黄龙病发病芦柑根际土壤细菌群落组成与多样性特征

doi:10.13560/j.cnki.biotech.bull.1985.2023-0837

摘要/Abstract

摘要：

【目的】本研究旨在研究福建南平地区芦柑（Citrus reticulata）植株根际土壤细菌群落，并分析黄龙病对其细菌群落的影响，为南平地区芦柑黄龙病的防控提供科学依据。【方法】采用五点法采集了健康和黄龙病发病的芦柑植株根际土壤样本，通过扩增子测序技术和生物信息学方法，深入研究了黄龙病发病芦柑根际土壤细菌群落的多样性和组成变化，分析了土壤理化因子与细菌群落的关联性。【结果】研究结果表明，黄龙病发病的芦柑植株根际土壤细菌群落的多样性略高于健康植株。黄龙病发病芦柑根际土壤中的变形菌门相对丰度明显升高，而放线菌门相对丰度显著降低。在属水平上，与健康植株相比，黄龙病发病芦柑植株根际土壤细菌群落中蔷薇属和嗜酸性杆菌属的相对丰度显著增加，与速效钾、有效磷以及有机质呈现显著负相关，而与pH值呈现显著正相关。相反地，康奈斯氏杆菌属和褚氏杆菌属的相对丰度明显低于健康植株，并与速效钾、有效磷以及有机质呈现显著正相关，同时与pH值呈现显著负相关。【结论】黄龙病菌感染改变了柑橘土壤理化特性，并显著减少根际土壤中有益细菌的相对丰度，从而导致了芦柑植株根际细菌群落的多样性和组成出现显著变化。与此同时，黄龙病发病的芦柑植株可能通过招募固氮和促进植物根际生长的有益细菌来对抗病原菌的侵害。

关键词: 黄龙病, 柑橘, 高通量测序, 根际土壤, 细菌群落, 差异物种, 环境因子

Abstract:

【Objective】 This study aims to investigate the rhizosphere soil bacterial community of Citrus reticulata plants in the Nanping region of Fujian province, China. The objective is to analyze the impact of Huanglongbing（HLB）disease on the bacterial community and provide a scientific basis for HLB management strategies in the Nanping region.【Method】 We collected root-zone soil samples from both healthy and HLB-affected C. reticulata plants using the five-point sampling method. We used amplicon sequencing technology and bioinformatics methods to investigate the changes in diversity and composition of the bacterial community in the root-zone soil of HLB-affected C. reticulata plants. Additionally, we analyzed the correlation between soil physicochemical factors and the bacterial community.【Result】 The findings of the study demonstrate that the diversity of the bacterial community in the root-zone soil of C. reticulata plants affected by HLB was slightly higher when compared to the microbiota of healthy plants. Specifically, there was a significant increase in the relative abundance of the Proteobacteria, and a significant decrease in the relative abundance of the Actinobacteriota in the root-zone soil of HLB-affected C. reticulata plants. At the genus level, it was observed that the relative abundance of the Roseiarcus and Acidiphilium significantly increased in the bacterial community of HLB-affected C. reticulata plants. Furthermore, these genera presented a significant negative correlation with available potassium, available phosphorus, and organic matter, while showing a significantly positive correlation with pH values. Conversely, the relative abundance of the Conexibacter and Chujaibacter were significantly lower in HLB-affected plants than healthy ones. Additionally, these genera demonstrated a significantly positive correlation with available potassium, available phosphorus, and organic matter, but a significant negative correlation with pH values.【Conclusion】 The infection of the HLB pathogen altered the physicochemical properties of citrus soil and significantly reduced the abundance of beneficial bacteria in the rhizosphere soil. This, in turn, leads to considerable changes in both the diversity and composition of the rhizosphere bacterial community in C. reticulata plants. Concurrently, HLB-infected C. reticulata plants appear to counteract pathogen invasion by recruiting beneficial bacteria that not only fix nitrogen but also promote plant growth in the rhizosphere.

Key words: Huanglongbing, citrus, high-throughput sequencing, rhizosphere soil, bacterial community, differential species, environmental factors

雷美玲, 饶文华, 胡进锋, 岳琪, 吴祖建, 范国成. 黄龙病发病芦柑根际土壤细菌群落组成与多样性特征[J]. 生物技术通报, 2024, 40(2): 266-276.

