Article(id=1321852266265559753, tenantId=1146029695717560320, journalId=1316781310003478668, issueId=1321852178331979976, articleNumber=null, orderNo=null, doi=10.11983/CBB25002, pmid=null, cstr=32102.14.CBB25002, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1736352000000, receivedDateStr=2025-01-09, revisedDate=null, revisedDateStr=null, acceptedDate=1743523200000, acceptedDateStr=2025-04-02, onlineDate=1793083739104, onlineDateStr=2026-10-27, pubDate=1762704000000, pubDateStr=2025-11-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793083739104, onlineIssueDateStr=2026-10-27, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793083739104, creator=13701087609, updateTime=1793083739104, updator=13701087609, issue=Issue{id=1321852178331979976, tenantId=1146029695717560320, journalId=1316781310003478668, year='2025', volume='60', issue='6', pageStart='859', pageEnd='1035', issueExtLink='null', onlineDate='null', pubDate='1762704000000', pubDateStr='2025-11-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1793083718140, creator='13701087609', updateTime=1793083787009, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1321852467248223111, tenantId=1146029695717560320, journalId=1316781310003478668, issueId=1321852178331979976, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1321852467252417416, tenantId=1146029695717560320, journalId=1316781310003478668, issueId=1321852178331979976, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=1005, endPage=1016, ext={EN=ArticleExt(id=1321852266462692042, articleId=1321852266265559753, tenantId=1146029695717560320, journalId=1316781310003478668, language=EN, title=Development and Application of 3D Reconstruction Technology at Different Scales in Plant Research, columnId=1317485676040409248, journalTitle=Chinese Bulletin of Botany, columnName=SPECIAL TOPICS, runingTitle=null, highlight=null, articleAbstract=

3D reconstruction technology involves using computer graphics and image processing technologies to extract the geometric and topological information of the target object from the two-dimensional image data. This information is then used to create a three-dimensional mathematical model that can be processed by a computer, enabling the virtual reconstruction of the target object. In plant science research, the construction of three-dimensional models has become an effective way to study plant growth and development, morphological structure and functional mechanism. These models provide robust support for multi-scale imaging, measurement and analysis, demonstrating significant application potential in the field of agriculture and forestry. In recent years, advancements in plant 3D reconstruction technology have led to diverse applications in botanical research, covering plant morphological structure modeling, growth and development dynamic monitoring, and plant breeding. In this paper, we summarize the development process of 3D reconstruction technology and its application in plant studies across different scales (from organs and tissues to cells). We focus on the basic principles and applications of these technologies, aiming to provide theoretical and technical support for multimodal cross-scale imaging and plant phenotypic and functional research. Additionally, this work offers a novel approach to understand the principles of plant growth and development and the mechanisms underlying their responses to environmental changes.

, authors=Mengsha Huang, Lingdie Kong, Miao Yu, Chang Liu, Siqin Wang, Ruohan Wang*, authorsList=Mengsha Huang, Lingdie Kong, Miao Yu, Chang Liu, Siqin Wang, Ruohan Wang, authorCompany=null, correspAuthors=Ruohan Wang, authorNote=null, correspAuthorsNote=
* E-mail:
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三维重建技术(3D reconstruction)是利用计算机图形学和图像处理技术, 从二维图像数据中提取目标物体的几何和拓扑信息, 构建计算机可处理的三维数学模型, 从而实现物体的虚拟重建。在植物学研究中, 三维模型的构建已成为研究植物生长发育、形态结构和功能机制的有效手段, 为多尺度成像、测量和分析提供了有力支撑, 并在农业和林业领域展现出巨大的应用潜力。近年来, 随着植物三维重建技术的不断完善, 其在植物学研究中衍生出不同的应用方向, 涵盖植物形态结构建模、生长发育动态监测以及植物育种等多方面。该文综述了三维重建技术的发展历程及常见的三维重建成像技术在植物不同尺度(从器官、组织到细胞)研究中的应用, 重点阐述这些技术的基本原理及应用, 旨在为植物多模态跨尺度成像以及表型与功能研究提供理论和技术支撑, 为揭示植物生长发育规律及响应环境变化的机制提供新途径。

