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茶树miRNA的分离鉴定及其与茶尺蠖取食诱导的差异表达特征分析-茶学专业论文
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Abstract
MicroRNAs are a class of endogenous,non-coding single-stranded small RNAs that regulate numerous genes by guiding target mRNA cleavage in plant, and miRNAs play an important role in various aspects of plant growth and development process and environment stress responses. So it is becoming concerned focus about the regulation role of miRNA.. However, little attetion has been focused on the function of miRNA in plant defense mechanisms against pests, and a few tea miRNA sequences have been reported. So the paper performed the small RNA deep sequencing, miRNA microarray, qRT-PCR method etc and combined with bioinformatics analysis, isolated and identified miRNAs in tea (Camellia sinensis) by using these technologies, on this basis we found differently expressed miRNAs in tea plants after E.oblique attack to further confirm their characteristics and functions. The main results of paper were as follows:
Solexa high-throughput sequencing enabled the exploration of small RNA populations in tea plant, 8,796,700 effective sequences were obtained, accounting for 81.30% of all reads, 27% of all miRNA sequences were more than 100 times by sequences analyzing. We used qRT-PCR approach to detect the miRNAs of different expressed levels, the results were consistent with the high throughput sequencing results.
In all obtained miRNA sequences, 253 were coserved miRNA belonging to 30 miRNA families and 574 tea-specific sequences. The number of 16 conserved miRNA family members were hiher than other plant species (Arabidopsis etc). Bioinformatics analysis showed target genes of many tea miRNAs involved in plant growth and development, secondary metabolism, stress response process and signal transduction etc.
Microarray validated the sequencing results and obtained 169 miRNA sequences, included 130 conserved miRNAs and 39 tea-specific miRNAs. Further analysis revealed that 35 miRNAs were significantly with higher abundance in leaf, while 60 miRNAs were higher in root.
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