ARC微生物菌剂缓解黑暗胁迫下花生根瘤功能衰退的效应

doi:10.3864/j.issn.0578-1752.2025.22.005

中国农业科学 ›› 2025, Vol. 58 ›› Issue (22): 4617-4627.doi: 10.3864/j.issn.0578-1752.2025.22.005

• 耕作栽培·生理生化·农业信息技术 • 上一篇下一篇

ARC微生物菌剂缓解黑暗胁迫下花生根瘤功能衰退的效应

赵亚¹^,²(), 张文², 王督², 张良晓², 张奇², 韩琴²(), 王伟¹(), 李培武²()

¹ 中国农业大学工学院，北京 100083
² 中国农业科学院油料作物研究所/农业农村部油料作物生物学与遗传育种重点实验室/农业农村部生物毒素检测重点实验室/农业农村部油料产品质量安全风险评估实验室/农业农村部油料及制品质量监督检验测试中心，武汉 430062

收稿日期:2025-05-15 接受日期:2025-06-30 出版日期:2025-11-16 发布日期:2025-11-21
通信作者:
韩琴，E-mail：hanqin@caas.cn。
王伟，E-mail：playerwxw@cau.edu.cn。
李培武，E-mail：peiwuli@oilcrops.cn。
联系方式: 赵亚，E-mail：zhaoya1109@163.com。
基金资助:
国家自然科学基金(32441047); 国家自然科学基金(32441046); 国家自然科学基金(32471627); 中国农业科学院科技创新工程(CAAS-ZDRW202416); 湖北省科技重大专项(2023BBA002)

Effects of ARC Microbial Agent on Alleviating Functional Decline of Peanut Root Nodules Under Dark Stress

ZHAO Ya¹^,²(), ZHANG Wen², WANG Du², ZHANG LiangXiao², ZHANG Qi², HAN Qin²(), WANG Wei¹(), LI PeiWu²()

¹ College of Engineering, China Agricultural University, Beijing 100083
² Oil Crops Research Institute, Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture and Rural Affairs /Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture and Rural Affairs/Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture and Rural Affairs/Quality Inspection and Test Center for Oilseeds Products, Ministry of Agriculture and Rural Affairs, Wuhan 430062

Received:2025-05-15 Accepted:2025-06-30 Published:2025-11-16 Online:2025-11-21

摘要/Abstract

摘要：

【目的】根瘤衰老是制约豆科作物共生固氮效率的重要因素，延缓根瘤衰老、延长其功能期是提升作物产量及营养品质的有效途径。针对花生结瘤少固氮效率低，且易受黄曲霉毒素污染等难题，团队研制出ARC微生物菌剂实现了高效结瘤与提质增产。本研究旨在明确ARC对花生根瘤黑暗胁迫的缓解效应，探讨其在延长根瘤寿命、维持固氮能力及活性方面的作用。【方法】在开花下针期对植株进行黑暗胁迫处理，构建快速衰老模型，并在不同胁迫时间（0、6、12、24、36、48和72 h），利用YOLOv8s算法动态提取图像中根瘤表型特征，利用乙炔还原法测定固氮酶活性，从表型与功能活性方面评价ARC对根瘤快速衰老的调控效果。【结果】与对照组相比，ARC处理显著改善根瘤表型结构和活性：平均直径增大6.34%，每克固氮酶活性提高117.11%；根瘤表面平均亮度和色度分别提高6.32%、3.05%。RGB信息进一步表明，处理组根瘤在各通道颜色强度上均显著高于对照组，红、绿、蓝通道平均强度分别提高6.32%、8.06%和9.35%，整体颜色更加鲜活、明亮。且在黑暗胁迫诱导的快速衰老过程中，ARC处理在不同阶段均可以有效减缓根瘤表型颜色及功能活性的衰退趋势。在胁迫初期（0—6 h），对照组根瘤平均亮度和色度分别降低8.36%、16.85%，固氮酶活性下降幅度高达82.65%，表明根瘤快速进入衰老状态。而ARC处理组亮度和色度仅下降1.26%、11.31%，固氮酶活性下降63.99%，显著减缓了早期衰退进程，体现出良好的干预和保护效果。胁迫后期（72 h），处理组根瘤亮度和色度降幅分别为4.85%和20.96%，显著低于对照组的13.82%和23.15%，有效减缓了颜色劣变趋势。同时，其固氮酶活性仍为初始值的22.36%，而对照组根瘤在黑暗胁迫48 h时已失活，进一步验证了ARC在根瘤衰老调控中的持效作用。此外，采用皮尔逊相关系数分析了根瘤表面RGB三通道强度与固氮酶活性之间的关系，红色通道与酶活性呈显著正相关关系（r=0.573，P=0.0003），绿色通道相关性较弱，而蓝色通道无显著相关性。色度作为综合表征颜色变化的指标，与固氮酶活性之间的相关性最强（r=0.736，P<0.001）。且随着固氮酶活性的逐渐衰退，根瘤呈现由鲜红向暗红、褐色渐变的趋势。【结论】ARC微生物菌剂不仅促进花生结瘤固氮，还能够有效延缓黑暗胁迫条件诱导的花生根瘤快速衰老，维持根瘤结构的稳定性及固氮功能的持续性，提高豆科作物共生固氮效率。此外，根瘤色度能够反映根瘤生理活性变化，可作为根瘤功能状态劣变的早期预警指标，为深入研究豆科作物结瘤固氮提供新途径和新方法。

