Project description:Kharchia local is an Indian tall landrace wheat cultivar. It is native to sodic-saline soils of Kharchia tehsil of the Pali district of Rajasthan, and is a line developed from selections from farmer's fields. It is the most salt tolerant wheat genotype found in India. No systematic study has been carried out in this direction so far. The gaps in understanding of the mechanism underlying salt tolerance limit our ability to improve the salt tolerance in other crop plants. Transcriptome analysis of Kharchia Local under salt stress will provide the insight into the genes involved in salinity tolerance.
Project description:A comparative RNA-Seq analysis was done in root and shoot of Najran wheat cultivar between plants grown under two conditions: control (0 mM NaCl) and salt treatment (200 mM NaCl). The current study revealed differentially expressed genes and various associated biological pathways involved in plant responses to salt stress between the two conditions in the root and shoot plant tissues, providing important insights into the molecular mechanisms underlying salt tolerance in wheat.
Project description:Comparative expression analysis between two highly salt-tolerant wheat lines, their parental lines, and a salt-sensitive line. The Chinese Spring (CS) line is salt sensitive. AJDAj5 and PhI are the two parental lines (with Chinese Spring background), and both have some salt-tolerance. W4909 and W4910 are two lines derived from crossing AJDAj5 and PhI, and both are more salt tolerant than either parental line.
Project description:Purpose: To characterize the functional implication of autophagy in the wheat response to stress, the key genes contributing in mediated salt tolerance of wheat seedlings through 3-MA were identified in normal or salt stress conditions in the presence or absence of added 3-MA by the transcriptome profiles. Methods: Four days after NaCl and 3-MA treatment, the roots and the third leaves were collected respectively with every 10 of them being mixed as one biological replicate for each treatment. Every treatment had four biological replicates. The wheat root and leaves mRNA profiles were generated by deep sequencing, in triplicate, using Illumina GAIIx. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays. Results: The RNA-Seq data had high quality and reliable results were obtained from the transcriptome assembly. A high correlation between biological replicates was observed for all treatments, which indicated that the four biological replicates were reliable in this study. Based on the principal component analysis (PCA), a clear separation between the NaCl-treated group and controls could be observed. The Q30 percentage (sequences with sequencing error rate lower than 0.1%) was over 94%, and the average GC content of the RNA-seq reads was 55.46%. After removing the adaptor and low-quality sequence, each library received 68310810-83844286 clean reads. These clean reads were mapped to the reference genome with match ratios in the range of 93.6%-95.9%, and 120744 genes predicted from the genome were found to be expressed (with FPKM > 0), including 25180 annotated genes in wheat genome. 3-MA treatment shifted the transcriptome a salt-stressed wheat seedling. The up-regulated DEGs and DEMs were increased, and the down-regulated DEGs and DEMs were decreased in 3-MA-added plants under NaCl stress condition. The study may help us understand the mechanism for 3-MA mediated salt tolerance and provide a theoretical basis for autophagy regulated salt response in wheat seedlings. Conclusions: 3-MA treatment shifted the transcriptome a salt-stressed wheat seedling. The up-regulated DEGs and DEMs were increased, and the down-regulated DEGs and DEMs were decreased in 3-MA-added plants under NaCl stress condition. The study may help us understand the mechanism for 3-MA mediated salt tolerance and provide a theoretical basis for autophagy regulated salt response in wheat seedlings.
Project description:Soil salinity is a major environmental stress that restricts crop growth and yield. Here, crucial proteins and biological pathways were investigated under salt-stress and recovery conditions in Tritipyrum “Y1805” to explore its salt-tolerance mechanism. In total, 44 and 102 differentially expressed proteins (DEPs) were identified in “Y1805” under salt-stress and recovery conditions, respectively. A proteome-transcriptome-associated analysis revealed that the expression patterns of 13 and 25 DEPs were the same under salt-stress and recovery conditions, respectively. “Response to stimulus”, “antioxidant activity”, “carbohydrate metabolism”, “amino acid metabolism”, “signal transduction”, “transport and catabolism” and “biosynthesis of other secondary metabolites” were present under both conditions in “Y1805”. In addition, “energy metabolism” and “lipid metabolism” were recovery-specific pathways, while “antioxidant activity”, and “molecular function regulator” under salt-stress conditions, and “virion” and “virion part” during recovery, were “Y1805”-specific compared with the salt-sensitive wheat “Chinese Spring”. “Y1805” contained 83 specific DEPs related to salt-stress responses. The strong salt tolerance of “Y1805” could be attributed to the strengthened cell walls, reactive oxygen species scavenging, osmoregulation, phytohormone regulation, transient growth arrest, enhanced respiration, ammonium detoxification, transcriptional regulation and error information processing. These data will facilitate an understanding of the molecular mechanisms of salt tolerance and aid in the breeding of salt-tolerant wheat.
