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[1]张俊涛,黄熙瀛,李蓉,等.四川蔬菜尾菜可培养乳酸菌多样性及优良菌株筛选[J].应用与环境生物学报,2019,25(06):1428-1433.[doi:10.19675/j.cnki.1006-687x.2019.02027]
 ZHANG Juntao,HUANG Xiying,LI Rong,et al.Diversity of culturable lactic acid bacteria and screening of silage strains in vegetable waste from Sichuan province, China[J].Chinese Journal of Applied & Environmental Biology,2019,25(06):1428-1433.[doi:10.19675/j.cnki.1006-687x.2019.02027]
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四川蔬菜尾菜可培养乳酸菌多样性及优良菌株筛选
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
25卷
期数:
2019年06期
页码:
1428-1433
栏目:
研究论文
出版日期:
2019-12-30

文章信息/Info

Title:
Diversity of culturable lactic acid bacteria and screening of silage strains in vegetable waste from Sichuan province, China
作者:
张俊涛黄熙瀛李蓉向泉桔张小平辜运富
四川农业大学资源学院应用微生物学系 成都 611130
Author(s):
ZHANG Juntao HUANG Xiying LI Rong XIANG Quanju ZHANG Xiaoping & GU Yunfu**
Department of Applied Microbiology, College of Resource Science and Technology, Sichuan Agricultural University, Chengdu 611130, China
关键词:
蔬菜尾菜乳酸菌多样性分离筛选16S rRNA
Keywords:
vegetable waste lactic acid bacteria diversity isolation and screening 16S rRNA
分类号:
S182
DOI:
10.19675/j.cnki.1006-687x.2019.02027
摘要:
为了解四川地区蔬菜尾菜中乳酸菌(Lactic acid bacteria,LAB)的多样性,筛选适用于尾菜青贮发酵的高效乳酸菌菌株,运用纯培养法、16S rRNA-RFLP和系统发育分析对乳酸菌遗传多样性进行研究,并对分离菌株的产酸能力、耐酸能力、耐高温能力和抗菌能力等4个指标进行测定. 结果显示,分离得到的120株乳酸菌中,有18株(占总分离菌株的15.0%)产酸能力较强,10株(8.3%)能耐受50 ℃高温,11株(9.2%)耐酸能力较强(pH = 2.5),13株(10.8%)表现出较强的抗菌能力,而菌株C20(Lactobacillus plantarum)综合发酵特性最优. 基于16S rRNA-RFLP聚类图选取10株代表菌株进行16S rRNA基因测序和系统发育分析,发现120株乳酸菌分属于明串珠菌属(Leuconostoc)、片球菌属(Pediococcus)、乳杆菌属(Lactobacillus)和肠球菌属(Enterococcus)4个属,其中明串珠菌属为优势菌. 综上,四川地区蔬菜尾菜中蕴含着较为丰富多样的乳酸菌资源,这些乳酸菌在蔬菜尾菜的青贮饲料化利用方面表现出潜在价值. (图2 表2 参30)
Abstract:
The aim of this study was to explore the diversity of lactic acid bacteria (LAB) present in vegetable waste samples from the Sichuan province, China. In this study, we screened potentially suitable isolates from vegetable waste silage and investigated the genetic diversity of the LAB isolates through bacterial culture, 16S rRNA-RFLP, and 16S rRNA gene sequencing. We also evaluated the LAB isolates according to the following four indices: acid production, acid resistance, high-temperature resistance, and antibacterial activity. According to the test results, 18 out of the 120 isolated LAB strains (15.0% of the total isolates) had strong acid-producing ability, 10 strains (8.3%) were able to withstand temperatures of 50 ℃, 11 strains (9.2%) could withstand an acidic environment (pH 2.5), and 13 strains (10.8%) showed strong antibacterial ability. The comprehensive fermentation characteristic of strain C20 (Lactobacillus plantarum) was the best. According to the 16S rRNA-RFLP patterns and colony morphologies, ten typical strains were selected for 16S rRNA gene sequencing and phylogeny analyses. Phylogeny analyses showed that the strains were phylogenetically affiliated with the following groups: Leuconostoc, Pediococcus, Lactobacillus, and Enterococcus. Out of these groups, Leuconostoc bacteria were the predominant species. In conclusion, diverse species of LAB were present in vegetable waste samples from the Sichuan province, and they are potentially valuable for the silage of vegetable waste.

