Effects of Microbial Inoculants on the Fermentation Quality and Microbial Community Diversity of Alfalfa Silage

doi:10.13560/j.cnki.biotech.bull.1985.2021-0831

Abstract

Abstract:

In order to develop microbial inoculants suitable for alfalfa silage,the relationship between the microbial community of silage and the quality of alfalfa silage was investigated. The experiment was conducted to isolate and screen lactic acid strains with high acid production capacity from the alfalfa fermented green juice. The 16S rDNA gene sequence homology analysis was used to identify and then to prepare silage bacteria GSSW in compound. Silage samples were divided into 4 groups,adding with fermented green juice（aFGJ）,sila-max silage（YB）,GSSW bacteria,and no addition as control（CK）. A high throughput sequencing technique was used to analyze the microbial community diversity and detect physicochemical fermentation indexes. The results showed that MXLZ-1,MXLZ-2 and MXLZ-4 isolated from the fermented green juice were identified as Pediococcus pentosaceus,Lactobacillus plantarum and Pediococcus parvulus. Compared with CK group,dry matter and crude protein in the other 3 groups significantly increased（P<0.05）;neutral detergent fiber,acid detergent fiber and lignin significantly decreased（P<0.05）;pH and ammonia nitrogen/total nitrogen significantly decreased（P<0.05）,lactic acid significantly increased（P<0.05）. Lactobacillis and Pediococcus were the genera with high relative abundance of silage samples in the 4 groups. The abundance of Lactobacillis was higher than that of Pediococcus group with microbial inoculants,while that of CK was the opposite. Lactobacillus plantarum,Pediococcus pentosaceus,Lactobacillus brevis,and Enterococcus mundti of silage samples in the 4 groups were significantly different. Adding three kinds of microbial inoculants increased the abundance of L. plantarum and decreased the abundance of P. pentosaceus and E. mundtii. Adding silage inoculants increased the number of beneficial bacteria,reduced the number of harmful bacteria,and improved the fermentation quality of alfalfa silage. The GSSW can be used in alfalfa silage.

Key words: fermented green juice, alfalfa silage, high-throughput sequencing, microbial community, fermentation quality

MAO Ting, NIU Yong-yan, ZHENG Qun, YANG Tao, MU Yong-song, ZHU Ying, JI Bin, WANG Zhi-ye. Effects of Microbial Inoculants on the Fermentation Quality and Microbial Community Diversity of Alfalfa Silage[J]. Biotechnology Bulletin, 2021, 37(9): 86-94.

Figures/Tables 7

References 29

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项目 Index	数值 Numeric value
干物质 Dry matter/（g·kg^-1 FM）	382±0.42
pH	6.43±0.25
酸性洗涤纤维Acid detergent fiber/（g·kg^-1 DM）	345±1.20
中性洗涤纤维Neutral detergent fiber/（g·kg^-1 DM）	452±0.35
粗蛋白Crude protein/（g·kg^-1 DM）	206±0.32
可溶性碳水化合物 Water soluble carbohydrate/（g·kg^-1 DM）	68.3±0.25

项目 Index	数值 Numeric value
干物质 Dry matter/（g·kg^-1 FM）	382±0.42
pH	6.43±0.25
酸性洗涤纤维Acid detergent fiber/（g·kg^-1 DM）	345±1.20
中性洗涤纤维Neutral detergent fiber/（g·kg^-1 DM）	452±0.35
粗蛋白Crude protein/（g·kg^-1 DM）	206±0.32
可溶性碳水化合物 Water soluble carbohydrate/（g·kg^-1 DM）	68.3±0.25

时间 Time/d	添加剂 Silage additive	干物质 DM/（g·kg^-1 FM）	营养成分/（g·kg^-1 DM）
时间 Time/d	添加剂 Silage additive	干物质 DM/（g·kg^-1 FM）	粗蛋白 CP	可溶性碳水化和物 WSC	中性洗涤纤维NDF	酸性洗涤纤维ADF	木质素 Lignin
30	YB	324±0.40^e	253±0.31^e	61.5±0.03^cd	333±0.26^c	286±0.30^b	86±0.52^ab
	aFGJ	323±0.26^e	225±0.25^c	59.7±0.02^ab	359±0.79^e	304±0.15^f	82±0.10^ab
	GSSW	305±0.58^ab	274±0.06^h	62.8±0.00^d	356±0.30^e	291±0.35^cd	87±0.06^b
	CK	304±0.21^ab	216.±0.29^b	60.4±0.05^bc	404±0.44^h	300±0.21^e	86±0.13^ab
50	YB	315±0.12^d	246±0.23^f	58.9±0.06^ab	325±0.17^b	263±0.41^a	83±0.05^ab
	aFGJ	311±0.21^cd	215±0.10^ab	58.6±0.03^a	330±0.06^bc	294±0.12^d	82±0.11^a
	GSSW	304±0.25ab	269±0.25^j	60.7±0.02^bc	343±0.15^d	287±0.16^bc	86±0.15^ab
	CK	299±0.12^a	213±0.29^ab	60.1±0.03^bcd	397±0.25^j	292±0.25^cd	87±0.13^ab
70	YB	317±0.20^d	237±0.25^d	62.5±0.01^d	323±0.32^b	260±0.27^a	83±0.14^ab
	aFGJ	315±0.35^d	210±0.12^a	60.6±0.03^bc	333±0.15^c	294±0.31^d	82±0.22^a
	GSSW	303±0.29^ab	245±0.25^f	67.8±0.21^f	314±0.21^a	284±0.09^b	83±0.16^ab
	CK	296±0.21^a	208±0.21^a	59.9±0.02^bcd	387±0.31^f	285±0.03^b	86±0.11^ab

