Project description:MicroRNAs (miRNAs) are a type of small non-coding RNAs, which play important roles in plant growth, development and stress responses. Tea (Camellia sinensis) prepared from tea tree is the oldest and most popular nonalcoholic beverages in the world, and has large economic, medicinal and cultural significance. Nevertheless, there are a few studies on the miRNAs and their functions in Camellia sinensis. We sequenced 9 small RNA libraries and 9 RNA-Seq libraries from roots, leaves and flowers tissues. Through comprehensive computational analyses of 9 small RNA profiles, we identified 200 conserved miRNAs of which 138 have not been reported, and 56 novel miRNAs with 33 have not been reported. Nearly, two thousands genes have significantly different expression levels in tissues. In order to identify targets of miRNAs, we sequenced two degradome profiles from leaves and roots, respectively. Totally, more than 3,000 putative targets of conserved miRNAs were identified in both degradome profiles by using the SeqTar algorithm. These results clearly enhanced our understanding about small RNA guided gene regulations in Camellia sinensis.
Project description:MicroRNAs (miRNAs) are a type of small non-coding RNAs, which play important roles in plant growth, development and stress responses. Tea (Camellia sinensis) prepared from tea tree is the oldest and most popular nonalcoholic beverages in the world, and has large economic, medicinal and cultural significance. Nevertheless, there are a few studies on the miRNAs and their functions in Camellia sinensis. We sequenced 9 small RNA libraries and 9 RNA-Seq libraries from roots, leaves and flowers tissues. Through comprehensive computational analyses of 9 small RNA profiles, we identified 200 conserved miRNAs of which 138 have not been reported, and 56 novel miRNAs with 33 have not been reported. Nearly, two thousands genes have significantly different expression levels in tissues. In order to identify targets of miRNAs, we sequenced two degradome profiles from leaves and roots, respectively. Totally, more than 3,000 putative targets of conserved miRNAs were identified in both degradome profiles by using the SeqTar algorithm. These results clearly enhanced our understanding about small RNA guided gene regulations in Camellia sinensis.
Project description:MicroRNAs (miRNAs) are a type of small non-coding RNAs, which play important roles in plant growth, development and stress responses. Tea (Camellia sinensis) prepared from tea tree is the oldest and most popular nonalcoholic beverages in the world, and has large economic, medicinal and cultural significance. Nevertheless, there are a few studies on the miRNAs and their functions in Camellia sinensis. We sequenced 9 small RNA libraries and 9 RNA-Seq libraries from roots, leaves and flowers tissues. Through comprehensive computational analyses of 9 small RNA profiles, we identified 200 conserved miRNAs of which 138 have not been reported, and 56 novel miRNAs with 33 have not been reported. Nearly, two thousands genes have significantly different expression levels in tissues. In order to identify targets of miRNAs, we sequenced two degradome profiles from leaves and roots, respectively. Totally, more than 3,000 putative targets of conserved miRNAs were identified in both degradome profiles by using the SeqTar algorithm. These results clearly enhanced our understanding about small RNA guided gene regulations in Camellia sinensis.
Project description:Tea (Camellia sinensis (L.) O. Kuntze) is an important non-alcoholic commercial beverage crop. Tea tree is a perennial plant, and winter dormancy is its part of biological adaptation to environmental changes. We recently discovered a novel tea tree cultivar that can generate tender shoots in winter, but the regulatory mechanism of this ever-growing tender shoot development in winter is not clear. In this study, we conducted a proteomic analysis for identification of key genes and proteins differentially expressed between the winter and spring tender shoots, to explore the putative regulatory mechanisms and physiological basis of its ever-growing character during winter.
Project description:Anthracnose disease is caused by Colletotrichum gloeosporioides, and is common in leaves of the tea plant Camellia sinensis. MicroRNAs (miRNAs) have been known as key modulators of gene expression in defense responses; however, the role of miRNAs in tea plant during defensive responses to C. gloeosporioides remains unexplored. Six miRNA sequencing data sets and two degradome data sets were generated from C. gloeosporioides-inoculated and control tea leaves. A total of 485 conserved and 761 novel miRNAs were identified. Of those, 239 known and 369 novel miRNAs exhibited significantly differential expression under C. gloeosporioides stress. 1134 and 596 mRNAs were identified as targets of 389 and 299 novel and conserved miRNAs by degradome analysis, respectively. The expression levels of twelve miRNAs and their targets were validated by quantitative real-time PCR. The predicted targets of five interesting miRNAs were further validated through 5'RLM-RACE. Furthermore, Gene Ontology and metabolism pathway analysis revealed that most of the target genes were involved in translation, carbohydrate metabolism and signal transduction pathways. This study enriches the resources of defense-responsive miRNAs and their targets in C. sinensis, and thus, provides novel insights into the miRNA-mediated regulatory mechanisms underlying immunity responses to biotic stress in tea plant.