, authors=黄梦莎, 孔令蝶, 于淼, 刘畅, 王思钦, 王若涵*, authorsList=黄梦莎, 孔令蝶, 于淼, 刘畅, 王思钦, 王若涵, authorCompany=null, correspAuthors=王若涵, authorNote=null, correspAuthorsNote=
* 王若涵, 北京林业大学教授, 博士生导师。主持国家自然科学基金、“十三五”及“十四五”重点研发计划及国家林草局子课题等国家级项目。主编规划教材2部, 部分代表性成果发表在《科学通报》《中国科学-生命科学》、Plant Physiology及The Plant Journal等国内外权威期刊。荣获北京市优秀青年科技论文一等奖等。E-mail:
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(in Chinese), articleTitle=Development and frontier of electron microscopy 3D reconstruction, refAbstract=null), Reference(id=1321852290227618660, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, doi=null, pmid=null, pmcid=null, year=2010, volume=26, issue=null, pageStart=533, pageEnd=559, url=null, language=null, rfNumber=[77], rfOrder=94, authorNames=张凯, 张艳, 胡仲军, 季刚, 孙飞, journalName=生物物理学报, refType=null, unstructuredReference=张凯, 张艳, 胡仲军, 季刚, 孙飞 (2010). 电子显微三维重构技术发展与前沿. 生物物理学报 26, 533-559., articleTitle=电子显微三维重构技术发展与前沿, refAbstract=null), Reference(id=1321852290332476262, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, doi=null, pmid=null, pmcid=null, year=2021, volume=64, issue=null, pageStart=1392, pageEnd=1422, url=null, language=null, rfNumber=[78], rfOrder=95, authorNames=Zhang X, Man Y, Zhuang XH, Shen JB, Zhang Y, Cui YN, Yu M, Xing JJ, Wang GC, Lian N, Hu ZJ, Ma LY, Shen WW, Yang SY, Xu HM, Bian JH, Jing YP, Li XJ, Li RL, Mao TL, Jiao YL, Sodmergen, Ren HY, Lin JX, journalName=Sci China Life Sci, refType=null, unstructuredReference=Zhang X, Man Y, Zhuang XH, Shen JB, Zhang Y, Cui YN, Yu M, Xing JJ, Wang GC, Lian N, Hu ZJ, Ma LY, Shen WW, Yang SY, Xu HM, Bian JH, Jing YP, Li XJ, Li RL, Mao TL, Jiao YL, Sodmergen, Ren HY, Lin JX (2021). Plant multiscale networks: charting plant connectivity by multi-level analysis and imaging techniques. Sci China Life Sci 64, 1392-1422., articleTitle=Plant multiscale networks: charting plant connectivity by multi-level analysis and imaging techniques, refAbstract=null), Reference(id=1321852290680603495, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, doi=null, pmid=null, pmcid=null, year=2023, volume=10, issue=null, pageStart=2461, pageEnd=2466, url=null, language=null, rfNumber=[79], rfOrder=96, authorNames=Zhou N, Sun JS, Zhang RN, Ye R, Li JJ, Bai ZD, Zhou S, Chen Q, Zuo C, journalName=ACS Photonics, refType=null, unstructuredReference=Zhou N, Sun JS, Zhang RN, Ye R, Li JJ, Bai ZD, Zhou S, Chen Q, Zuo C (2023). Quasi-isotropic high-resolution Fourier ptychographic diffraction tomography with opposite illuminations. ACS Photonics 10, 2461-2466., articleTitle=Quasi-isotropic high-resolution Fourier ptychographic diffraction tomography with opposite illuminations, refAbstract=null), Reference(id=1321852290760295272, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, doi=null, pmid=null, pmcid=null, year=2023, volume=16, issue=null, pageStart=1, pageEnd=14, url=null, language=null, rfNumber=[80], rfOrder=97, authorNames=Zhu YY, journalName=Int J Inf Technol Syst Approach, refType=null, unstructuredReference=Zhu YY (2023). 3D reconstruction of ancient building structure scene based on computer image recognition. Int J Inf Technol Syst Approach 16, 1-14., articleTitle=3D reconstruction of ancient building structure scene based on computer image recognition, refAbstract=null)], funds=[Fund(id=1321852272691233540, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, awardId=32370391, language=CN, fundingSource=国家自然科学基金(32370391), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1321852267465130704, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, xref=null, ext=[AuthorCompanyExt(id=1321852267473519313, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, companyId=1321852267465130704, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=State Key Laboratory of Tree Genetics and Breeding, College of Biological Science and Technology, Beijing Forestry University, Beijing 100083, China), AuthorCompanyExt(id=1321852267481907922, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, companyId=1321852267465130704, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=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)])], figs=[ArticleFig(id=1321852271814623996, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, language=EN, label=Figure 1, caption=Trends in the application scenarios of 3D reconstruction technology in botanical research

The node represents the application scenarios of 3D reconstruction technology in the field of botany, and the flow data is derived from 484 articles on plant 3D reconstruction published from 2011 to 2024, with the diversions indicating shifts in application scenarios over time.

, figureFileSmall=cmHz7MfZPuZm5ywxDAyILg==, figureFileBig=C9xAxydf9nNNg2rkIVXt8w==, tableContent=null), ArticleFig(id=1321852271894315773, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, language=CN, label=图1, caption=三维重建技术在植物学研究中的应用场景趋势分析

节点表示三维重建技术在植物学中的应用场景, 流量数据基于2011-2024年植物三维重建相关的484篇文献, 分流表示应用场景的变化。

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The 3D reconstruction based on micro-computed tomography (Micro-CT) imaging technology is used as an example at the organ scale, and the 3D reconstruction based on confocal microscopy is used as an example at the tissue and cell scales.

, figureFileSmall=B3MlP9s5qSLY65K0pRWQzA==, figureFileBig=0AFRLoDfA8nNxDXRTVZFEA==, tableContent=null), ArticleFig(id=1321852272196305663, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, language=CN, label=图2, caption=不同尺度样本的三维重建流程(Jackson et al., 2017; Tracy et al., 2017; Gómez-Felipe and de Folter, 2019)

器官尺度以基于显微CT成像技术(Micro-CT)的三维重建为例, 组织和细胞尺度以基于共聚焦显微成像技术的三维重建为例。

, figureFileSmall=B3MlP9s5qSLY65K0pRWQzA==, figureFileBig=0AFRLoDfA8nNxDXRTVZFEA==, tableContent=null), ArticleFig(id=1321852272263414528, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, language=EN, label=Table 1, caption=

Three-dimensional reconstruction imaging methods and applications of plants at different scales (organ, tissue, and cell)

