虎杖根系细菌群落组成及其与有效成分含量相关性研究

doi:10.13560/j.cnki.biotech.bull.1985.2024-0375

摘要/Abstract

摘要：

【目的】通过分析虎杖（Polygonum cuspidatum）根系细菌群落组成及其与主要有效成分含量之间的相关性，探讨虎杖根系细菌群落对其药材品质的影响。【方法】收集一、二、三年生虎杖非根际土、根际土和根样品，利用高通量测序对细菌群落组成进行分析，并测定白藜芦醇苷、白藜芦醇、大黄素和大黄素甲醚等有效成分的含量，利用皮尔逊相关性分析探究细菌群落与有效成分含量间的相关性。【结果】生长年限对虎杖有效成分含量的影响显著（P<0.05）。非根际土和根际土细菌群落的优势属为假单胞菌属、鞘氨醇单胞菌属，而根内生细菌优势属为假单胞菌属、Chloroplast、游动放线菌属。根际土中鞘氨醇单胞菌属丰度逐年降低，根内假单胞菌属丰度逐年降低，根内Chloroplast丰度逐年升高。不同生长年限土壤样本和根样本间细菌群落α多样性存在显著差异，非根际土和根际土显著高于根（P<0.05）。非根际土、根际土与根之间细菌群落结构差异极显著（P<0.001），一、二、三年生虎杖间细菌群落结构差异显著（P<0.05）。非根际土与根际土细菌群落结构受生长年限影响显著（P<0.05），而根内生菌结构受生长年限影响不显著（P>0.05）。白藜芦醇苷含量与根际中微枝形杆菌属、Gaiella丰度显著正相关，与根内假单胞菌属丰度呈显著正相关，与根内游动放线菌属、类诺卡氏菌属丰度呈显著负相关；白藜芦醇含量与根内游动放线菌属、类诺卡氏菌属丰度呈显著负相关；大黄素含量与根际中节细菌属丰度呈显著正相关，与根内分枝杆菌属丰度呈显著正相关，与根内假单胞菌属丰度呈显著负相关；大黄素甲醚含量与根际中节细菌属丰度呈显著正相关，与根内分枝杆菌属丰度呈显著正相关，与根内游动放线菌属丰度呈显著负相关。【结论】虎杖根系细菌群落组成与有效成分之间存在关联，为虎杖品质改良提供科学依据。

关键词: 虎杖, 高通量测序, 根系细菌, 群落组成, 有效成分

Abstract:

【Objective】This work aims to explore the effect of root-associated bacterial microbiomes on the quality of Polygonum cuspidatum by analyzing the correlation between the bioactive compounds and microbiomes. 【Method】The bulk soil, rhizosphere soil and roots of P. cuspidatum in the first, second and third years were collected, high-throughput sequencing technology was used to analyze bacterial microbiome compositions, and the contents of bioactive compounds were determined, including polydatin, resveratrol, emodin and physcion. The relationship between the root-associated bacteria and the bioactive components was explored through Pearon correlation analysis.【Result】Bioactive ingredients were distinct across different cultivation years（P < 0.05）. Pseudomonas and Sphingomonas were the most abundant genus of bulk soil and rhizosphere microbiome, while Pseudomonas, Chloroplast and Actinoplanes were the most dominant in the root endophyte. The abundance of Sphingomonas in the rhizosphere soil was decreasing year by year, the abundance of Pseudomonas in the root was decreasing year by year, while the abundance of Chloroplast in root was increasing year by year. The alpha diversity significantly varied along with the soil and root sample with different growth years, the abundance and diversity of bacteria in the bulk soil and rhizosphere were higher than that in root（P < 0.05）. The microbiome composition among the bulk soil, rhizosphere soil and roots extremely significantly differed（P < 0.001）, it also significantly differed across 1-year, 2-year, and 3 -year ones（P < 0.05）. The microbiome composition among the bulk soil and rhizosphere soil was significantly affected by cultivation years（P < 0.05）, while the root endosphere microbiome was not significantly affected by the growth years（P > 0.05）. The content of polydatin was significantly positively correlated with the relative abundance of Microvirga and Gaiella in rhizosphere, significantly positively correlated with the relative abundance of Pseudomonas in the root, and significantly negatively correlated with the relative abundance of Actinoplanes and Nocardioides in the root. The content of resveratrol was significantly negatively correlated with the relative abundance of Actinoplanes and Nocardioides in the root. The content of emodin was significantly positively correlated with the relative abundance of Arthrobacter in the rhizosphere, significantly positively correlated with the relative abundance of Mycobacterium in root, and significantly negatively correlated with the relative abundance of Pseudomonas in root. The content of physcion was significantly positively correlated with the relative abundance of Arthrobacter in the rhizosphere, significantly positively correlated with the relative abundance of Mycobacterium in the root, as well as significantly negatively correlated with the relative abundance of Actinoplanes in the root. 【Conclusion】This study provides insights into the interaction networks between P. cuspidatum root-associated bacteria and bioactive compounds, giving us a new opportunity to manipulate the production of bioactive compounds in future.

