拉曼测定木质素强度Improving sample preparation to investigate lignin inte...
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Vol.:(0123456789)
1 3
J. For. Res. (2021) 32(5):2135–2142
https://doi.org/10.1007/s11676-020-01244-1
ORIGINAL PAPER
Improving sample preparation toinvestigate lignin
intensity ofxylem atthecellular level byconfocal Raman
microspectroscopy ofPopulus tomentosa
BingWang
2
· MeiLuo
2
· YadiLiu
2
· XiaoruiGuo
2
·
XiatongLiu
2
· ChongZhang
2
· ZhijingZhao
2
· DiLiu
2
·
HuiLi
1,2,3
· HaiLu
1,2,3
Received: 12 May 2020 / Accepted: 21 July 2020 / Published online: 4 November 2020
© Northeast Forestry University 2020
detailed difference in lignin intensity at the cellular level.
Thus, CRM proved to be a useful insitu method to rapidly
analyze the spatial variation of lignin content in the xylem
of woody plants.
Keywords Cell wall· Confocal Raman
microspectroscopy· Lignin· Paraffin section· Populus
tomentosa
Introduction
Raman spectroscopy has been widely applied for chemical
mapping and imaging biological and biomimetic samples
(Evans and Xie 2008; Roeffaers etal. 2011; Sergo etal.
2013; De Bleye etal. 2014; Nima etal. 2014). It has emerged
as a powerful approach to analyze the chemical composition
of the cell wall in a particular area of plants (Gorzsas 2017;
Heiner etal. 2018). The combination of microscopy and
Raman spectroscopy has enabled descriptions of the cellu-
lar structure and composition of individual features of plant
tissues in their native state. These advanced micro-Raman
spectroscopy techniques include confocal Raman micro-
spectroscopy (CRM), coherent anti-Stokes Raman scattering
microspectroscopy, and stimulated Raman scattering micro-
spectroscopy (Zeng etal. 2010; Larsen and Barsberg 2010;
Lelie etal. 2012; D’Arco etal. 2016). Micro-Raman spec-
troscopy can be used to quantify the chemical distribution of
the cell wall in plants insitu, and has provided remarkable
insight into visualizing the cell wall (Gierlinger etal. 2008,
2012; Pohling etal. 2014).
Lignin is the major component of the secondary cell wall
of the vascular system in woody plants, and has enabled
plants to adapt to land from the aquatic environment (Boer-
jan etal. 2003; Popper etal. 2014; Hofte and Voxeur 2017).
Abstract Confocal Raman microspectroscopy (CRM)
is an important tool for analyzing the compositional dis-
tribution of cell walls insitu. In this study, we improved
the sample preparation method using paraffin-embedded
sections combined with hexane dewaxing to obtain high
resolution Raman images. We determined that the cell wall
components of fiber cells were different from those of ray
cells and vessel cells in the xylem of Populus tomentosa.
Acetyl bromide and CRM methods produced similar trends
when the difference in lignin intensity in the xylem region
was compared between transgenic PtrLac4 and wild-type P.
tomentosa. However, CRM proved more useful to analyze
the lignin distribution in each cell type and distinguished the
Bing Wang, Mei Luo and Yadi Liu contributed equally to this
work.
Project funding: This work was funded by the Fundamental
Research Funds for the Central Universities (Grant No.
2019ZY30) and National Natural Science Foundation of China
(Grant No. 31971618, Grant No. 31570582).
The online version is available at http://www.sprin gerli nk.com.
Corresponding editor: Yanbo Hu.
* Hui Li
lihui830@bjfu.edu.cn
1
Beijing Advanced Innovation Center forTree Breeding
By Molecular Design, Beijing Forestry University,
Beijing100083, People’sRepublicofChina
2
College ofBiological Sciences andBiotechnology,
Beijing Forestry University, Beijing100083,
People’sRepublicofChina
3
National Engineering Laboratory forTree Breeding,
Beijing Forestry University, Beijing100083,
People’sRepublicofChina
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