CN118048259A - Endophytic bacterium for promoting growth of tea seedlings and improving tea polyphenol content and application - Google Patents
Endophytic bacterium for promoting growth of tea seedlings and improving tea polyphenol content and application Download PDFInfo
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- CN118048259A CN118048259A CN202410166503.XA CN202410166503A CN118048259A CN 118048259 A CN118048259 A CN 118048259A CN 202410166503 A CN202410166503 A CN 202410166503A CN 118048259 A CN118048259 A CN 118048259A
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- tea
- strain
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- seedlings
- promoting
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Links
- 241001122767 Theaceae Species 0.000 title claims abstract description 104
- 230000012010 growth Effects 0.000 title claims abstract description 40
- 150000008442 polyphenolic compounds Chemical class 0.000 title claims abstract description 28
- 235000013824 polyphenols Nutrition 0.000 title claims abstract description 28
- 230000001737 promoting effect Effects 0.000 title claims abstract description 22
- 241001576300 endophytic bacterium Species 0.000 title description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims abstract description 42
- 229910052698 phosphorus Inorganic materials 0.000 claims abstract description 42
- 239000011574 phosphorus Substances 0.000 claims abstract description 42
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 claims abstract description 24
- 239000002689 soil Substances 0.000 claims abstract description 22
- 239000003617 indole-3-acetic acid Substances 0.000 claims abstract description 13
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 12
- 241000894006 Bacteria Species 0.000 claims abstract description 11
- 241000588912 Pantoea agglomerans Species 0.000 claims abstract description 11
- 230000000813 microbial effect Effects 0.000 claims abstract description 9
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 6
- 238000009629 microbiological culture Methods 0.000 claims abstract description 3
- 230000001965 increasing effect Effects 0.000 claims description 13
- 239000003337 fertilizer Substances 0.000 claims description 9
- 239000001506 calcium phosphate Substances 0.000 claims description 8
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 claims description 8
- 229910000391 tricalcium phosphate Inorganic materials 0.000 claims description 7
- 235000019731 tricalcium phosphate Nutrition 0.000 claims description 7
- 229940078499 tricalcium phosphate Drugs 0.000 claims description 7
- 108020004465 16S ribosomal RNA Proteins 0.000 claims description 6
- WPEXVRDUEAJUGY-UHFFFAOYSA-B hexacalcium;(2,3,4,5,6-pentaphosphonatooxycyclohexyl) phosphate Chemical compound [Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])(=O)OC1C(OP([O-])([O-])=O)C(OP([O-])([O-])=O)C(OP([O-])([O-])=O)C(OP([O-])([O-])=O)C1OP([O-])([O-])=O WPEXVRDUEAJUGY-UHFFFAOYSA-B 0.000 claims description 6
- 238000010521 absorption reaction Methods 0.000 claims description 4
- 239000002054 inoculum Substances 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 235000016709 nutrition Nutrition 0.000 claims description 3
- 230000035764 nutrition Effects 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 2
- 230000001580 bacterial effect Effects 0.000 abstract description 19
- 230000000694 effects Effects 0.000 abstract description 7
- 235000015097 nutrients Nutrition 0.000 abstract description 5
- 239000000126 substance Substances 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 229910019142 PO4 Inorganic materials 0.000 abstract description 2
- 238000004090 dissolution Methods 0.000 abstract description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 abstract description 2
- 239000010452 phosphate Substances 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 18
- 241000196324 Embryophyta Species 0.000 description 12
- 239000001963 growth medium Substances 0.000 description 10
- 235000009024 Ceanothus sanguineus Nutrition 0.000 description 9
- 240000003553 Leptospermum scoparium Species 0.000 description 9
- 235000015459 Lycium barbarum Nutrition 0.000 description 9
- 230000029087 digestion Effects 0.000 description 9
- 238000011282 treatment Methods 0.000 description 9
- 239000008223 sterile water Substances 0.000 description 8
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 238000002835 absorbance Methods 0.000 description 6
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 description 6
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 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 4
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 4
- WYWFMUBFNXLFJK-UHFFFAOYSA-N [Mo].[Sb] Chemical compound [Mo].[Sb] WYWFMUBFNXLFJK-UHFFFAOYSA-N 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 238000002386 leaching Methods 0.000 description 4
- 239000012086 standard solution Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 239000002028 Biomass Substances 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 238000003794 Gram staining Methods 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 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 3
- 229930006000 Sucrose Natural products 0.000 description 3
- 241001052560 Thallis Species 0.000 description 3
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 3
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 3
