CN115702667B - Mixed silage method for sesbania and sweet sorghum - Google Patents
Mixed silage method for sesbania and sweet sorghum Download PDFInfo
- Publication number
- CN115702667B CN115702667B CN202110907045.7A CN202110907045A CN115702667B CN 115702667 B CN115702667 B CN 115702667B CN 202110907045 A CN202110907045 A CN 202110907045A CN 115702667 B CN115702667 B CN 115702667B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus
- silage
- sesbania
- buchneri
- fermentation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000004460 silage Substances 0.000 title claims abstract description 118
- 244000138286 Sorghum saccharatum Species 0.000 title claims abstract description 52
- 235000011684 Sorghum saccharatum Nutrition 0.000 title claims abstract description 52
- 238000000034 method Methods 0.000 title claims abstract description 29
- 244000275012 Sesbania cannabina Species 0.000 title description 57
- 238000000855 fermentation Methods 0.000 claims abstract description 57
- 241000186679 Lactobacillus buchneri Species 0.000 claims abstract description 54
- 230000004151 fermentation Effects 0.000 claims abstract description 54
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 37
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 37
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 37
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 34
- 241000186660 Lactobacillus Species 0.000 claims abstract description 31
- 229940039696 lactobacillus Drugs 0.000 claims abstract description 31
- 241001468157 Lactobacillus johnsonii Species 0.000 claims abstract description 25
- 238000002360 preparation method Methods 0.000 claims abstract description 25
- 239000002131 composite material Substances 0.000 claims abstract description 22
- 239000004310 lactic acid Substances 0.000 claims abstract description 17
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 17
- 239000000203 mixture Substances 0.000 claims abstract description 17
- 241000219782 Sesbania Species 0.000 claims abstract description 15
- 241000894006 Bacteria Species 0.000 claims abstract description 12
- 150000001875 compounds Chemical class 0.000 claims abstract description 11
- 239000002994 raw material Substances 0.000 claims abstract description 10
- 238000002156 mixing Methods 0.000 claims abstract description 8
- 239000007787 solid Substances 0.000 claims abstract description 7
- 239000004480 active ingredient Substances 0.000 claims abstract description 5
- 239000002068 microbial inoculum Substances 0.000 claims description 10
- 238000004321 preservation Methods 0.000 claims description 8
- 238000009629 microbiological culture Methods 0.000 claims description 6
- 241000196324 Embryophyta Species 0.000 abstract description 11
- 230000000813 microbial effect Effects 0.000 abstract description 11
- 241000186685 Lactobacillus hilgardii Species 0.000 abstract description 10
- 210000002421 cell wall Anatomy 0.000 abstract description 3
- 150000004676 glycans Chemical class 0.000 abstract description 3
- 229920001282 polysaccharide Polymers 0.000 abstract description 3
- 239000005017 polysaccharide Substances 0.000 abstract description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 24
- 230000001580 bacterial effect Effects 0.000 description 19
- 239000000047 product Substances 0.000 description 14
- 239000002054 inoculum Substances 0.000 description 12
- 239000001963 growth medium Substances 0.000 description 10
- DNIAPMSPPWPWGF-GSVOUGTGSA-N (R)-(-)-Propylene glycol Chemical compound C[C@@H](O)CO DNIAPMSPPWPWGF-GSVOUGTGSA-N 0.000 description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 235000011054 acetic acid Nutrition 0.000 description 8
- DNIAPMSPPWPWGF-UHFFFAOYSA-N monopropylene glycol Natural products CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 8
- 229960004063 propylene glycol Drugs 0.000 description 8
- 235000013772 propylene glycol Nutrition 0.000 description 8
- 230000000694 effects Effects 0.000 description 7
- 235000019750 Crude protein Nutrition 0.000 description 6
- 239000003795 chemical substances by application Substances 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 239000002504 physiological saline solution Substances 0.000 description 6
- 241000894007 species Species 0.000 description 6
- SRBFZHDQGSBBOR-SOOFDHNKSA-N D-ribopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@@H]1O SRBFZHDQGSBBOR-SOOFDHNKSA-N 0.000 description 5
- 108020004414 DNA Proteins 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 4
- 229930006000 Sucrose Natural products 0.000 description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 4
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 4
- 229910052799 carbon Inorganic materials 0.000 description 4
- 238000009630 liquid culture Methods 0.000 description 4
- 238000012163 sequencing technique Methods 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 239000005720 sucrose Substances 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 3
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 3
- 230000025938 carbohydrate utilization Effects 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- -1 polyethylene Polymers 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- 244000063299 Bacillus subtilis Species 0.000 description 2
- 235000014469 Bacillus subtilis Nutrition 0.000 description 2
- GUBGYTABKSRVRQ-CUHNMECISA-N D-Cellobiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-CUHNMECISA-N 0.000 description 2
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 2
- QWIZNVHXZXRPDR-UHFFFAOYSA-N D-melezitose Natural products O1C(CO)C(O)C(O)C(O)C1OC1C(O)C(CO)OC1(CO)OC1OC(CO)C(O)C(O)C1O QWIZNVHXZXRPDR-UHFFFAOYSA-N 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 229930091371 Fructose Natural products 0.000 description 2
- 239000005715 Fructose Substances 0.000 description 2
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 2
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 2
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 2
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 2
- 239000004698 Polyethylene Substances 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 2
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 2
- DLRVVLDZNNYCBX-ZZFZYMBESA-N beta-melibiose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)O1 DLRVVLDZNNYCBX-ZZFZYMBESA-N 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 229930182830 galactose Natural products 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000012165 high-throughput sequencing Methods 0.000 description 2