LEI Mei-ling, RAO Wen-hua, HU Jin-feng, YUE Qi, WU Zu-jian, FAN Guo-cheng. Bacterial Diversity and Structure in Rhizosphere Soil of Citrus Infested with Huanglongbing[J]. Biotechnology Bulletin, 2024, 40(2): 266-276.

图/表 9

表1 柑橘黄龙病病原菌qPCR检测结果

Table 1 qPCR detection of citrus HLB pathogen

分组Group	发病组HLB group					健康组Healthy group
分组Group	1	2	3	4		1	2	3	4
黄龙病菌滴度值CLas Ct	26.62±0.20	23.03±0.20	20.56±0.05	22.56±0.25		None	None	None	None

图1 田间芦柑植株和叶片 A：黄龙病发病芦柑植株；B：健康芦柑植株；C：黄龙病发病芦柑植株叶片；D：健康芦柑植株叶片

Fig. 1 Plant and leaf of C. reticulata in field A: Citrus reticulata with Huanglongbing（HLB）. B: Healthy C. reticulata.C: Huanglongbing leaf of C. reticulata with HLB. D: Healthy C. reticulata leaf

图2 健康组与发病组在OTU水平的Alpha多样性指数分析 Healthy：健康芦柑根际土壤样品组；HLB：黄龙病发病芦柑根际土壤样品组。下同

Fig. 2 Alpha diversity index of healthy group and HLB group at the OTU level Healthy: Rhizosphere soil sample group of healthy C. reticulata; HLB: rhizosphere soil sample group of C. reticulata with HLB. The same below

图3 健康组与发病组在OTU水平的非度量多维尺度分析

Fig. 3 NMDS analysis of healthy group and HLB group at the OTU level

图4 发病组与健康组样品在门、科和属水平上共有和独有的物种情况图

Fig. 4 Common and unique species of HLB group and healthy group at the phylum, family and genus level

图5 健康组（A）与发病组（B）细菌群落丰度在门水平上的相对丰度比较

Fig. 5 Comparison of bacterial community abundance at the phylum level between healthy group（A）and HLB group（B）

表2 健康组与发病组柑橘根际部分细菌群在属水平上的相对丰度比较

Table 2 Comparative study on the relative abundance of rhizosphere part of the bacteria between healthy group and HLB group at the genus level

属Genus	相对丰度的差异Abundance difference	健康组的相对丰度Healthy group abundance	发病组的相对丰度HLB group abundance
芽孢杆菌属Bacillus	-0.476	2.264	1.788
假单胞菌属 Pseudomonas	-0.123	0.150	0.027
慢生根瘤菌属Bradyrhizobium	-0.126	1.458	1.332
伯克霍尔德菌属Bur- kholderia-Caballero- nia-Paraburkholderi	-0.174	0.557	0.383
鞘氨醇单胞菌属Sphingomonas	-0.207	0.298	0.091
中根瘤菌属 Mesorhizobium	-0.004	0.017	0.013
铜菌属Cupriavidus	0.002	4.386×10^-3	6.280×10^-3
细胞弧菌属Cellvibrio	0.002	1.595×10^-3	3.589×10^-3
变异菌属Variovorax	0.047	0.027	0.074

图6 健康组与发病组细菌群落在门（A）、属（B）水平相对丰度显著差异分析 X轴表示发病组和健康组，Y轴表示发病组和健康组中物种的平均相对丰度。*：P<0.05，**：P<0.01，***：P<0.001，下同

Fig. 6 Significant difference in relative abundance of bacteria at the phylum（A）and genus（B）level between healthy and HLB groups X axis represents HLB group and healthy group, and Y axis represents the average relative abundance of a species in HLB group and healthy group. *: P < 0.05, **: P < 0.01, ***: P < 0.001. The same below

图7 属水平健康与黄龙病发病芦柑植株根际土壤细菌群落和环境因子之间的相关性 X轴和Y轴分别为环境因子和物种，通过计算获得相关性R值和P值。R值在图中以不同颜色展示，右侧图例是不同R值的颜色区间

Fig. 7 Correlation between bacterial community and environmental factors in the rhizosphere soil of healthy and HLB-infected citrus plants at the genus level The X-axis and Y-axis are environmental factors and species, respectively. The R values are shown in different colors, and the right legend is a color range of different R values

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