Analysis of Non-Cellulosic Monosaccharide Composition of Poplar Cell Wall by HPAEC-PAD

doi:10.7525/j.issn.1673-5102.2023.05.015

Abstract

Abstract:

To establish an ion exchange chromatography-pulsed amperometric detection （HPAEC-PAD） method for non-cellulosic monosaccharides in poplar（Populus spp.） cell walls， the 9 main cell wall monosaccharides could be base-line separated in one run by optimizing the separation conditions. Dionex CarbiPac^TM PA100 （4 mm×250 mm） with sodium hydroxide analytical column was used for gradient elution at flow rate is 1.0 mL·min^-1 and the column temperature of 30 ℃ was used for the determination of monosaccharide fraction in the cell wall of different poplar species. The results showed that the 9 monosaccharides had a good linear relationship in the linear range of 0.5-150.0 mg·L^-1， the correlation coefficient R² was 0.999 0-0.999 3， the reproducibility（RSD） of the method was 1.09%-3.96%， and the recovery rate by standard addition was 91.32%-109.25%. The detection limit was 1.57×10^-3-1.41×10^-2 mg·L^-1. Galacturonic acid was the highest composition followed by xylose， and the glucuronic acid content was the lowest monosaccharide. The method developed in this study was widely applicable and simple， and could be used for the detection of monosaccharides in the cell walls of poplar and other species.

Key words: poplar, cell wall, ion chromatography, neutral sugar, uronic acid

CLC Number:

S792.11

Xiaoling JIN, Huimin WU, Chaofeng YANG, Jin ZHANG, Mengzhu LU, Wei ZENG. Analysis of Non-Cellulosic Monosaccharide Composition of Poplar Cell Wall by HPAEC-PAD[J]. Bulletin of Botanical Research, 2023, 43(5): 787-793.

