Response of endophytes in root nodules of Arachis hypogaea ‘Qicai’ to nitrogen addition and its relationship with plant stoichiometry characteristics

doi:10.17521/cjpe.2024.0002

Abstract

Abstract:

Aims The root nodules of leguminous plants are the site of biological nitrogen fixation by rhizobia, and nodules also harbor diverse endophytes, which play important roles in plant growth and nutrient status. However, how root nodule endophytes respond to nitrogen input and the associations of endophyte composition with plant ecological stoichiometric characteristics remain largely unkown.

Methods We planted Arachis hypogaea‘Qicai’ in a field experiment with three levels of nitrogen treatment (N0: 0 kg·hm^-2, N1: 140 kg·hm^-2, N2: 280 kg·hm^-2). We also quantified the structures of endophyte communities and the contents of carbon (C), nitrogen (N) and phosphorus (P) in leaves, then the endophytic functions were predicted by PICRUST 2 software.

Important findings The results showed that endophytes are extremely diverse, with a total of 546 families and 1 049 genera. Among them, Bradyrhizobium accounted for 27.83% in relative abundance, while other genera accounted for more than 70%. The average proportion of Burkkholderia-Caballeronia-Paraburkholderia and Enterobacter was more than 10%. Nitrogen application decreased the diversity of endophytes and shifted the community composition, and the relative abundance of microorganisms from Bradyrhizobium, Enterobacter, Kosakonia and Pantoea increased significantly, while those from Allorhizobium-Neorhizobium-Pararhizobium- Rhizobium, Mesorhizobium, Mycobacterium and Burkholderia-Caballeronia-Paraburkholderia were significantly decreased. The relative abundance of part of genera was closely correlated with the stoichiometric characteristics of the plants. The results of co-occurrence network analysis show that some modules are significantly correlated with the contents and stoichiometric characteristics of C, N, and P in the leaves. Meanwhile, the results of PICRUSt 2 functional prediction show that the functions of microbial C, N, and P metabolic enzymes are significantly correlated with the contents and stoichiometric characteristics of C, N, and P in leaves, indicating that there is a close association of the microbial community structure with the contents and balance of plant nutrients. In summary, our study shows that the composition of endophytes in the root nodules of the Arachis hypogaea‘Qicai’ is extremely diverse, and the community structures and functions are sensitive to nitrogen addition. In future studies, the functions of nitrogen-tolerant and sensitive endophytes can be further explored through microbial isolation and synthetic microbial community techniques.

Key words: Arachis hypogaea ‘Qicai’, nitrogen fertilization, root nodule endophyte, stoichiometric characteristics, PICRUSt 2

LI Lin, SUN Yi, YANG Xiao-Qiong, FANG Hai-Dong, YAN Bang-Guo. Response of endophytes in root nodules of Arachis hypogaea ‘Qicai’ to nitrogen addition and its relationship with plant stoichiometry characteristics[J]. Chin J Plant Ecol, 2024, 48(10): 1374-1384.

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Figures/Tables 5

Fig. 1 Changes of carbon (C), nitrogen (N) and phosphorus (P) contents and stoichiometric characteristics of Arachis hypogaea ‘Qicai’ leaves under different nitrogen addition treatments. The same lowercase letters indicate no significant differences between treatments (p > 0.05). N0, 0 kg·hm-2; N1, 140 kg·hm-2; N2, 280 kg·hm-2.

Fig. 2 Diversity (A), changes in the relative abundance of dominant genera (B), principal coordination analysis (PCoA) (C) and volcano plot illustrating changes in the relative abundance (D) of endophytes in root nodules under different nitrogen addition treatments of Arachis hypogaea ‘Qicai’. Different lowercase letters in A indicate significant differences among treatments (p < 0.05). Each point represents an amplicon sequence variant (ASV) in D. Each red point indicate an enriched ASV while each blue point represents a depleted ASV. N0, 0 kg·hm-2; N1, 140 kg·hm-2; N2, 280 kg·hm-2. *, p < 0.05.

Table 1 Relationship between relative abundance of Arachis hypogaea ‘Qicai’ root nodule endophytes and leaf carbon (C), nitrogen (N) and phosphorus (P) stoichiometric characteristics

相关性系数 Correlation coeffficient	C	N	P	C:N	C:P	N:P
根瘤菌属 Allorhizobium-Neorhizobium- Pararhizobium-Rhizobium	0.00	-0.35^*	-0.11	0.37^**	0.13	-0.12
慢生根瘤菌属 Bradyrhizobium	0.01	0.17	0.27	-0.19	-0.23	-0.08
伯克霍尔德菌属 Burkholderia-Caballeronia- Paraburkholderia	0.37^**	0.16	-0.03	-0.08	0.06	0.10
肠杆菌属 Enterobacter	-0.22	0.18	-0.05	-0.23	0.02	0.18
科萨克氏菌属 Kosakonia	-0.27	-0.09	-0.04	0.03	-0.01	-0.03
分枝杆菌属 Mycobacterium	0.03	-0.16	0.13	0.18	-0.11	-0.20
新鞘氨醇菌属 Novosphingobium	0.18	-0.21	-0.04	0.24	0.09	-0.09
泛菌属 Pantoea	0.08	-0.02	-0.04	0.04	0.04	0.01
糖单孢菌 Saccharimonadales	-0.04	-0.21	0.03	0.17	-0.03	-0.15

Fig. 3 Response of root nodule endophyte enzyme functions (predicted by PICRUSt 2) to nitrogen addition treatments and their association with leaf stoichiometric characteristics. A, Changes in enzyme activities (mean ± SE). B, Principal component analysis (PCoA) of enzyme activity. C, Relationships between enzyme activities and leaf stoichiometric characteristics. N0, 0 kg·hm-2; N1, 140 kg·hm-2; N2, 280 kg·hm-2. C, carbon; N, nitrogen; P, phosphorus。*, p ≤ 0.05; **, p ≤ 0.01. Different lowercase letters in A indicate significant differences among treatments (p < 0.05).

Fig. 4 Co-occurrence network analysis of endophytic communities in root nodules of Arachis hypogaea ‘Qicai’. A, Cooccurrence network of root nodule endophytes. B, Associations between the relative abundance of different clusters with leaf stoichiometric characteristics. C, Main bacterial genus composition of each module. N0, 0 kg·hm-2; N1, 140 kg·hm-2; N2, 280 kg·hm-2. C, carbon; N, nitrogen; P, phosphorus. *, p ≤ 0.05; **, p ≤ 0.01.

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