Transcription profiling of parental lines and bulked salt sensitive and salt tolerant RILs derived from 2 rice varieties
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ABSTRACT: The aim of this study was to minimize the number of candidate genes responsible for salt tolerance between a pair of rice varieties (CSR27 and MI48) with contrasting level of salt tolerance by bulked segregant analysis of their recombinant inbred lines. Microarray analysis of RNA extracted from the tolerant and susceptible parents without and with stress showed 798 and 2407 differentially expressed genes, respectively. The number of differentially expressed genes was drastically reduced to 70 and 30, by pooling the RNAs from ten extreme tolerant and ten extreme susceptible RILs due to normalization of irrelevant differentially expressed genes between the parents. RNA from CSR27, MI48, tolerant bulk and susceptible bulk grown under control and salt stress conditions were analysed in two different biological replications (A and B) making total sixteen samples
Project description:The aim of this study was to minimize the number of candidate genes responsible for salt tolerance between a pair of rice varieties (CSR27 and MI48) with contrasting level of salt tolerance by bulked segregant analysis of their recombinant inbred lines. Microarray analysis of RNA extracted from the tolerant and susceptible parents without and with stress showed 798 and 2407 differentially expressed genes, respectively. The number of differentially expressed genes was drastically reduced to 70 and 30, by pooling the RNAs from ten extreme tolerant and ten extreme susceptible RILs due to normalization of irrelevant differentially expressed genes between the parents.
Project description:A submergence tolerant indica rice cultivar FR13A, was also reported to withstand salt stress and proven in our experiments. The mechanism of tolerance is yet to be studied by forward genetics approach. However, it is known that salt stress tolerance is governed by several QTLs and not by a single gene. To understand the mechanism of such a complex mechanism of salt tolerance we selected, two indica rice genotypes namely, I) FR13A, a tolerant indica variety and ii) IR24, a susceptible genotype for this study. We used the 22K rice Oligoarray from Agilent technologies to study the transcript profile in the leaves of the two contrasting rice genotypes under constitutive and salt stress conditions at seedling stage. Experiment Overall Design: We used Agilent rice gene chips (G4138A) to investigate the transcript level changes in rice plant tissues during salt stress treatment. We used two contrasting rice genotypes (FR13A tolerant and IR24 susceptible) differing in salt stress response. Plants were grown in growth chambers and treated with 150 mM salt concentration at 14th DAS. Sampling was done in both constitutive and treated plants at 3 time points. Two replications of microarray experiments were carried out by hybridizing the RNA from tolerant samples against the susceptible lines on the same slide.
Project description:A submergence tolerant indica rice cultivar FR13A, was also reported to withstand salt stress and proven in our experiments. The mechanism of tolerance is yet to be studied by forward genetics approach. However, it is known that salt stress tolerance is governed by several QTLs and not by a single gene. To understand the mechanism of such a complex mechanism of salt tolerance we selected, two indica rice genotypes namely, I) FR13A, a tolerant indica variety and ii) IR24, a susceptible genotype for this study. We used the 22K rice Oligoarray from Agilent technologies to study the transcript profile in the leaves of the two contrasting rice genotypes under constitutive and salt stress conditions at seedling stage. Keywords: Mechanism of salt tolerance
Project description:To detect salt-tolerance-related miRNAs, comparative analysis of miRNA expression profiles was performed between the salt-tolerant and -sensitive cotton cultivars in control and salt-stressed conditions (treated with 300 mM NaCl for 24 h) using microRNA microarray
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:To detect salt-tolerance-related miRNAs, comparative analysis of miRNA expression profiles was performed between the salt-tolerant and -sensitive cotton cultivars in control and salt-stressed conditions (treated with 300 mM NaCl for 24 h) using microRNA microarray Total RNA was extracted from (1) the seedling of salt-tolerant cotton cultivar in normal growth conditions, (2) the seedling of salt-tolerant cotton cultivar in salt-stressed growth conditions, (3) the seedling of salt-sensitive cotton cultivar in normal growth conditions, and (4) the seedling of salt-sensitive cotton cultivar in salt-stressed growth conditions. Then, the low-molecular-weight RNA (LMW-RNA) was isolated using the PEG solution precipitation method and used to hybridization.
Project description:The N6-methyladenosine (m6A) modification is the most common internal post-transcriptional modification, with important regulatory effects on RNA export, splicing, stability,and translation. However, the effects of m6A modifications on the resistance of sweet sorghum to salt stress remain unclear. In this study, we mapped the m6A modifications in two sorghum inbred lines (salt-tolerant M-81E and salt-sensitive Roma) that differ regarding salt tolerance. Dynamic changes to m6A modifications in sweet sorghum were identified in response to salt stress. Our data suggest that the differences in the m6A modifications between salt-tolerant and salt-sensitive sweet sorghum might contribute to the diversity in salt tolerance.
Project description:Salt stress has become one of the main abiotic stress factors restricting agricultural production worldwide. Sweet sorghum is an important salt and drought tolerant feed and energy crop. Its salt tolerance mechanism has not been widely studied. With the development of transcriptome sequencing technology, it is possible to study the molecular mechanism of sweet sorghum salt tolerance. The purpose of this study was to further reveal the potential salt-tolerant molecular mechanisms of sweet sorghum through high-throughput sequencing analysis of the transcriptome. Finally, through high-throughput sequencing, we read approximately 54.4G of raw base and 53.7G of clean base in total, and used FastQC to assign a quality score (Q) to each base in the read using a similar phred algorithm, Analysis shows that the data is highly credible. We conclude that RNA-based transcriptome characterization will accelerate the study of genetics and molecular biology of sweet sorghum salt tolerance mechanisms and provide a framework for this.
Project description:Over the years, many traditional rice varietites of India were sourced and studied owing to their ability to withstand abiotic pressures like excessive salinity in the soil and water. These cultivars growing in specific areas of the country represent a rich gene pool from where a deeper understanding of the processes underlying tolerance to abiotic stress can be gained. Indigenous varieties like Nonabokra and Pokkali are known salt tolerant varieties and are being studied in great detail. In the present study, we have analyzed the transcriptomes of the contrasting cultivars; Nonabokra (tolerant), Pokkali (tolerant) and IR29 (susceptible) in order to decipher the differences in their responses to salinity stress by utilizing microarray.
Project description:Sprobolus virginicus is a halophytic C4 grass found in worldwide from tropical to warm temperate regions. A Japanese genotype showed a salinity tolerance up to 1,500 mM NaCl, a three-fold higher concentration than seawater salinity. To identify key genes involved in the regulation of salt tolerance in S. virginicus, random cDNA libraries were constructed from salt-treated leaves, and were introduced into Arabidopsis for salt tolerant plant screening. Eight independent transgenic lines were found to be more salt tolerant than wild type from the screen of 3011 lines on the medium containing 175 mM NaCl. Among the selected lines, two contained cDNAs encoding glycine-rich RNA-binding proteins (GRPs). To identify transcriptomic change in the GRP-transgenic line, we performed microarray analysis of the transgenic line and WTunder salt stress.