Bacterial Diversity and Structure in Rhizosphere Soil of Citrus Infested with Huanglongbing

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

Abstract

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

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.

Figures/Tables 9

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

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

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

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

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

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

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

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

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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