| 朱妍峰，王 倩，赵惠贤.病毒诱导的全新 tae-miR9663的沉默影响叶片发育和籽粒大小[J].麦类作物学报,2017,(4):465 |
| 病毒诱导的全新 tae-miR9663的沉默影响叶片发育和籽粒大小 |
| Silencing of tae-miR9663 in Wheat by VIGS Affects Leaf Development and Seed Size |
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| DOI：10.7606/j.issn.1009-1041.2017.04.06 |
| 中文关键词: 小麦 tae-miR9663 STTM 病毒诱导的基因沉默（VIGS）技术 |
| 英文关键词:Wheat tae-miR9663 STTM Virus-induced gene silencing (VIGS) technology |
| 基金项目:国家自然科学基金青年基金项目（31501295）；国家自然科学基金面上项目（31471482） |
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| 中文摘要: |
| MicroRNA（miRNA）是一类长度为18~24 nt的非编码小分子RNA，参与植物各种发育进程。 tae-miR9663是新发现的在小麦幼苗、旗叶和籽粒中高表达的miRNA，但其生物学功能未知。为了探索 tae-miR9663的功能，通过人工合成 tae-miR9663的小串联模拟靶标（short tandem target mimic, STTM），将其构建到大麦条斑花叶病毒（barley stripe mosaic virus, BSMV）载体上，利用病毒介导的基因沉默（virus-inducing gene silencing, VIGS）技术转染小麦宁春16五叶一心期的第5片叶，转染20 d后观察叶片表型并取旗叶进行实时定量PCR，成熟时观察种子大小。叶片表型观察结果表明，与BSMV:00相比，接种BSMV:STTM- tae-miR9663的9株幼苗中出现4种叶片表型，即第6片叶有白点或白条纹，旗叶（第7片叶）有白点或白条纹，第6片叶边缘有锯齿状，旗叶叶尖处有皱缩。实时定量PCR分析结果表明，STTM- tae-miR9663过表达植株的 tae-miR9663表达丰度下降，说明BSMV-VIGS技术可通过过表达STTM有效地沉默内源miRNA。成熟种子大小观察结果表明，与BSMV:00比较，接种BSMV:STTM- tae-miR9663的植株种子的长和宽均减小。 |
| 英文摘要: |
| MicroRNA (miRNA), which is a class of non-coding small RNA, with 18 to 24 nt in length, regulates various developmental processes in plants. tae-miR9663 is a novel miRNA highly expressing in seedling, flag leaf and seed in wheat, but its biological function is unknown. In order to explore the function of tae-miR9663, STTM (short tandem target mimic) of tae-miR9663 was synthesized, which was connected to BSMV (barley stripe mosaic virus) vector. The fifth leaves were inoculated by VIGS (virus-inducing gene silencing) in Ningchun 16. The phenotypes of leaves were observed at 20 d post-inoculation. tae-miR9663 expression in flag leaves was detected by real-time quantitative PCR and size of mature seeds was observed. Phenotypic observation results on leaves indicated that compared with BSMV:00,there are four phenotypes including dots or white stripes appeared on the sixth leaf,dots or white stripes appeared on flag leaf(the seventh leaf),serrate margins appeared in the sixth leaf,and shrinking existed on flag leaf blade tips.Real-time quantitative PCR results indicated that the expression of tae-miR9663 was decreased, showing that BSMV-VIGS could silence endogenous miRNA by overexpression of STTM effectively. Comparison of seeds from BSMV:STTM- tae-miR9663 with those from control plants, the lengths and widths of seeds from BSMV:STTM- tae-miR9663 inoculated plants were decreased. |
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