- 235000011130 ammonium sulphate Nutrition 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 229940074391 gallic acid Drugs 0.000 description 3
- 235000004515 gallic acid Nutrition 0.000 description 3
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 3
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 229930000044 secondary metabolite Natural products 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000005303 weighing 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
- UFBJCMHMOXMLKC-UHFFFAOYSA-N 2,4-dinitrophenol Chemical compound OC1=CC=C([N+]([O-])=O)C=C1[N+]([O-])=O UFBJCMHMOXMLKC-UHFFFAOYSA-N 0.000 description 2
- 239000004382 Amylase Substances 0.000 description 2
- 102000013142 Amylases Human genes 0.000 description 2
- 108010065511 Amylases Proteins 0.000 description 2
- 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 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 2
- 239000012880 LB liquid culture medium Substances 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
- 229930195725 Mannitol Natural products 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 2
- 235000019418 amylase Nutrition 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- REKUHYGYKMBCCW-UHFFFAOYSA-N azane;hydrochloride;hydrofluoride Chemical compound [NH4+].[F-].Cl REKUHYGYKMBCCW-UHFFFAOYSA-N 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
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- RYYVLZVUVIJVGH-UHFFFAOYSA-N caffeine Chemical compound CN1C(=O)N(C)C(=O)C2=C1N=CN2C RYYVLZVUVIJVGH-UHFFFAOYSA-N 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 229940050906 magnesium chloride hexahydrate Drugs 0.000 description 2
- DHRRIBDTHFBPNG-UHFFFAOYSA-L magnesium dichloride hexahydrate Chemical compound O.O.O.O.O.O.[Mg+2].[Cl-].[Cl-] DHRRIBDTHFBPNG-UHFFFAOYSA-L 0.000 description 2
- 239000000594 mannitol Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000004570 mortar (masonry) Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000008635 plant growth Effects 0.000 description 2
- 239000001103 potassium chloride Substances 0.000 description 2
- 235000011164 potassium chloride Nutrition 0.000 description 2
- 238000004382 potting Methods 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241001453380 Burkholderia Species 0.000 description 1
- 241000589989 Helicobacter Species 0.000 description 1
- LPHGQDQBBGAPDZ-UHFFFAOYSA-N Isocaffeine Natural products CN1C(=O)N(C)C(=O)C2=C1N(C)C=N2 LPHGQDQBBGAPDZ-UHFFFAOYSA-N 0.000 description 1
- 241001467578 Microbacterium Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 241000520272 Pantoea Species 0.000 description 1
- 241000983368 Pantoea sp. Species 0.000 description 1
- 239000006004 Quartz sand Substances 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000005708 Sodium hypochlorite Substances 0.000 description 1
- 108010046334 Urease Proteins 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 238000010876 biochemical test Methods 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 229960001948 caffeine Drugs 0.000 description 1
- VJEONQKOZGKCAK-UHFFFAOYSA-N caffeine Natural products CN1C(=O)N(C)C(=O)C2=C1C=CN2C VJEONQKOZGKCAK-UHFFFAOYSA-N 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000010977 jade Substances 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000000643 oven drying Methods 0.000 description 1
- 239000010451 perlite Substances 0.000 description 1
- 235000019362 perlite Nutrition 0.000 description 1
- 235000011007 phosphoric acid Nutrition 0.000 description 1
- 238000013081 phylogenetic analysis Methods 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 230000008121 plant development Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000024053 secondary metabolic process Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000000779 smoke Substances 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 239000010455 vermiculite Substances 0.000 description 1
- 235000019354 vermiculite Nutrition 0.000 description 1
- 229910052902 vermiculite Inorganic materials 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P21/00—Plant growth regulators
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Virology (AREA)
- Environmental Sciences (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Forests & Forestry (AREA)
- Ecology (AREA)
- General Engineering & Computer Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Botany (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Chemical & Material Sciences (AREA)
- Dentistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Agronomy & Crop Science (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides endophytic bacteria for promoting the growth of tea seedlings and improving the content of tea polyphenol and application thereof in preparing microbial agents, wherein the endophytic bacteria are bacterial strain SJ1, classified and named as pantoea agglomerans Pantoea agglomerans, and are preserved in China general microbiological culture collection center (CGMCC) No.29420. The invention also provides a microbial inoculum and application thereof. The strain SJ1 has the characteristics of phosphate dissolution, indoleacetic acid production and the like, can remarkably promote the growth of tea seedlings, increase the content of tea polyphenol in the tea, improve the phosphorus content of rhizosphere soil, root and leaf parts of the tea seedlings, and has important effects of improving the utilization of phosphorus nutrient substances by the tea seedlings, promoting the growth of the tea seedlings and improving the quality of the tea.
