Project description:Transcriptomic analysis of Col-0 and AITF1 overexpressing transgenic plants was used to screen the differentially expressed genes between AITF1 overexpressing transgenic plants and Col-0. Further, transcriptomic analysis of Col-0 and aitf1-1asr3-1 plants was used to screen the differentially expressed genes between aitf1-1asr3-1 and Col-0 after flg22 treatment for indicated times, we analyzed the ASR3/AITF1-regulated target genes to further elucidate the function of ASR3 and AITF1 in plant immune response.
Project description:We examined global expression profiles of 7-days old 35S-TrAP transgenic plants compared to Col-0 wild-type using an Affymetrix ATH1 GeneChip and identified 586 genes that are differentially expressed in the 35S-TrAP transgenic plants (q<0.005). Of these, 295 transcripts were elevated whereas 291 were reduced (Figure 2E). We performed real-time PCR and RNA blot assays to validate the microarray results for the differentially expressed genes (DEGs).
Project description:Solanum lycopersicoides (LA2408), collected at higher altitudes (up to 3600 meters) than any of other solanum species, is a wild nightshade distant-allied to cultivated tomato. Many traits of Solanum lycopersicoides including cold tolerance, resistance to virus diseases and insect pests were previously confirmed. Thus, it is an ideal candidate plant for isolating cold-tolerance-related genes. In this study, we successfully cloned the full-length cDNA of the CBF1 gene from Solanum lycopersicoides which was designated as SsCBF1. In order to investigate the possible functions of SsCBF1 in plant growth and stress responses, we generated transgenic Arabidopsis overexpressing SsCBF1. We employed the RNA-seq approach to identify the differentially expressed genes between the two genotypes. Processing of RNA samples on the Illumina HiSeq 2000 system produced more than 20 million reads, each 100bp in length, encompassing 2.0 Gb of sequence data for each sample which was then mapped to the reference genome. The statistical analysis identified a total of 338 differentially expressed genes between Col-0 (WT) and transgenic Arabidopsis overexpressing SsCBF1 with the criteria of Q-value < 0.001 and fold change >2, among which 120 (35.5%) were up-regulated and 218 (64.5%) were down-regulated. RNA-sequencing was carried out using one transgenic line (35S:SsCBF1-11) and the Col-0 (WT) plants. Total RNA was isolated with Trizol reagent (Invitrogen, USA) from the aerial parts of 4-week-old seedlings grown in parallel under unstressed conditions. Materials from 20 plants of each genotype were pooled for RNA isolation.
Project description:Inorganic phosphate (Pi) is an essential nutrient, which is often served as a limiting factor in plant growth. It has been reported that SPL family members, such as SPL3, regulate Pi deficiency responses by controlling the expression of Pi deficiency responsive genes. To elucidate whether SPL9 respond to low phosphorus stress, we investigated the phenotypes and conduct RNA sequencing analysis in transgenic Arabidopsis thaliana with overexpressing SPL9 (R9) under conditions of both normal and low Pi availability. Compared with wild-type plants, R9 showed decreased anthocyanin accumulation and increased Pi contents in shoots under Pi deficiency. Through RNA-seq analysis compared with wild-type plants, we detected 217 genes significantly differentially expressed in conditions of Pi sufficiency, and 121 genes differentially expressed in conditions of Pi deficiency in R9 plants. Under Pi deficiency, these genes included multiple protein kinases, jasmonic acid response genes and genes related to salt stress responses. Genes associated with hydrolase and transferase activity were also differentially regulated by Pi deficiency, such as cytochrome P450 monooxygenases. Of particular note, the transcription factor AP2-EREBP and members of the bHLH family were among the most significantly differentially regulated genes identified under both Pi sufficient and Pi deficient conditions.