- BJRNKVDFDLYUGJ-RMPHRYRLSA-N hydroquinone O-beta-D-glucopyranoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=C(O)C=C1 BJRNKVDFDLYUGJ-RMPHRYRLSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- QWIZNVHXZXRPDR-WSCXOGSTSA-N melezitose Chemical compound O([C@@]1(O[C@@H]([C@H]([C@@H]1O[C@@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O)CO)CO)[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O QWIZNVHXZXRPDR-WSCXOGSTSA-N 0.000 description 2
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 2
- 229950006780 n-acetylglucosamine Drugs 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 229920000573 polyethylene Polymers 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000004064 recycling Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- OMDQUFIYNPYJFM-XKDAHURESA-N (2r,3r,4s,5r,6s)-2-(hydroxymethyl)-6-[[(2r,3s,4r,5s,6r)-4,5,6-trihydroxy-3-[(2s,3s,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]methoxy]oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O[C@H]2[C@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)[C@H](O)[C@H](O)[C@H](O)O1 OMDQUFIYNPYJFM-XKDAHURESA-N 0.000 description 1
- XUCIJNAGGSZNQT-JHSLDZJXSA-N (R)-amygdalin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O[C@@H](C#N)C=2C=CC=CC=2)O1 XUCIJNAGGSZNQT-JHSLDZJXSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 108091093088 Amplicon Proteins 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- HEBKCHPVOIAQTA-NGQZWQHPSA-N D-Arabitol Natural products OC[C@H](O)C(O)[C@H](O)CO HEBKCHPVOIAQTA-NGQZWQHPSA-N 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- HEBKCHPVOIAQTA-QWWZWVQMSA-N D-arabinitol Chemical compound OC[C@@H](O)C(O)[C@H](O)CO HEBKCHPVOIAQTA-QWWZWVQMSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 229920000926 Galactomannan Polymers 0.000 description 1
- 229920002907 Guar gum Polymers 0.000 description 1
- HEBKCHPVOIAQTA-IMJSIDKUSA-N L-arabinitol Chemical compound OC[C@H](O)C(O)[C@@H](O)CO HEBKCHPVOIAQTA-IMJSIDKUSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 102100030569 Nuclear receptor corepressor 2 Human genes 0.000 description 1
- 101710153660 Nuclear receptor corepressor 2 Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- MUPFEKGTMRGPLJ-RMMQSMQOSA-N Raffinose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 MUPFEKGTMRGPLJ-RMMQSMQOSA-N 0.000 description 1
- 241000122973 Stenotrophomonas maltophilia Species 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- MUPFEKGTMRGPLJ-UHFFFAOYSA-N UNPD196149 Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC2C(C(O)C(O)C(CO)O2)O)O1 MUPFEKGTMRGPLJ-UHFFFAOYSA-N 0.000 description 1
- 229920002310 Welan gum Polymers 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 241001148470 aerobic bacillus Species 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 229940089837 amygdalin Drugs 0.000 description 1
- YZLOSXFCSIDECK-UHFFFAOYSA-N amygdalin Natural products OCC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC(C#N)c3ccccc3 YZLOSXFCSIDECK-UHFFFAOYSA-N 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229960000271 arbutin Drugs 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 238000005422 blasting Methods 0.000 description 1
- 230000005200 bud stage Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000024346 drought recovery Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- YGHHWSRCTPQFFC-UHFFFAOYSA-N eucalyptosin A Natural products OC1C(O)C(O)C(CO)OC1OC1C(OC(C#N)C=2C=CC=CC=2)OC(CO)C(O)C1O YGHHWSRCTPQFFC-UHFFFAOYSA-N 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000665 guar gum Substances 0.000 description 1
- 229960002154 guar gum Drugs 0.000 description 1
- 235000010417 guar gum Nutrition 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000015784 hyperosmotic salinity response Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 229920005610 lignin Polymers 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000010871 livestock manure Substances 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000005272 metallurgy Methods 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 238000005065 mining Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- BJRNKVDFDLYUGJ-UHFFFAOYSA-N p-hydroxyphenyl beta-D-alloside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C(O)C=C1 BJRNKVDFDLYUGJ-UHFFFAOYSA-N 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000007639 printing Methods 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000012113 quantitative test Methods 0.000 description 1
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for mixed ensiling of sesbania and sweet sorghum. The invention discloses a method for mixed ensiling of sesbania and sweet sorghum, which comprises the following steps: mixing silage raw materials with a composite lactobacillus preparation for solid anaerobic fermentation, and collecting fermentation products to obtain sesbania silage; silage is a mixture composed of sesbania aerial parts and sweet sorghum aerial parts; the active ingredients of the compound lactobacillus preparation are lactobacillus plantarum (Lactobacillus plantarum), lactobacillus johnsonii (Lactobacillus hilgardii) and lactobacillus buchneri (Lactobacillus buchneri). The method can remarkably prolong the aerobic stability of sesbania and sweet sorghum, fully decompose plant cell walls, maintain silage pH and temperature by utilizing a more polysaccharide source, maintain the relative abundance of lactic acid bacteria in microbial flora and reduce the loss of dry matters.
Description
Technical Field
The invention relates to a method for mixed ensiling of sesbania and sweet sorghum in the technical field of agricultural product processing.
Background
Sesbania (Sesbania cannabina) is a leguminous plant. Is widely distributed in south areas such as China, hainan, jiangsu, zhejiang, jiangxi, fujian, guangxi, yunnan and the like, and cultivated in northern areas such as Shandong and the like. Is often used in paddy fields, ditches and other wet low lands. Sesbania has strong adaptability, salt tolerance, waterlogging tolerance, barren tolerance, drought tolerance, warmth and moisture preference, and is pioneer crop for improving saline-alkali soil and restoring soil.
The sesbania has wide application range, is an excellent green manure crop with various soil types, can produce sesbania gum from seed endosperm, is a novel galactomannan gum discovered in the 70 th century, and can replace imported guar gum. Sesbania gum has good water solubility and is widely applied to industries such as oil extraction in oil fields, mineral processing metallurgy, mining blasting, textile printing and dyeing, medicines and pesticides, daily chemical industry and the like. Stems and leaves of sesbania can be used as livestock feed, but the research is less at present.
