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    许欣, 卢惠君, 王玉成, 王智博, 及晓宇. 刚毛柽柳SAIR6长链非编码RNA耐盐功能初探[J]. 北京林业大学学报, 2021, 43(3): 36-43. DOI: 10.12171/j.1000-1522.20200235
    引用本文: 许欣, 卢惠君, 王玉成, 王智博, 及晓宇. 刚毛柽柳SAIR6长链非编码RNA耐盐功能初探[J]. 北京林业大学学报, 2021, 43(3): 36-43. DOI: 10.12171/j.1000-1522.20200235
    Xu Xin, Lu Huijun, Wang Yucheng, Wang Zhibo, Ji Xiaoyu. Salt stress tolerance analysis of SAIR6 long non-coding RNA in Tamarix hispida[J]. Journal of Beijing Forestry University, 2021, 43(3): 36-43. DOI: 10.12171/j.1000-1522.20200235
    Citation: Xu Xin, Lu Huijun, Wang Yucheng, Wang Zhibo, Ji Xiaoyu. Salt stress tolerance analysis of SAIR6 long non-coding RNA in Tamarix hispida[J]. Journal of Beijing Forestry University, 2021, 43(3): 36-43. DOI: 10.12171/j.1000-1522.20200235

    刚毛柽柳SAIR6长链非编码RNA耐盐功能初探

    Salt stress tolerance analysis of SAIR6 long non-coding RNA in Tamarix hispida

    • 摘要:
        目的  长链非编码RNA(lncRNA)是一类长度大于200个核苷酸、无蛋白质编码能力或编码能力极低的转录本,已有研究表明,lncRNA是植物胁迫反应中关键的调控因子之一。本研究拟对刚毛柽柳SAIR6长链非编码RNA是否具有提高刚毛柽柳的耐盐能力进行分析,为阐明刚毛柽柳lncRNA响应盐胁迫的分子调控机制奠定基础,进一步丰富木本植物lncRNA响应逆境胁迫分子机制的研究。
        方法  本研究从盐胁迫下刚毛柽柳转录组中筛选出一条差异表达的lncRNA-224223.1,将其命名为ThSAIR6。利用实时荧光定量PCR(qRT-PCR)技术分析盐胁迫下刚毛柽柳叶组织中ThSAIR6的表达模式,初步鉴定其是否响应盐胁迫。为了进一步分析ThSAIR6的抗逆功能,构建其植物过表达载体(pROKII-ThSAIR6),利用农杆菌介导的高效遗传瞬时转化体系,获得ThSAIR6瞬时过表达及对照刚毛柽柳植株。在盐胁迫下分别对ThSAIR6瞬时过表达及对照植株的耐盐相关生理指标进行测定,判断ThSAIR6是否能提高刚毛柽柳的耐盐能力。
        结果  qRT-PCR结果表明,在盐胁迫24 h后,ThSAIR6在刚毛柽柳植株中的表达量显著上升(P < 0.05),说明该lncRNA能响应盐胁迫。抗逆生理指标的测定结果表明,ThSAIR6在刚毛柽柳中过表达能够使H2O2 \rmO_\rm2^ - \text• 的含量显著降低(P < 0.05),使POD、SOD的酶活性显著增强(P < 0.05);同时降低刚毛柽柳组织中的死亡细胞数量、电解质渗透率及失水率。
        结论  刚毛柽柳SAIR6长链非编码RNA能响应盐胁迫;在盐胁迫下,ThSAIR6过表达显著减轻了植物组织细胞的受损程度,增强了POD、SOD酶活性,降低了植株内H2O2 \rmO_\rm2^ - \text• 的含量,提高了活性氧(ROS)清除能力,有效提高了刚毛柽柳的耐盐能力。

       

      Abstract:
        Objective  Long non-coding RNA (lncRNA) is a class of transcripts, which are larger than 200 nt in length and have extremely low protein-coding ability or without protein-coding ability. LncRNA is one of the key regulators of plant stress response. In this study, we studied the salt-tolerant physiological indexes under the salt stress of transiently overexpressing lncRNA of Tamarix hispida, and analyzed whether it had the ability to improve the salt tolerance and enrich the molecular mechanism of woody plant lncRNA in response to stress. It laid a foundation for the molecular regulation mechanism of lncRNA in T. hispida in response to salt stress.
        Method  In this study, a differential expression lncRNA-224223.1 was selected and named ThSAIR6 from transcriptomes of T. hispida under salt stress. The quantitative real-time PCR (qRT-PCR) was used to analyze the expression pattern of ThSAIR6 in leaf tissues of wild type T. hispida under salt stress. It could initially identify whether it responded to salt stress or not. Overexpression vectors (pROKII-ThSAIR6) were constructed in order to further study the stress tolerance. Overexpression (OE) and control T. hispida plants were obtained by agrobacterium-mediated high-efficient transient transformation system. Physiologic indexes related to salt tolerance of OE and control T. hispida plants under salt stress were measured in order to judge whether it could improve the salt tolerance of T. hispida plants.
        Result  The results of qRT-PCR showed that the expression quantity of ThSAIR6 in wild type plants significantly increased after salt stress for 24 h (P < 0.05), indicating that it could response to salt stress. The results showed that overexpression of ThSAIR6 in T. hispida plants significantly decreased the contents of H2O2 and \rmO_\rm2^ - \text• (P < 0.05), enhanced the POD and SOD activities (P < 0.05), and lowered the amount of dead cells, the electrolyte leakage and water loss rate of T. hispida tissues at the same time.
        Conclusion  In conclusion, lncRNA ThSAIR6 of T. hispida can response to salt stress. Overexpression of ThSAIR6 in plants significantly decreases the contents of H2O2 and \rmO_\rm2^ - \text• , enhances the POD and SOD activities, improves ROS scavenging, and reduces cell damage of plant tissues under salt stress, thus effectively improves its salt tolerance.

       

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