毛竹小RNA高通量测序及病毒分析
Small RNA Analysis Using High-Throughput Sequencing and Virus Investigation in Bamboo (Phyllostachys edulis)
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摘要: 以毛竹叶片为材料，采用小RNA高通量测序结合生物信息学对小RNA数据库进行组装，进一步分析了毛竹中存在的病毒和类病毒，并采用RT-PCR和RACE进行验证。结果表明：在竹子样品中存在水稻东格鲁病毒（RTBV），覆盖率达到91.0%。在毛竹样品中扩增得到1 992 bp RTBV病毒类似序列，占其基因组的24.9%。RTBV病毒在多个毛竹样品中存在且不存在多态性。RTBV病毒可能是一个古老的植物病毒，在进化过程中禾本科植物将其序列整合到基因组中来防御RTBV病毒的浸染。Abstract: The leaves of Phyllostachys edulis were harvested for RNA extraction, a sRNA library was constructed and sequenced by using the Solexa high-throughput sequencing technology. Bioinformatics was performed to identify the sRNAs and viruses. RT-PCR and RACE techniques were explored to identify and confirm the viruses or viroid in bamboo. The assembly of sRNA in all samples revealed that a sequence which covered 91.0% of rice tungro bacilliform virus (RTBV) genome existed widely in bamboo samples. Meanwhile, 1 992bp DNA sequence which covered 24.9% of RTBV completed genome was amplified and no polymorphism was observed in this samples. It concluded that RTBV virus may exist in planta in ancient time and was late integrated into the plant genomes against the RTBV infection.
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Key words:
- Phyllostachys edulis
- / high-throughput sequencing
- / small RNA
- / RTBV
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