3,5-二硝基水杨酸法的优化及日本落叶松木质部发育中阿拉伯半乳聚糖的含量变异

doi:10.11707/j.1001-7488.LYKX20230597

摘要/Abstract

摘要：

目的: 建立3,5-二硝基水杨酸法最佳检测条件和最佳水解条件，以便准确测定日本落叶松木质部中阿拉伯半乳聚糖的含量及其株内变异，为深入了解阿拉伯半乳聚糖在落叶松木质部中的分布、开发与生物合成提供技术支撑。方法: 从显色剂体积、反应时间和反应温度对3,5-二硝基水杨酸法进行分析，确定最佳的落叶松阿拉伯半乳聚糖检测条件，同时利用正交试验从盐酸浓度、盐酸体积和水解时间对总糖水解条件进行优化，分别测定总糖和还原糖含量，从而计算出多糖含量。结果: 3,5-二硝基水杨酸法的最佳检测条件：3,5-二硝基水杨酸显色剂体积为5 mL，反应时间为9 min，反应温度为90 ℃；最佳水解条件：盐酸浓度为6 mol·L^?1，盐酸体积为15 mL，水解时间为40 min。在日本落叶松单株中，轴向上木质部中阿拉伯半乳聚糖含量随着树木高度的增加而逐渐降低；径向上心材含量远高于边材，总体上心材区含量随着年轮向外逐渐增加，边材区随着年轮向内逐渐增加，在边材和心材过渡区含量最高，同一年轮中早材含量高于晚材，过渡区早材中阿拉伯半乳聚糖含量远高于晚材。结论: 本研究建立了简便、稳定的不依赖于高质量样本即可准确检测阿拉伯半乳聚糖含量的方法，同时发现阿拉伯半乳聚糖在日本落叶松树干基部含量最高，并且在心材中高于边材，在早材中高于晚材，过渡区含量最高，暗示了阿拉伯半乳聚糖可能于过渡区合成，与心材的形成有关。

关键词: 日本落叶松, 阿拉伯半乳聚糖, 3,5-二硝基水杨酸法, 含量变异

Abstract:

Objective: This study aims to establish the optimal detection and hydrolysis conditions for the 3,5-dinitrosalicylic acid method, in order to accurately determine the content and its variation of arabinogalactan in the xylem of Larix kaempferi, and provide technical support for deeply understanding the distribution, development, and biosynthesis of arabinogalactan in the xylem of L. kaempferi. Method: The optimal detection conditions of 3,5-dinitrosalicylic acid method were determined by analyzing the three aspects of coloration reagent volume, reaction time, and reaction temperature. At the same time, an orthogonal test was used to optimize the hydrolysis conditions of total sugar from three aspects of hydrochloric acid concentration, hydrochloric acid volume and hydrolysis time. The contents of total sugar and reducing sugar were measured, respectively, with which the polysaccharide content was calculated. Result: The optimum detection conditions of the 3,5-dinitrosalicylic acid method were established. The coloration reagent volume, reaction time, and reaction temperature were 5 mL, 9 min, and 90 ℃, respectively. The optimal hydrolysis conditions were hydrochloric acid concentration of 6 mol·L^?1, hydrochloric acid volume of 15 mL, and hydrolysis time of 40 min. In individual L. kaempferi trees, the content of arabinogalactan in xylem gradually decreased with the increase of tree height in the axial direction, and was much lower in the sapwood than that in the heartwood in the radial direction. Overall, the arabinogalactan content in heartwood area gradually increased outward with the growth ring, while the arabinogalactan content in the sapwood area gradually increased inward the growth ring. The arabinogalactan content was highest in the transition zone between sapwood and heartwood, the arabinogalactan content in springwood was much higher than that in summerwood within each growth ring, and the arabinogalactan content in earlywood was much higher than that in latewood within the transition zone. Conclusion: In this study, a simple and stable method for accurately detecting the content of arabinogalactan without relying on high-quality samples has been established. At the same time, it has been found that arabinogalactan content is highest in the basal trunk, and is higher in the heartwood than in the sapwood, and higher in the springwood than in the summerwood, and highest in the transition zone, suggesting that the arabinogalactan may be synthesized in the transition zone and related to the formation of heartwood.