, figureFileSmall=null, figureFileBig=null, tableContent=
尺度 植物物种 方法 应用部位 参考文献
器官 水稻(Oryza sativa) Micro-CT 茎 吴迪, 2019
银杏(Ginkgo biloba)、拟南芥(Arabidopsis thaliana)和榆树(Ulmus pumila) Micro-CT、LSFM、Auto CUTS-SEM和X-CT 种子 马灵玉, 2021; 祁晓红, 2022; Ma et al., 2024
柑橘(Citrus reticulata ‘Bu Zhi Huo’)、苹果(Malus pumila)、梨(Pyrus spp.)、茄子(Solanum melongena)、萝卜(Raphanus sativus)和葡萄(Vitis vinifera) X-CT 果实 Herremans et al., 2014; Nugraha et al., 2019; Janssen et al., 2020; Van de Looverbosch et al., 2020; 蔡健荣等, 2024
拟南芥(A. thaliana)和大麦(Hordeum vulgare) X-CT 花 Ijiri et al., 2014; Tracy et al., 2017; Xiao et al., 2021
组织 雪松(Cedrus deodara)、苹果(M. pumila)和月季(Rosa hybrida) FIB-SEM和X-CT 输导组织 Herppich et al., 2015; Herremans et al., 2015; Nugraha et al., 2019; 王静等, 2022
拟南芥(A. thaliana) CLSM 分生组织 Gómez-Felipe and de Folter, 2019
橡树(Quercus palustris) X-CT 保护组织 Morisset et al., 2012
橡树(Q. palustris) CLSM和X-CT 营养组织 Fink, 2006; Morisset et al., 2012
细胞 拟南芥(A. thaliana)、狗尾草(Setaria viridis)、青杄(Picea wilsonii)、红松(Pinus densiflora)和黑松(P. thunbergii) ODT、ssTEM、 FIB-SEM和CLSM 叶肉细胞、花粉粒细胞、下胚轴细胞和雄蕊细胞 Jackson et al., 2017; Kim et al., 2018; Cui et al., 2019; Wang et al., 2020; Silveira et al., 2022; Zechmann et al., 2022; Guo et al., 2023; Yamakawa et al., 2023; Collevatti et al., 2024; Hu et al., 2024
), ArticleFig(id=1321852272359883521, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, language=CN, label=表1, caption=

植物不同尺度(器官、组织和细胞)三维重建成像方法与应用

, figureFileSmall=null, figureFileBig=null, tableContent=
尺度 植物物种 方法 应用部位 参考文献
器官 水稻(Oryza sativa) Micro-CT 茎 吴迪, 2019
银杏(Ginkgo biloba)、拟南芥(Arabidopsis thaliana)和榆树(Ulmus pumila) Micro-CT、LSFM、Auto CUTS-SEM和X-CT 种子 马灵玉, 2021; 祁晓红, 2022; Ma et al., 2024
柑橘(Citrus reticulata ‘Bu Zhi Huo’)、苹果(Malus pumila)、梨(Pyrus spp.)、茄子(Solanum melongena)、萝卜(Raphanus sativus)和葡萄(Vitis vinifera) X-CT 果实 Herremans et al., 2014; Nugraha et al., 2019; Janssen et al., 2020; Van de Looverbosch et al., 2020; 蔡健荣等, 2024
拟南芥(A. thaliana)和大麦(Hordeum vulgare) X-CT 花 Ijiri et al., 2014; Tracy et al., 2017; Xiao et al., 2021
组织 雪松(Cedrus deodara)、苹果(M. pumila)和月季(Rosa hybrida) FIB-SEM和X-CT 输导组织 Herppich et al., 2015; Herremans et al., 2015; Nugraha et al., 2019; 王静等, 2022
拟南芥(A. thaliana) CLSM 分生组织 Gómez-Felipe and de Folter, 2019
橡树(Quercus palustris) X-CT 保护组织 Morisset et al., 2012
橡树(Q. palustris) CLSM和X-CT 营养组织 Fink, 2006; Morisset et al., 2012
细胞 拟南芥(A. thaliana)、狗尾草(Setaria viridis)、青杄(Picea wilsonii)、红松(Pinus densiflora)和黑松(P. thunbergii) ODT、ssTEM、 FIB-SEM和CLSM 叶肉细胞、花粉粒细胞、下胚轴细胞和雄蕊细胞 Jackson et al., 2017; Kim et al., 2018; Cui et al., 2019; Wang et al., 2020; Silveira et al., 2022; Zechmann et al., 2022; Guo et al., 2023; Yamakawa et al., 2023; Collevatti et al., 2024; Hu et al., 2024
), ArticleFig(id=1321852272456352514, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, language=EN, label=Table 2, caption=

Comparative analysis of six common 3D reconstruction imaging methods in plants

, figureFileSmall=null, figureFileBig=null, tableContent=
方法 分辨率 成像速度 样品制备 定位水平 参考文献
LSCM 200-500 nm 秒级/分级, 快 需荧光标记和透明化处理 细胞、亚细胞和分子水平 李叶等, 2015; 李成辉等, 2020; 路姣等, 2023
X-CT 250 μm以下 分级, 较快 需稳定固定 器官、组织、细胞和亚细胞水平 Du et al., 2019; Piovesan et al., 2021; Karahara et al., 2023
Micro-CT 50 μm以下 分级/小时级, 较快, 越精细越慢且越清晰 采用冷冻或化学固定 器官、组织、细胞和亚细胞水平 Wang et al., 2018; Karahara et al., 2023
ODT 200-600 nm 秒级/分级, 快 无需荧光标记, 透明化处理 器官、组织、细胞和亚细胞水平 He et al., 2023; Zhou et al., 2023
FIB-SEM 1-10 nm 小时级, 慢 制备方法复杂, 常温树脂包埋方法、高压冷冻及冷冻等 细胞及亚细胞水平 Heymann et al., 2006; Wei et al., 2012; 贾星等, 2022
TEM 0.1-2 nm 小时级, 慢 化学固定、常温脱水、包埋、切片、染色后观察 细胞、亚细胞、分子和原子水平 Drummy et al., 2004; 李斗星, 2004
), ArticleFig(id=1321852272577987331, tenantId=1146029695717560320, journalId=1316781310003478668, articleId=1321852266265559753, language=CN, label=表2, caption=

