鄂尔多斯地区酸粥细菌多样性分析及乳酸菌的分离鉴定

史学敏; 刘珊娜; 杜茜

doi:10.13386/j.issn1002-0306.2022060261

鄂尔多斯地区酸粥细菌多样性分析及乳酸菌的分离鉴定

doi: 10.13386/j.issn1002-0306.2022060261

史学敏^1,,
刘珊娜^{1, 2,*, ,},
杜茜¹

1.
天津农学院食品科学与生物工程学院，天津 300392
2.
天津市农副产品深加工技术工程中心，天津 300392

详细信息

作者简介:
史学敏（1997−），女，硕士研究生，研究方向：食品加工与安全，E-mail：2395568435@qq.com

通讯作者:
刘珊娜（1984−），女，博士，副教授，研究方向：食品生物技术，E-mail：lsn_nxy@aliyun.com

中图分类号: TS201.3
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出版历程
- 收稿日期: 2022-06-28
- 刊出日期: 2023-05-01

Bacterial Diversity Analysis of Sour Porridge from Ordos and Isolation and Identification of Lactic Acid Bacteria

SHI Xuemin^1
,,
LIU Shanna^{1, 2,*
, ,},
DU Xi¹

1.
College of Food Science and Bioengineering, Tianjin Agricultural University, Tianjin 300392, China
2.
Tianjin Engineering and Technology Research Center of Agricultural Products Processing, Tianjin 300392, China

摘要

摘要: 以采集自内蒙古鄂尔多斯地区的8个酸粥样品为研究对象，利用Illumina MiSeq高通量测序技术，对样品进行细菌菌群结构及多样性研究，分离鉴定其优势菌株。结果表明：8个样品中的细菌主要来自厚壁菌门（Firmicutes）和变形菌门（Proteobacteria），乳酸杆菌属（Lactobacillus）和醋酸杆菌属（Acetobacter）为优势菌属。乳酸杆菌属在各样品中占比最高，为42.63%~97.49%。SZ1、SZ2样品与其他样品的菌群结构差异较大，可能与不同采集时间有关。对上述样品中的乳酸菌进行分离纯化，通过生理生化试验结合分子生物学方法，鉴定获得1株干酪乳杆菌（Lactobacillus casei），1株发酵乳杆菌（Lactobacillus fermentum），2株植物乳杆菌（Lactobacillus plantarum）和4株副干酪乳杆菌（Lactobacillus paracasei）。上述研究结果为酸粥工业化生产提供菌种资源和理论依据。
- 酸粥 /
- 高通量测序 /
- 细菌多样性 /
- 乳酸菌 /
- 分离鉴定
Abstract: The bacterial community structure and diversity of eight sour porridge samples collected from Ordos, Inner Mongolia were studied by Illumina Miseq high-throughput sequencing technology, and the dominant strains were isolated and identified. The results showed that the bacteria in the eight samples mainly belonged to Firmicutes and Proteobacteria. Lactobacillus and Acetobacter were the dominant genera. Lactobacillus was the highest abundance genus in all samples accounting for 42.63% to 97.49%. The structures of bacterial community in SZ1 and SZ2 differed from those in other samples, which was probably related to different sampling time. One strain was identified as Lactobacillus casei, one strain was identified as Lactobacillus fermentum, two strains were identified as Lactobacillus plantarum and four strains were identified as Lactobacillus paracasei by physiological and biochemical experiments as well as molecular biological method. The above research results provide strain resources and theoretical basis for the industrialized production of sour porridge.
- sour porridge /
- high-throughput sequencing /
- bacterial diversity /
- lactic acid bacteria /
- isolation and identification

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图 1 酸粥样品的稀释性曲线（A）和香农曲线（B）

Figure 1. Rarefaction curves (A) and Shannon index curves (B) of sour porridge samples

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图 2 在门水平（A）和属水平（B）上的细菌群落结构

Figure 2. Bacterial community structure at phylum level (A) and genus level (B)

