Repression of ARF10 by microRNA160 plays an important role in the mediation of leaf water loss
Repression of ARF10 by microRNA160 plays an important role in the mediation of leaf water loss
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microRNA160 对 ARF10 的抑制在介导叶片失水中发挥重要作用
DOI：
10.1007/s11103-016-0514-3
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发表时间：
2016-10-01
影响因子：
5.1
通讯作者：
Li, Tianlai
中科院分区：
文献类型：
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作者：
Liu, Xin;Dong, Xiufen;Li, Tianlai
Solanum lycopersicumauxin response factor 10（SlARF10）受ysl - mir160转录后调控。过表达aSl-miR160-resistantSlARF10（mSlARF10）导致小叶叶片变窄，气孔变大，但密度降低。35S：与野生型（WT）相比，切除叶片较窄的mslarf10 -6植株水分流失较大。进一步分析发现，脱水处理下35S: mslarf10 -6和WT的实际失水与计算的气孔失水不一致，说明水力导度存在差异。脱落酸（ABA）和hgcl2预处理证实35s:mSlARF10具有较高的水力导度，这与较大的气孔大小和较高的水通道蛋白（AQPs）活性有关。在外源ABA处理下，35S: mslarf10 -6表现出更强的敏感性，气孔迅速关闭。RNA测序筛选结果显示，5个aqp相关基因、14个ABA生物合成/信号基因和3个气孔发育基因在35s: mslarf10 -6植株中发生了显著改变，并通过qRT-PCR验证了这一结果。启动子分析显示，上调的AQPs中含有AuxRE和ABRE，这意味着这些元件可能是35s:mSlARF10-6中AQPs高表达的原因。选择三个上调最多的aqp （SlTIP1-1-like, SlPIP2；4和slnip -type-like）来确认AuxRE和ABRE的功能。启动子瞬时表达表明，slpip2；4 andslnip-type-likeauxres andSlPIP2;4和sltip1 -1 like abres可显著增强GUS报告基因在35s:mSlARF10-6中的表达，证实AuxRE和ABRE可能是诱导AQPs表达的主要因子。此外，35s中两个上调的转录因子：mSlARF10-6、slarf10和slabi5 - like3在电迁移性转移实验和酵母单杂交实验中被证明直接与这些元件结合。此外，在番茄叶片中瞬时表达下调的arf10或上调的abi5表明，arf10是诱导35s:mSlARF10-6水分流失的直接因子。本研究表明，slarf10通过调节气孔孔径增加ABA合成/信号响应，减轻水分流失，但slarf10还通过影响气孔发育和AQP表达，影响水分运输，并共同控制番茄叶片水分流失。因此，本研究揭示了一个以前未被认识的叶片水分流失调节因子，以及在这一重要过程中协调生长素和ABA信号的网络。在进化背景下，mir160调节arf10维持叶片水分平衡，从而保证植物正常发育和环境适应。
Solanum lycopersicumauxin response factor 10(SlARF10) is post-transcriptionally regulated bySl-miR160. Overexpression of aSl-miR160-resistantSlARF10(mSlARF10) resulted in narrower leaflet blades with larger stomata but lower densities.35S:mSlARF10-6plants with narrower excised leaves had greater water loss, which was in contrast to the wild type (WT). Further analysis revealed that the actual water loss was not consistent with the calculated stomatal water loss in 35S:mSlARF10-6and the WT under the dehydration treatment, indicating that there is a difference in hydraulic conductance. Pretreatment with abscisic acid (ABA) and HgCl2confirmed higher hydraulic conductance in35S:mSlARF10, which is related to the larger stomatal size and higher activity of aquaporins (AQPs). Under ABA treatment,35S:mSlARF10-6showed greater sensitivity, and the stomata closed rapidly. Screening by RNA sequencing revealed that five AQP-related genes, fourteen ABA biosynthesis/signal genes and three stomatal development genes were significantly altered in35S:mSlARF10-6plants, and this result was verified by qRT-PCR. The promoter analysis showed that upregulated AQPs contain AuxRE and ABRE, implying that these elements may be responsible for the high expression levels of AQPs in35S:mSlARF10-6. The three most upregulated AQPs (SlTIP1-1-like, SlPIP2;4andSlNIP-type-like) were chosen to confirm AuxRE and ABRE function. Promoters transient expression demonstrated that theSlPIP2;4andSlNIP-type-likeAuxREs andSlPIP2;4andSlTIP1-1-likeABREs could significantly enhance the expression of the GUS reporter in35S:mSlARF10-6, confirming that AuxRE and ABRE may be the main factors inducing the expression of AQPs. Additionally, two upregulated transcription factors in35S:mSlARF10-6, SlARF10andSlABI5-likewere shown to directly bind to those elements in an electromobility shift assay and a yeast one-hybrid assay. Furthermore, transient expression of down-regulatedARF10or up-regulatedABI5in tomato leaves demonstrated thatARF10is the direct factor for inducing the water loss in35S:mSlARF10-6. Here, we show that althoughSlARF10increased the ABA synthesis/signal response by regulating stomatal aperture to mitigate water loss,SlARF10also influenced stomatal development and AQP expression to affect water transport, and both act cooperatively to control the loss of leaf water in tomato. Therefore, this study uncovers a previously unrecognized leaf water loss regulatory factor and a network for coordinating auxin and ABA signalling in this important process. In an evolutionary context,miR160regulatesARF10to maintain the water balance in the leaf, thus ensuring normal plant development and environmental adaptation.