title: Comparative microbiota assessment of wilted Italian ryegrass, whole crop corn, and wilted alfalfa silage using denaturing gradient gel electrophoresis and next-generation sequencing.
abstract: The microbiota of pre-ensiled crop and silage were examined using denaturing gradient gel electrophoresis (DGGE) and next-generation sequencing (NGS). Wilted Italian ryegrass (IR), whole crop corn (WC), and wilted alfalfa (AL) silages stored for 2 months were examined. All silages contained lactic acid as a predominant fermentation product. Across the three crop species, DGGE detected 36 and 28 bands, and NGS identified 253 and 259 genera in the pre-ensiled crops and silages, respectively. The NGS demonstrated that, although lactic acid bacteria (LAB) became prevalent in all silages after 2 months of storage, the major groups were different between crops: Leuconostoc spp. and Pediococcus spp. for IR silage, Lactobacillus spp. for WC silage, and Enterococcus spp. for AL silage. The predominant silage LAB genera were also detected by DGGE, but the presence of diverse non-LAB species in pre-ensiled crops was far better detected by NGS. Likewise, good survival of Agrobacterium spp., Methylobacterium spp., and Sphingomonas spp. in IR and AL silages was demonstrated by NGS. The diversity of the microbiota described by principal coordinate analysis was similar between DGGE and NGS. Our finding that analysis of pre-ensiled crop microbiota did not help predict silage microbiota was true for both DGGE and NGS.
authors.affiliation: Department of Animal Science, Graduate School of Environmental and Life Science, Okayama University, Okayama, 700-8530, Japan.|Henan Provincial Key Laboratory of Ion Beam Bio-engineering, Zhengzhou University, Zhengzhou, China.
authors.forename: Kuikui
authors.initials: K
authors.lastname: Ni
authors.affiliation: Department of Animal Science, Graduate School of Environmental and Life Science, Okayama University, Okayama, 700-8530, Japan.
authors.forename: Tang Thuy
authors.initials: TT
authors.lastname: Minh
authors.affiliation: Department of Animal Science, Graduate School of Environmental and Life Science, Okayama University, Okayama, 700-8530, Japan.
authors.forename: Tran Thi Minh
authors.initials: TT
authors.lastname: Tu
authors.affiliation: Department of Animal Science, Graduate School of Environmental and Life Science, Okayama University, Okayama, 700-8530, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Tsuruta
authors.affiliation: Henan Provincial Key Laboratory of Ion Beam Bio-engineering, Zhengzhou University, Zhengzhou, China.
authors.forename: Huili
authors.initials: H
authors.lastname: Pang
authors.affiliation: Department of Animal Science, Graduate School of Environmental and Life Science, Okayama University, Okayama, 700-8530, Japan. j1oufeed@okayama-u.ac.jp.
authors.forename: Naoki
authors.identifier: http://orcid.org/0000-0001-6105-8124
authors.initials: N
authors.lastname: Nishino
mesh_terms: D058645:Denaturing Gradient Gel Electrophoresis / Q000379:methods*; D005285:Fermentation / Q000502:physiology; D019344:Lactic Acid / Q000378:metabolism; D008129:Lolium / Q000378:metabolism*; D000455:Medicago sativa / Q000378:metabolism*; D064307:Microbiota / Q000502:physiology; D012820:Silage / Q000382:microbiology*; D003313:Zea mays / Q000378:metabolism*
chemical_list: D019344:Lactic Acid
keywords: Denaturing gradient gel electrophoresis; Fermentation; Microbiota; Next-generation sequencing; Silage