Description
Technical Field
The invention relates to the technical field of agricultural microorganisms, in particular to endophytic bacteria for promoting growth of tea seedlings and improving tea polyphenol content and application thereof.
Background
Tea is one of three beverages in the world and is also an important economic crop in China. The tea contains a large amount of secondary metabolites beneficial to human health, including tea polyphenol, caffeine, free amino acids and the like. Wherein, the tea polyphenol is the main component for determining the color, aroma, taste and efficacy of tea, and has various physiological activities such as antioxidation, radiation protection, aging resistance, blood lipid reduction, blood sugar reduction, bacteriostasis and the like.
To increase tea leaf yield and tea polyphenol content, fertilizer is applied in a conventional manner. At present, simple chemical fertilizers and compound fertilizers are mainly applied to tea gardens, and if the chemical fertilizers are applied for a long time, the problems of hardening of tea garden soil, change of the micro-ecological environment of the soil, reduction of tea yield and quality and the like can occur. Therefore, the microbial fertilizer or the microbial inoculum is utilized to produce the organic green pollution-free tea, and the microbial fertilizer or the microbial inoculum has important significance for improving the yield and the quality of the tea and protecting the ecological environment.
Endophytic bacteria, which are long-term present in plants, generally form a stable reciprocal relationship with plants, can affect the growth and development of plants and the synthesis of secondary metabolites. Endophytes can promote plant growth and development by mainly producing indoleacetic acid (IAA), fixing nitrogen, dissolving phosphorus, and enhancing the absorption of soil nutrients by plants. At present, studies on separation and diversity of tea tree endophytes have been reported in literature, and tea tree endophytes mainly include helicobacter, microbacterium, immobilized bacteria, bacillus, burkholderia and the like, but studies on promotion of growth of tea seedlings by microbial species and quality of tea leaves are not much.
The tea tree endophytic bacteria with growth promoting effect are screened, the corresponding microbial inoculum is developed, the dependence on chemical fertilizers in tea garden management is reduced, the growth of tea trees is improved, the utilization of nutrient elements is promoted, meanwhile, the regulation potential of endophytes on plant secondary metabolism can be fully exerted, the synthesis of secondary metabolites in tea trees is effectively promoted, and an innovative agricultural management strategy is provided for sustainable development of tea industry.
Disclosure of Invention
The invention aims to provide endophytic bacteria for promoting growth of tea seedlings and improving tea polyphenol content and application thereof.
The invention adopts the following technical scheme to solve the technical problems:
an endophyte promoting the growth of tea seedlings and increasing the content of tea polyphenols, which endophyte is a strain SJ1, classified and named pantoea agglomerans (Pantoea agglomerans), deposited and presented as surviving in the chinese microbiological bacterial culture collection center at day 27, 12 of 2023, address: the preservation number is CGMCC No.29420 in the Korean region North Star, west Lu No. 1, 3 of Beijing city.
The bacterial strain SJ1 is circular in colony, yellow, smooth and opaque in surface and neat in edge on an LB culture medium; the strain has gram staining negative, amylase negative, contact enzyme positive, mannitol, starch, sucrose, maltose and glucose. The growth is good at 20-35 ℃ after the resistance to 7% sodium chloride.
As one of the preferred modes of the invention, the 16S rDNA sequence of the strain SJ1 is shown as SEQ ID NO. 1.
As one of the preferred modes of the invention, the strain SJ1 is used for dissolving insoluble inorganic phosphorus, organic phosphorus and producing indoleacetic acid.
As one of the preferred modes of the present invention, the poorly soluble inorganic phosphorus and organic phosphorus include tricalcium phosphate and calcium phytate.
As one of the preferable modes of the invention, the strain SJ1 is used for promoting the growth of tea seedlings and increasing the tea polyphenol content of tea leaves.
The application of the endophyte for promoting the growth of tea seedlings and improving the content of tea polyphenol in preparing microbial fertilizer.
A microbial inoculum is prepared from endophytic bacteria for promoting growth of tea seedlings and improving tea polyphenol content.
