in Guiyan1 but unaltered in Yunyan2, and 17 miRNA families were up-regulated only in Yunyan2 under Cr stress. Only 1 family, miR6149, was down-regulated in Yunyan2 but remained unchanged in Guiyan1. Of the 29 novel miRNA families, 14 expressed differently in the 2 genotypes under Cr stress. Based on a high-throughput degradome sequencing homology search, potential targets were predicted for the 41 conserved and 14 novel Cr-responsive miRNA families. Clusters of Orthologous Groups functional category analysis revealed that some of these predicted target transcripts of miRNAs are responsive to biotic and abiotic stresses. Furthermore, the expression patterns of many Cr-responsive miRNAs were validated by stem-loop real-time transcription polymerase chain reaction. The results of the present study provide valuable information and a framework for understanding the function of miRNAs in Cr tolerance.
Greenhouse hydroponic experiments were carried out using three different heavy metal accumulation tobacco genotypes to evaluate how different genotypes responded to chromium (Cr) toxicity in the presence of 24-epibrassinolide (EBR; a biologically active brassinosteroid). The results showed that Cr stress caused a marked reduction in plant biomass, chlorophyll content, chlorophyll fluorescence, and photosynthesis parameters but induced malondialdehyde accumulation and ultrastructure damage, with 2010-38 (L) less affected. Foliar application of 24-epibrassinolide (0.1 μM) on Cr-stressed plants greatly alleviated Cr-induced inhibition of growth and photosynthesis, oxidative stress and ultrastructure damage, decreased Cr accumulation in different parts of leaves and roots, with the exception of the upper and lower of leaves of genotype L, and maintained ion homeostasis. Regarding genotypes, L was more tolerant than M and H, as it absorbed less Cr and also performed better in all of the studied parameters. These findings suggest a potential role for 24-epibrassinolide in Cr stress alleviation and the utilization of elite genetic resources in future breeding programs to develop low Cr accumulation genotypes. These results advocate a positive role for 24-epibrassinolide in reducing pollutant residues from health point of view.
MicroRNAs (miRNAs) are known to play important roles in plant growth and stress response. Heat stress is a severe abiotic stresses by adversely affecting plant growth and yield. To identify heat-responsive miRNAs at the genome-wide level in rice (Oryza sativa), we constructed two small RNA libraries from young panicles treated or not with heat conditions. Ion torrent sequencing of the two libraries identified 294 known miRNAs and 539 novel miRNAs. Differential expression analysis showed that 26 miRNAs were downregulated and 21 miRNAs were upregulated in response to heat stress. Among them, five heat-responsive miRNAs, including miR162b, miR529a-p5, PC-5P-62245-9, miR171b and miR169n, were validated by quantitative real-time polymerase chain reaction. A total of 44 target genes of the differentially expressed miRNAs were predicted. These target genes are most significantly overrepresented in the cell growth process. The results demonstrated that rice miRNAs play critical roles in the heat stress response. This study opens up a new avenue for understanding the regulatory mechanisms of miRNAs involvement in the heat stress response in rice.
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