Key words: Larix kaempferi, arabinogalactan, 3,5-dinitrosalicylic acid method, content variation

中图分类号:

S718.43

张恺恺,谢允慧,孙晓梅. 3,5-二硝基水杨酸法的优化及日本落叶松木质部发育中阿拉伯半乳聚糖的含量变异[J]. 林业科学, 2025, 61(2): 113-121.

Kaikai Zhang,Yunhui Xie,Xiaomei Sun. Optimization of 3,5-Dinitrosalicylic Acid Method and Variation of Arabinogalactan Content in the Developing Xylem of Larix kaempferi[J]. Scientia Silvae Sinicae, 2025, 61(2): 113-121.

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试验号 No.	因素 Factors			方案 Programs
试验号 No.	A 水解时间 Hydrolyzing time/min	B 盐酸浓度 HCl concentration/ (mol·L^?1)	C 盐酸体积 HCl volume/ mL	方案 Programs
1	20 (1)	6 (1)	5 (1)	A1B1C1
2	20 (1)	9 (2)	10 (2)	A1B2C2
3	20 (1)	12 (3)	15 (3)	A1B3C3
4	30 (2)	6 (1)	10 (2)	A2B1C2
5	30 (2)	9 (2)	15 (3)	A2B2C3
6	30 (2)	12 (3)	5 (1)	A2B3C1
7	40 (3)	6 (1)	15 (3)	A3B1C3
8	40 (3)	9 (2)	5 (1)	A3B2C1
9	40 (3)	12 (3)	10 (2)	A3B3C2

试验号 No.	因素 Factors			方案 Program	吸光值 Absorbance
试验号 No.	A 水解时间 Hydrolyzing time/min	B 盐酸浓度 HCl concentration/ (mol·L^?1)	C 盐酸体积 HCl volume/mL	方案 Program	吸光值 Absorbance
1	20	6	5	A1B1C1	0.004 0
2	20	9	10	A1B2C2	0.000 3
3	20	12	15	A1B3C3	0.017 3
4	30	6	10	A2B1C2	0.014 7
5	30	9	15	A2B2C3	0.003 7
6	30	12	5	A2B3C1	0.008 3
7	40	6	15	A3B1C3	0.025 0
8	40	9	5	A3B2C1	0.020 3
9	40	12	10	A3B3C2	0.017 7
k₁	0.007 2	0.014 6	0.010 9
k₂	0.008 9	0.008 1	0.010 9
k₃	0.021 0	0.014 4	0.015 3
R	0.013 8	0.006 4	0.004 4

项目Item	1	2	3	4	5	6	平均值 Mean	相对标准偏差 RSD (%)
总糖Total sugar	0.198	0.198	0.199	0.199	0.199	0.198	0.199	0.252
还原糖Reducing sugar	0.115	0.115	0.116	0.115	0.116	0.116	0.116	0.433

项目Item	样本1 Sample 1	样本2 Sample 2	样本3 Sample 3	样本4 Sample 4	相对标准偏差 RSD (%)
总糖Total sugar	0.134	0.123	0.127	0.128	2.917
还原糖Reducing sugar	0.104	0.102	0.099	0.100	1.821

项目Item	时间 Time/min										相对标准偏差 RSD (%)
项目Item	0	10	20	30	40	50	60	70	80	90	相对标准偏差 RSD (%)
总糖Total sugar	0.199	0.203	0.212	0.200	0.208	0.207	0.208	0.207	0.206	0.208	1.763
还原糖Reducing sugar	0.155	0.119	0.119	0.117	0.120	0120	0.119	0.119	0.120	0.119	1.280