Generally, leguminous plants have higher protein content and lower soluble sugar content, cannot meet the basic requirement of silage, and how to silage leguminous plants is a technical problem to be solved at present.
Disclosure of Invention
The technical problem to be solved in the aspect is how to silage leguminous plants, in particular to silage sesbania.
In order to solve the technical problems, the invention firstly provides a method for preparing sesbania silage, which comprises the following steps: mixing silage raw materials with a compound lactobacillus preparation to obtain a pre-fermentation mixture, carrying out solid anaerobic fermentation on the pre-fermentation mixture, and collecting fermentation products to obtain sesbania silage;
the silage raw material is a mixture composed of sesbania aerial parts and sweet sorghum aerial parts;
the active ingredients of the compound lactobacillus preparation are lactobacillus plantarum (Lactobacillus plantarum), lactobacillus johnsonii (Lactobacillus hilgardii) and lactobacillus buchneri (Lactobacillus buchneri).
In the above method, the lactobacillus plantarum (Lactobacillus plantarum) can be lactobacillus plantarum (Lactobacillus plantarum) WQ-01;
the lactobacillus johnsonii (Lactobacillus hilgardii) can be lactobacillus johnsonii (Lactobacillus hilgardii) 60TS-2;
the lactobacillus buchneri (Lactobacillus buchneri) can be lactobacillus buchneri (Lactobacillus buchneri) NX205, and the lactobacillus buchneri (Lactobacillus buchneri) NX205 is a strain with a preservation number of CGMCC No.16534 in the China general microbiological culture Collection center.
In the compound lactic acid bacteria preparation according to the above method, the ratio of the viable count of the lactobacillus plantarum (Lactobacillus plantarum), the lactobacillus johnsonii (Lactobacillus hilgardii) and the lactobacillus buchneri (Lactobacillus buchneri) may be 1:1:1.
in one embodiment of the invention, the complex lactobacillus preparation is composed of the lactobacillus plantarum (Lactobacillus plantarum), the lactobacillus johnsonii (Lactobacillus hilgardii), the lactobacillus buchneri (Lactobacillus buchneri) and physiological saline.
In the method, the mass percentage of the sesbania overground part in the silage raw material can be 30-70%.
In the method, the aerial parts of sesbania and the aerial parts of sweet sorghum are small sections with the length of 2-4 cm.
In the pre-fermentation mixture according to the above method, the ratio of silage to lactobacillus plantarum (Lactobacillus plantarum), lactobacillus johnsonii (Lactobacillus hilgardii) and lactobacillus buchneri (Lactobacillus buchneri) in the compound lactobacillus preparation may be 1:1:1.
in the above method, the fermentation may be carried out at 15 to 25℃such as 20 to 25 ℃. The fermentation time may be 14-60 days.
The fermentation can be carried out under vacuum, sealing and light-shielding conditions, and in particular can be carried out in a sealed polyethylene bag.
The sesbania silage prepared by the method for preparing the sesbania silage also belongs to the protection scope of the invention.
The compound lactobacillus preparation also belongs to the protection scope of the invention.
The invention also provides a composition for preparing sesbania silage, which consists of the composite lactobacillus preparation and the silage raw material.
Lactobacillus buchneri (Lactobacillus buchneri) NX205 also belongs to the protection scope of the invention, and the preservation number of the Lactobacillus buchneri (Lactobacillus buchneri) NX205 in the China general microbiological culture Collection center is CGMCC No.16534.
The invention also provides a microbial inoculum, and an active ingredient of the microbial inoculum is the lactobacillus buchneri (Lactobacillus buchneri) NX205.
The application of the method for preparing sesbania silage, the composite lactobacillus preparation or the composition, or the lactobacillus buchneri (Lactobacillus buchneri) NX205 or the microbial inoculum in preparing animal feed also belongs to the protection scope of the invention.
The invention utilizes a composite lactobacillus preparation for mixed silage of sesbania and sweet sorghum (which contains lactobacillus plantarum (Lactobacillus plantarum) WQ-01, lactobacillus johnsonii (Lactobacillus hilgardii) 60TS-2 and lactobacillus buchneri (Lactobacillus buchneri) NX 205) to carry out silage fermentation on the sesbania and sweet sorghum, can remarkably prolong the aerobic stability of the sesbania and the sweet sorghum, fully decompose plant cell walls, and maintain silage pH and temperature by utilizing a more polysaccharide source, maintain the relative abundance of lactobacillus in microbial flora and reduce dry matter loss.
Description of biological Material preservation
Classification naming: lactobacillus buchneri (Lactobacillus buchneri)
Strain number: NX205
Preservation unit name: china general microbiological culture Collection center (China Committee for culture Collection of microorganisms)
The preservation unit is abbreviated as: CGMCC
Deposit unit address: beijing, chaoyang, north Chen West Lu 1, 3, postal code: 100101
Preservation date: 2018-9-26
Accession numbers of the preservation center: CGMCC No.16534
Drawings
FIG. 1 is a graph showing the effect of a composite silage inoculant on pH of a sesbania and sweet sorghum mixed silage sample.
FIG. 2 is a graph showing the effect of a composite silage inoculant on lactic acid content of a sesbania and sweet sorghum mixed silage sample.
FIG. 3 is a graph showing the effect of a composite silage inoculant on the mixed silage acetic acid content of sesbania and sweet sorghum samples.
FIG. 4 is an effect of a composite silage inoculant on the content of 1, 2-propanediol in a mixed silage of sesbania and sweet sorghum.
FIG. 5 is an effect of a composite silage inoculant on the culturable lactic acid bacteria content of a sesbania and sweet sorghum mixed silage sample.
FIG. 6 is an effect of a composite silage inoculant on the culturable yeast content of a sesbania and sweet sorghum mixed silage sample.
