木质素单体跨膜转运的技术研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2024-0312

生物技术通报 ›› 2024, Vol. 40 ›› Issue (10): 198-207.doi: 10.13560/j.cnki.biotech.bull.1985.2024-0312

木质素单体跨膜转运的技术研究进展

毛馨艺¹^,²^,³^,⁴(), 兰芸¹^,²^,³^,⁴, 张准¹^,²^,³^,⁴, 张叶卓¹^,²^,³^,⁴, 金琪¹^,²^,³^,⁴, 赵美棋¹^,²^,³^,⁴, 曾子柽¹^,²^,³^,⁴, 李晔¹^,²^,³^,⁴()

1.北京林业大学生物科学与技术学院林木遗传育种全国重点实验室，北京 100083
2.北京林业大学生物科学与技术学院林木育种与生态修复国家工程研究中心，北京 100083
3.北京林业大学生物科学与技术学院林木、花卉遗传育种教育部重点实验室，北京 100083
4.北京林业大学生物科学与技术学院树木花卉育种生物工程国家林业和草原局重点实验室，北京 100083

收稿日期:2024-03-31 出版日期:2024-10-26 发布日期:2024-11-20
通讯作者: 李晔，女，博士，副教授，研究方向：细胞壁形成机制，物质的跨膜转运机制；E-mail: liye0223@bjfu.edu.cn
作者简介:毛馨艺，女，研究方向：细胞壁形成机制；E-mail: maoxinyi2003@bjfu.edu.cn
基金资助:
北京林业大学“5·5工程”科研创新团队项目(BLRC2023A01);国家重点研发计划(2022YFF0712500);北京林业大学大学生创新创业训练计划(X202310022101)

Advances in Technology for Transport of Monolignols across Membrane

MAO Xin-yi¹^,²^,³^,⁴(), LAN Yun¹^,²^,³^,⁴, ZHANG Zhun¹^,²^,³^,⁴, ZHANG Ye-zhuo¹^,²^,³^,⁴, JIN Qi¹^,²^,³^,⁴, ZHAO Mei-qi¹^,²^,³^,⁴, ZENG Zi-cheng¹^,²^,³^,⁴, LI Ye¹^,²^,³^,⁴()

1. State Key Laboratory of Tree Genetics and Breeding, College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083
2. National Engineering Laboratory for Tree Breeding, College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083
3. Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083
4. The Tree and Ornamental Plant Breeding and Biotechnology Laboratory of National Forestry and Grassland Administration, College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083

Received:2024-03-31 Published:2024-10-26 Online:2024-11-20

摘要/Abstract

摘要：

木质素（lignin）作为一种重要的可再生资源，主要集中在植物的次生细胞壁中，在植物的营养物质运输、机械支持和病原体防御等生理过程中发挥着关键作用。木质素的形成过程包括3个主要环节：单体的胞内合成、跨膜转运以及胞外聚合。深入探究木质素单体跨膜转运的分子机制，对揭示细胞壁形成分子机制及木材改良具有重要意义。基于此，本综述从转录组学、基因工程、点击化学、荧光显微成像、分子模拟等技术角度，对木质素单体跨膜转运的相关进展和最新技术进行了梳理和总结，阐述了木质素单体跨膜转运相关研究的新技术和存在的技术瓶颈，并对比分析了不同技术之间的优缺点，最后提出了木质素单体跨膜转运机制研究中仍存在的问题和面临的挑战。相信随着成像技术、结构生物学、人工智能等科技的飞速发展，将为进一步阐明木质素单体跨膜转运的分子机理提供有效的技术手段。期望本综述为木质素单体的相关研究提供新的思路和方法。

关键词: 木质素单体, 跨膜转运, 点击化学, 分子模拟, 荧光显微成像

Abstract:

As an important renewable resource, lignin is mainly deposited in the secondary cell wall of vascular plants and plays an important role in physiological processes such as nutrient transport, mechanical support, and plant pathogen defense. The formation of lignin can be divided into three processes: Intracellular synthesis, transmembrane transport and extracellular polymerization of monolignols. Exploring the transmembrane transport mechanism of monolignols is of great significance for revealing the mechanism of cell wall formation and wood improvement.. In this review, we sorted and summarized the relevant progress and latest technologies of monolignols transmembrane transport from the perspectives of transcriptomics, genetic engineering, click chemistry, fluorescence microscopy, and molecular simulation. Then we described the new technologies and existing technical bottlenecks in the research on monolignol transmembrane transport. In addition, we compared and analyzed the advantages and disadvantages of different technologies. Finally, we proposed the issues and challenges in the study of the mechanism of monolignols transmembrane transport. It is believed that the rapid development of imaging technology, structural biology, artificial intelligence and other technologies will provide new tools for further investigating the transmembrane transport mechanism of monolignols. This review is expected to provide new methods for the study of monolignols.

Key words: monolignols, membrane transport, click chemistry, molecular simulation, fluorescence microscopy

毛馨艺, 兰芸, 张准, 张叶卓, 金琪, 赵美棋, 曾子柽, 李晔. 木质素单体跨膜转运的技术研究进展[J]. 生物技术通报, 2024, 40(10): 198-207.

MAO Xin-yi, LAN Yun, ZHANG Zhun, ZHANG Ye-zhuo, JIN Qi, ZHAO Mei-qi, ZENG Zi-cheng, LI Ye. Advances in Technology for Transport of Monolignols across Membrane[J]. Biotechnology Bulletin, 2024, 40(10): 198-207.

