Project description:Tea is a popular and natural non-alcoholic beverage, and is produced from fresh leaves of Camellia sinensis. Tea leaves contain many bioactive compounds that have significant health benefits. We constructed a high quality bacterial artificial chromosome (BAC) library by using the fresh petals of C. sinensis ‘Shuchazao’ for genome sequencing and improvement of genomic assembly.
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:Next-generation sequencing (NGS) was performed to identify genes changed in tea plant cultivar Zhongcha 108 upon Colletotrichum camelliae infection. The goal of the work is to find interesting genes involved in tea plant in response to fungi infection. The object is to reveal the molecular mechanism of tea plant defense.
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.
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:To investigate the large-scale gene expression in different tea clones, a custom oligo microarray was developed using sequences from RNA-seq for probe designing. We succeeded in developing a tea oligo microarray resource which can be successfully used to analyze gene expression in any tea clones without the need for prior sequence knowlege.
Project description:In this study, we performed deep sequencing and bioinformatics analyses of tea plant leaves to identify and characterize known and novel miRNAs. A total of 26,876,261 raw reads were produced from 2 libraries. We detected 422 known miRNAs belonging to 125 families, and 68 putative novel miRNAs.
Project description:Anthracnose, caused by Colletotrichum gloeosporioides is one of the most serious diseases of tea plant [Camellia sinensis (L.) O. Kuntze]. MicroRNAs are key modulators of gene expression in defense responses and plant immunity; although, foliar application of exogenous caffeine in anthracnose disease control management has proven to be effective, miRNA-mediated regulatory mechanisms underlying caffeine-induced plant defense response to C. gloeosporioides remain unexplored in tea plant. Using high-throughput-sequencing, 24 miRNA sequencing data sets and 8 degradome data sets were generated from the susceptible cultivar Longjing43 (LJ43) and the resistant cultivar Zhongcha108 (ZC108) leaves treated with CK (Water), C. gloeosporioides-inoculation (CgI), exogenous caffeine (CN) and CgI + CN. Using sRNA sequencing, 424 conserved miRNAs and 417 novel miRNAs were identified; of these, 146 and 130 miRNAs were differentially expressed under CgI + CN treatment in the LJ43 and ZC108, respectively. Degradome sequencing identified 599 targets predicted to be cleaved by 210 conserved and 70 novel miRNAs. Majority of the annotated targets were found to involve in regulation of transcription factors, oxidation-reduction and metabolic process for plant growth and development as well as stress responses in tea plant against C. gloeosporioides stress. The expression pattern of eight miRNAs and their targets were validated by qRT-PCR, and correlation analysis of csn-miR164a_R+1_1ss21AG/NAC-17 and csn-miR396b-5p/GRF-1 showed highly significant negative R-value at 7th dpi under CgI + CN in the LJ43. This study provides important insights into the novel approach of exogenous caffeine-induced miRNAs dynamically exerts its fungicidal activity through regulating JA/ET signaling pathway, thereby accurately switch on LJ43 susceptibility nature to resistance activity against C. gloeosporioides infection.
