外源添加L-脯氨酸对棉花黄萎病发生及其根际土壤微生物群落的影响

doi:10.3864/j.issn.0578-1752.2024.11.008

Abstract

Abstract:

【Objective】Root exudates are signal mediators for the interaction between plants and soil microorganisms, which have important regulatory functions for plant disease occurrence and plant growth. The objective of this study is to clarify the microecological mechanism of L-proline in root exudates of cotton against the occurrence of Verticillium wilt, reveal the relationship of L-proline-mediated interaction between rhizosphere microorganisms and the occurrence of cotton Verticillium wilt, and to provide a new perspective for the construction of beneficial bacterial communities for the biological control of soil-borne diseases.【Method】Through greenhouse pot experiments, different concentrations of L-proline (0, 50, 100, 200, and 400 mmol·L^-1) were set as experimental treatments, and real-time fluorescence quantitative PCR and metagenomic sequencing techniques were used to determine the DNA copy numbers of Verticillium dahliae and microbial community structure and function of soil samples treated with L-proline. The microbial community structures of rhizosphere soils under different concentrations of L-proline were compared using principal component analysis. Redundancy analysis was used to study the correlations between soil nutrient factors and microbial community structure, and Spearman correlation analysis was focused on the relationships between microbial community structure and functional metabolic pathways.【Result】Compared with the blank control, the occurrence of cotton Verticillium wilt could not reduce under low concentration (50 mmol·L^-1) L-proline treatment, while the disease indexes under high-concentration (100, 200, and 400 mmol·L^-1) L-proline treatments were decreased by 22.51%, 60.23%, and 64.23%, respectively. The results of qPCR showed that L-proline treatment did not significantly decrease the copy numbers of V. dahliae in soil. The metagenomic sequencing analysis showed that the Shannon index of bacterial diversity significantly increased after L-proline treatment, while the Shannon index of fungal diversity showed a downward trend. At the genus level, the relative abundances of Nocardioides, Lysobacter, Arthrobacter, Phycicoccus, Pseudomonas, and Mucor were increased after treatment with L-proline. Linear discriminant analysis (LDA) showed that the enrichment of microbial KEGG pathways in rhizosphere soil changed after exogenous addition of L-proline, except for L-proline at the concentration of 100 mmol·L^-1. Redundancy analysis showed that bacterial community composition was significantly influenced by pH, conductivity, nitrate nitrogen, ammonium nitrogen, and organic matter (OM), while fungal community composition was significantly correlated with ammonium nitrogen. Spearman correlation analysis showed that the bacterial KEGG pathways were negatively correlated with pH, OM and ammonium nitrogen, while positively correlated with conductivity and nitrate nitrogen. Most of the KEGG pathways of fungi had poor correlation with soil nutrients.【Conclusion】The exogenous addition of an appropriate amount of L-proline can influence the occurrence of cotton Verticillium wilt through altering the structure and function of soil bacterial communities, increasing the relative abundance of beneficial microorganisms, but it cannot change the DNA copy numbers of pathogen. Meanwhile, the composition and function of bacterial community are related to soil nutrients.

Key words: L-proline, rhizospheric microorganism, cotton Verticillium wilt, Verticillium dahliae, soil nutrient

ZHAO WeiSong, GUO QingGang, CUI NaQi, LU XiuYun, LI SheZeng, MA Ping. Effects of Exogenous Addition of L-Proline on the Occurrence of Cotton Verticillium Wilt and Its Soil Microbial Community in Rhizosphere[J].Scientia Agricultura Sinica, 2024, 57(11): 2143-2160.