Key words: Polygonum cuspidatum, high-throughput sequencing, root-associated bacteria, community composition, active ingredient

杜洁, 黄选怡, 张岩, 姜晴春, 余知和, 王允, 柳忠玉. 虎杖根系细菌群落组成及其与有效成分含量相关性研究[J]. 生物技术通报, 2024, 40(12): 208-217.

DU Jie, HUANG Xuan-yi, ZHANG Yan, JIANG Qing-chun, YU Zhi-he, WANG Yun, LIU Zhong-yu. Composition of Root-associated Bacteria of Polygonum cuspidatum and Their Relationship with the Bioactive Ingredients[J]. Biotechnology Bulletin, 2024, 40(12): 208-217.

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生长年限 Cultivation year	白藜芦醇苷 Polydatin/（mg·g^-1）	白藜芦醇 Resveratrol/（mg·g^-1）	大黄素 Emodin/（mg·g^-1）	大黄素甲醚 Physcion/（mg·g^-1）
一年生（1 Y）	21.623±1.174c	1.753±0.081c	1.886±0.051b	3.785±0.207c
二年生（2 Y）	32.738±1.530b	3.346±0.056b	4.520±0.160a	7.656±0.529a
三年生（3 Y）	48.147±1.416a	3.907±0.050a	2.110±0.038b	5.167±0.308b
F值	92.929***	305.202***	216.261***	49.952***

生长年限 Cultivation year	白藜芦醇苷 Polydatin/（mg·g^-1）	白藜芦醇 Resveratrol/（mg·g^-1）	大黄素 Emodin/（mg·g^-1）	大黄素甲醚 Physcion/（mg·g^-1）
一年生（1 Y）	21.623±1.174c	1.753±0.081c	1.886±0.051b	3.785±0.207c
二年生（2 Y）	32.738±1.530b	3.346±0.056b	4.520±0.160a	7.656±0.529a
三年生（3 Y）	48.147±1.416a	3.907±0.050a	2.110±0.038b	5.167±0.308b
F值	92.929***	305.202***	216.261***	49.952***

样本类型 Sample type	Shannon指数Shannon index	Observed species指数Observed species index
1年生非根际土 1-year old bulk soil（BS1）	5.858 4±2.063 4bc	1 496.93±418cd
1年生根际土 1-year old rhizosphere（RS1）	8.956 8±0.496 2ab	1 753.53±226.34cd
1年生根 1-year old root（R1）	5.372±0.592 9c	809.67±220.22d
2年生非根际土 2-year old bulk soil（BS2）	9.829 7±0.332 7a	3 569.5±608.87a
2年生根际土 2-year old rhizosphere（RS2）	9.550 7±0.169 0a	2 350.17±343.23bc
2年生根 2-year old root（R2）	4.183 7±1.979 5c	983.9±554.14d
3年生非根际土 3-year old bulk soil（BS3）	8.812 3±0.832 6ab	2 083.3±306.84cd
3年生根际土 3-year old rhizosphere（RS3）	10.256±0.140 0a	335 4.6±428.67ab
3年生根3-year old root（R3）	3.104 6±0.677 0c	834.2±67.89d
F值	6.508***	7.054***

样本类型 Sample type	Shannon指数Shannon index	Observed species指数Observed species index
1年生非根际土 1-year old bulk soil（BS1）	5.858 4±2.063 4bc	1 496.93±418cd
1年生根际土 1-year old rhizosphere（RS1）	8.956 8±0.496 2ab	1 753.53±226.34cd
1年生根 1-year old root（R1）	5.372±0.592 9c	809.67±220.22d
2年生非根际土 2-year old bulk soil（BS2）	9.829 7±0.332 7a	3 569.5±608.87a
2年生根际土 2-year old rhizosphere（RS2）	9.550 7±0.169 0a	2 350.17±343.23bc
2年生根 2-year old root（R2）	4.183 7±1.979 5c	983.9±554.14d
3年生非根际土 3-year old bulk soil（BS3）	8.812 3±0.832 6ab	2 083.3±306.84cd
3年生根际土 3-year old rhizosphere（RS3）	10.256±0.140 0a	335 4.6±428.67ab
3年生根3-year old root（R3）	3.104 6±0.677 0c	834.2±67.89d
F值	6.508***	7.054***

部位 Compartment	分组 Group	ADONIS分析 ADONIS analysis		ANOSIM分析ANOSIM analysis
部位 Compartment	分组 Group	R²	P	R	P
非根际土 Bulk soil（BS）	一年生vs. 二年生vs. 三年生 1-year old vs. 2-year old vs.3-year old	0.474*	0.021	0.621*	0.015
根际土 Rhizosphere soil（RS）	一年生vs. 二年生vs. 三年生 1-year old vs. 2-year old vs.3-year old	0.571**	0.005	0.687**	0.005
根Root（R）	一年生vs. 二年生vs. 三年生 1-year old vs. 2-year old vs.3-year old	0.397	0.111	0.251	0.111