FIG. 7 is an effect of a composite silage inoculant on the culturable aerobic content of a sesbania and sweet sorghum mixed silage sample.
FIG. 8 dynamic changes in microbial flora of mixed silage samples of horizontal sesbania and sweet sorghum. D3, D7, D14, D30, D60 represent day 3, day 7, day 14, day 30 and day 60 of silage fermentation, respectively.
Wherein, SS100, SS70, SS50 and SS30 respectively represent sesbania proportions of 100%, 70%, 50% and 30%.
Detailed Description
The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.
The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents, instruments and the like used in the examples described below are commercially available unless otherwise specified. The quantitative tests in the following examples were all set up in triplicate and the results averaged.
Example 1 preparation of Mixed silage from sesbania and sweet sorghum
The embodiment provides a composite lactobacillus preparation for mixed silage of sesbania and sweet sorghum, which comprises lactobacillus plantarum (Lactobacillus plantarum) WQ-01 (called lactobacillus plantarum WQ-01 for short), lactobacillus johnsonii (Lactobacillus hilgardii) 60TS-2 (called lactobacillus johnsonii 60TS-2 for short) and lactobacillus buchneri (Lactobacillus buchneri) NX205 (called lactobacillus buchneri NX205 for short), and can remarkably prolong the aerobic stability of sesbania and sweet sorghum, fully decompose plant cell walls, maintain silage pH and temperature by using a more polysaccharide source, maintain the relative abundance of lactobacillus in microbial flora and reduce dry matter loss.
Among them, lactobacillus plantarum (Lactobacillus plantarum) WQ-01 and Lactobacillus johnsonii (Lactobacillus hilgardii) 60TS-2 are described in Chinese patent application 202010258231.8 (application publication No. CN 111534456A).
The lactobacillus buchneri NX205 is a strain NX205 separated from sweet sorghum in Ningxia welan county by Luo Aiqun, and the separation and identification steps are as follows:
1) 10.00g of sweet sorghum sample is accurately weighed by a balance, fully mixed with 90mL of sterile water, shaken for 30min at room temperature by a shaking table of 150rpm, filtered by four layers of gauze to obtain leaching solution, and uniformly coated on an MRS solid culture medium by a sterilization coating rod after gradient dilution, and placed at 37 ℃ for anaerobic culture for 48h.
2) After the MRS solid culture medium grows out of the bacterial colony, a sterilized inoculating loop is used for picking out white, smooth and convex bacterial colonies with regular edges, and the bacterial colonies are inoculated into 3mL MRS liquid culture medium and placed at 37 ℃ for standing anaerobic culture for 48 hours.
3) After bacterial cells grow out of the MRS liquid medium, transferring the bacterial liquid into a sterilized centrifuge tube by using a 1mL pipettor, centrifuging at 12000rpm for 3min, collecting the bacterial cells, and extracting genomic DNA (bacterial genomic DNA rapid extraction kit, bomeide, product number: DL 111-02) was used as a template, using the universal primer (27F: 5'-AGAGTTTGATCCTGGCTCAG-3',1492R: 5'-TACGGCTACCTTGTTACGACTT-3') the 16S DNA was amplified by PCR and detected by 1% agarose gel electrophoresis. And (3) purifying and recycling the PCR amplified product by using a PCR product purifying and recycling kit (Bomadder, product number: DH 102-02), and sequencing, wherein the sequence of the strain NX205 is sequence 1 in a sequence table.
Strain NX205 was rod-shaped. The NCBI is used for comparing the sequenced sequences, the strain NX205 is identified as Lactobacillus buchneri (Lactobacillus buchneri), and is recorded as Lactobacillus buchneri (Lactobacillus buchneri) NX205, and the strain is preserved in China general microbiological culture Collection center (CGMCC) No. 16534) at 9 and 26 of 2018.
1. Strain characteristics and sugar utilization characteristics
Lactobacillus plantarum WQ-01, lactobacillus johnsonii 60TS-2 and Lactobacillus buchneri NX205 were grown in MRS broth and in MRS solid medium (1.5% agar powder added to MRS broth) and anaerobically cultured at 37 ℃. The characteristics of the strains are shown in Table 1, and the three are gram-positive bacteria, rod-shaped structures, catalase negative and oxidase negative. Lactobacillus plantarum WQ-01 is a homotype fermentation strain, can grow in the pH range of 3.5-8.5, has weaker growth at the pH of 2.0, grows at the temperature of 15-45 ℃, and tolerates 3.0% and 6.5% NaCl concentration; lactobacillus johnsonii 60TS-2 is a heterofermentative strain, grows in the pH range of 3.5-8.0, grows weakly at pH2.0, grows at 15-40 ℃, grows weakly at 45 ℃, and tolerates 3.0% and 6.5% NaCl concentration; lactobacillus buchneri NX205 is a heterofermentative strain and can grow in the range of pH 4.0-4.5, weaker at pH2.0 and pH3.5, at 15-40 ℃, weaker at 45 ℃, tolerating 3.0% NaCl concentration and weaker at 6.5% NaCl concentration.
The sugar utilization conditions are shown in Table 2, and lactobacillus plantarum WQ-01 has the utilization capacity for ribose, galactose, glucose, fructose, mannose, mannitol, sorbitol, N-acetyl-glucosamine, amygdalin, arbutin, salitol, cellobiose, maltose, lactose, melibiose, sucrose, trehalose, melezitose, raffinose, trepanose and L-arabitol and has weak growth in a culture medium respectively taking glycerol and D-arabitol as the sole carbon sources; lactobacillus johnsonii 60TS-2 has the ability to utilize L-arabinose, ribose, D-xylose and sucrose, and has weak growth in a culture medium which takes mannose, N-acetyl-glucosamine, cellobiose and sucrose as sole carbon sources respectively; lactobacillus buchneri NX205 has the utilization capacity for L-arabinose, ribose, D-xylose, glucose, maltose, melibiose, sucrose and melezitose, and has weak growth in a culture medium respectively taking galactose and fructose as the only carbon sources. In Table 2, "control" means that the medium does not contain a carbon source.
