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首页> 外文期刊>Journal of Experimental Botany >Redefining nitric oxide production in legume nodules through complementary insights from electron paramagnetic resonance spectroscopy and specific fluorescent probes
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Redefining nitric oxide production in legume nodules through complementary insights from electron paramagnetic resonance spectroscopy and specific fluorescent probes

机译：通过电子顺磁共振光谱和特定荧光探针的互补见解重新定义豆类结节中的一氧化氮生产

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Nitric oxide (NO) is a signaling molecule with multiple functions in plants. Given its critical importance and reactivity as a gaseous free radical, we have examined NO production in legume nodules using electron paramagnetic resonance (EPR) spectroscopy and the specific fluorescent dye 4,5-diaminofluorescein diacetate. Also, in this context, we critically assess previous and current views of NO production and detection in nodules. EPR of intact nodules revealed that nitrosyl-leghemoglobin (Lb(2+)NO) was absent from bean or soybean nodules regardless of nitrate supply, but accumulated in soybean nodules treated with nitrate that were defective in nitrite or nitric oxide reductases or that were exposed to ambient temperature. Consequently, bacteroids are a major source of NO, denitrification enzymes are required for NO homeostasis, and Lb(2+)NO is not responsible for the inhibition of nitrogen fixation by nitrate. Further, we noted that Lb(2+)NO is artifactually generated in nodule extracts or in intact nodules not analyzed immediately after detachment. The fluorescent probe detected NO formation in bean and soybean nodule infected cells and in soybean nodule parenchyma. The NO signal was slightly decreased by inhibitors of nitrate reductase but not by those of nitric oxide synthase, which could indicate a minor contribution of plant nitrate reductase and supports the existence of nitrate-and arginine-independent pathways for NO production. Together, our data indicate that EPR and fluorometric methods are complementary to draw reliable conclusions about NO production in plants.
机译：一氧化氮（NO）是具有多种植物功能的信号分子。鉴于其作为气体自由基的至关重要和反应性，我们在使用电子顺磁共振（EPR）光谱和特定荧光染料4,5-二氨基荧光素二乙酸酯的豆类结节中没有在豆科结节中进行生产。此外，在这种情况下，我们批判性地评估了结节中没有生产和检测的先前和目前的视图。完整结节的EPR显示豆类或大豆结节中亚硝基 - leghemoglobin（LB（2+）NO）无论硝酸盐供给，但在用硝酸盐处理的大豆结节中积累，所述硝酸盐或一氧化氮还原酶还是暴露的硝酸盐环境温度。因此，菌体是否的主要来源，没有稳态需要脱氮酶，并且LB（2+）不负责抑制通过硝酸盐的氮固定。此外，我们注意到，在结节提取物中或在脱离后立即未分析的完整结节中，LB（2+）否在结节提取物中产生。荧光探针检测到豆类和大豆结节感染细胞中没有形成，并在大豆结节薄膜中。硝酸盐还原酶的抑制剂略微降低了NO信号，但不是一氧化氮合酶的抑制剂，这可能表明植物硝酸盐还原酶的少量贡献，并支持硝酸盐和真生无关的途径没有生产。在一起，我们的数据表明EPR和荧光测定方法是互补的，以绘制植物中没有生产的可靠结论。

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