Figures/Tables 8

Fig.1

Fig.2

Fig.3

Table 1

Table 2

Table 3

Table 4

Fig.4

References 19

1	喻谨，岳永德，汤锋，等.不同地区竹叶中多糖的提取测定［J］.生物质化学工程，2014，48（3）：40-43.
	YU J， YUE Y D， TANG F，et al.Determination of polysaccharides from bamboo leaves in different regions［J］.Biomass Chemical Engineering，2014，48（3）：40-43.
2	LIU W， LIU Y M， ZHU R，et al.Structure characterization，chemical and enzymatic degradation，and chain conformation of an acidic polysaccharide from Lycium barbarum L.［J］.Carbohydrate Polymers，2016，147：114-124.
3	柴银，王虹，刘雅萌，等.离子色谱测定多糖的单糖组成的方法学研究及其应用［J］.药物生物技术，2016，23（4）：332-336.
	CHAI Y， WANG H， LIU Y M，et al.Methodology study of monosaccharide composition by HPAEC-PAD and its application［J］.Pharmaceutical Biotechnology，2016，23（4）：332-336.
4	TUSKAN G A， DIFAZIO S， JANSSON S，et al.The genome of black cottonwood，Populus trichocarpa（Torr.& Gray）［J］.Science，2006，313（5793）：1596-1604.
5	王烨.毛白杨速生纸浆林地下滴灌施肥效应研究［D］.北京：北京林业大学，2015.
	WANG Y.Research on effects of nitrogen fertigation on tree-growth and its mechanisms of action in populus tomentosa plantation［D］.Beijing：Beijing Forestry University，2015.
6	丁莉萍，王宏芝，魏建华.杨树转基因研究进展及展望［J］.林业科学研究，2016，29（1）：124-132.
	DING L P， WANG H Z， WEI J H.Progress and prospect of research in transgenic poplar［J］.Forest Research，2016，29（1）：124-132
7	KEEGSTRA K.Plant cell walls［J］.Plant Physiology，2010，154（2）：483-486.
8	BAR-PELED M， O’NEILL M A.Plant nucleotide sugar formation，interconversion，and salvage by sugar recycling［J］.Annual Review of Plant Biology，2011，62（1），127-155.
9	HAAS M， LAMOUR S， TRAPP O.Development of an advanced derivatization protocol for the unambiguous identification of monosaccharides in complex mixtures by gas and liquid chromatography［J］.Journal of Chromatography，2018，1568：160-167.
10	乐胜锋，王尉，王雅萱，等.离子色谱-脉冲安培法测定芦荟多糖中7种单糖的含量［J］.色谱，2019，37（3）：319-324.
	LE S F， WANG W， WANG Y X，et al.Determination of seven monosaccharides in aloe polysaccharide using ion chromatography with pulsed amperometric detector［J］.Chinese Journal of Chromatography，2019，37（3）：319-324.
11	曾荣香，张清华，管咏梅，等.柱前衍生化高效液相色谱法测定地稔多糖中单糖组成［J］.中国实验方剂学杂志，2015，21（10）：73-76.
	ZENG R X， ZHANG Q H， GUAN Y M，et al.Analysis of monosaccharide composition in polysaccharides extract from Melastoma dodecandrum by precolumn derivatization HPLC［J］.Chinese Journal of Experimental Traditional Medical，2015，21（10）：73-76
12	张强，季红福，周燕，等.离子色谱法测毛竹竹叶单糖和糖醛酸组成［J］.生物质化学工程，2019，53（6）：33-38.
	ZHANG Q， JI H F， ZHOU Y，et al.Monosaccharide and alduronic acid composition of bamboo leaves by ion chromatography［J］.Biomass Chemical Engineering，2019，53（6）：33-38.
13	O’NEILL M A， BLACK I， URBANOWICZ B，et al.Locating methyl-etherified and methyl-esterified uronic acids in the plant cell wall pectic polysaccharide rhamnogalacturonan II［J］.SLAS Technology，2020，25（4）：329-344.
14	BIDLACK J E， DASHEK W V.Plant cell walls［M］.Plant Cells and Their Organelles，2016：209-238.
15	程小乔，李科，陈雪梅，等.若干双子叶与单子叶植物细胞壁果胶结构单糖组成特征研究［J］.北京林业大学学报，2012，34（5）：44-49.
	CHENG X Q， LI K， CHEN X M，et al.Comparison of pectin structural monosaccharides in cell wall of dicotyledon and monocotyledon［J］.Journal of Beijing Forestry University，2012，34（5）：44-49.
16	CAFFALL K H， MOHNEN D.The structure，function，and biosynthesis of plant cell wall pectic polysaccharides［J］.Carbohydrate Research，2009，344（14）：1879-1900.
17	RATKE C， PAWAR P M A， BALASUBRAMANIAN V K，et al.Populus GT43 family members group into distinct sets required for primary and secondary wall xylan biosynthesis and include useful promoters for wood modification［J］.Plant Biotechnology Journal，2015，13（1）：26-37.
18	KANEDA M， RENSING K， SAMUELS L.Secondary cell wall deposition in developing secondary xylem of poplar［J］.Journal of Integrative Plant Biology，2010，52（2）：234-243.
19	PETTOLINO F A， WALSH C， FINCHER G B，et al.Determining the polysaccharide composition of plant cell walls［J］.Nature Protocols，2012，7（9）：1590-1607.

标准品 Standards	线性范围 Linear range /（mg·L^-1）	线性方程 Linear equation	相关系数 R²	检出限 Limit of detection /（mg·L^-1）	定量限 Limit of quantification /（mg·L^-1）	重现性 RSD（n=6） /%	保留时间 Retention time /min
岩藻糖 L-fucose	0.5~5.0	y=0.901 9x+0.067 5	0.999 2	1.57×10^-3	4.71×10^-3	1.09	5.43
鼠李糖 L-rhamnose	0.5~5.0	y=0.507 3x+0.024 2	0.999 3	5.77×10^-3	1.73×10^-2	1.76	11.72
阿拉伯糖 L-arabinose	5.0~25.0	y=0.865 4x+1.697 3	0.999 1	3.21×10^-3	9.63×10^-3	2.36	12.37
半乳糖 D-galactose	5.0~30.0	y=1.011 7x+1.608 6	0.999 0	2.14×10^-3	6.42×10^-3	1.64	16.00
葡萄糖 D-glucose	2.0~15.0	y=1.143 1x+0.676 7	0.999 1	3.49×10^-3	1.05×10^-2	1.75	18.07
木糖 D-xylose	5.0~40.0	y=0.7050x+3.079 2	0.999 0	6.28×10^-3	1.88×10^-2	2.11	21.68
甘露糖 D-mannose	0.5~5.0	y=0.803 7x-0.026 2	0.999 2	1.41×10^-2	4.22×10^-2	3.07	22.76
半乳糖醛酸 D-galacturonic acid	40.0~150.0	y=0.724 8x-4.107 8	0.999 1	7.76×10^-3	2.32×10^-2	3.84	33.07
葡萄糖醛酸 D-glucuronic acid	0.5~5.0	y=0.903 4x-0.101 8	0.999 3	5.17×10^-3	1.55×10^-2	3.96	35.89