植物中6种常见的三维重建成像方法比较分析

, figureFileSmall=null, figureFileBig=null, tableContent=
方法 分辨率 成像速度 样品制备 定位水平 参考文献
LSCM 200-500 nm 秒级/分级, 快 需荧光标记和透明化处理 细胞、亚细胞和分子水平 李叶等, 2015; 李成辉等, 2020; 路姣等, 2023
X-CT 250 μm以下 分级, 较快 需稳定固定 器官、组织、细胞和亚细胞水平 Du et al., 2019; Piovesan et al., 2021; Karahara et al., 2023
Micro-CT 50 μm以下 分级/小时级, 较快, 越精细越慢且越清晰 采用冷冻或化学固定 器官、组织、细胞和亚细胞水平 Wang et al., 2018; Karahara et al., 2023
ODT 200-600 nm 秒级/分级, 快 无需荧光标记, 透明化处理 器官、组织、细胞和亚细胞水平 He et al., 2023; Zhou et al., 2023
FIB-SEM 1-10 nm 小时级, 慢 制备方法复杂, 常温树脂包埋方法、高压冷冻及冷冻等 细胞及亚细胞水平 Heymann et al., 2006; Wei et al., 2012; 贾星等, 2022
TEM 0.1-2 nm 小时级, 慢 化学固定、常温脱水、包埋、切片、染色后观察 细胞、亚细胞、分子和原子水平 Drummy et al., 2004; 李斗星, 2004
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不同尺度三维重建技术在植物研究中的发展及应用
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黄梦莎 , 孔令蝶 , 于淼 , 刘畅 , 王思钦 , 王若涵 *
植物学报 | 专题论坛 2025,60(6): 1005-1016
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植物学报 |专题论坛 2025 , 60 (6) : 1005 -1016
不同尺度三维重建技术在植物研究中的发展及应用
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黄梦莎, 孔令蝶, 于淼, 刘畅, 王思钦, 王若涵*
作者信息
  • 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
通讯作者:
* 王若涵, 北京林业大学教授, 博士生导师。主持国家自然科学基金、“十三五”及“十四五”重点研发计划及国家林草局子课题等国家级项目。主编规划教材2部, 部分代表性成果发表在《科学通报》《中国科学-生命科学》、Plant Physiology及The Plant Journal等国内外权威期刊。荣获北京市优秀青年科技论文一等奖等。E-mail:
Development and Application of 3D Reconstruction Technology at Different Scales in Plant Research
Mengsha Huang, Lingdie Kong, Miao Yu, Chang Liu, Siqin Wang, Ruohan Wang*
Affiliations
  • State Key Laboratory of Tree Genetics and Breeding, College of Biological Science and Technology, Beijing Forestry University, Beijing 100083, China
  • Corresponding Author:
出版时间: 2025-11-10 doi: 10.11983/CBB25002
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三维重建技术(3D reconstruction)是利用计算机图形学和图像处理技术, 从二维图像数据中提取目标物体的几何和拓扑信息, 构建计算机可处理的三维数学模型, 从而实现物体的虚拟重建。在植物学研究中, 三维模型的构建已成为研究植物生长发育、形态结构和功能机制的有效手段, 为多尺度成像、测量和分析提供了有力支撑, 并在农业和林业领域展现出巨大的应用潜力。近年来, 随着植物三维重建技术的不断完善, 其在植物学研究中衍生出不同的应用方向, 涵盖植物形态结构建模、生长发育动态监测以及植物育种等多方面。该文综述了三维重建技术的发展历程及常见的三维重建成像技术在植物不同尺度(从器官、组织到细胞)研究中的应用, 重点阐述这些技术的基本原理及应用, 旨在为植物多模态跨尺度成像以及表型与功能研究提供理论和技术支撑, 为揭示植物生长发育规律及响应环境变化的机制提供新途径。

三维重建  /  不同尺度  /  成像技术  /  模型  /  植物

3D reconstruction technology involves using computer graphics and image processing technologies to extract the geometric and topological information of the target object from the two-dimensional image data. This information is then used to create a three-dimensional mathematical model that can be processed by a computer, enabling the virtual reconstruction of the target object. In plant science research, the construction of three-dimensional models has become an effective way to study plant growth and development, morphological structure and functional mechanism. These models provide robust support for multi-scale imaging, measurement and analysis, demonstrating significant application potential in the field of agriculture and forestry. In recent years, advancements in plant 3D reconstruction technology have led to diverse applications in botanical research, covering plant morphological structure modeling, growth and development dynamic monitoring, and plant breeding. In this paper, we summarize the development process of 3D reconstruction technology and its application in plant studies across different scales (from organs and tissues to cells). We focus on the basic principles and applications of these technologies, aiming to provide theoretical and technical support for multimodal cross-scale imaging and plant phenotypic and functional research. Additionally, this work offers a novel approach to understand the principles of plant growth and development and the mechanisms underlying their responses to environmental changes.

