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用于高盐胁迫下番茄叶片脱落酸检测的传感器研制及试验

Development and test of sensor to detect abscisic acid in tomato leaves under high-salinity stress

  • 摘要: 脱落酸(abscisic acid, ABA)是平衡植物内源激素和有关生长活性物质代谢的关键因子。针对盐胁迫下番茄叶片缺乏高灵敏、准确且能实现ABA原位检测方法的问题,该研究构建了一种基于碘氧化铋/g-氮化碳(BiOI/g-C3N4) II型异质结的光电化学(Photoelectrochemical, PEC)适配体传感器。首先,利用g-C3N4作为高效载体和光敏化剂,大幅增强了BiOI的光电响应。得益于II型异质结的能带结构对光生电荷分离的促进效果,该复合材料的光电流强度较纯 BiOI 提升了约13倍。在此基础上,进一步耦合适配体(Aptamer, Apt)的靶向识别功能构建了一种“信号增强”型识别界面,实现了1 pmol/L~300 nmol/L范围内ABA的定量检测,检出限为0.33 pmol/L。该传感器对100倍浓度共存干扰物仍保持较好的选择性,储存14 d后光电流保持率为91.8%,在15~35 ℃范围内响应稳定。将其用于盐胁迫下番茄叶片ABA的原位监测,结果显示ABA表观浓度随胁迫时间先升高后降低,24 h达到0.896 nmol/L,72 h降至0.417 nmol/L。研究结果表明,该传感器可用于植物叶片ABA的快速、原位检测,并为作物逆境诊断和精准农业传感提供技术参考。

     

    Abstract: Abscisic acid (ABA) is one of the most important endogenous hormones to regulate seed dormancy, stomatal movement, root architecture, and plant responses to abiotic stresses. Therefore, it is often required to rapidly and accurately monitor ABA in living leaves for crop stress status in precision agriculture. However, conventional high-performance liquid chromatography, liquid chromatography–mass spectrometry, and enzyme-linked immunosorbent assay generally require the type-II band alignment, resulting in suppression of electron-hole recombination. A signal-enhanced PEC sensing interface has been constructed to immobilize an ABA-specific aptamer on the BiOI/g-C3N4/ITO electrode. The binding between ABA and aptamer has induced conformational rearrangement of the recognition layer, reduced the interfacial charge-transfer resistance, and further promoted electron transport, leading to an increased cathodic photocurrent. However, the aptasensor shared a better linear relationship between photocurrent intensity and the logarithm of sample pretreatment, expensive instruments, and laboratory operation, thereby limiting their use for on-site and dynamic analysis. In this study, a photoelectrochemical (PEC) aptasensor was developed for in-situ detection of ABA in tomato leaves under salt stress, according to a bismuth oxyiodide/graphitic carbon nitride (BiOI/g-C3N4) type-II heterojunction. BiOI was selected as the visible-light-responsive semiconductor, while g-C3N4 was introduced as the carrier and photosensitizer to improve the separation and transfer of photogenerated charge carriers. The BiOI/g-C3N4 composite was prepared by simple physical mixing and stirring. The sample was characterized by scanning electron microscopy, transmission electron microscopy, X-ray diffraction, X-ray photoelectron spectroscopy, and ultraviolet-visible diffuse reflectance spectroscopy. The results showed that g-C3N4 nanosheets were tightly coupled with flower-like BiOI microspheres, indicating an effective heterojunction interface with enhanced visible-light absorption and interfacial charge transport. Compared with pure BiOI, the BiOI/g-C3N4-modified electrode exhibited a 13-fold higher photocurrent response, which was attributed to ABA concentration in the range of 1 pmol/L to 300 nmol/L, with a correlation coefficient of 0.992 and a detection limit of 0.33 pmol/L. The sensor also exhibited high selectivity against common coexisting substances, including inorganic ions, amino acids, sugars, and other plant hormones, even when the interferents were present at 100-fold higher concentrations. In addition, stable responses were maintained in the temperature range of 15-35 °C, and 91.8% of the initial signal was retained after 14 days of storage at 4 °C. Recovery tests showed that tomato leaf extracts shared recoveries of 93%-106% with relative standard deviations of 1.1%-2.1%, indicating acceptable accuracy in complex plant matrices. Finally, the PEC sensing system was applied to the in-situ monitoring of ABA in tomato leaves during salt stress. The ABA level increased rapidly at the early stage of stress, reached 0.896 nmol/L at 24 h, and then decreased to 0.417 nmol/L at 72 h. This trend was consistent with the physiological role of ABA in early osmotic stress adaptation and subsequent metabolic adjustment. The BiOI/g-C3N4 PEC aptasensor can provide a sensitive, selective, and practical approach for dynamic ABA monitoring in plant leaves. The findings can offer a useful sensing strategy for crop stress diagnosis in precision agriculture.

     

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