分析物 Analytes	样品质量浓度 Found/（mg·L^-1）	加标量 Add/（mg·L^-1）	测定值 Measured value/（mg·L^-1）	加标回收率 Recovery/%	重现性 RSD（n=3）/%
岩藻糖 L-fucose	2.40	2.5	5.03	105.52	1.90
鼠李糖 L-rhamnose	1.81	2.0	3.97	109.25	4.86
阿拉伯糖 L-arabinose	8.42	10.0	17.70	91.51	2.92
半乳糖 D-galactose	13.74	15.0	29.61	105.80	1.69
葡萄糖 D-glucose	4.73	5.0	10.00	105.83	2.36
木糖 D-xylose	13.60	10.0	22.42	91.32	3.83
甘露糖 D-mannose	0.78	0.8	1.56	98.01	2.00
半乳糖醛酸 D-galacturonic acid	72.05	70.0	140.60	97.98	1.69
葡萄糖醛酸 D-glucuronic acid	1.57	1.5	3.15	105.19	3.96

分析物 Analytes	84K杨 84K poplar /（mg·L^-1）	山新杨 Shanxin poplar /（mg·L^-1）	银中杨 Yinzhong poplar /（mg·L^-1）	717杨 717 poplar /（mg·L^-1）	南林895杨 Nanlin 895 poplar /（mg·L^-1）
岩藻糖 L-fucose	3.40	3.28	2.48	3.37	2.40
鼠李糖 L-rhamnose	1.15	1.87	2.06	1.39	1.81
阿拉伯糖 L-arabinose	12.72	13.60	12.04	6.65	8.41
半乳糖 D-galactose	15.93	19.59	15.62	17.56	13.74
葡萄糖 D-glucose	9.04	8.71	6.95	9.48	4.73
木糖 D-xylose	35.37	20.03	14.69	30.19	13.60
甘露糖 D-mannose	1.73	1.45	1.94	1.81	0.78
半乳糖醛酸 D-galacturonic acid	79.90	84.71	81.87	44.85	72.05
葡萄糖醛酸 D-glucuronic acid	2.74	2.25	2.07	0.86	1.57

杨树种类 Poplar varieties	岩藻糖 L-fucose /%	鼠李糖 L-rhamnose /%	阿拉伯糖 L-arabinose /%	半乳糖 D-galactose /%	葡萄糖 D-glucose /%	木糖 D-xylose /%	甘露糖 D-mannose /%	半乳糖醛酸 D-galacturonic acid /%	葡萄糖醛酸 D-glucuronic acid /%
84K杨 84K poplar	2.44	0.74	9.99	5.92	5.92	27.78	1.13	44.41	1.66
山新杨 Shanxin poplar	2.60	1.33	11.77	6.28	6.29	17.33	1.05	51.86	1.51
银中杨 Yinzhong poplar	1.95	1.62	10.36	11.20	4.98	12.64	1.39	54.48	1.38
717杨 717 poplar	2.42	1.26	7.33	8.70	8.71	33.25	1.66	34.96	0.73
南林895杨 Nanlin 895 poplar	2.22	1.67	8.51	11.58	3.99	13.76	0.66	56.37	1.23