3D reconstruction  /  different scales  /  imaging techniques  /  models  /  plants
黄梦莎, 孔令蝶, 于淼, 刘畅, 王思钦, 王若涵. 不同尺度三维重建技术在植物研究中的发展及应用. 植物学报, 2025 , 60 (6) : 1005 -1016 . DOI: 10.11983/CBB25002
Mengsha Huang, Lingdie Kong, Miao Yu, Chang Liu, Siqin Wang, Ruohan Wang. Development and Application of 3D Reconstruction Technology at Different Scales in Plant Research[J]. Chinese Bulletin of Botany, 2025 , 60 (6) : 1005 -1016 . DOI: 10.11983/CBB25002
据统计, 全球植物种类约30-35万种, 其中包括苔藓植物1.6万余种, 蕨类植物1.3万余种, 裸子植物1 000余种, 而有花植物最为庞大, 约26万种(蒋志刚, 2016), 这些多样性丰富的植物类型构成了森林、草原、湿地等不同尺度的生态系统。从宏观的生物个体到微观的蛋白质分子, 植物世界涵盖不同尺度的元素和结构, 进一步增加了研究的复杂性(Zhang et al., 2021)。传统切片法容易破坏组织器官的完整性和细胞内亚细胞结构的连续性, 从而导致表型特征采集不完整或组织器官成像分辨率低, 甚至部分细胞信息缺失(Hériché et al., 2022)。这些局限性使得传统方法通用性差, 难以捕捉到植物复杂的立体细节。而三维重建技术可提供解决样本无损完整成像、高精度捕捉内部几何细节的有效方案, 不仅可实现对植物整体器官形态的解读, 还能获取组织细胞的位置、拓扑参数和发育等多维度信息。鉴于自然界中植物的多样性和尺度差异显著, 开展多尺度、多层次的形态发生过程三维重建, 对于揭示植物生长发育规律非常必要, 这在植物表型、结构功能建模乃至农林等生物领域具有较大的应用价值(Luo et al., 2023; Arshad et al., 2024; Wen et al., 2024)。
三维重建技术利用计算机图形学和图像处理技术, 从1个或多个视角的图像中提取二维物体的几何拓扑特征, 构建符合计算机表达、用于处理三维对象的数学模型, 从而实现三维信息的重建(Zhu, 2023)。这成为通过虚拟现实来表征客观存在的重要手段。Roberts (1963)编写了一项可将照片处理成线条图, 将线条图转换为三维模型的计算机程序, 进而实现从任何角度显示去除所有隐藏线的三维结构。Cooley和Tukey (1965)提出的快速傅里叶变换(fast Fourier transform, FFT)算法极大地促进了数字信号处理技术的发展。利用该算法可高效计算离散傅里叶变换(discrete Fourier transform, DFT), 将信号从空间域转换到频率域, 不同频率揭示出不同图像特征, 包括边缘、纹理和亮度水平。FFT算法的应用, 促使计算机视觉技术开始执行图像过滤、降噪和特征提取等基本任务, 这对于增强图像清晰度和对象识别准确率以及提高各种计算机视觉应用的整体性能至关重要。随后, David Marr于1982年基于数学、神经生理学和计算机图形学等学科, 开创性地汇集整理出一系列计算机视觉理论(Stevens, 2012)。此后利用视觉进行三维建模逐渐成为主流。自20世纪80年代起, 研究者开始用四元数进行匹配研究, 并由Besl和Mcaky (1992)提出了迭代最近点(iterative closest point, ICP)算法, 该算法包括筛选、匹配、权重、去除、误差度量和最小化等步骤。现有的三维重建视觉系统, 通常先建立相对基本的图像数据框架, 再基于特定视角点构建2.5D图像数据(即该视角可观测到的图像数据), 最终通过整合这些2.5D数据实现三维图像重建(陈雪娇, 2015)。
随着计算机视觉理论不断完善, 三维重建技术在生物学研究中迅速发展, 并在植物学研究中衍生出多种应用场景。这些场景包括器官结构、生长和发育监测、组织结构、植物病理结构检测、生理指标、植物育种、细胞3D重建和体积提取(图1), 呈现出精细化和多元化的发展趋势, 涵盖从宏观到微观的多个研究层面。目前已开发出基于相机的成像系统和重建算法等多种用于植物3D重建的方法(Montenegro-Johnson et al., 2015; Gao et al., 2021; Ovečka et al., 2022)。采用显微CT成像技术(micro-computed tomography, Micro-CT)或X射线断层扫描技术(X-ray computed tomography, X-CT)获得的多视角成像图有效应用于细胞或组织水平各种器官的三维重建(Wang et al., 2017; Qi et al., 2022; Trueba et al., 2022); 采用激光共聚焦扫描显微技术(laser scanning confocal microscopy, LSCM)和光片荧光显微技术(light sheet fluorescence microscopy, LSFM)可对透明植物组织或器官进行三维重建(Truernit et al., 2008; Maizel et al., 2011; Janes et al., 2018), 扩展了植物多尺度分析与重建技术的应用。
近年来, 植物形态结构研究已从细胞和组织扩展到器官等多尺度, 涵盖参数测量、几何结构解析、三维模型构建、结构功能关系模拟以及三维模型的真实感渲染等方面。随着激光共聚焦扫描显微技术、显微CT技术、光学衍射断层扫描技术(optical diffraction tomography, ODT)、聚焦离子束扫描电子显微技术(scanning electron microscope-focused ion beam, FIB-SEM)和透射电子显微技术(transmission electron microscope, TEM)的快速更新, 利用这些先进成像技术进行植物内外形态结构测量和表型功能分析, 已成为加速实现研究数据高精度、多维度、可复现的关键。因此, 精研图像识别和深度学习等AI算法, 结合多种先进的光谱成像三维重建技术, 研发多模态跨尺度植物分析与重建技术, 从而为当今植物科研“数智”赋能, 已成为国际植物研究领域竞相争夺的战略高地。