TABLE 1 characterization of Lactobacillus plantarum WQ-01, lactobacillus his 60TS-2 and Lactobacillus buchneri NX205 strains
In Table 1, "W" indicates weak growth.
TABLE 2 sugar utilization by Lactobacillus plantarum WQ-01, lactobacillus johnsonii 60TS-2 and Lactobacillus buchneri NX205
In Table 2, "W" indicates weak growth.
2. Preparation of composite silage microbial inoculum
Culturing lactobacillus plantarum WQ-01 with MRS liquid culture medium (product number M8540-250g, beijing Soy Bao technology Co., ltd.), collecting culture solution, centrifuging, collecting thallus, washing with physiological saline for three times, and collecting thallus to obtain lactobacillus plantarum WQ-01 microbial inoculum.
Culturing Lactobacillus johnsonii 60TS-2 with MRS liquid culture medium (product number M8540-250g, beijing Soilebao technology Co., ltd.), collecting culture solution, centrifuging, collecting thallus, washing with physiological saline for three times, and collecting thallus to obtain Lactobacillus johnsonii 60TS-2 microbial inoculum.
Culturing Lactobacillus buchneri NX205 with MRS liquid culture medium (product number M8540-250g, beijing Soy Bao technology Co., ltd.), collecting culture solution, centrifuging, collecting thallus, washing with physiological saline for three times, and collecting thallus to obtain Lactobacillus buchneri NX205 microbial inoculum.
Mixing lactobacillus plantarum WQ-01 bacterial agent, lactobacillus johnsonii 60TS-2 bacterial agent and lactobacillus buchnsonii NX205 bacterial agent according to a viable bacterial ratio of 1:1:1, and adding physiological saline to obtain a composite silage bacterial agent, wherein the viable bacterial contents of the lactobacillus plantarum WQ-01, lactobacillus johnsonii 60TS-2 and lactobacillus buchnsonii NX205 in the obtained composite silage bacterial agent are respectively 10 8 cfu/g、10 8 cfu/g、10 8 cfu/g。
3. Preparation of sesbania and sweet sorghum mixed silage
Harvesting the aerial parts of fresh sesbania in the bud stage and the aerial parts of sweet sorghum in the finishing stage, and cutting the aerial parts of sesbania and the aerial parts of sweet sorghum respectively, wherein the cutting length is 2-4 cm (Kaiyu mechanical company, model 9 ZP-3.6).
Mixing and grouping the cut sesbania and sweet sorghum according to the table 3 respectively to obtain fresh plant samples; then adding a composite silage bacterial agent into each group of fresh plant samples to obtain a pre-fermentation mixture, wherein the viable bacteria content of lactobacillus plantarum WQ-01, lactobacillus johnsonii 60TS-2 and lactobacillus buchneri NX205 in the pre-fermentation mixture is 10 8 CFU/g, the total mass of fresh plant samples was 300g.
The pre-fermentation mixtures of each group are filled into polyethylene bags, the total mass of fresh plant samples in each bag is 300g, each group is repeated 4 times, silage fermentation is carried out at room temperature (20-25 ℃) in a dark place after vacuumizing, sampling is carried out on days 0, 3, 7, 14, 30 and 60 of silage fermentation respectively, and Dry Matter (DM) and silage fermentation products are detected.
TABLE 3 grouping and mixing ratio of sesbania and sweet sorghum
4. Sample collection and detection
1. Dry Matter (DM) detection
Dry matter detection was performed on day 0, day 3, day 7, day 14, day 30 and day 60 of silage fermentation with bag opening: weighing 100g of sample, drying in a constant temperature dryer at 65 ℃ to constant weight (72 h), naturally cooling to room temperature, weighing, and recording the weight m of the sample, wherein the dry matter content (DM) =m/100×100%; the samples were then assayed for Crude Protein (CP), neutral wash fiber (NDF), acid wash fiber (ADF), acid wash lignin (ADL), and soluble carbohydrate (WSC). The detection method is based onhttps://chinese.foragelab.com/Resources/Lab-ProceduresSaid method.
2. Determination of silage fermentation products
On days 0, 3, 7 and 14 of silage fermentationBags were opened at day 30 and day 60 for pH, lactic acid, acetic acid and 1, 2-propanediol detection: taking 10g of silage material, adding 90mL of physiological saline, uniformly mixing, and placing in a refrigerator at 4 ℃ for overnight; the pH, lactic acid, acetic acid and 1, 2-propanediol were then measured in a 50mL centrifuge tube using a 4-layer gauze filter. The pH was measured using a pH meter (pH 213; HANNA; italy). Lactic acid, acetic acid and 1, 2-propanediol were detected by HPLC (1200, agilent, america) equipped with a UV detector (210 nm) and a column (ICSep COREGEL-87H), with a mobile phase of 0.005M H at 55deg.C 2 SO 4 The flow rate of the aqueous solution was 0.6mL/min.
3. Culturable microbial flora detection
Opening bags on the 0 th, 3 rd, 7 th, 14 th, 30 th and 60 th days of silage fermentation, taking 10g silage samples, adding 90mL of sterile PBS, oscillating for 30min, filtering the solution by 4 layers of gauze, collecting filtrate (namely bacterial suspension) and placing the filtrate into a 50mL sterile centrifuge tube; the bacterial suspension is diluted and coated, and is respectively coated in MRS, LB and PDA solid culture media for about 36-48 hours for microorganism counting.