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处理 Treatment	指标Index
处理 Treatment	株高 Plant height (cm)	地径 Ground diameter (cm)	地上鲜重 Shoot fresh weight (g)	地上干重 Shoot dry weight (g)	地下鲜重 Root fresh weight (g)	地下干重 Root dry weight (g)
CK	53.48±0.99a	3.04±0.21a	8.73±0.73a	3.12±0.34a	1.02±0.15a	0.40±0.06a
L50	56.73±6.14ab	3.01±0.20a	10.03±1.99a	2.82±0.23a	0.90±0.23a	0.52±0.29a
L100	63.34±6.82b	2.97±0.17a	9.79±0.63a	2.83±0.29a	1.05±0.45a	0.39±0.21a
L200	59.85±6.80ab	3.04±0.21a	10.35±1.57a	2.94±0.61a	0.82±0.26a	0.35±0.09a
L400	55.53±5.08ab	2.80±0.09a	9.73±0.95a	2.83±0.22a	0.85±0.20a	0.26±0.02b

指标 Index	处理Treatment
指标 Index	CK	L50	L100	L200	L400
速效钾含量AK content (mg·kg^-1)	157.40±1.27a	155.99±5.75a	154.45±2.45a	157.96±6.58a	140.38±3.19b
速效磷含量AP content (μmol·g^-1)	1.39±0.12a	1.44±0.02a	1.58±0.11a	1.44±0.15a	1.40±0.22a
铵态氮含量NH₄⁺-N content (mg·kg^-1)	23.85±2.79a	21.33±1.98a	24.26±0.58a	18.17±0.35b	17.27±2.33b
硝态氮含量NO₃^--N content (μg·g^-1)	11.87±2.86b	4.06±0.32d	6.75±0.79c	12.80±2.55b	375.76±7.31a
有机质含量OM content (%)	4.66±0.02b	4.71±0.11b	4.98±0.07a	4.93±0.04a	4.99±0.03a
pH	7.89±0.01ab	7.90±0.02a	7.93±0.01a	7.85±0.04b	7.75±0.02c
电导率Conductivity (μm·cm^-1)	215.67±31.94b	197.37±4.38b	191.20±3.08b	212.67±3.06b	316.00±1.00a

处理 Treatment	细菌Bacteria		真菌Fungi
处理 Treatment	Shannon	ACE	Shannon	ACE
CK	4.57±0.01c	2806.70±16.17a	4.60±0.11a	388.67±12.67a
L50	4.59±0.01ab	2791.00±17.78a	4.55±0.04a	367.00±7.00a
L100	4.60±0.01a	2808.00±14.53a	4.52±0.10a	382.50±9.19a
L200	4.60±0.01a	2796.00±13.53a	4.46±0.11ab	369.00±54.56a
L400	4.59±0.01ab	2794.00±8.66a	4.32±0.11b	366.00±7.55a

微生物 Microorganism	指标 Index	RDA1	RDA2	决定系数 Coefficients of determination (R²)	P值 P value
细菌Bacteria	pH	0.9613	-0.2756	0.7921	0.001
	电导率Conductivity	-0.9959	0.0907	0.7810	0.004
	硝态氮含量NO₃^--N content	-0.9977	0.0685	0.8673	0.005
	铵态氮含量NH₄⁺-N content	0.8755	-0.4833	0.7556	0.001
	速效磷含量AP content	0.6975	-0.7166	0.0443	0.760
	速效钾含量AK content	0.9999	0.0116	0.6554	0.002
	有机质含量OM content	0.3713	-0.9285	0.5679	0.005
真菌Fungi	pH	-0.8198	-0.5723	0.0988	0.532
	电导率Conductivity	0.4810	0.8767	0.0553	0.729
	硝态氮含量NO₃^--N content	0.3452	0.9385	0.0345	0.827
	铵态氮含量NH₄⁺-N content	-0.8095	-0.5872	0.5188	0.015
	速效磷含量AP content	-0.6515	0.7587	0.0989	0.543
	速效钾含量AK content	-0.5847	-0.8113	0.0306	0.850
	有机质含量OM content	-0.9769	0.2136	0.3668	0.064

Effects of Exogenous Addition of L-Proline on the Occurrence of Cotton Verticillium Wilt and Its Soil Microbial Community in Rhizosphere

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