[1]	Jing ZHANG, Weixi ZHANG, Changjun DING, Zhengsai YUAN, Lirui DAI, Xiaohua SU, Yingbai SHEN, Guanzheng QU. Comparative Analysis of Growth, Photosynthetic Physiology and Root Tip Ion Flow Characteristics of Five Poplar Varieties [J]. Bulletin of Botanical Research, 2024, 44(1): 96-106.
[2]	Huiling YAN, Xinxin ZHANG, Xiyang ZHAO, Guanzheng QU, Zhaoning WANG, Rui HAN. Effects of Light Signal on Anthocyanin Biosynthesis and Gene Expression of AmRosea1 Overexpressed 84K Poplar [J]. Bulletin of Botanical Research, 2023, 43(6): 815-825.
[3]	Qing-Yan GAI, Zi-Ying WANG, Yu-Jie FU, Jiao JIAO, Hui-Mei WANG, Yao LU, Jing LIU, Jin-Xian FU, Xiao-Jie XU, Lan YAO. Effects of UV-B Radiation on Phenolic Acid Accumulation and Physiological and Biochemical Characters in 84K Poplar [J]. Bulletin of Botanical Research, 2021, 41(6): 878-887.
[4]	Dan CAO, Shi-Zhen WANG, Mei HAN. Effect of the Environment-friendly Nutrient Synergist Polyaspartic Acid on the Growth of Poplar Seedlings [J]. Bulletin of Botanical Research, 2021, 41(5): 712-720.
[5]	QIAO Bin-Jie, WANG De-Qiu, GAO Hai-Yan, LI Zhao-Min, GE Li-Li, DING Wen-Ya, ZHAO Xi-Yang. Photosynthetic and Stomatal Morphological Variation of Poplar Clones in Seedling Stage under Drought Stress [J]. Bulletin of Botanical Research, 2020, 40(2): 177-188.
[6]	YUAN Jia-Qiu, TIAN Ye, FU Xiang-Xiang. Genetic Identification and Relationship Analysis of Main Southern-type Poplar Clones with EST-SSR [J]. Bulletin of Botanical Research, 2019, 39(5): 770-778.
[7]	LIU Zhao-Ying, JIANG Chuan-Ying, ZHAI Tong-Tong, CHANG Yuan, YAO Zhi-Hong, LIU Zhi-Hua, ZHANG Rong-Shu. Effects of Trichoderma asperellum Combined Application on Growth and Photosynthesis Characteristics of Populus davidiana×P.alba var. pyramidlis [J]. Bulletin of Botanical Research, 2018, 38(1): 64-74.
[8]	WANG Li-Na, WANG Yu-Cheng, YANG Chuan-Ping. The Comparative study of Callus and Direct Differation Regenaration System of 84K Poplar [J]. Bulletin of Botanical Research, 2017, 37(4): 542-548.
[9]	LI Xi-Yan, ZHAO Kai, ZHANG Xue-Mei, ZHOU Bo-Ru. Expression Analysis of Poplar MYB Transcription Factor Gene Family in Response to Salt Stress [J]. Bulletin of Botanical Research, 2017, 37(3): 424-431.
[10]	SI Dong-Jing, ZHANG Xin-Xin, HAN Dong-Hui, JIN Yun-Zhe, DONG Yuan-Hai, ZHAO Guang-Hao, LENG Wei-Wei, ZHAO Xi-Yang. Variance Analysis and Growth Model Building of H and DBH of Poplar Clones at Different Sites [J]. Bulletin of Botanical Research, 2016, 36(4): 588-595.
[11]	YAN Dong, LIU Dian-Kun, SI Dong-Jing, ZHENG Mi, ZHAO Xi-Yang, QU Guan-Zheng, LI Ying. Optional Protein Extracting Methods of Poplar Leaves for Two-dimensional Gel Electrophoresis [J]. Bulletin of Botanical Research, 2016, 36(3): 469-475.
[12]	LIANG Jing1,2；WANG Qing-Cheng1*；XU Li-Juan1；WU Wen-Juan1. Effect of Thinning on Respiration Rate and Soil Carbon Density of Young-age Poplar-birch Stands in Western Changbai Mountains [J]. Bulletin of Botanical Research, 2015, 35(1): 110-116.
[13]	WANG Xiu-Wei1；JIA Gui-Mei2；MAO Zi-Jun2*；SUN Tao2；YUAN Xian-Lei2. Effect of NaCl Tolerance on Photosynthetic Physiology and Growth of Seedlings of Three Poplar Clones [J]. Bulletin of Botanical Research, 2015, 35(1): 27-33.
[14]	YANG Feng-Jian;LIU Wei;LI De-Wen;ZU Yuan-Gang*. Effect of Poplar Plantation on Soil Amelioration in Severe Saline-alkali Soil [J]. Bulletin of Botanical Research, 2012, 32(3): 339-342.
[15]	YAN Yong-Qing;WANG Wen-Jie;ZHU Hong;SHI Xi-Chan;LIU Xing-Liang;ZU Yuan-Gang*. Growth and Physiological Adaptability of Three Hybrid Poplars Planted in Different Saline-alkali Soil [J]. Bulletin of Botanical Research, 2009, 29(4): 433-438.