4. Microbial analysis method based on high-throughput sequencing
On days 0, 3, 7, 14, 30 and 60 of silage fermentation, bags were opened, 10.0g samples were taken for DNA extraction, SMRT sequencing was performed, and the 16S rDNA full length sequence was determined. Microbial DNA was analyzed by paired-end sequencing (2×300 bp) using Illumina MiSeq platform. Bacterial 16S rDNA amplicon sequencing used V1-V9F (5'-AGAGTTTGATCCTGGCTCAG-3'), V1-V9R (5 '-GNTACCTTGTTACGACTT-3') universal primers. High quality sequences were obtained using the QIIME quality control procedure and chimeric sequences were detected and deleted using the UCHIME algorithm. And obtaining an OUT table, selecting species with the highest abundance ranking of each group on each classification level (Phylum, class, order, family, genus, species) by adopting a maximum ranking method according to the species annotation result, and generating a species relative abundance column accumulation map so as to visually check the microorganism composition, relative abundance and proportion of each group on different classification levels.
5. Results and analysis
1. Influence of the composite silage inoculant on sesbania and sweet sorghum silage chemical components:
the results are shown in table 4, where the mixing ratio of sesbania and sweet sorghum together with silage fermentation time affects DM, CP, NDF, ADF, ADL and WSC of silage. The DM content of the individual combined silage at each time point remained relatively stable throughout the sesbania and sweet sorghum mixed silage process. The CP content and ADL content of each combination silage decreased significantly (P < 0.05) with increasing sweet sorghum addition, and each combination silage CP content and ADL content decreased continuously with fermentation time. The NDF content and ADF content of each combination were not significantly related to the sweet sorghum addition, and each combination remained relatively stable at different time points. The WSC content of each combined silage increased significantly (P < 0.05) with increasing sweet sorghum addition, and the WSC content of each combined silage decreased continuously with fermentation time, which also conforms to silage law.
TABLE 4 variation of DM, CP, NDF, ADF, ADL and WSC content during Mixed silage fermentation of sesbania and sweet sorghum
In Table 4, different lowercase letters indicate that the nutritional quality of the same sample at the same fermentation time is significantly different (p < 0.05), and different uppercase letters indicate that the nutritional quality of the same sample at different fermentation times is significantly different (p < 0.01).
2. Influence of the composite silage inoculant on sesbania and sweet sorghum silage fermentation products:
as shown in fig. 1 and table 5, the pH values of the SS70 combination, the SS50 combination and the SS30 combination all showed a significant tendency to drop within 60 days of silage fermentation, indicating that even if lactic acid bacteria were added, the pH value of the individual sesbania silage fermentation was difficult to drop, whereas the mixed addition of sesbania to sweet sorghum can well exert the advantages of lactic acid bacteria, effectively maintaining the pH of the mixed silage of sesbania and sweet sorghum.
As shown in fig. 2 and table 5, the lactic acid content of the SS30, SS50 and SS70 combinations was higher than that of the SS100 combinations for each time period, and the lactic acid content of the SS30 combination was highest overall, followed by the SS50 combination, within 60 days of silage fermentation.
As shown in fig. 3 and table 5, the silage from SS100 group (100% sesbania group) had the highest acetic acid content at four time points within 30 days of fermentation, followed by SS30 combination. The acetic acid content of the combined silage of SS70, SS50 and SS30 is slightly changed within 14 days of silage fermentation, and the silage is kept stable. After 14 days of silage fermentation, the acetic acid content of the SS100 combined silage was gradually reduced, and the acetic acid content of the SS70, SS50 and SS30 combined silage was rapidly increased.
As shown in fig. 4 and table 5, the 1, 2-propanediol content was detected at day 14 of fermentation, but as the fermentation time increased, the 1, 2-propanediol content of the SS100 combined silage was always 0, the 1, 2-propanediol content of the SS30 combined silage increased rapidly, and the 1, 2-propanediol content of the SS30 combined silage with a high sweet sorghum addition ratio was highest.
TABLE 5 influence of silage fermentation 60 days of Compound silage inoculant on sesbania and sweet sorghum silage fermentation products
/>
Different lowercase letters indicate that the fermentation quality of different samples at the same fermentation time has a significant difference (p < 0.05), and different uppercase letters indicate that the fermentation quality of the same sample at different fermentation times has a significant difference (p < 0.01).
3. Influence of the Complex silage inoculant on sesbania and sweet sorghum silage culturable microbial flora:
as shown in FIG. 5, the amounts of the four combinations of culturable lactic acid bacteria varied approximately equally over 60 days of silage, wherein the amounts of the SS100 combinations of culturable lactic acid bacteria were always highest at each time point.
As shown in FIG. 6, the amounts of the four combinations of culturable yeasts varied approximately equally over 60 days of silage, with the amounts of the SS100 combinations of culturable yeasts being highest at each time point.
As shown in FIG. 7, the tendency of the culturable aerobic bacteria content of the four combinations was approximately the same within 60 days of silage, with the culturable yeast content of the SS100 combination being highest at each time point.
4. Influence of the composite silage inoculant on sesbania and sweet sorghum silage microorganism high-throughput sequencing flora:
as shown in fig. 8, tables 6-8, a bar graph of species relative abundance (species level) of sesbania and sweet sorghum mixed silage bacterial community. Before silage, fresh sesbania samples and fresh sweet sorghum samples are relatively complex in flora, the relative abundance of Bacillus subtilis in the fresh sesbania samples is 41.29%, and the fresh sesbania samples are the most dominant strains, and the relative abundance of Stenotrophomonas maltophilia is 6.06%. Bacillus subtilis in the fresh sweet sorghum sample has a relative abundance of 31.83%, and is the most dominant strain. After silage, all strains in the composite silage microbial inoculum exert dominant advantages, and only have different action time. The homotype fermentation lactobacillus plantarum is dominant in the early fermentation period (before 14 days), the homotype fermentation lactobacillus shi and the lactobacillus buchneri are dominant in the late fermentation period (after 14 days) after replacing the homotype fermentation lactobacillus, and the trend of the above rules in the SS70, SS50 and SS30 combined silage is obvious. Whereas the trend was less pronounced in SS100 combined silage. And after silage fermentation in SS100 combined silage for 60 days, the relative abundance of the three lactobacillus added strains of lactobacillus johnsonii, lactobacillus buchneri and lactobacillus plantarum is 10.63%, 7.29% and 11.54% respectively. The relative abundance of lactobacillus johnsonii in the combined silage of SS70, SS50 and SS30 was 33.13%, 28.75% and 67.58%, respectively, and the relative abundance of lactobacillus buchnsonii in the combined silage of SS70, SS50 and SS30 was 62.46%, 65.93% and 29.47%, respectively.
