盐胁迫下氮素对花生种子萌发和种子际细菌菌群结构的调控

doi:10.13560/j.cnki.biotech.bull.1985.2023-0753

摘要/Abstract

摘要：

【目的】盐胁迫影响花生种子萌发和植株生长，阐明盐胁迫下适量施肥提高种子萌发率和花生产量的内在调控机制，并解析该过程与种子际土壤细菌菌群结构的关系，为通过改良种子际土壤微生物环境，提高花生出苗健苗率、耐盐抗逆性和花生生产能力提供理论和技术依据。【方法】以耐盐花生品种（花育25号，HY25）为试验材料，设置3个氮素水平0、90和180 kg/hm²，采用盆栽试验和高通量测序技术，阐明氮肥施用对盐胁迫下花生种子际微生物菌群结构、花生发芽出苗和产量的影响。【结果】施加氮肥可有效提高花生种子在盐胁迫下的发芽率和最终产量，并以施氮量90 kg/hm²最适。16S rRNA测序分析显示，种子际的土壤细菌以变形菌门（Proteobacteria）、放线菌门（Actinobacteria）、厚壁菌门（Firmicutes）、酸杆菌门（Acidobacteria）、拟杆菌门（Bacteroidetes）、绿弯菌门（Chloroflexi）及芽孢杆菌门（Gemmatimonadetes）等为优势菌门。在属水平上，盐胁迫虽然提高了有益菌属拟杆菌属（Bacteroides）的相对丰度，但同时导致有害的链球菌属（Streptococcus）增多，最终降低了有益菌属芽孢杆菌属（Bacillus）、鞘脂单胞菌属（Sphingomonas）和溶杆菌属（Lysobacter）的相对丰度。盐胁迫下施氮可以显著改善种子际的土壤微环境，提高有益菌属拟杆菌属、芽孢杆菌属、鞘脂单胞菌属的相对丰度，对土壤修复和地力提升有一定的帮助，同时还能增强花生抗逆性。【结论】盐胁迫下适量施氮可提高种子际有益菌属的相对丰度，从而提高花生种子的发芽率和耐盐性，最终促进盐胁迫下的花生增产。

关键词: 花生, 盐胁迫, 发芽率, 产量, 种子际, 细菌菌群结构

Abstract:

【Objective】 Salt stress affects seed germination and peanut growth, and the internal regulatory mechanism of increasing seed germination rate and peanut yield by appropriate fertilization under salt stress was elucidated, and the relationship between the process and the bacterial structure of peanut spermosphere soil was analyzed, so as to provide theoretical and technical basis for improving peanut emergence and health rate, salt tolerance and stress resistance and peanut production capacity by regulating the microenvironment in spermosphere soil.【Method】 We set up three nitrogen levels of 0, 90, and 180 kg/hm² with a salt-tolerant peanut variety（Huayu 25, HY25）as experimental materials. Potted experiments and high-throughput sequencing techniques were used to study the effects of nitrogen fertilizer application on the spermosphere bacterial community structure, germination and yield of peanut under salt stress.【Result】 The application of nitrogen fertilizer increased the germination rate and peanut yield, and the optimal nitrogen application was 90 kg/hm². Proteobacteria, Actinobacteria, Firmicutes, Acidobacteria, Bacteroidetes, Chloroflexi, and Gemmatimonadetes were the dominant phyla in peanut spermosphere soil of different groups via 16S rRNA sequence. At the genus level, salt stress increased the relative abundance of beneficial bacteria Bacteroides, but induced a large number of harmful Streptococcus, whereas the relative abundance of the beneficial bacteria genera Bacillus, Sphingomonas, and Lysobacter decreased. Nitrogen application under salt stress may improve the spermosphere soil microenvironment and increase the relative abundance of beneficial soil bacteria Bacteroides, Bacillus, and Sphingomonas, which is helpful to soil restoration and fertility improvement, and can also enhance the stress resistance of peanuts.【Conclusion】 Appropriate nitrogen application under salt stress increases the relative abundances of beneficial bacteria in the spermosphere soil, thus which may improve the germination rate and salt tolerance, and finally increase the peanut yield under salt stress.

Key words: peanut（Arachis hypogaea L.）, salt stress, germination rate, yield, spermosphere, bacterial community structure

徐扬, 张瑞英, 戴良香, 张冠初, 丁红, 张智猛. 盐胁迫下氮素对花生种子萌发和种子际细菌菌群结构的调控[J]. 生物技术通报, 2024, 40(2): 253-265.

