Isolation and Identification of Lactic Acid Bacteria Strain from Pennisetum purpureum × P. americanum ‘Reyan No.4’ Silage

doi:10.11733/j.issn.1007-0435.2023.08.032

Acta Agrestia Sinica ›› 2023, Vol. 31 ›› Issue (8): 2537-2544.DOI: 10.11733/j.issn.1007-0435.2023.08.032

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Isolation and Identification of Lactic Acid Bacteria Strain from Pennisetum purpureum × P. americanum ‘Reyan No.4’ Silage

DANG Jia-hao¹, LONG Shi-he¹, LI Xue-feng², PAN Jun-xin¹, WANG Jian¹

1. College of Animal Science and Technology, Hainan University, Haikou, Hainan Province 570228, China;
2. College of forestry, Hainan University, Haikou, Hainan Province 570228, China

Received:2023-01-04 Revised:2023-02-11 Online:2023-08-15 Published:2023-09-01

‘热研4号’王草青贮饲料中优良乳酸菌的分离与鉴定

党佳皓¹, 龙仕和¹, 李雪枫², 潘俊歆¹, 王坚¹

1. 海南大学动物科技学院, 海南海口 570228;
2. 海南大学林学院, 海南海口 570228

通讯作者: 王坚,E-mail:wangjian901@163.com
作者简介:党佳皓(2000-),男,汉族,山西运城人,硕士研究生,主要从事饲草料调制与加工利用研究,E-mail:1003449925@qq.com
基金资助:
国家自然科学基金项目(31860682)；海南省自然科学基金项目(321RC471，317054)资助

Abstract

Abstract: The objective of this experiment was to explore the main species and properties of excellent lactic acid bacteria in Pennisetum purpureum×P. americanum 'Reyan No. 4' silage. Pennisetum purpureum×P. americanum 'Reyan No. 4' was taken as the research material,and the lactic acid bacteria attached to Pennisetum purpureum×P. americanum 'Reyan No. 4' silage were isolated,cultured,screened and identified by the combining methods of traditional microbial culture,physiological and biochemical characteristics and 16S rRNA gene sequence analysis. The results showed that one hundred and seventeen strains of lactic acid bacteria were isolated from the Pennisetum purpureum×P. americanum 'Reyan No. 4' silage,of which the four strains was able to rapidly multiply and produce acid within 8 hours,no lag period for acidproduction and reduce the pH of the fermentation broth to 3.89~4.22 after 24 hours of culture. The four strains were Pectorococcus pentosus (K1 and K2),Pectorococcus lactis (K3),and Enterococcus faecalis(K4),respectively. The K1,K2 and K4 lactic acid bacteria strains can grow well in MRS medium with NaCl concentration of 3.5% and 7.5%,pH of 3.5~8.0,and temperature of 15~45℃,along with using at least eight types of carbon sources. The K4 strain was not only use fifteen kinds of carbon sources,but also withstand 50℃ high temperature. In conclusion,Pediococcus pentosus (K1,K2) and Enterococcus faecalis (K4) stains had a high potential to be used as excellent silage additives in Pennisetum purpureum×P. americanum 'Reyan No. 4' silage making.

Key words: King grass, Silage, Lactic acid bacteria, Isolation and identification, Temperature

摘要： 为探究王草(Pennisetum purpureum×P. americana)青贮饲料中优质乳酸菌的主要种类及其性质，本研究以'热研4号'王草为对象，用传统微生物培养、生理生化特征分析与16S r RNA基因序列分析相结合的方法对王草青贮饲料上附着的乳酸菌进行了分离、培养、筛选和鉴定。研究结果表明：王草青贮饲料中分离得到了117株乳酸菌，能在8 h内快速繁殖和产酸，没有迟滞期，培养24 h可使发酵液pH值降至3.89~4.22的菌株有4株，它们分别为戊糖片球菌(K1和K2)、乳酸片球菌(K3)和粪肠球菌(K4)；但只有K1，K2和K4这3株菌株在培养基的NaCl浓度为3.5%和7.5%，pH值为3.5~8.0，15~45℃的环境中均能良好生长，均能利用至少8种碳源；其中K4菌株不但能利用15种碳源，还能耐受50℃高温。综合以上考虑，本试验筛选出的戊糖片球菌(K1，K2)和粪肠球菌(K4)具有成为王草青贮饲料生产使用的优质乳酸菌添加剂的潜力。

关键词: 王草, 青贮饲料, 乳酸菌, 分离鉴定, 温度

CLC Number:

S816.5+3

DANG Jia-hao, LONG Shi-he, LI Xue-feng, PAN Jun-xin, WANG Jian. Isolation and Identification of Lactic Acid Bacteria Strain from Pennisetum purpureum × P. americanum ‘Reyan No.4’ Silage[J]. Acta Agrestia Sinica, 2023, 31(8): 2537-2544.

党佳皓, 龙仕和, 李雪枫, 潘俊歆, 王坚. ‘热研4号’王草青贮饲料中优良乳酸菌的分离与鉴定[J]. 草地学报, 2023, 31(8): 2537-2544.

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Isolation and Identification of Lactic Acid Bacteria Strain from Pennisetum purpureum × P. americanum ‘Reyan No.4’ Silage

‘热研4号’王草青贮饲料中优良乳酸菌的分离与鉴定

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