Table 6 relative abundance of microbial flora before and after silage of mixed sesbania and sweet sorghum
TABLE 7 relative abundance of microbial flora before and after ensiling of Mixed sesbania and sweet sorghum
TABLE 8 relative abundance of microbial flora before and after ensiling of Mixed sesbania and sweet sorghum
The present invention is described in detail above. It will be apparent to those skilled in the art that the present invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with respect to specific embodiments, it will be appreciated that the invention may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The application of some of the basic features may be done in accordance with the scope of the claims that follow.
<110> institute of microorganisms at national academy of sciences
<120> a method for mixed silage of sesbania and sweet sorghum
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1521
<212> DNA
<213> Lactobacillus buchneri (Lactobacillus buchneri)
<220>
<221> misc_feature
<222> (1)..(2)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (136)..(136)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (154)..(154)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (197)..(197)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (225)..(225)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (231)..(233)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (272)..(272)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (371)..(373)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (387)..(387)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (562)..(563)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (582)..(582)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (648)..(648)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (772)..(773)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (777)..(777)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (779)..(779)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (969)..(969)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (1003)..(1004)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (1026)..(1026)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (1424)..(1425)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (1438)..(1440)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (1485)..(1485)
<223> n is a, c, g, or t
<400> 1
nntaagatga gagtttgatc ctggctcagg acgaacgctg gcggcgtgcc taatacatgc 60
aagtcgaacg cgtctccgtt ratgayttta rgtgcttgca yttgaaagat ttaacattga 120
gacgagtggc gaactngtga gtaacacgtg ggtnacctgc ccttgaagta ggggataaca 180
cttggaaaca ggtgctnata ccgtataaca accaaaacct acctngtttt nnnttaaaag 240
acggcttcgg ctgtcacttt aggatggacc cncggcgtat tagcttgttg gtaaggtaac 300
ggcctaccaa ggcgatgata cgtagccgac ctgagagggt aatcggccac attgggactg 360
agacacggcc nnnactccta cgggagncag cagtagggaa tcttccacaa tggacgaaag 420
tctgatggag caacgccgcg tgagtgatga agggtttcgg ctcgtaaaac tctgttgttg 480
gagaagaaca ggtgtcagag yaactgttga catcttgacg gtatccaacc agaaagccac 540
ggctaactac gtgccagcag cnncggtaat acgtaggtgg cnagcgttgt ccggatttat 600
tgggcgtaaa gcgagcgcag gcggtttttt aggtctgatg tgaaagcntt cggcttaacc 660
ggagaagtgc atcggaaacc gggaggcttg agtgcagaag aggacagtgg aactccatgt 720
gtagcggtga aatgcgtaga tatatggaag aacaccagtg gcgaaggcgg cnntctngnc 780
tgtaactgac gctgaggctc gaaagcatgg gtagcgaaca ggattagata ccctggtagt 840
ccatgccgta aacgatgagt gctaagtgtt ggagggtttc cgcccttcag tgctgcagct 900
aacgcattaa gcactccgcc tggggagtac gaccgcaagg ttgaaactca aaggaattga 960
cgggggccng cacaagcggt ggagcatgtg gtttaattcg atnntacgcg aagaacctta 1020
ccaggncttg acatcttctg ccaacctaag agattaggcg ttcccttcgg ggacagaatg 1080
acaggtggtg catggttgtc gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac 1140
gagcgcaacc cttattgtta gttgccagca ttcagttggg cactctagca agactgccgg 1200
tgacaaaccg gaggaaggtg gggatgacgt caaatcatca tgccccttat gacctgggct 1260
acacacgtgc tacaatggac ggtacaacga gtcgcgaaac cgcgaggtca agctaatctc 1320
ttaaagccgt tctcagttcg gattgtaggc tgcaactcgc ctacatgaag ttggaatcgc 1380
tagtaatcgt ggatcagcat gccacggtga atacgttccc gggnnttgta cacaccgnnn 1440
gtcacaccat gagagtttgt aacacccaaa gccggtgagg taacnttcgg ggaccagccg 1500
tctaaggtgg gacagatgat t 1521
Claims (13)
1. A method of preparing sesbania silage comprising: mixing silage raw materials with a compound lactobacillus preparation to obtain a pre-fermentation mixture, carrying out solid anaerobic fermentation on the pre-fermentation mixture, and collecting fermentation products to obtain sesbania silage;
the silage raw material is a mixture composed of sesbania aerial parts and sweet sorghum aerial parts;
the active ingredient of the composite lactobacillus preparation is lactobacillus plantarumLactobacillus plantarum) Lactobacillus johnsoniiLactobacillushilgardii) And Lactobacillus buchneri @Lactobacillus buchneri)；
The Lactobacillus buchneri is @Lactobacillus buchneri) Is Lactobacillus buchneriLactobacillus buchneri) NX205, the Lactobacillus buchneriLactobacillus buchneri) NX205 is a strain with a preservation number of CGMCC No.16534 in China general microbiological culture Collection center.