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图 3 细菌群落PCoA分析

Figure 3. PCoA analysis of bacterial community

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图 4 乳酸菌菌落形态

Figure 4. Colony morphology of lactic acid bacteria

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图 5 乳酸菌革兰氏染色结果

Figure 5. Gram staining of lactic acid bacteria

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表 1 酸粥样品高通量测序结果

Table 1. High-throughput sequencing data of sour porridge samples

样品名称	原始序列数	高质量序列数	有效序列数	样品平均序列长度（bp）	GC（%）	Q20（%）	Q30（%）
SZ1	79915	79554	75123	423	52.76	98.92	95.70
SZ2	80121	79763	78951	416	52.77	99.00	95.89
SZ3	80129	79938	76016	429	51.70	99.29	96.80
SZ4	79925	79767	78243	422	53.00	99.38	97.23
SZ5	79652	79492	78233	428	52.14	99.31	96.95
SZ6	80224	80047	77922	419	53.40	99.39	97.21
SZ7	80359	80179	76030	428	51.99	99.30	96.90
SZ8	80173	80019	76593	422	51.97	99.40	97.20

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表 2 酸粥样品的Alpha多样性指数分析

Table 2. Analysis of Alpha diversity indexes of sour porridge samples

样品名称	OTU数	ACE	Chao1	Simpson	Shannon	覆盖率（%）
SZ1	60	128.64	93.33	0.69	1.91	99.97
SZ2	59	123.70	84.30	0.70	2.04	99.97
SZ3	171	187.89	192.75	0.44	1.46	99.96
SZ4	203	205.35	205.40	0.83	3.21	99.99
SZ5	203	205.44	207.50	0.17	0.95	99.99
SZ6	210	210.16	210.00	0.75	3.04	100.00
SZ7	189	197.01	199.06	0.57	1.80	99.97
SZ8	171	204.95	202.89	0.69	1.92	99.94

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表 3 酸粥pH及乳酸菌的菌落总数

Table 3. The pH of sour porridge samples and the total number of lactic acid bacteria colonies

样品名称	pH	乳酸菌菌落总数（lg CFU/mL）
SZ1	3.49±0.02^c	6.02±0.56^d
SZ2	3.51±0.01^c	6.08±0.47^d
SZ3	4.03±0.02^b	6.65±0.38^c
SZ4	4.01±0.03^b	6.78±0.26^a
SZ5	4.52±0.03^a	4.18±0.71^e
SZ6	3.98±0.01^b	5.20±0.64^e
SZ7	4.01±0.03^b	6.70±0.39^b
SZ8	4.01±0.01^b	5.51±0.46^e
注：同列不同小写字母表示差异显著（P<0.05）。

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表 4 乳酸菌的生理生化鉴定结果

Table 4. Physiological and biochemical identification results of lactic acid bacteria

菌株名称	来源	七叶苷	纤维二糖	麦芽糖	甘露醇	水杨苷	山梨醇	蔗糖	棉子糖	菊糖	乳糖
L140	SZ1	+	+	+	+	+	+	+	−	+	+
L24	SZ2	+	+	+	+	+	+	+	+	+	+
L218	SZ2	+	+	+	+	+	+	+	+	+	+
L353	SZ3	+	+	+	+	+	+	+	−	+	+
L520	SZ5	+	+	+	+	+	+	+	−	+	+
L74	SZ7	+	+	+	+	+	+	+	−	+	+
L813	SZ8	+	+	+	−	+	−	+	−	+	+
A117	SZ1	+	+	+	+	+	+	+	−	+	+
注：“+”表示结果呈阳性；“−”表示结果呈阴性。

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表 5 乳酸菌的16S rDNA序列同源性分析

Table 5. 16S rDNA sequence homology analysis of lactic acid bacteria

菌株名称	鉴定结果	同源性（%）	NCBI登录号
L140	Lactobacillus casei	99.79	9W0ZB1H3016
L24	Lactobacillus plantarum	99.79	EHFNJHFC016
L218	Lactobacillus plantarum	99.93	9W1GPU97016
L353	Lactobacillus paracasei	99.79	EHG55D2H013
L520	Lactobacillus paracasei	99.97	9W1YJJPB01R
L74	Lactobacillus paracasei	99.86	9W22CWAC016
L813	Lactobacillus fermentum	99.73	EHGAFDAN013
A117	Lactobacillus paracasei	100	9W2A3P4501R

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