克雷伯菌作为α-生育酚源促进植物乳杆菌发酵水稻秸秆青贮饲料

doi:10.1016/j.jia.2023.11.036

Journal of Integrative Agriculture ›› 2024, Vol. 23 ›› Issue (12): 4186-4202.DOI: 10.1016/j.jia.2023.11.036

克雷伯菌作为α-生育酚源促进植物乳杆菌发酵水稻秸秆青贮饲料

收稿日期:2023-03-13 接受日期:2023-10-07 出版日期:2024-12-20 发布日期:2024-11-18

Klebsiella as an α-tocopherol source facilitating Lactobacillus plantarum fermentation in rice straw silage

Cheng Zong^1*, Lu Tang^{4, 5*}, Tao Shao¹, Yu Xiao¹,Zhongyong Huang¹, Wanqi Jiang¹, Jiugang Zhu¹, Zhihao Dong¹, Mao Li^3#, Qinhua Liu^{1, 2#}

¹ Institute of Ensiling and Processing of Grass, College of Agro-grassland Science, Nanjing Agricultural University, Nanjing 210095, China
² Yunnan Provincial Key Laboratory of Animal Nutrition and Feed, Faculty of Animal Science and Technology, Yunnan AgriculturalUniversity, Kunming 650201, China
³ Tropical Crops Genetic Resource Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China
⁴ Chongqing Sericulture Science and Technology Research Institute, Chongqing 400700, China
⁵ Chongqing Academy of Animal Sciences, Chongqing 402460, China

Received:2023-03-13 Accepted:2023-10-07 Online:2024-12-20 Published:2024-11-18
About author:Cheng Zong, E-mail: zongchengnj@163.com; Lu Tang, E-mail: 15281737065@163.com; #Correspondence Qinhua Liu, E-mail: liuqinhua@njau.edu.cn; Mao Li, E-mail: limaohn@163.com * These authors contributed equally to this study.
Supported by:
This work was supported by the Yunnan Revitalization Talents Support Plan, China (XDYC-QNRC-2023-0408), the Yunnan Revitalization Talents Support Plan, China (XDYC-CYCX-2022-0036), the Key Research and Development Projects of Hainan Province, China (ZDYF2022XDNY153), the Natural Science Foundation of Chongqing City, China (cstc2019jcyj-msxmX0066), and the Fundamental Research Funds or the Central Universities, China (KYYZ2023002).

摘要/Abstract

摘要：

青贮饲料是反刍家畜重要的日粮成分和维生素E来源，发挥着促进反刍家畜健康养殖的作用。α-生育酚即维生素E，具有强抗氧化特性，可在青贮饲料中富集增加，但富集增加的机制尚未被研究透彻。本研究通过设置添加剂（空白对照、苯甲酸钠、植物乳杆菌、细胞壁降解酶、植物乳杆菌与细胞壁降解酶的组合）和控制青贮窖空气残留的抽真空时间（5，8，11和14 s）双因素随机区组处理试验，利用单分子实时测序、产α-生育酚细菌显色鉴定和细菌共培养技术，探究了α-生育酚在水稻秸秆青贮饲料中增加的机制。结果表明，细胞壁降解酶和植物乳杆菌处理组在最短抽真空时间（5 s）的青贮饲料中具有最高的α-生育酚含量 (P<0.05)。相反，苯甲酸钠处理组在较短抽真空时间（8 s）的青贮饲料具有最低的α-生育酚含量 (P<0.05)。与长的抽真空时间（14 s）相比，最短抽真空时间(5 s)的青贮饲料具有高pH值，且产生α-生育酚的克雷伯菌的丰度较高 (P<0.05)。克雷伯菌的耐酸能力弱，但在植物乳杆菌处理的水稻秸秆青贮饲料中未被灭绝，且能增加青贮饲料的α-生育酚含量。将克雷伯菌与植物乳杆菌共培养时发现，α-生育酚促进了植物乳杆菌生长。与克雷伯菌单独培养相比，在其与植物乳杆菌共培养的培养液中，对克雷伯菌起保护作用的荚膜多糖含量仍保持高浓度水平。综上所述，克雷伯菌是水稻秸秆青贮饲料α-生育酚增加的来源。克雷伯菌供应α-生育酚促进植物乳杆菌增殖的同时，高浓度水平的荚膜多糖保护其免于被产酸的植物乳杆菌灭亡。本研究初步探明了α-生育酚在水稻青贮饲料中富集增加的机制，为应用植物乳杆菌替代化学抑菌剂提高秸秆青贮饲料的卫生质量和抗氧化特性提供了科学依据和理论支撑。

Abstract:

Silage serves as the indispensable diet of ruminants, the increasing mechanism of α-tocopherol during silage making is unclear. Rice straw lacks chlorophyll after harvesting the grain, this can eliminate the impact of tocopherols formed by the breakdown of chlorophyll. Here, we explored the α-tocopherol source, its influencing factor, and its relationship with dominant lactic acid bacteria in rice straw silage treated without or with different additives (sodium benzoate, Lactobacillus plantarum, cell wall degrading enzymes, the combination of L. plantarum and cell wall degrading enzymes) and vacuum times (5, 8, 11, and 14 s) after ensiling for 42 d. We found that the pathogenic Klebsiella was traced as the source of increased α-tocopherol in rice straw silage. The residue air in the silo, pH value, and additive variety had impacts on Klebsiella activity, which was strongly active at levels of residue air in the silo and pH that were high. As an acidic niche creator, L. plantarum was more effective than sodium benzoate in restraining Klebsiella. Despite having a low acidity tolerance, Klebsiella was still present in rice straw silage treated with L. plantarum. The relationship between Klebsiella and L. plantarum was that Klebsiella could afford α-tocopherol to the multiplication of L. plantarum and residue capsular polysaccharide protected Klebsiella from escaping the extinction in rice straw silage.

Key words: α-tocopherol , Klebsiella , Lactobacillus plantarum , rice straw silage

Cheng Zong, Lu Tang, Tao Shao, Yu Xiao, Zhongyong Huang, Wanqi Jiang, Jiugang Zhu, Zhihao Dong, Mao Li, Qinhua Liu. 克雷伯菌作为α-生育酚源促进植物乳杆菌发酵水稻秸秆青贮饲料[J]. Journal of Integrative Agriculture, 2024, 23(12): 4186-4202.

Cheng Zong, Lu Tang, Tao Shao, Yu Xiao, Zhongyong Huang, Wanqi Jiang, Jiugang Zhu, Zhihao Dong, Mao Li, Qinhua Liu. Klebsiella as an α-tocopherol source facilitating Lactobacillus plantarum fermentation in rice straw silage[J]. Journal of Integrative Agriculture, 2024, 23(12): 4186-4202.

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克雷伯菌作为α-生育酚源促进植物乳杆菌发酵水稻秸秆青贮饲料

Klebsiella as an α-tocopherol source facilitating Lactobacillus plantarum fermentation in rice straw silage

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