当前植物中应用较多的三维重建成像技术主要包括激光共聚焦扫描显微技术、X射线断层扫描成像技术、显微CT成像技术、光学衍射断层扫描技术、聚焦离子束扫描电子显微技术和透射电子显微技术。下文将详细阐述三维重建成像方法在植物不同尺度(器官、组织和细胞)研究中的应用(表1)以及6种常见的三维成像方法的优缺点(表2)。
传统荧光显微镜的激发光均匀地投射到整个视野, 激发该区域内的样品发出发射光, 使用图像采集设备采集该区域的图像, 同时收集视野内样品发出的所有发射信号, 导致来自焦平面的发射信号被来自非焦平面的信号严重干扰, 尤其在厚样品成像中, 图像模糊问题尤为突出。激光共聚焦扫描显微镜则采用激光束作为点光源激发样品, 显著减小了激发区域, 降低了相邻样品间的信号串扰。此外, LSCM通过在检测器前的共轭位置加装针孔(pinhole), 仅允许焦平面发射光通过, 有效阻挡了非焦平面的反射光信号, 提高了图像的信噪比。
为了获得完整图像, LSCM利用XYZ扫描振镜实现逐点扫描(李叶等, 2015; 李成辉等, 2020; 路姣等, 2023)。通过控制载物台的Z轴移动, LSCM可进行光学切片, 获取一系列高分辨率、高对比度的二维光学图像投影。这些图像经LSCM软件处理后, 可进行三维重建, 直观呈现样品的立体结构以及亚细胞结构的空间分布关系。
X射线计算机断层扫描具备良好的穿透性与高空间分辨率, 能快速获取厘米至纳米级物体内部微观结构的详细空间及结构异质性信息(Du et al., 2019; Piovesan et al., 2021)。在CT成像过程中, X射线源发射的射线穿透样本后, 由检测器从不同角度接收信号(Davies et al., 2011)。由于样本各部位对X射线吸收率不同, 这些信号经计算机数位几何处理, 可重建出断层影像。通过堆叠这些断层图像, 最终形成样品的三维立体影像。
微计算机断层扫描技术也称为显微CT、微焦点CT或者微型CT。它利用微焦点X射线球管扫描样品的各个层面, 探测器接收透过的X射线, 并将其依次转化为可见光和电信号, 再经模拟数字转换为数字信号, 最终由计算机重建图像。Micro-CT不仅提供几何信息(如尺寸、体积和空间坐标), 还能获取结构基础信息(衰减值、密度和多孔性等材料学信息) (Wang et al., 2018; Karahara et al., 2023)。
CT成像技术的发展离不开重建算法的革新。1917年, CT成像重建算法始于Radon变换与逆变换的提出, 奠定了CT图像重建的数学基础; 1971-1974年, 滤波反投影算法的提出开启了图像精确重建的进程(Masters et al., 2009); 1980年, FDK算法开启了图像三维重建的新纪元; 2002年, Katsevich (2002)解决了锥形束螺旋CT图像精确重建的轴向截断问题; 2004年, 反投影滤波算法解决了数据横向截断问题(李亮等, 2006); 2019年, 基于人工智能技术/深度学习技术的智能重建方法推动了CT重建算法革新(李青等, 2023)。
光学衍射层析成像技术从全息图获取物体振幅与相位, 通过不同照明角度采集复振幅, 经频域层析叠加, 获取三维频谱信息, 再转化至空域, 最终得到物体三维折射率(refractive index, RI)。在光学成像领域, 对于弱散射样品或相位物体, 可通过测量其在不同照明角度下的多个二维全息图, 重建三维折射率分布。通过亥姆霍兹方程与滤波后的反投影算法, 获得物体在不同照明角度下多个二维X射线图像的测量值, 最终实现物体三维分布的重建。从算法角度来看, ODT的重建方法主要分为衍射算法与投影算法, 二者均为逆散射解, 区别在于对光衍射影响的考量。一般来说, ODT采用衍射算法重建三维RI图谱。也有一些研究使用投影算法来研究细胞(Jin et al., 2017)。生物细胞内部结构复杂, 光衍射现象尤为显著, 因此衍射算法在精度上更具优势。
电子显微镜(electron microscope, EM)包括多种技术, 大致分为扫描电子显微镜(scanning electron microscopy, SEM)和透射电子显微镜。
聚焦离子束扫描电子显微镜结合了聚焦镓离子束和带二级电子探测器的电子束。其中, 镓光束可用于成像, 可磨铣感兴趣的区域, 通过持续暴露表面, 清晰呈现组织与细胞特征; 或者制备高质量的样品切片, 用于透射电子显微镜观察。扫描电镜光束类似于传统的扫描电镜, 能可视化感兴趣的区域。当两种光束结合在一起时, FIB-SEM能同步实现对样品的切片与可视化操作(Wei et al., 2012), 获取物体内部结构的三维图像。
在生物组织中, FIB-SEM主要有2种应用方式。(1) 使用FIB-SEM设备对样品进行嵌入、切片和成像(Heymann et al., 2006); (2) 在某些情况下, 利用FIB功能制备TEM成像的高质量切片(Wei et al., 2012)。目前, FIB-SEM已广泛应用于多种生物样品的成像, 包括动物细胞(Bushby et al., 2011)、微生物(Heymann et al., 2006; Wei et al., 2012)、植物细胞中的淀粉颗粒(Crumpton-Taylor et al., 2012)和花粉细胞(House and Balkwill, 2013)。
透射电子显微镜(TEM)的工作原理是利用高能电子束穿透极薄的样品, 电子束与样品原子相互作用, 进而揭示出样品内部结构的精细特征(Kim et al., 2015; Masyutin et al., 2023)。由于电子束的波长比可见光和紫外光短得多, 且电子束的波长与发射电子束的电压平方根成反比, 因此TEM能够以极高的分辨率成像。