参考文献/References:

1 邵建宁, 彭章普, 张文齐, 麻和平, 刘彩云, 王洁, 赵昊星. 尾菜液体青贮菌剂制备及应用[J]. 中国酿造, 2016, 35 (10): 95-98 [Shao JN, Peng ZP, Zhang WQ, Ma HP, Liu CY, Wang J, Zhao HX. Preparation and application of bacterial inoculants for vegetable wastes silage [J]. China Brew, 2016, 35 (10): 95-98]
2 王亚利, 杨光, 熊才耘, 谢尚宏, 李银生, 曹林奎, 赵琦. 蔬菜废弃物蚯蚓堆肥对鸡毛菜生长的影响[J]. 农业环境科学学报, 2017, 36 (10): 2129-2135 [Wang YL, Yang G, Xiong CY, Xie SH, Li YS, Cao LK, Zhao Q. Effect of vegetable waste vermicompost on the growth of Brassica chinensis [J]. J Agro-Environ Sci, 2017, 36 (10): 2129-2135]
3 刘荣厚, 王远远, 孙辰, 梅晓岩. 蔬菜废弃物厌氧发酵制取沼气的试验研究[J]. 农业工程学报, 2008, 24 (4): 209-213 [Liu RH, Wang YY, Sun C, Mei XY. Experimental study on biogas production from vegetable waste by anaerobic fermentation [J]. Trans Chin Soc Agric Eng, 2008, 24 (4): 209-213]
4 Li D, Ran Y, Chen L, Cao Q, Li ZD, Liu XF. Instability diagnosis and syntrophic acetate oxidation during thermophilic digestion of vegetable waste [J]. Water Res, 2018, 139: 263-271
5 Sarkar S, Pal S, Chanda S. Optimization of a vegetable waste composting process with a significant thermophilic phase [J]. Procedia Environ Sci, 2016, 35: 435-440
6 Gulhane M, Khardenavis AA, Karia S, Pandit P, Kanade GS, Lokhande S, Vaidya AN, Purohit HJ. Biomethanation of vegetable market waste in an anaerobic baffled reactor: effect of effluent recirculation and carbon mass balance analysis [J]. Bioresour Technol, 2016, 215: 100-109
7 Varma VS, Kalamdhad AS. Evolution of chemical and biological characterization during thermophilic composting of vegetable waste using rotary drum composter [J]. Int J Environ Sci Technol, 2014, 12 (6): 2015-2024
8 任海伟, 王聪, 马延琴, 李志忠, 范文广, 王昱, 张丙云, 秦州州, 余倩倩. 接种不同乳酸菌对干玉米秸秆与白菜废弃物混贮品质的影响[J]. 应用与环境生物学报, 2018, 24 (3): 547-556 [Ren HW, Wang C, Ma YQ, Li ZZ, Fan WG, Wang Y, Zhang BY, Qin ZZ, Yu QQ. Effects of inoculating different lactic acid bacteria on the mixed storage quality of dry maize straw and cabbage waste [J]. Chin J Appl Environ Biol, 2018, 24 (3): 547-556]
9 林龙镇, 邹卫玲, 李安章, 朱红惠. 产酸、耐酸乳酸菌的分离鉴定及益生特性[J]. 华南农业大学学报, 2018, 39 (2): 95-102 [Lin LZ, Zou WL, Li AZ, Zhu HH. Isolation, identification and probiotic characteristics of acid-producing and acid-resistant Lactobacillus strains [J]. J South Chin Agric Univ, 2018, 39 (2): 95-102]
10 敖晓琳. 四川自然发酵酸奶及牧场环境中乳酸菌生物多样性及其在青贮饲料中的应用[D]. 雅安: 四川农业大学, 2011 [Ao XL. Biodiversity of LAB isolated from naturally fermented yak milk and ranch environment and their application in silage fermentation [D]. Ya’an: Sichuan Agricultural University, 2011]
