<br>烟草柱头分泌物中的过氧化氢影响花粉管中的花粉蛋白质组和膜电位,Plant Biology

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烟草柱头分泌物中的过氧化氢影响花粉管中的花粉蛋白质组和膜电位
Plant Biology ( IF 4.2 ) Pub Date : 2021-03-20 , DOI: 10.1111/plb.13255
M Breygina ₁ , E Klimenko ₁ , E Shilov ₂ , A Podolyan ₁ , A Mamaeva ₃ , V Zgoda ₄ , I Fesenko ₃

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众所周知，ROS 积聚在不同物种的柱头中，并且可能发挥对植物繁殖重要的不同功能。在这里，我们测试了这样的假设：它们的功能之一是控制膜电位并激发发芽花粉中独特蛋白质的合成。
我们使用分光荧光法和分光光度法检测柱头分泌物中的 H ₂ O ₂ ，使用花粉管的定量荧光显微镜和花粉原生质体的流式细胞术揭示对膜电位的影响，并使用无标记定量方法研究 H ₂ O 后花粉蛋白质组的变化₂治疗。
我们发现，在生长的花粉管和花粉原生质体渗出物中都会引起类似于 H ₂ O ₂引起的质膜超极化。这种效应可通过过氧化氢酶处理和 ROS 猝灭剂 MnTMPP 消除。抑制分析表明Ca ²⁺ - 和K ^{+ -}传导通道可能参与观察到的超极化。为了更深入地了解花粉反应，我们分析了 H ₂ O ₂处理的花粉粒中蛋白质组的变化。我们发现了 50 种独特的蛋白质和 20 种不同积累的蛋白质，它们主要涉及细胞代谢、能量学、蛋白质合成和折叠。
观察到的超极化和蛋白质组改变与之前报道的 H ₂ O ₂刺激花粉萌发以及 Ca ²⁺ - 和 K ^{+ -}传导通道对此 ROS 的敏感性非常吻合。因此，H ₂ O ₂是烟草柱头渗出物中的活性物质之一，其刺激花粉萌发中的各种生理过程。

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Hydrogen peroxide in tobacco stigma exudate affects pollen proteome and membrane potential in pollen tubes

ROS are known to be accumulated in stigmas of different species and can possibly perform different functions important for plant reproduction. Here we tested the assumption that one of their functions is to control membrane potential and provoke synthesis of unique proteins in germinating pollen.
We used spectrofluorometry and spectrophotometry to detect H₂O₂ in stigma exudate, quantitative fluorescent microscopy of pollen tubes and flow cytometry of pollen protoplasts to reveal effects on membrane potential, and a label-free quantification approach to study pollen proteome changes after H₂O₂ treatment.
We found that in both growing pollen tubes and pollen protoplasts exudate causes plasmalemma hyperpolarization similar to that provoked by H₂O₂. This effect is abolished by catalase treatment and the ROS quencher, MnTMPP. Inhibitory analysis indicates probable participation of Ca²⁺- and K⁺-conducting channels in the observed hyperpolarization. For a deeper understanding of pollen response, we analysed proteome alterations in H₂O₂-treated pollen grains. We found 50 unique proteins and 20 differently accumulated proteins that are mainly involved in cell metabolism, energetics, protein synthesis and folding.
Observed hyperpolarization and proteome alterations agree well with previously reported stimulation of pollen germination by H₂O₂ and sensitivity of Ca²⁺- and K⁺-conducting channels to this ROS. Thus, H₂O₂ is one of the active substances in tobacco stigma exudate that stimulates various physiological processes in germinating pollen.

更新日期：2021-03-20

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