TEM三维重建的基本原理基于中央截面定理, 即物体投影的傅里叶变换等价于物体傅里叶空间中垂直于电子束方向的中心截面。通过采集大量不同方向的颗粒投影(单颗粒分析)或同一物体不同倾角的投影(断层成像)实现三维重构。首先, 确定每张投影的取向和中心, 如利用等价线或投影匹配方法。随后, 将每张投影进行傅里叶变换, 并根据其取向填充到三维傅里叶空间的对应截面上, 利用插值算法计算完整的三维傅里叶空间数据。最后, 进行反傅里叶变换得到物体的三维结构(张凯等, 2010; 沈若涵, 2021)。
三维重建技术广泛应用于植物器官研究, 为深入理解植物生长发育提供了强大的工具。利用三维重建技术不仅能量化早期器官形态特征, 还能揭示复杂器官重建过程中的发育动力学(Serral et al., 2022)。LSFM技术克服了传统共聚焦显微镜的局限, 实现了植物器官的微创、多色和三维成像, 可用于探究侧根的昼夜生长节律(Maizel et al., 2011)。
在植物茎秆研究中, 高分辨率三维模型对于探究其生长、抗逆性和生物力学特性至关重要。Micro-CT结合改进的生成式对抗网络(generative adversarial networks, GAN)可高效构建水稻(Oryza sativa)幼苗茎秆微观三维模型, 有效降低实验成本(吴迪, 2019)。
传统上, 植物种子的内部结构研究依赖于二维切片, 通过三维重建技术可获得更全面的信息。马灵玉(2021)使用Micro-CT、LSFM和连续超薄切片扫描电镜成像(Auto CUTS-SEM)技术结合软件分析, 重建了银杏(Ginkgo biloba)和拟南芥(Arabidopsis thaliana)种子的三维结构, 并深入探究了种胚及分泌腔的三维发育过程。此外, 祁晓红(2022)将Micro-CT与LSFM等三维重建技术、转录组测序及质谱成像相结合, 解析了榆树(Ulmus pumila)种子内部构造、基因表达和代谢物变化之间的关系。林金星团队使用改进染色方法的Micro-CT技术研究种子胚胎发育过程中的结构和形态动力学, 为揭示种子起源、进化和适应性机制提供了证据(马灵玉等, 2022; Ma et al., 2024)。
传统方法利用肉眼识别果实生理指标, 而X-CT技术可无损获得果实生理指标, 揭示果实内部特征和生理状态。有研究利用该技术分析了苹果(Malus pumila)、梨(Pyrus spp.)、茄(Solanum melongena)和萝卜(Raphanus sativus)孔隙率的变化(Nugraha et al., 2019; Janssen et al., 2020), 以此检测果实内部缺陷, 如梨的褐变(Van de Looverbosch et al., 2020)和苹果的水核(Herremans et al., 2014)。此外, 针对传统的无损检测技术难以定量分析厚皮柑橘(Citrus reticulata ‘Bu Zhi Huo’)可食率的问题, 蔡健荣等(2024)研发了基于线阵X射线成像和三维重建技术的厚皮柑橘可食率检测方法。
CT技术也广泛应用于植物形态学和解剖学, 特别是对于脆弱结构(如娇嫩的花朵和柔软的浆果)的无损成像。Ijiri等(2014)结合X-CT和主动轮廓模型, 开发出针对薄器官(如花朵)的三维重建方法, 显著提高了三维重建的精度。Xiao等(2021)利用Micro-CT研究葡萄(Vitis vinifera)器官形态及其内部空隙分布的异质性, 深化了对葡萄生殖器官结构和功能的理解。此外, Micro-CT在作物产量研究中也有重要应用, 可提供非破坏性三维表型信息, 用于精确分析花发育(Tracy et al., 2017), 如拟南芥和大麦(Hordeum vulgare)的花期判定及环境胁迫下的花粉发育研究。
三维重建技术不仅可实现对完整器官的无损成像, 还能从组织层面分析多种特定区域的空间结构差异, 并深入解析其发生机理。例如, 利用X-CT揭示橡树(Quercus palustris)营养组织髓与保护组织树皮表皮之间的强相关性, 并量化表皮发育的不同阶段(Morisset et al., 2012)。还有研究者利用X-CT观察苹果果实内部输导组织和孔隙网络的三维形态, 解析了其发育过程中的微结构异质性(Herremans et al., 2015; Nugraha et al., 2019)。此外, Herppich等(2015)应用同步辐射X射线计算机断层扫描技术(synchrotron radiation X-ray computed tomography, SXCT)研究了月季(Rosa hybrida) 3个品种(Akito、Milva和Red Giant)花梗输导组织的内部结构差异, 并探究其弯颈敏感性机制。FIB-SEM结合Imaris软件可实现对雪松(Cedrus deodara)木质部具缘纹孔微纳结构的精准成像, 揭示了木质部水分输导、栓塞形成、扩散以及疲劳机制, 为木材干燥和保护提供了科学依据(王静等, 2022)。CLSM技术结合组织透明化、细胞壁染色技术以及水浸镜片, 提高了植物组织(如拟南芥花组织)的成像质量(Gómez-Felipe and de Folter, 2019)。上述研究表明, 高分辨率三维成像技术为深入研究复杂植物组织结构和功能提供了有力工具。
在细胞尺度上, 电子显微镜和电子断层扫描能够提供细胞和细胞器在亚细胞分辨率下的3D超结构数据, 如细胞异质微结构、细胞间空间(形状、体积、表面积、孔隙密度、孔隙率、曲折度及连通性等)及间隙(Cui et al., 2019; Wang et al., 2020; Guo et al., 2023)。连续切片透射电子显微镜(serial section transmission electron microscopy, ssTEM)、FIB-SEM和CLSM可将厚切片转化为数张光学切片, 从而对植物细胞进行3D重建和体积提取, 以揭示细胞大小、数量和细胞器组成的差异(Zechmann et al., 2022)。Yamakawa等(2023)对狗尾草(Setaria viridis)叶肉细胞和叶绿体的3D重建量化了其在聚集运动前后的结构或细胞内位置, 显示聚集位置与叶绿体体积或表面积无关, 且叶绿体在运动前后未改变形状。Kim等(2018)使用Auto CUTS-SEM重建了青杄(Picea wilsonii)花粉粒的内部细胞组成。此外, 他们采用ODT这一无标记快速三维成像技术, 获取红松(Pinus densiflora)和黑松(P. thunbergii)花粉粒的三维折射率及形态特征, 并对不同亚结构进行了清晰的区分和分析, 为解析花粉粒生理特性提供了新视角。Collevatti等(2024)结合CLSM技术、图像分析软件和机器学习, 识别并分析了现代及化石花粉粒, 通过形状和外显子定量及定性参数, 揭示了花粉发育的重要变量。