11 Schillinger U, Lücke, FK. Antibacterial activity of Lactobacillus sake isolated from meat [J]. Appl Environ Microbiol, 1989, 55 (8): 1901
12 Zhou J, Bruns MA, Tiedje JM. DNA recovery from soils of diverse composition [J]. Appl Environ Microb, 1996, 62 (2): 316-322
13 Zhu H, Qu F, Zhu LH. Isolation of genomic DNAs from plants, fungi and bacteria using benzyl chloride [J]. Nucleic Acids Res, 1993, 21 (22): 5279-5280
14 Ponnusamy K, Jose S, Savarimuthu I, Michael GP, Redenbach M. Genetic diversity study of Chrornobacterium violaceum isolated from Kolli hills by amplified ribosomal DNA restriction analysis (ARDRA) and random amplified polymorphic DNA (RAPD) [J]. Lett Appl Microb, 2011, 53 (3): 341-349
15 Yu J, Gao W, Qing MJ, Sun ZH, Wang WH, Liu WJ, Pan L, Sun T, Wang HM, Bai N, Zhang HP. Identification and characterization of lactic acid bacteria isolated from traditional pickles in Sichuan, China [J]. J Gen Appl Microb, 2012, 58 (3): 163-172
16 王彦苏, 张一凡, 严振亚, 丁成龙, 戴传超. 水稻秸秆青贮饲料中可培养微生物多样性分析及优势乳酸菌的分离鉴定[J]. 草地学报, 2014, 22 (3): 586-592 [Wang YS, Zhang YF, Yan ZY, Ding CL, Dai CC. Isolation and identification of lactic acid bacteria, analysis of cultivable microbial diversity in rice straw silage [J]. Acta Agrestia Sin, 2014, 22 (3): 586-592]
17 陈静, 李斌, 曾庆宾, 罗定棋, 王昌全, 陈强, 刘松青, 辜运富. 攀枝花地区烤烟可培养内生固氮菌的多样性[J]. 微生物学通报, 2017, 44 (2): 428-437 [Chen J, Li B, Zeng QB, Luo DQ, Wang CQ, Chen Q, Liu SQ, Gu YF. Diversity of culturable endophytic diazotrophic bacteria in flue-cured tobacco in Panzhihua [J]. Microb China, 2017, 44 (2): 428-437]
18 Yuan XJ, Guo G, Wen AY, Desta ST, Wang J, Wang Y, Shao T. The effect of different additives on the fermentation quality, in vitro, digestibility and aerobic stability of a total mixed ration silage [J]. Anim Feed Sci Technol, 2015, 207 (6): 41-50
19 Castillo MFA, Balciunas EM, Salgado JM, Domínguez GJM, Converti a, Oliveira RPDS. Lactic acid properties, applications and production: a review [J]. Trends Food Sci Technol, 2013, 30 (1): 70-83
20 Wang C, Nishino N. Effects of storage temperature and ensiling period on fermentation products, aerobic stability and microbial communities of total mixed ration silage [J]. J Appl Microb, 2013, 114 (6): 1687-1695
21 刘秦华, 李湘玉, 李君风, 白晰, 张建国, 邵涛, 吴琳, 赵新国, 田佳鹭. 温度和添加剂对象草青贮发酵品质、α-生育酚和β-胡萝卜素的影响[J]. 草业学报, 2015, 24 (7): 116-122 [Liu QH, Li XY, Li JF, Bai X, Zhang JG, Shao T, Wu L, Zhao XG, Tian JL. Effect of temperature and additives on fermentation and α-tocopherol and β-carotene content of Pennisetum purpureum silage [J]. Acta Pratacult Sin, 2015, 24 (7): 116-122]
