Project description:We analyzed global gene expression in the crown tip of 2 pearl millet (Pennisetum glaucum) inbred lines with high (line 249) and low (line 220) root soil aggregation using RNAseq. The obtective was to identify genes potentially associated with changes in rhizosheath formation.
Project description:We analysed global gene expression in the primary root tip of 2 pearl millet (Pennisetum glaucum) inbred lines with high (line 249) and low (line 337) primary root growth using RNAseq. The objective was to identify genes potentially associated with changes in root growth.
Project description:Screening cDNA clones from SSH library by hybridization with cDNA used to construct the library. Treated samples were drought-stressed cowpea plants, and control samples were cowpea plants subjected to a standard watering regime. cDNA clones from forward and reverse libraries are spotted on the same array, but data from each library were analysed separately after normalization using SSHscreen software (http://microarray.up.ac.za/SSHscreen ) to calculate Enrichment Ratio 3 (ER3) and Enrichment Ratio 2 (ER2) values for each clone. ER3 is a measure of differential expression, and was determined using a set of hybridizations with unsubtracted treated (UT) and unsubtracted control (UC) cDNA. ER2 is a measure of the relative abundance of a clone's transcript in the original tester sample, relative to other transcripts in the sample prior to the SSH process. ER2 for the forward library was determined using a set of hybridizations with subtracted treated (ST) and unsubtracted treated (UT) cDNA. ER2 for the reverse library is determined using a set of hybridizations with subtracted control (SC) and unsubtracted control (UC) cDNA.
Project description:The magnocellullar neurons (MCNs) of the supraoptic nucleus (SON) of the hypothalamus are the principle source of the neuropeptide hormone vasopressin (VP), which has a crucial role in osmoregulation. Physiological activation of the SON by dehydration results is a massive release of VP from stores in posterior pituitary axon terminals into the general circulation. By binding to V2-type receptors located in the kidney, VP decrease the amount of water lost in urine. Osmotic activation of the SON is accompanied by a dramatic morphological and functional remodelling. We have sought to understand the mechanistic basis of this plasticity in terms of the differential expression of genes. To identify such genes, we adopted a completely unbiased and global approach based on Suppressive Subtractive Hybridisation-Polymerase Chain Reaction (SSH-PCR) Using this method, we generated a library of clones putatively differentially expressed in control vs dehydrated SON. In order to rapidly screen this library, 1152 of these clones were subjected to microarray analysis, resulting in the identification of 459 differentially expressed clones, 56 of which were sequenced. Many of these clones are expressed sequences that are new to science. Four of these were shown by in situ hybridisation (ISH) to be significantly up- or down-regulated in the SON following dehydration. These genes may represent novel effectors or mediators of SON physiological remodelling. Keywords: dehydrated, SON, clone
Project description:Screening cDNA clones from SSH library by hybridization with cDNA used to construct the library. Treated samples were drought-stressed cowpea plants, and control samples were cowpea plants subjected to a standard watering regime. cDNA clones from forward and reverse libraries are spotted on the same array, but data from each library were analysed separately after normalization using SSHscreen software (http://microarray.up.ac.za/SSHscreen ) to calculate Enrichment Ratio 3 (ER3) and Enrichment Ratio 2 (ER2) values for each clone. ER3 is a measure of differential expression, and was determined using a set of hybridizations with unsubtracted treated (UT) and unsubtracted control (UC) cDNA. ER2 is a measure of the relative abundance of a clone's transcript in the original tester sample, relative to other transcripts in the sample prior to the SSH process. ER2 for the forward library was determined using a set of hybridizations with subtracted treated (ST) and unsubtracted treated (UT) cDNA. ER2 for the reverse library is determined using a set of hybridizations with subtracted control (SC) and unsubtracted control (UC) cDNA. Direct comparison of UT and UC cDNAs to calculate ER3 values for F and R library, including dye swaps and replicate arrays. Direct comparison of ST and UT cDNAs to calculate ER2 values for F library, including dye swaps and replicate arrays. Direct comparison of SC and UC cDNAs to calculate ER2 values for R library, including dye swaps and replicate arrays.
Project description:High salinity is one of the most serious threats to crop production. To 1 better understand the molecular basis of plant responses to salt stress, we combined suppression subtractive hybridization (SSH) and microarray approaches to identify the potential important or novel genes involved in salt tolerance. First, SSH libraries were constructed for two cultivated tomato (Solanum lycopersicum) genotypes: LA2711, a salt tolerant cultivar, and ZS-5, a salt sensitive cultivar, to compare salt treatment and non-treatment plants. Then a subset of clones from these SSH libraries were used to construct a tomato cDNA array and microarray analysis was carried out to verify the expression changes of this set of clones upon salt treatment at various time points compared to the corresponding non-treatment controls. A totalof 201 non-redundant genes differentially expressed upon 30 min of salt stress treatment either in LA2711 or ZS-5 were identified from microarray analysis, most of which were not previously associated with salt stress. The diversity of the putative functions of these genes indicated that salt stress resulted in a complex response in tomato plants. Keywords: gene expression, genotype, microarray, salt stress, SSH, tomato
Project description:The magnocellullar neurons (MCNs) of the supraoptic nucleus (SON) of the hypothalamus are the principle source of the neuropeptide hormone vasopressin (VP), which has a crucial role in osmoregulation. Physiological activation of the SON by dehydration results is a massive release of VP from stores in posterior pituitary axon terminals into the general circulation. By binding to V2-type receptors located in the kidney, VP decrease the amount of water lost in urine. Osmotic activation of the SON is accompanied by a dramatic morphological and functional remodelling. We have sought to understand the mechanistic basis of this plasticity in terms of the differential expression of genes. To identify such genes, we adopted a completely unbiased and global approach based on Suppressive Subtractive Hybridisation-Polymerase Chain Reaction (SSH-PCR) Using this method, we generated a library of clones putatively differentially expressed in control vs dehydrated SON. In order to rapidly screen this library, 1152 of these clones were subjected to microarray analysis, resulting in the identification of 459 differentially expressed clones, 56 of which were sequenced. Many of these clones are expressed sequences that are new to science. Four of these were shown by in situ hybridisation (ISH) to be significantly up- or down-regulated in the SON following dehydration. These genes may represent novel effectors or mediators of SON physiological remodelling. 4 chips were hybridised with control SON and compared to 4 chips hybridised with dehydrated SON
Project description:Pearl millet (Pennisetum glaucum) is a cereal crop well-adapted to arid and high-temperature environments. Its root system exhibits a high degree of complexity, comprising distinct root types with specific anatomical and functional characteristics. These include the primary root, which is established during embryogenesis; crown roots, which develop at the base of the stem post-embryonically; and lateral roots, which emerge from both the primary and crown roots. In this study, we conducted a comparative analysis of gene expression profiles across these distinct root types to better understand their functional specialization.