植物生长和分化的复杂过程一直是植物学领域研究的难点, 但高分辨率三维成像技术的应用为揭示多细胞生物的发育机制开辟了新的研究方向。基于共聚焦显微成像技术的三维重建对拟南芥下胚轴细胞图谱的拓扑重建和雄蕊发育的全面定量表征, 解析了下胚轴和雄蕊发育定量图谱的细胞生长动力学(Jackson et al., 2017; Silveira et al., 2022)。基于大体积全自动细胞重建(large-volume full automated cell reconstruction, LVACR)框架, 通过植物中的三维细胞学特征和细胞连接, 构建了不同植物组织的细胞图谱, 为揭示多细胞生物的精确空间排列和细胞行为提供了新视角(Hu et al., 2024)。
植物的多尺度三维重建对于深入理解其复杂结构和功能至关重要(图2)。植物由小分子、生物大分子、亚细胞组分、多细胞结构、组织器官及系统架构等不同尺度的元件构成, 这些不同尺度的元件相互连接, 形成复杂的网络, 共同调控植物的各种生命活动。然而, 构建完整的多尺度三维模型仍然面临诸多挑战。
当前, 尽管已经开发出许多先进的三维重建技术、深度学习分割算法以及各类植物细胞图谱(Li et al., 2017; Zhang et al., 2021; 施般若等, 2022; Hu et al., 2024; Ma et al., 2024), 但在重建植物多细胞特异性以及组织系统连接方面仍具挑战性。(1) 活体植物细胞的原位无损成像及动态行为无损捕捉技术有待突破, 以便用于研究瞬时、小概率的生理活动和发育过程; (2) 基于三维可视化仪的重建方法成本高且耗时, 在未来的研究中, 应进一步开发高效准确的多细胞模型, 以实现自动化、智能化重建植物多细胞特征及组织系统连接; (3) 高等植物的生命活动复杂, 目前已开发了很多新的细胞图谱, 揭示了局部多细胞结构的生长发育动力学过程, 但是由于开发难度大, 通常只能用于某种条件下, 对整体多细胞之间的活动缺乏全局性了解, 未来可从单个细胞出发, 深入解析组织特异性及器官尺度的植物体系。
此外, 植物器官的大规模高分辨率3D重建亦具有挑战性。(1) 大容量全自动细胞重建效率低, 需要进一步优化细胞分割和聚集算法; (2) 器官尺度的成像分辨率有待提升, 一般物体尺度越小, 分辨率越高, 未来需要开发更高分辨率的成像技术和先进算法; (3) 目前植物多尺度网络分析框架较为单一, 需构建统一、全面且随时可用的框架, 以支持系统级的计算建模, 并应用于更广泛的植物学研究领域。
随着智能算法和成像技术的不断发展, 结合多尺度三维成像数据和拓扑分析方法, 将有助于深入解析植物发育过程中的三维时空信息。此外, 深度学习等先进算法可提高图像处理和特征提取效率, 实现细胞尺度的精细重建。显微图像三维重建模型的构建与优化进一步推动了三维重建技术在植物微观尺度的应用, 将为植物科学研究和实际应用提供更强大的工具。
展望未来, 发展跨尺度活体生物样品多模态三维重建成像技术至关重要, 高时空分辨率成像方式以及生物大分子高分辨率成像等方面的新型多模态跨尺度生物成像设施也在推陈出新, 这将有助于拓展三维重建技术在植物生长发育和生态功能研究中的应用, 为生态保护、农林业管理和生物研究提供技术保障。
黄梦莎: 文献收集整理, 作图, 撰写和修改文章; 孔令蝶, 于淼: 修改文章, 提供指导和建议; 刘畅, 王思钦: 提供指导和建议; 王若涵: 制定论文提纲, 修改文章, 提供指导、修改建议及资金支持。
  • 国家自然科学基金(32370391)
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2025年第60卷第6期
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doi: 10.11983/CBB25002
  • 接收时间:2025-01-09
  • 首发时间:2026-10-27
  • 出版时间:2025-11-10
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  • 收稿日期:2025-01-09
  • 录用日期:2025-04-02
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国家自然科学基金(32370391)
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    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

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* 王若涵, 北京林业大学教授, 博士生导师。主持国家自然科学基金、“十三五”及“十四五”重点研发计划及国家林草局子课题等国家级项目。主编规划教材2部, 部分代表性成果发表在《科学通报》《中国科学-生命科学》、Plant Physiology及The Plant Journal等国内外权威期刊。荣获北京市优秀青年科技论文一等奖等。E-mail:
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2种不同金属材料的力学参数
科
Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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