Project description:Basic region/leucine zipper (bZIP) transcription factors play vital roles in the abiotic stress response of plants. However, little is known about the function of bZIP genes in Camellia sinensis. Here, we show that CsbZIP6 is induced during cold acclimation in tea plant. Constitutive overexpression of CsbZIP6 in Arabidopsis lowered the plants’ tolerance to freezing stress and ABA exposure during seedling growth. Compared to wildtype (WT) plants, CsbZIP6 overexpression (OE) lines exhibited increased levels of electrolyte leakage (EL) and malondialdehyde (MDA) contents and reduced levels of total soluble sugars (TSS) under cold stress conditions. Microarray analysis of transgenic Arabidopsis revealed that many differentially expressed genes (DEGs) between OE lines and WT plants could be mapped to ‘response to cold’ and ‘response to water deprivation’ terms based on GO analysis. Interestingly, CsbZIP6 overexpression repressed most of the cold- and drought-responsive genes as well as the starch metabolism under cold stress conditions. Taken together, our data suggests that CsbZIP6 functions as a negative regulator of the cold stress response in Arabidopsis thaliana, potentially by down-regulating cold-responsive genes. To obtain insights into the molecular mechanisms by which CsbZIP6 mediates senstivity to cold stress in Arabidopsis plants, gene expression profiles in leaves of two CsbZIP6 OE lines and WT plants under normal (22ºC) and cold (4ºC) conditions were compared. The Agilent Arabidopsis Gene Expression (4×44K, Design ID: 021169) was used in this experiment.
Project description:Solexa sequencing technology was used to perform high throughput sequencing of the small RNA library from the cold treatment of tea leaves. Subsequently, aligning these sequencing date with plant known miRNAs, we characterized 112 C. sinensis conserved miRNAs. In addition, 215 potential candidate miRNAs were found; among them, 131 candidates with star sequence were chosen as novel miRNAs. There are both congruously and differently regulated miRNAs, and line-specific miRNAs were identified by microarray-based hybridization in response to cold stress. The miRNA chip included 3228 miRNA probes corresponding to miRNA transcripts listed in Sanger miRBase release 19.0 and 283 novel miRNAs probes founding in tea plant. In the study presented here, two tea plant cultivars, ‘Yingshuang’ (YS, a cold-tolerant tea plant cultivar) and ‘Baiye 1’ (BY, a cold-sensitive tea plant cultivar), were kept at 4°C for 4,12, 24 h, respectively, and 28°C for as control. These samples were used to acquire expression profiles of a total of 3,511 unique genes, leading to the successful construction of supervised
Project description:We discovered 31 up-regulated miRNAs and 43 down-regulated miRNAs in ‘Yingshuang’, and 46 up-regulated miRNA and 45 down-regulated miRNAs in ‘Baiye 1’ in response to cold stress, respectively. A total of 763 related target genes were detected by degradome sequencing. The RLM-5’RACE procedure was successfully used to map the cleavage sites in six target genes of C. sinensis. These findings reveal important information about the regulatory mechanism of miRNAs in C. sinensis, and promote the understanding of miRNA functions during the cold response. The miRNA genotype-specific expression model might explain the distinct cold sensitivities between tea lines.
Project description:Tea plants (Camellia sinensis) present an excellent system to study evolution and diversification of specialized metabolites due to their abundance in classes, numbers and contents. A large number of tea cultivars have been cultivated throughout the world not only because of their adaption to different environments but of selection for specific flavors. The chemical and genetic basis for unique taste and aroma of different tea cultivars remains largely unknown, but is critical for guiding genetic breeding of new cultivars. Using transcriptomic data from 136 representative tea accessions in China, we obtain 925,854 high-quality single-nucleotide polymorphisms (SNPs) useful for marker-assisted breeding. Phylogenetic and population structure analyses separate sampled tea accessions into five major groups. Different major alleles are identified on 1183 SNP sites for the two major types of tea, C. sinensis var. assamica (CSA) and C. sinensis var. sinensis (CSS), reflecting fixation of these alleles after population divergence. Non-targeted metabolomic analyses detect 2,818 and 2,311 metabolic features in tea samples in positive and negative ionization modes, respectively, including 355 and 286 metabolites respectively that are differentially accumulated in different tea groups. Each phylogenetic group contains signature metabolites. In particular, CSA tea accessions are featured with high accumulation of diverse classes of flavonoid compounds, such as flavanols, flavonol mono-/di-glycosides and proanthocyanidin dimers. Comparisons of gene expression profiles of different tea groups identify hundreds of differentially expressed genes with some involved in the biosynthesis of characteristic tea metabolites, reflecting a combinational effect of genetic and environmental factors. Taken together, our study provides new insights into the phylogenetic relationships, molecular markers, metabolite compositions, and gene expression profiles of representative cultivated tea accessions in China, which are beneficial for targeted tea breeding and improvement.