2. The method according to claim 1, characterized in that: the lactobacillus plantarum is [ ]Lactobacillus plantarum) Is Lactobacillus plantarumLactobacillus plantarum) WQ-01；
The Lactobacillus johnsonii is @ theLactobacillushilgardii) Is Lactobacillus johnsoniiLactobacillushilgardii）60TS-2。
3. The method according to claim 1 or 2, characterized in that: in the composite lactobacillus preparation, the lactobacillus plantarum is preparedLactobacillus plantarum) The Lactobacillus johnsonii isLactobacillushilgardii) And the Lactobacillus buchneriLactobacillus buchneri) The ratio of viable count of (2) is 1:1:1.
4. the method according to claim 1 or 2, characterized in that: the mass percentage of the sesbania overground part in the silage raw material is 30-70%.
5. The method according to claim 1 or 2, characterized in that: in the mixture before fermentation, the silage raw material and the lactobacillus plantarum in the compound lactobacillus preparation are mixedLactobacillus plantarum) The Lactobacillus johnsonii isLactobacillushilgardii) The Lactobacillus buchneri is @Lactobacillus buchneri) The ratio is 1:1:1.
6. the method according to claim 1 or 2, characterized in that: the fermentation is carried out at 15-25 ℃;
and/or the fermentation time is 14-60 days.
7. Sesbania silage prepared by the method of any one of claims 1-6.
8. A complex lactic acid bacteria preparation, which is the complex lactic acid bacteria preparation of any one of claims 1 to 3.
9. A composition for preparing sesbania silage, consisting of the compound lactic acid bacteria preparation of any one of claims 1-3 and the silage raw material of any one of claims 1-4.
10. Lactobacillus buchneri @Lactobacillus buchneri) NX205 has a preservation number of CGMCC No.16534 in China general microbiological culture Collection center.
11. A microbial inoculum comprising the Lactobacillus buchneri as claimed in claim 10 as an active ingredientLactobacillus buchneri)NX205。
12. Use of the method according to any one of claims 1-6 for the preparation of animal feed.
13. The sesbania silage of claim 7, or the compound lactic acid bacteria preparation of claim 8, the composition of claim 9, or the lactobacillus buchneri of claim 10Lactobacillus buchneri) Use of NX205, or the microbial inoculum of claim 11, in the preparation of an animal feed.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110907045.7A CN115702667B (en) | 2021-08-09 | 2021-08-09 | Mixed silage method for sesbania and sweet sorghum |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110907045.7A CN115702667B (en) | 2021-08-09 | 2021-08-09 | Mixed silage method for sesbania and sweet sorghum |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115702667A CN115702667A (en) | 2023-02-17 |
| CN115702667B true CN115702667B (en) | 2024-03-08 |
Family
ID=85179403
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110907045.7A Active CN115702667B (en) | 2021-08-09 | 2021-08-09 | Mixed silage method for sesbania and sweet sorghum |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115702667B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108935972A (en) * | 2018-05-14 | 2018-12-07 | 中国科学院微生物研究所 | A kind of method of feeding soybean and the mixed storage of sugar grass |
-
2021
- 2021-08-09 CN CN202110907045.7A patent/CN115702667B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108935972A (en) * | 2018-05-14 | 2018-12-07 | 中国科学院微生物研究所 | A kind of method of feeding soybean and the mixed storage of sugar grass |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115702667A (en) | 2023-02-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106754557B (en) | Bacillus subtilis YBM-4 and application thereof in preventing and treating tobacco black shank and promoting growth | |
| CN110591988B (en) | Lactobacillus rhamnosus753 and application thereof, silage additive and silage | |
| CN111876351A (en) | Bacillus belgii and application thereof in relieving apple continuous cropping obstacle | |
| CN110643544B (en) | Lactobacillus paracasei761, application thereof, silage additive and silage | |
| CN110591987B (en) | Lactobacillus salivarius358 and application thereof, silage additive and silage | |
| CN102409014A (en) | Ziziphus jujube mill photosphere rhizosphere bacillus sp. capable of promoting growth | |
| CN113215010A (en) | Bacillus belgii ZF128 and application thereof in preventing and treating potato wilt | |
| CN112080435A (en) | Purple lilac spore strain and application thereof in degrading chicken feather | |
| CN111534456A (en) | Composite lactic acid bacteria agent and application thereof in silage of sugarcane tail leaves | |
| CN112725214B (en) | Biogas residue aerobic compost composite microbial agent and application thereof | |
| CN112553103B (en) | Bullwhip grass silage lactobacillus additive and preparation method thereof | |
| CN112877238A (en) | Low-temperature-resistant pediococcus pentosaceus OL77 and application thereof | |
| CN106635926B (en) | Bacillus safensis YJC-4 and application thereof in prevention and treatment of tobacco black shank and growth promotion | |
| CN108060102A (en) | A kind of cibarium Wei Si Salmonellas and its application, screening calibration method | |
| CN114908017B (en) | Lactobacillus plantarum and application thereof in silage | |
| CN115702667B (en) | Mixed silage method for sesbania and sweet sorghum | |
| CN111909877A (en) | Application of Siamese bacillus B11 in prevention and/or treatment of chestnut branch blight | |
| CN113604399B (en) | Sphingolipid bacteria with growth promoting function of garden plants and application thereof | |
| CN106591176B (en) | Lactobacillus pentosus and application thereof | |
| CN114908018A (en) | Lactobacillus brevis and application thereof in silage | |
| CN112715758B (en) | Lactobacillus buchneri BL4 and method for preparing silage by using same | |
| CN115702668A (en) | Method for mixed ensiling of cyperus esculentus stem leaves and rape straws | |
| CN113832071A (en) | Brevibacillus halotolerans strain and application thereof in preparation of biocontrol microbial inoculum | |
| CN111040969A (en) | Compound lactobacillus agent and application thereof in buffalo silage | |
| CN113943667B (en) | Lactobacillus plantarum isolated from camel rumen and application of lactobacillus plantarum in silage |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |