Project description:The study of salt tolerance mechanisms in halophyte plants can provide valuable information for crop breeding and plant engineering programs. The aim of the present study was to investigate whole transcriptome analysis of Aeluropus littoralis in response to salinity stress (200 and 400 mM NaCl) by de novo RNA-sequencing. To assemble the transcriptome, Trinity v2.4.0 and Bridger tools, were comparatively used with two k-mer sizes (25 and 32 bp). The de novo assembled transcriptome by Bridger (k-mer 32) was chosen as final assembly for subsequent analysis. In general, 103290 transcripts were obtained. The differential expression analysis (log2FC > 1 and FDR < 0.01) showed that 1861 transcripts expressed differentially, including169 up and 316 down-regulated transcripts in 200 mM NaCl treatment and 1035 up and 430 down-regulated transcripts in 400 mM NaCl treatment compared to control. In addition, 89 transcripts were common in both treatments. The most important over-represented terms in the GO analysis of differentially expressed genes (FDR < 0.05) were chitin response, response to abscisic acid, and regulation of jasmonic acid mediated signaling pathway under 400 mM NaCl treatment and cell cycle, cell division, and mitotic cell cycle process under 200 mM treatment. In addition, the phosphatidylcholine biosynthetic process term was common in both salt treatments. Interestingly, under 400 mM salt treatment, the PRC1 complex that contributes to chromatin remodeling was also enriched along with vacuole as a general salinity stress responsive cell component. Among enriched pathways, the MAPK signaling pathway (ko04016) and phytohormone signal transduction (ko04075) were significantly enriched in 400 mM NaCl treatment, whereas DNA replication (ko03032) was the only pathway that significantly enriched in 200 mM NaCl treatment. Finally, our findings indicate the salt-concentration depended responses of A. littoralis, which well-known salinity stress-related pathways are induced in 400 mM NaCl, while less considered pathways, e.g. cell cycle and DNA replication, are highlighted under 200 mM NaCl treatment.

Project description:BackgroundThe use of stably expressed genes as normalizers has crucial role in accurate and reliable expression analysis estimated by quantitative real-time polymerase chain reaction (qPCR). Recent studies have shown that, the expression levels of common housekeeping genes are varying in different tissues and experimental conditions. The genomic DNA contamination in RNA samples is another important factor that also influence the interpretation of the data obtained from qPCR. It is estimated that the gDNA contamination in gene expression analysis lead to an overestimation of the RNA transcript level. The aim of this study was to validate the most stably expressed reference genes in two different tissues of Aeluropus littoralis-halophyte grass at salt stress and recovery condition. Also, a qPCR-based approach for monitoring contamination with gDNA was conducted.ResultsTen candidate reference genes participating in different biological processes were analyzed in four groups of samples including root and leaf tissues, salt stress and recovery condition. To determine the most stably expressed reference genes, three statistical methods (geNorm, NormFinder and BestKeeper) were applied. According to results obtained, ten candidate reference genes were ranked based on the stability of their expression. Here, our results show that a set of four housekeeping genes (HKGs) e.g. RPS3, EF1A, GTF and RPS12 could be used as general reference genes for the all selected conditions and tissues. Also, four set of reference genes were proposed for each tissue and condition including: RPS3, EF1A and UBQ for salt stress and root samples; RPS3, EF1A, UBQ as well as GAPDH for recovery condition; U2SURP and GTF for leaf samples. Additionally, for assessing DNA contamination in RNA samples, a set of unique primers were designed based on the conserved region of ribosomal DNA (rDNA). The universality, specificity and sensitivity of these primer pairs were also evaluated in Poaceae.ConclusionsOverall, the sets of reference genes proposed in this study are ideal normalizers for qPCR analysis in A. littoralis transcriptome. The novel reference gene e.g. RPS3 that applied this study had higher expression stability than commonly used housekeeping genes. The application of rDNA-based primers in qPCR analysis was addressed.

Project description:Understanding the response variation of morphological parameters and biomass allocation of plants in heterogeneous saline environments is helpful in evaluating the internal correlation between plant phenotypic plasticity mechanism and biomass allocation. The plasticity of plants alters the interaction among individuals and their environment and consequently affects the population dynamics and aspects of community and ecosystem functioning. The current study aimed to assess the plasticity of Aeluropus lagopoides traits with variation in saline habitats. Understanding the habitat stress tolerance strategy of A. lagopoides is of great significance since it is one of the highly palatable forage grass in the summer period. Five different saline flat regions (coastal and inland) within Saudi Arabia were targeted, and the soil, as well as the morphological and physiological traits of A. lagopoides, were assessed. Comprehensive correlation analyses were performed to correlate the traits with soil, region, or among each other. The soil analysis revealed significant variation among the five studied regions for all measured parameters, as well as among the soil layers showing the highest values in the upper layer and decreased with the depth. Significant differences were determined for all tested parameters of the morphological and reproductive traits as well as for the biomass allocation of A. lagopoides, except for the leaf thickness. In the highly saline region, Qaseem, A. lagopoides showed stunted aerial growth, high root/shoot ratio, improved root development, and high biomass allocation. In contrast, the populations growing in the low saline region (Jizan) showed the opposite trend. Under the more stressful condition, like in Qaseem and Salwa, A. lagopoides produce low spikes in biomass and seeds per plant, compared to the lowest saline habitats, such as Jouf. There was no significant difference in physiological parameters except stomatal conductance (gs), which is highest in the Jizan region. In conclusion, the population of A. lagopoides is tolerant of harsh environments through phenotypic plasticity. This could be a candidate species to rehabilitate the saline habitats, considering saline agriculture and saline soil remediation.

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Aeluropus lagopoides

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