关键词: 花生, 黑暗胁迫, 快速衰老, ARC微生物菌剂, 根瘤表型及活性

Abstract:

【Objective】Root nodule senescence is a major factor limiting the symbiotic nitrogen fixation efficiency of leguminous crops. Delaying nodule senescence and prolonging its functional period is considered an effective strategy to improve crop yield and nutritional quality. To address the challenges of low nodulation efficiency, poor nitrogen fixation, and susceptibility to aflatoxin contamination in peanuts, our team has developed the ARC microbial agent, which effectively enhances nodulation while improving yield and quality. This study aims to elucidate the alleviating effects of the ARC microbial agent on peanut root nodules under dark stress, and to investigate its role in extending nodule lifespan and maintaining nitrogenase activity and functional stability. 【Method】A rapid senescence model was established by applying dark stress to peanut plants at the flowering and pegging stage. Root nodule phenotypic traits were dynamically extracted at different stress time points (0, 6, 12, 24, 36, 48, and 72 h) using the YOLOv8s algorithm, while nitrogenase activity was determined via the acetylene reduction assay. Based on both phenotypic and functional parameters, the regulatory effects of the ARC microbial agent on the rapid senescence of root nodules were comprehensively evaluated. 【Result】Compared with the control group, treatment with the ARC microbial agent significantly improved both the structural and functional traits of root nodules: the average diameter increased by 6.34%, and nitrogenase activity per gram increased by 117.11%. In addition, the average surface brightness and chroma of nodules increased by 6.32% and 3.05%, respectively. RGB color analysis further showed that nodules in the treatment group exhibited significantly higher color intensity across all channels, with mean increases of 6.32%, 8.06%, and 9.35% in the red, green, and blue channels, respectively, indicating a more vivid and brighter color appearance. During the dark stress-induced rapid senescence process, the ARC microbial agent effectively mitigated the decline in both nodule color phenotype and nitrogen fixation activity across different time points. In the early stress stage (0-6 h), the average brightness and chroma of control group nodules decreased by 8.36% and 16.85%, respectively, and nitrogenase activity dropped by 82.65%, indicating rapid onset of senescence. In contrast, nodules in the ARC treatment group showed only 1.26% and 11.31% decreases in brightness and chroma, respectively, with nitrogenase activity decreasing by 63.99%, reflecting a clear delay in early senescence and a strong protective effect. By the late stage of stress (72 h), the ARC treatment group exhibited brightness and chroma reductions of only 4.85% and 20.96%, significantly lower than the 13.82% and 23.15% declines observed in the control group, effectively alleviating the trend of color deterioration. At the same time, nitrogenase activity in the treatment group remained at 22.36% of its initial level, while nodules in the control group had already lost activity by 48 h, further confirming the sustained regulatory effect of ARC on nodule senescence. In addition, Pearson correlation analysis revealed that the red channel intensity was significantly positively correlated with nitrogenase activity (r=0.573, P=0.0003), while the green channel showed a weaker correlation and the blue channel showed no significant correlation. Notably, chroma, as a composite indicator of color variation, exhibited the strongest correlation with nitrogenase activity (r=0.736, P<0.001). Furthermore, with the progressive decline in nitrogenase activity, root nodules showed a gradual color transition from bright red to dark red and brown. 【Conclusion】The ARC microbial agent not only promotes peanut nodulation and nitrogen fixation, but also effectively delays the rapid senescence of root nodules induced by dark stress, thereby maintaining structural stability and sustaining nitrogen-fixing function. This contributes to enhanced symbiotic nitrogen fixation efficiency in leguminous crops. Moreover, nodule chroma was found to reflect changes in physiological activity and can serve as an early warning indicator of functional decline. These findings provide new perspectives and methodological support for advancing the study of nodulation and nitrogen fixation in legumes.

Key words: peanut, dark stress, rapid senescence, ARC microbial agent, nodule phenotype and activity

赵亚, 张文, 王督, 张良晓, 张奇, 韩琴, 王伟, 李培武. ARC微生物菌剂缓解黑暗胁迫下花生根瘤功能衰退的效应[J]. 中国农业科学, 2025, 58(22): 4617-4627.

ZHAO Ya, ZHANG Wen, WANG Du, ZHANG LiangXiao, ZHANG Qi, HAN Qin, WANG Wei, LI PeiWu. Effects of ARC Microbial Agent on Alleviating Functional Decline of Peanut Root Nodules Under Dark Stress[J]. Scientia Agricultura Sinica, 2025, 58(22): 4617-4627.

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ARC微生物菌剂缓解黑暗胁迫下花生根瘤功能衰退的效应

Effects of ARC Microbial Agent on Alleviating Functional Decline of Peanut Root Nodules Under Dark Stress

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