时间 Time/d	添加剂 Silage additive	干物质 DM/（g·kg^-1 FM）	营养成分/（g·kg^-1 DM）
时间 Time/d	添加剂 Silage additive	干物质 DM/（g·kg^-1 FM）	粗蛋白 CP	可溶性碳水化和物 WSC	中性洗涤纤维NDF	酸性洗涤纤维ADF	木质素 Lignin
30	YB	324±0.40^e	253±0.31^e	61.5±0.03^cd	333±0.26^c	286±0.30^b	86±0.52^ab
	aFGJ	323±0.26^e	225±0.25^c	59.7±0.02^ab	359±0.79^e	304±0.15^f	82±0.10^ab
	GSSW	305±0.58^ab	274±0.06^h	62.8±0.00^d	356±0.30^e	291±0.35^cd	87±0.06^b
	CK	304±0.21^ab	216.±0.29^b	60.4±0.05^bc	404±0.44^h	300±0.21^e	86±0.13^ab
50	YB	315±0.12^d	246±0.23^f	58.9±0.06^ab	325±0.17^b	263±0.41^a	83±0.05^ab
	aFGJ	311±0.21^cd	215±0.10^ab	58.6±0.03^a	330±0.06^bc	294±0.12^d	82±0.11^a
	GSSW	304±0.25ab	269±0.25^j	60.7±0.02^bc	343±0.15^d	287±0.16^bc	86±0.15^ab
	CK	299±0.12^a	213±0.29^ab	60.1±0.03^bcd	397±0.25^j	292±0.25^cd	87±0.13^ab
70	YB	317±0.20^d	237±0.25^d	62.5±0.01^d	323±0.32^b	260±0.27^a	83±0.14^ab
	aFGJ	315±0.35^d	210±0.12^a	60.6±0.03^bc	333±0.15^c	294±0.31^d	82±0.22^a
	GSSW	303±0.29^ab	245±0.25^f	67.8±0.21^f	314±0.21^a	284±0.09^b	83±0.16^ab
	CK	296±0.21^a	208±0.21^a	59.9±0.02^bcd	387±0.31^f	285±0.03^b	86±0.11^ab

时间Time /d	添加剂 Silage additive	pH	氨氮/总氮 Ammonia nitrogen/Total nitrogen	挥发性脂肪酸 Volatile fatty acid/（g·kg^-1DM）
时间Time /d	添加剂 Silage additive	pH	氨氮/总氮 Ammonia nitrogen/Total nitrogen	乳酸 Lactic acid	乙酸 Acetic acid	丁酸 Butyric acid
30	YB	5.02±0.02^ef	60.5±0.03^a	42.3±0.01^e	12.4±0.04^c
	aFGJ	5.13±0.01^fg	63.1±0.03^c	32.9±0.06^a	11.1±0.01^b	0.1±0.01^a
	GSSW	5.08±0.07^e	59.7±0.05^a	43.2±0.04^f	11.2±0.02^b	--
	CK	5.57±0.07ⁱ	69.5±0.07^f	33.4±0.05^a	10.5±0.04^a	0.9±0.12^d
50	YB	4.56±0.02^c	60.3±0.11^a	44.6±0.02^h	18.9±0.04^d	--
	aFGJ	4.82±0.01^d	63.3±0.01^c	40.2±0.02^d	12.5±0.02^c	0.6±0.05^c
	GSSW	4.50±0.02^b	61.9±0.06^b	59.8±0.06^g	11.2±0.03^b	--
	CK	5.21±0.02^h	72.5±0.12^g	33.2±0.06^a	10.2±0.02^a	1.2±0.04^f
70	YB	4.25±0.01^a	67.2±0.05^e	44.5±0.03^h	20.2±0.03^e	--
	aFGJ	4.80±0.02^d	65.1±0.01^d	38.7±0.06^c	13.9±0.05^f	0.6±0.02^c
	GSSW	4.20±0.01^a	64.5±0.03^d	52.3±0.03^k	13.2±0.03^g	--
	CK	5.17±0.05^gh	78.5±0.12^h	35.5±0.04^b	11.2±0.01^b	2.0±0.01^ab