图/表 3

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技术 Technology	原理 Principle	优势 Advantage	劣势 Disadvantage	参考文献 Reference
转录组学 Transcriptomics	检测细胞的基因表达情况，识别具有相似表达模式的基因，从整体水平上研究基因转录情况以及调控规律	根据不同细胞的转录谱表达，通过相关性分析可对潜在转运蛋白进行筛选，为研究木质素单体跨膜转运的分子机理提供有价值的信息，是进一步实现功能研究的基础	林木庞大的基因组为基因表达分析带来了挑战	[14,25⇓⇓⇓⇓⇓⇓⇓⇓⇓⇓ -36]
基因工程 Genetic engineering	使用生物技术直接操纵有机体基因组，改变细胞的遗传物质，可以进行基因导入、敲除以及替换	可以获得蛋白定位和互作信息，为特定基因产物的功能分析提供直接证据，是研究相关转运蛋白的功能以及木质素单体跨膜转运分子机理的一个重要手段	ABC转运蛋白的冗余性可能影响基因敲除实验的结果	[14-15,35,37⇓⇓⇓⇓-42]
点击化学 Click chemistry	利用细胞自身的生物代谢合成机制，将生物正交化学报告基团整合到目标生物分子中以达到标记目的	具基因标记的简单性、抗体标记的特异性以及小分子探针的多功能性；操作简单，几乎不改变标记对象的构象，对植物细胞毒害性小，并能高度还原木质素单体转运过程中的生物过程	植物细胞壁的存在增加了对标记类似物的代谢吸收难度，限制代谢标记效率	[43⇓⇓⇓⇓-48]
荧光显微技术 Fluorescent microscopy	木质素单体中存在苯环，能够产生自发荧光，苯环上的不同侧链以及环或侧链上不同位置取代会导致发色团的变化，据此可以识别不同类型的木质素及其单体	可以在不进行染色的情况下观察和分析样品，简化操作；可以追踪木质素单体在植物细胞中的动态变化	受自身分辨率、成像时间等限制，且木质素的复杂结构会对成像结果造成一定干扰	[50⇓⇓⇓⇓-55]
分子模拟 Molecular simulation	基于物理原理（量子力学和统计热力学），利用计算机构建原子水平的分子模型，进而研究分子结构和行为、分子的性质	相比于实验手段，可以在原子水平上提供详细信息，有助于理解单体跨膜以及与转运蛋白互作的动力学机制	模拟体系可能与细胞内真实的环境有差异，只能作为参考，不能完全反映木质素跨膜转运的真实状态	[15,56⇓⇓⇓⇓⇓⇓ -63]

技术 Technology	原理 Principle	优势 Advantage	劣势 Disadvantage	参考文献 Reference
转录组学 Transcriptomics	检测细胞的基因表达情况，识别具有相似表达模式的基因，从整体水平上研究基因转录情况以及调控规律	根据不同细胞的转录谱表达，通过相关性分析可对潜在转运蛋白进行筛选，为研究木质素单体跨膜转运的分子机理提供有价值的信息，是进一步实现功能研究的基础	林木庞大的基因组为基因表达分析带来了挑战	[14,25⇓⇓⇓⇓⇓⇓⇓⇓⇓⇓ -36]
基因工程 Genetic engineering	使用生物技术直接操纵有机体基因组，改变细胞的遗传物质，可以进行基因导入、敲除以及替换	可以获得蛋白定位和互作信息，为特定基因产物的功能分析提供直接证据，是研究相关转运蛋白的功能以及木质素单体跨膜转运分子机理的一个重要手段	ABC转运蛋白的冗余性可能影响基因敲除实验的结果	[14-15,35,37⇓⇓⇓⇓-42]
点击化学 Click chemistry	利用细胞自身的生物代谢合成机制，将生物正交化学报告基团整合到目标生物分子中以达到标记目的	具基因标记的简单性、抗体标记的特异性以及小分子探针的多功能性；操作简单，几乎不改变标记对象的构象，对植物细胞毒害性小，并能高度还原木质素单体转运过程中的生物过程	植物细胞壁的存在增加了对标记类似物的代谢吸收难度，限制代谢标记效率	[43⇓⇓⇓⇓-48]
荧光显微技术 Fluorescent microscopy	木质素单体中存在苯环，能够产生自发荧光，苯环上的不同侧链以及环或侧链上不同位置取代会导致发色团的变化，据此可以识别不同类型的木质素及其单体	可以在不进行染色的情况下观察和分析样品，简化操作；可以追踪木质素单体在植物细胞中的动态变化	受自身分辨率、成像时间等限制，且木质素的复杂结构会对成像结果造成一定干扰	[50⇓⇓⇓⇓-55]
分子模拟 Molecular simulation	基于物理原理（量子力学和统计热力学），利用计算机构建原子水平的分子模型，进而研究分子结构和行为、分子的性质	相比于实验手段，可以在原子水平上提供详细信息，有助于理解单体跨膜以及与转运蛋白互作的动力学机制	模拟体系可能与细胞内真实的环境有差异，只能作为参考，不能完全反映木质素跨膜转运的真实状态	[15,56⇓⇓⇓⇓⇓⇓ -63]

木质素单体跨膜转运的技术研究进展

Advances in Technology for Transport of Monolignols across Membrane

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