Effects of Rhizobium Seed Dressing on the Soil Microbial Community of Grass-legume Mixtures in Alpine Regions

doi:10.13560/j.cnki.biotech.bull.1985.2021-1066

Abstract

Abstract:

Rhizobium seed dressing is one of the effective measures to improve the nitrogen fixation efficiency of plants. Our study investigated the effects of rhizobium seed dressing on the structure and diversity of soil microbial community in typical grass-legume mixtures in alpine regions，aiming to provide a theoretical basis for the application in forage production. Taking the mixed sowing of Medicago sativa cv.‘Beilin201’，Elymus sibiricus L.cv.‘Chuancao No.2’，and Elynus nutans Griseb.cv.‘Aba’ as the research object，2 treatments of rhizobium seed dressing and no seed dressing were set. The bulk and rhizosphere soil were collected，and the prokaryotic biodiversity and community structure were analyzed based on 16S rRNA gene high-throughput sequencing technology，the functional prediction was calculated by FAPROTAX tools and the classification and changes of prokaryotes microbial high-level phenotypes were compared by BugBase analysis. A total of 8 814 OTUs were detected from 32 soil samples，belonging to 3 577 genera in 6 phyla，while the dominant phyla were Actinobacteria（26.11%-46.20%）and Proteobacteria（20.61%-32.91%）. Rhizobium seed dressing changed prokaryotic community structure and decreased diversity（Observed_richness，Shannon index，and inverse Simpson index），but increased the abundance of digestion and denitrification taxa. The population belonging to aerobic，anaerobic，facultative anaerobic，gram-negative，and gram-positive prokaryotes in the rhizosphere soil of Gramineae increased after seed dressing，while the results of Legumes were the opposite. After seed dressing，the abundance of nifH gene in the legume rhizosphere significantly increased. In summary，seed dressing reduced the α-diversity of community，changed the structural composition，but increased the relative abundances of digesting and denitrifying populations，and increased the abundance of nifH genes. This study provides a scientific basis for the cultivation of rhizobia seed dressing in the grass-legume mixtures.

Key words: rhizobium with seed dressing, nifH gene, microbial diversity, structure of microbial community, function prediction

YAN Hui-lin, LU Guang-xin, DENG Ye, GU Song-song, YAN Cheng-liang, MA Kun, ZHAO Yang-an, ZHANG Hai-juan, WANG Ying-cheng, ZHOU Xue-li, DOU Sheng-yun. Effects of Rhizobium Seed Dressing on the Soil Microbial Community of Grass-legume Mixtures in Alpine Regions[J]. Biotechnology Bulletin, 2022, 38(10): 204-215.

Figures/Tables 11

References 31

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牧草种类 Forage species	混播草地 Mixed pasture	重复数 Repetition	标记 Remark
‘川草2号’老芒麦（Elymus sibiricus L.cv.‘Chuancao No.2’）	苜蓿‘北林201’（拌种） Medicago sativa cv.‘Beilin201’（Rhizobium）	3	A1
‘川草2号’老芒麦（Elymus sibiricus L.cv.‘Chuancao No.2’）	苜蓿‘北林201’Medicago sativa cv.‘Beilin201’	3	D1
‘阿坝’垂穗披碱草（Elynus nutans Griseb.cv.‘Aba’）	苜蓿‘北林201’（拌种）Medicago sativa cv.‘Beilin201’（Rhizobium）	3	A2
‘阿坝’垂穗披碱草（Elynus nutans Griseb.cv.‘Aba’）	苜蓿‘北林201’Medicago sativa cv.‘Beilin201’	3	D2

牧草种类 Forage species	混播草地 Mixed pasture	重复数 Repetition	标记 Remark
‘川草2号’老芒麦（Elymus sibiricus L.cv.‘Chuancao No.2’）	苜蓿‘北林201’（拌种） Medicago sativa cv.‘Beilin201’（Rhizobium）	3	A1
‘川草2号’老芒麦（Elymus sibiricus L.cv.‘Chuancao No.2’）	苜蓿‘北林201’Medicago sativa cv.‘Beilin201’	3	D1
‘阿坝’垂穗披碱草（Elynus nutans Griseb.cv.‘Aba’）	苜蓿‘北林201’（拌种）Medicago sativa cv.‘Beilin201’（Rhizobium）	3	A2
‘阿坝’垂穗披碱草（Elynus nutans Griseb.cv.‘Aba’）	苜蓿‘北林201’Medicago sativa cv.‘Beilin201’	3	D2