22 Missotten JAM, Goris J, Michiels J, Van CE, Herman L, De SS, Dierick NA, Heyndrickx M. Screening of isolated lactic acid bacteria as potential beneficial strains for fermented liquid pig feed production [J]. Anim Feed Sci Technol, 2009, 150 (1-2): 122-138
23 杨杨, 石超, 郭旭生. 高寒草甸魏斯氏乳酸菌的分离鉴定及理化特性研究[J]. 草地学报, 2014, 23 (1): 266-275 [Yang Y, Shi C, Guo XS. Characterization and identification of Weissella species isolated from Kobresia littledalei growing in alpine meadows [J]. Acta Pratacult Sin, 2014, 23 (1): 266-275]
24 李志忠, 窦俊伟, 任海伟, 李金平, 孙文斌, 黄娟娟, 李梦玉. 玉米秸秆和白菜尾菜混贮料的乳酸菌多样性及耐高温优良菌株筛选[J]. 草业科学, 2017, 34 (6): 1337-1346 [Li ZZ, Dou JW, Ren HW, Li JP, Sun WB, Huang JJ, Li MY. Lactic acid bacteria diversity analysis and screening of superior thermotolerant strains from corn stover and cabbage waste mixed-silages [J]. Pratacult Sci, 2017, 34 (6): 1337-1346]
25 张慧杰, 玉柱, 王林, 蔡义民, 李峰, 陶雅, 孙启忠. 青贮饲料中乳酸菌的分离鉴定及优良菌株筛选[J]. 草地学报, 2011, 19 (1): 137-141 [Zhang HJ, Yu Z, Wang L, Cai YM, Li F, Tao Y, Sun QZ. Isolation and identification of lactic acid bacteria from silage and filtering of excellent strains [J]. Acta Agrestia Sin, 2011, 19 (1): 137-141]
26 郗恩光, 谭海生, 杨劲松, 崔坤鹏, 张万昌, 鞠雪莉, 李晓雷, 张桂和. 香蕉植株上乳酸菌的分离鉴定、系统发育及抑菌特性分析[J]. 食品科学, 2018, 39 (14): 112-118 [Xi EG, Tan HS, Yang JS, Cui KP, Zhang WC, Ju XL, Li XL, Zhang GH. Separation, identification, phylogenetic analysis and antibacterial activity of Lactobacillus from banana plants [J]. Food Sci, 2018, 39 (14): 112-118]
27 Han LY, Zhou H. Effects of ensiling processes and antioxidants on fatty acid concentrations and compositions in corn silages [J]. J Anim Sci Biotech, 2013, 4 (2): 1-7
28 Pang HL, Zhang M, Qin GY, Tan ZF, Li ZW, Wang YP, Cai YM. Identification of lactic acid bacteria isolated from corn stovers [J]. Anim Sci J, 2011, 82 (5): 642-653
29 Yang J, Cao Y, Cai Y, Terada F. Natural populations of lactic acid bacteria isolated from vegetable residues and silage fermentation [J]. J Dairy Sci, 2010, 93 (7): 3136-3145
30 陶雅, 李峰, 高凤芹, 徐春城, 孙启忠. 籽粒苋与青贮玉米混贮品质及微生物特性研究[J]. 草业学报, 2016, 25 (12): 119-129 [Tao Y, Li F, Gao FQ, Xu CC, Sun QZ. Fermentation quality and microbial characteristics of Amaranthus hypochondriacus-corn mixed silage [J]. Acta Pratacult Sin, 2016, 25 (12): 119-129]

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更新日期/Last Update: 2019-12-25