Project description:Analysis of transcripts in response to salt treatment. In order to design the 22k wheat oligo-DNA microarray, a total of 148,676 expressed sequence tags of common wheat were collected from the database of the Wheat Genomics Consortium of Japan. These were grouped into 34,064 contigs, which were then used to design an oligonucleotide DNA microarray. Following a multi-step selection of the sense strand, 21,939 60-mer oligo DNA probes were selected for attachment on the microarray slide. This 22k oligo DNA microarray was used to examine the transcriptional response of wheat to salt stress. More than 95% of the probes gave reproducible hybridization signals when targeted with RNAs extracted from salt-treated wheat shoots and roots. With the microarray, we identified 1,811 genes whose expressions changed more than two-fold in response to salt. These included genes known to mediate the response to salt as well as unknown genes, and they were classified into 12 major groups by hierarchical clustering. These gene expression patterns were also confirmed by real-time reverse transcription (RT)-PCR. Many of the genes with unknown function were clustered together with genes known to be involved in the response to salt stress. Thus, analysis of gene expression patterns combined with gene ontology should help identify the function of the unknown genes. Also, functional analysis of these wheat genes should provide new insight into the response to salt stress. Finally, these results indicate that the 22k oligo DNA microarray is a reliable method for monitoring global gene expression patterns in wheat. Keywords: time cource, stress response
Project description:Global expression analysis of transcripts in response to salt treatment was carried out for common wheat using oligo-DNA microarrays. Microarrays have been designed from unique wheat genes classified from a large number of expressed sequence tags (ESTs). Two-week-old seedlings of common wheat were treated with 150 mM NaCl for 1, 6 and 24 hours and their roots and shoots were separately subjected to microarray analyses. Consequently, 5996 genes showed changes in expression of more than two-fold, and were classified into 12 groups according to correlations in gene expression patterns. These salt-responsive genes were assigned functions using Gene Ontology (GO) terms. Genes assigned to transcription factor, transcription-regulator activity and DNA binding functions were preferentially classified into early response groups. On the other hand, those assigned transferase and transporter activity were classified into late response groups. These data on gene expression suggest that multiple signal transduction pathways in response to salt treatment exist in wheat. Salt-responsive transcription factors (TFs), namely AP2/EREBP, MYB, NAC and WRKY, were selected and their expression patterns compared with those of rice. Most showed different expression patterns in wheat and rice in response to salt treatment. Furthermore, comparing the microarray data for wheat and rice, only a small number of genes were up- or down-regulated in common in response to salt treatment. These findings suggest that salt-responsive mechanisms distinct from rice might be present in wheat, and wheat genes can contribute to providing novel gene resources for breeding of salt-tolerant crops. Keywords: time cource, stress response
Project description:Hexaploid wheat is the most important cereal crop with the biggest planting area in the world, while its production is dramatically decreased by salt stress. Ethylene is a stress hormone, which improves salt tolerance in plants by regulating adaptive changes in gene expression level. Proteomic analysis provides an efficient method to excavate downstream functional genes in a large scale. In order to clarify the ethylene regulated salt response pathway in wheat, the roots and shoots 2-week-old wheat seedlings of cultivar ‘Qingmai 6’ treated with salt, salt and ethylene precursor ACC, and salt and ethylene signaling inhibitor 1-MCP were collected, respectively, and analyzed with untreated samples by proteomic analysis in this research.
Project description:The hypothesis whether CaCl2 seed treatment prior to sowing (osmopriming) can enhance drought stress tolerance, and/or alleviate the effect on plant growth was tested. Here we elucidate whether seed treatment prior to sowing (osmopriming) with CaCl2 can improve the drought stress tolerance, as it was previously reported for salt stress on wheat. Thus the effect of 50 mM CaCl2, previously chosen based on pilot experiment, versus water-treated control exposed to drought stress was investigated.