环境因子Environmental factors	变量Variables	分组Group
环境因子Environmental factors	变量Variables	DB	DN	HB	HN
理化性质 Soil physical-chemical property	总氮TN/%	1 805.37±293.06	1 814.36±152.19	1 676.88±248.44	1 777.86±137.70
	铵态氮NH₄^-N /（mg·kg^-1）	4.41±1.11	4.22±0.54	4.52±0.94	4.30±1.00
	硝态氮NO₃^-N /（mg·kg^-1）	38.87±2.85	39.97±1.70	41.84±4.35	40.06±2.08
	有机碳TOC/%	2.29±0.30	2.24±0.29	2.23±0.31	2.17±0.28
土壤性质 Soil property	温度MAT/℃	13.49±3.54	11.88±0.18	13.98±4.36	11.87±0.20
	水分SMC/%	9.81±1.06	10.63±1.31	10.63±1.23	10.70±1.45
	电导率EC/（us·cm^-1）	0.06±0.04	0.04±0.02	0.07±0.0.1	0.04±0.02

环境因子Environmental factors	变量Variables	分组Group
环境因子Environmental factors	变量Variables	DB	DN	HB	HN
理化性质 Soil physical-chemical property	总氮TN/%	1 805.37±293.06	1 814.36±152.19	1 676.88±248.44	1 777.86±137.70
	铵态氮NH₄^-N /（mg·kg^-1）	4.41±1.11	4.22±0.54	4.52±0.94	4.30±1.00
	硝态氮NO₃^-N /（mg·kg^-1）	38.87±2.85	39.97±1.70	41.84±4.35	40.06±2.08
	有机碳TOC/%	2.29±0.30	2.24±0.29	2.23±0.31	2.17±0.28
土壤性质 Soil property	温度MAT/℃	13.49±3.54	11.88±0.18	13.98±4.36	11.87±0.20
	水分SMC/%	9.81±1.06	10.63±1.31	10.63±1.23	10.70±1.45
	电导率EC/（us·cm^-1）	0.06±0.04	0.04±0.02	0.07±0.0.1	0.04±0.02

整体比较Global test			两两比较Pairwise composition
整体比较Global test			B	N	HB	HN	DB	DN
MRPP	0.415 8	B		0.6	0.048	0.011	0.006	0.025
δ（P-value）	0.001	N	0.291 6		0.004	0.002	0.004	0.008
		HB	0.462 1	0.461 0		0.75	0.504	0.946
		HN	0.359 8	0.317 9	0.513 3		0.049	0.232
		DB	0.362 1	0.320 3	0.516 1	0.392 8		0.368
		DN	0.430 4	0.392 0	0.582 7	0.469 4	0.471 7
ANOSIM	0.423 6	B		0.73	0.011	0.012	0.003	0.03
R（P-value）	0.001	N	-0.055		0.008	0.002	0.001	0.009
		HB	0.488 0	0.678 5		0.284	0.185	0.606
		HN	0.586 6	0.853 3	0.068 7		0.048	0.373
		DB	0.936	0.997 3	0.131 2	0.384		0.48
		DN	0.340 7	0.485 1	-0.06 7	0.018 6	-0.026
PERMANOVA	4.100 5	B		0.504	0.049	0.009	0.011	0.023
F（P-value）	0.001	N	0.962 6		0.016	0.001	0.002	0.008
		HB	3.041 3	4.806 7		0.861	0.424	0.909
		HN	6.581 5	11.153 1	0.541 9		0.045	0.286
		DB	12.739 4	19.585 4	1.021 9	2.477 6		0.316
		DN	4.704 1	7.176 4	0.416 5	1.189 4	1.150 2