XU Yang, ZHANG Rui-ying, DAI Liang-xiang, ZHANG Guan-chu, DING Hong, ZHANG Zhi-meng. Regulation of Nitrogen Application on Peanut Seed Germination and Spermosphere Bacterial Community Structure Under Salt Stress[J]. Biotechnology Bulletin, 2024, 40(2): 253-265.

图/表 9

图1 盐胁迫下不同氮肥施用对花生萌发的影响 CN：正常条件无氮处理；CM：正常条件施氮90 kg/hm2；CH：正常条件施氮180 kg/hm2；SN：盐胁迫下无氮处理；SM：盐胁迫下施氮90 kg/hm2；SH：盐胁迫下施氮180 kg/hm2。下同

Fig. 1 Effects of different nitrogen fertilizer applications on peanut germination under salt stress CN: No N-applied under normal condition; CM: 90 kg/hm2 N-applied under normal condition; CH: 180 kg/hm2 N-applied under normal condition; SN: no N-applied under salt stress; SM: 90 kg/hm2 N-applied under salt stress; SH: 180 kg/hm2 N-applied under salt stress. The same below

表1 不同处理条件下花生产量构成因素统计

Table 1 Statistics of yield components of peanut under different treatments

处理 Treatment	编号缩写 Abbreviation	单株荚果数 Number of pods per plant	百果重 100 pods weight/g	百仁重 100 seeds weight/g	出米率 Kernel percentage/%
正常 Normal condition	CN	20.84±0.30b	190.21±0.42b	77.13±0.45b	68.19±0.77b
	CM	22.87±0.54a	206.22±0.66a	82.97±0.67a	72.63±0.36a
	CH	21.95±0.054a	207.05±0.42a	84.05±0.59a	73.53±0.34a
盐胁迫 Salt stress	SN	17.08±0.19c	169.53±1.07d	69.28±0.85d	64.16±0.13c
	SM	17.94±0.19c	184.90±1.40c	75.42±0.65c	68.67±0.56b
	SH	17.48±0.12c	185.74±0.85c	75.44±0.63c	68.93±0.27b

图2 α多样性分析 A：稀释性曲线；B：等级丰度曲线；C：物种累计曲线。CK：空白土壤；CNS：正常条件无氮处理种子际土壤；CMS：正常条件施氮90 kg/hm2种子际土壤；CHS：正常条件施氮180 kg/hm2种子际土壤；SNS：盐胁迫下无氮处理种子际土壤；SMS：盐胁迫下施氮90 kg/hm2种子际土壤；SHS：盐胁迫下施氮180 kg/hm2种子际土壤

Fig. 2 α diversity analysis A: Rarefaction curve. B: Rank abundance curve. C: Species accumulation curves. CK: Bulk soil; CNS: no N-applied spermosphere soil under normal condition; CMS: 90 kg/hm2 N-applied spermosphere soil under normal condition; CHS: 180 kg/hm2 N-applied spermosphere soil under normal condition; SNS: no N-applied spermosphere soil under salt stress; SMS: 90 kg/hm2 N-applied spermosphere soil under salt stress; SHS: 180 kg/hm2 N-applied spermosphere soil under salt stress

图3 β多样性分析 A：主坐标分析；B：样本层次聚类分析；C：ANOIMS相似性分析

Fig. 3 β diversity analysis A: Principal component analysis. B: Hierarchical clustering tree analysis. C: ANOIMS analysis

图4 物种组成及样本间差异比较 A：门水平物种组成百分比统计；B：纲水平物种组成百分比统计；C：目水平物种组成百分比统计；D：科水平物种组成百分比统计

Fig. 4 Comparison of species composition and differences between diverse groups A: Percentage of species composition at the phylum level. B: Percentage of species composition at the class level. C: Percentage of species composition at the order level. D: Percentage of species composition at the family level

图5 属水平物种组成及样本间差异比较 A：属水平物种组成百分比统计；B：热图分析

Fig. 5 Comparison of bacterial composition and differences between diverse groups at the genus level A: Percentage of bacterial composition at the genus level; B: heatmap analysis

图6 Lefse多级物种层级树图

Fig. 6 Lefse hierarchical tree of species

图7 花生种子际细菌群落代谢功能KEGG分析

Fig. 7 KEGG analysis of metabolic functional features of the peanut spermosphere bacterial community

图8 RDA分析 A：门水平细菌群落与环境因子RDA分析；B：属水平细菌群落与环境因子RDA分析

Fig. 8 Redundancy analysis A: RDA analysis of bacterial communities and environmental factors at phylum level. B: RDA analysis of bacterial communities and environmental factors at genus level

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