基于高通量测序的甜高粱青贮饲料中微生物群落分析

doi:10.6048/j.issn.1001-4330.2023.11.018

摘要/Abstract

摘要：

【目的】 研究甜高粱青贮饲料的微生物多样性,为青贮饲料的生产和贮藏提供理论基础。【方法】 以甜高粱青贮饲料为研究对象,利用高通量测序技术分析甜高粱青贮饲料中微生物群落结构和多样性,研究添加剂对甜高粱青贮饲料中微生物群落的影响。【结果】 甜高粱青贮饲料中细菌优势菌门为厚壁菌门、变形菌门、拟杆菌门、蓝细菌门、酸杆菌门,优势菌属主要为乳酸杆菌属、肠杆菌属、乳球菌属、棒状杆菌;真菌群落优势菌门为子囊菌门。【结论】 不同处理的甜高粱青贮饲料之间微生物菌群组成相近,但相对丰度存在明显差异,青贮菌剂降低了甜高粱青贮饲料中微生物的相对丰度。

关键词: 甜高粱; 青贮饲料; 微生物多样性; 高通量测序

Abstract:

【Objective】 To clarify the microbial diversity of sweet sorghum silage in the hope of providing a theoretical basis for silage production and storage.【Methods】 In this study, the microbial diversity of silage from two varieties with or without the addition of silage additives were analyzed using high-throughput sequencing.【Results】 The dominant bacterial phyla in sweet sorghum silage were Firmicutes, Proteobacteria, Bacteroidetes, Cyanobacteria and Acidobacteria, and the dominant bacterial genera were Lactobacillus, Enterobacter, Lactococcus and Corynebacterium. Ascomycota was the dominant phylum in fungal community.【Conclusion】 The microbial flora composition of sweet sorghum silage is similar among different treatments, but the relative abundance is significantly different. Silage microbial agent reduces the relative abundance of microbial flora in sweet sorghum silage.

Key words: sweet sorghum; silage; microbial diversity; high throughput sequencing

中图分类号:

S182
S514

岳丽, 王卉, 山其米克, 再吐尼古丽·库尔班, 涂振东. 基于高通量测序的甜高粱青贮饲料中微生物群落分析[J]. 新疆农业科学, 2023, 60(11): 2742-2750.

YUE Li, WANG Hui, Shanqimike , Zaituniguli Kuerban, TU Zhendong. Microbial diversity analysis of sweet sorghum silage using High-Throughput sequencing[J]. Xinjiang Agricultural Sciences, 2023, 60(11): 2742-2750.

图/表 10

表1 青贮饲料样品细菌、真菌测序统计

Tab.1 Statistics of bacterial and fungal sequencing results of silage samples

微生物 Microorganism	样品 Sample	原始序列数 Raw reads	质控序列数 Clean reads	有效序列数 Effective reads	平均长度 Average length(bp)	GC (%)	OTU数量 Feature number
细菌	A	79 998	75 702	74 324	425	52.87	598
Bacteria	B	48 929	45 422	45 019	427	51.68	374
	C	48 228	44 844	44 062	426	52.56	505
	D	79 801	75 536	74 281	427	52.5	386
真菌	A	71 581	64 442	63 916	181	41.22	158
Fungus	B	172 836	49 376	48 549	224	46.55	67
	C	51 480	49 593	49 141	253	40.68	154
	D	79 728	77 351	77 191	178	39.85	29

图1 基于 OTU 水平的甜高粱青贮样品中细菌群落的 Venn 图

Fig.1 Venn plot of bacterial community in sweet sorghum silage samples based on OTU levels

表2 样品细菌的 Alpha 多样性指数

Tab.2 The Alpha diversity index of bacteria in samples

样品 Sample	Ace指数 Ace index	Chao1指数 Chao1 index	Simpson指数 Simpson index	Shannon指数 Shannon index	OTU 数量 Feature number	覆盖度 Coverage
A	629.01	634.25	0.90	5.07	598	0.998 3
B	407.90	425.11	0.73	3.10	374	0.998 5
C	536.29	548.83	0.84	4.56	505	0.998 4
D	470.95	470.73	0.51	2.31	386	0.997 4

图2 基于门水平的细菌组成及物种丰度聚类热图

Fig.2 Heat map of bacterial composition and species abundance clustering at the phylum level

图3 基于属水平的细菌组成及物种丰度聚类热图

Fig.3 Heat map of bacterial composition and species abundance clustering at the genus level

图4 细菌非度量多维尺度(a)和基于Beta多样性距离的样品层级聚类树(b)

Fig.4 Bacterial NMDS diagram (a) and sample hierarchical clustering tree based on Beta diversity distance (b)

图5 基于 OTU 水平的甜高粱青贮饲料中真菌群落的 Venn 图

Fig.5 Venn diagram of fungal community in sweet sorghum silage based on OTU level

表3 甜高粱青贮饲料中真菌的 Alpha 多样性

Tab.3 The Alpha diversity index of fungal in samples

样品 Sample	Ace指数 Ace index	Chao1指数 Chao1 index	Simpson指数 Simpson index	Shannon指数 Shannon index	OTU 数量 Feature number	覆盖度 Coverage
A	119.04	124.18	0.56	0.12	158	0.999 8
B	109.27	120.95	0.70	0.21	67	0.999 4
C	170.43	170.74	3.50	0.83	154	0.999 9
D	42.00	50.04	0.06	0.01	29	0.999 8

图6 基于门水平的甜高粱青贮饲料真菌组成及物种丰度聚类热图

Fig.6 Heat map of fungal composition and species abundance of sweet sorghum silage based on phylum level

图7 基于属水平的甜高粱青贮饲料真菌组成及物种丰度聚类热图

Fig.7 Heat map of fungal composition and species abundance in sweet sorghum silage based on genus level

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