香鳞毛蕨dfr-miR160a和靶基因DfARF10的生物信息学及表达模式分析
收稿日期: 2023-02-27
录用日期: 2023-08-29
网络出版日期: 2023-09-07
基金资助
国家自然科学基金(32270394)
Bioinformatic and Expression Pattern Analysis of dfr-miR160a and Target Gene DfARF10 in Dryopteris fragrans
Received date: 2023-02-27
Accepted date: 2023-08-29
Online published: 2023-09-07
为深入探究miRNA参与调控香鳞毛蕨(Dryopteris fragrans)生长发育的分子机理, 依据实验室前期建立的miRNA数据库, 筛选出与非生物胁迫相关的差异表达dfr-miR160a前体(dfr-pri-mir160a), 预测其靶基因为DfARF10, 并通过本氏烟草(Nicotiana benthamiana)瞬时共转化GUS以及双荧光素酶(LUC)系统验证dfr-miR160a和DfARF10的靶向关系。结果显示, 共注射dfr-pri-mir160a和DfARF10的本氏烟草叶片中GUS活性和LUC活性明显降低; qRT-PCR分析显示, dfr-miR160a及其靶基因DfARF10在香鳞毛蕨的根、配子体、叶柄、叶片和孢子囊中均有表达, 在叶片中表达量最高, 在根中表达量最低; 通过qRT-PCR分析干旱、盐(NaCl)、高温和低温胁迫对dfr-miR160a及其靶基因DfARF10表达的影响。结果表明, 在干旱和高温处理下, dfr-miR160a的表达均上调, 但在NaCl处理下, dfr-miR160a的表达下调; 低温处理下, dfr-miR160a的表达在0-1小时下调, 在3-48小时上调。在NaCl、高温以及低温处理下DfARF10表达均上调; 但在干旱处理下, DfARF10表达下调, 与dfr-miR160a呈现相反的表达趋势。综上, dfr-miR160a靶向DfARF10基因且二者均能响应非生物胁迫。研究结果为从分子层面揭示香鳞毛蕨非生物胁迫抗性机制提供了新的科学依据。
关键词： 香鳞毛蕨; dfr-miR160a; DfARF10; 非生物胁迫
仲昭暄 , 张冬瑞 , 李璐 , 苏颖 , 王黛宁 , 王泽冉 , 刘洋 , 常缨 . 香鳞毛蕨dfr-miR160a和靶基因DfARF10的生物信息学及表达模式分析[J]. 植物学报, 2024 , 59(1) : 22 -33 . DOI: 10.11983/CBB23025
To further understand the molecular mechanism underlying miRNA regulation of growth and development of Dryopteris fragrans, we screened the differentially expressed dfr-pri-mir160a through the miRNA database established earlier in the laboratory, and predicted its target gene as DfARF10. The target relationship between dfr-pri-mir160a and DfARF10 was verified by tobacco transient co-transformation, together with double luciferase (LUC) activity. The results showed that the GUS and LUC activity in tobacco leaves co-injected with dfr-pri-mir160a and DfARF10 decreased significantly. qRT-PCR analysis showed that dfr-miR160a and its target gene DfARF10 were expressed in the gametophytes, roots, petioles, leaves and sporangium of D. fragrans, with the highest expression in the leaves and the lowest in the roots. We analyzed the effects of drought, NaCl, high temperature and low temperature stress treatments on dfr-miR160a and its target gene DfARF10 through qRT-PCR. Under drought and high temperature treatment, the relative expression of dfr-miR160a was up-regulated, but under NaCl treatment, the expression of dfr-miR160a was down-regulated. Under low temperature treatment, the expression of dfr-miR160a was down-regulated at 0-1 h and was up-regulated at 3-48 h. The expression of DfARF10 was up-regulated under NaCl, high temperature and low temperature treatments. However, under drought treatment, the expression of DfARF10 decreased, distinct from dfr-miR160a. The above results indicated that target gene of dfr-miR160 was DfARF10, and both of them can respond to abiotic stress treatment. This study provides a new scientific basis for revealing the abiotic stress resistance mechanism of D. fragrans at the molecular level.
Key words： Dryopteris fragrans; dfr-miR160a; DfARF10; abiotic stress
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