As one of the preferred modes of the invention, the microbial inoculum is a biological liquid microbial inoculum, and the effective viable count of the strain SJ1 is more than 2 hundred million/ml.
The application of the microbial inoculum is used for promoting the growth of tea seedlings and/or increasing the content of tea polyphenol in tea leaves.
The application of the microbial inoculum is used for increasing the phosphorus content of soil and promoting the absorption and utilization of tea seedlings to phosphorus nutrition.
Compared with the prior art, the invention has the advantages that:
(1) The invention screens out a bacterial strain which can dissolve insoluble inorganic phosphorus and organic phosphorus, the capability of the bacterial strain SJ1 for dissolving tricalcium phosphate reaches 229.29mg/L, the capability for dissolving calcium phytate reaches 272.10mg/L, and the bacterial strain can produce plant growth hormone-indoleacetic acid.
(2) Compared with the unvaccinated control, the strain SJ1 can obviously improve the growth of tea seedlings, and the plant height, the overground fresh weight, the underground fresh weight, the overground dry weight and the underground dry weight are respectively improved by 10.94%,67.09%,35.40%,76.47% and 35.29%.
(3) The strain SJ1 can obviously improve the tea polyphenol content of tea leaves by 18.89%, and can improve the quality of tea leaves while promoting the growth of tea seedlings.
(4) The bacterial strain SJ1 of the invention is inoculated, so that the effective phosphorus content of the rhizosphere soil of the tea seedlings can be improved, the soil fertility can be increased, and the soil ecological environment can be improved.
(5) The total phosphorus content of leaves and roots of tea seedlings can be improved by inoculating the strain SJ 1.
Drawings
FIG. 1 is a diagram of normal growth colonies of strain SJ1 of example 2;
FIG. 2 is a staining microscopic image of strain SJ1 in example 2;
FIG. 3 is a phylogenetic tree of strain SJ1 in example 2;
FIG. 4 is a graph showing the growth states of the potted plants of the treatment group SJ1 and the control group CK in example 5;
fig. 5 is the effect of strain SJ1 on tea polyphenol content in example 5 (in the figure, "x" indicates that there was a significant difference between treatments, p < 0.05);
fig. 6 is the effect of strain SJ1 on soil available phosphorus content in example 6 (in the figure, "x" indicates significant differences between treatments, p < 0.01);
Fig. 7 is the effect of strain SJ1 on leaf phosphorus content of tea seedlings in example 6 (in the figure, "x" indicates significant differences between treatments, p < 0.05);
Fig. 8 is the effect of strain SJ1 on phosphorus content of tea roots in example 6 (in the figure, "x" indicates significant differences between treatments, p < 0.01).
Detailed Description
The following describes in detail the examples of the present invention, which are implemented on the premise of the technical solution of the present invention, and detailed embodiments and specific operation procedures are given, but the scope of protection of the present invention is not limited to the following examples.
Example 1: isolation of strains
The pantoea agglomerans strain SJ1 separated and preserved in the invention is obtained by separating and purifying from tea tree roots, and the specific separation steps are as follows:
The collected tea tree roots are firstly washed by tap water, the surfaces of the tea tree roots are rinsed by sterile water, and the tea tree roots are sheared into small sections with the length of 2 cm to 3cm by scissors. Then, sterilizing the surface of the material on an ultra-clean workbench, sequentially soaking the material in 70% ethanol solution for 1min, soaking the material in 3.5% sodium hypochlorite for 3-4 min, washing the material with sterile water for 3-4 times, and taking 100 mu L of sterile water washed for the last time to be coated on an LB plate as a blank control, wherein after the culture, no bacterial colony grows on the surface of the plate, and the root surface is completely sterilized.
Placing the sterilized in-vitro material into a sterile mortar, adding a little sterile quartz sand for grinding, and adding sterile water for gradient dilution by 10 -1、10-2、10-3 times; 100. Mu.L of each gradient was plated on LB plates and incubated in an incubator at 30℃with inversion. When colonies appear on the flat plate, single colonies with vigorous growth, different sizes and different forms are selected, streaked on a new LB solid culture medium flat plate, cultured for 48 hours at 30 ℃, repeated transfer operation is carried out for 3 times, after the sterility is determined, the single colonies are selected and connected into an LB liquid culture medium, and placed in a shaking table, and cultured for 28-36 hours at 28 ℃ at 180 r/min; finally, the strain is preserved at-80 ℃ with 40% glycerol suspension for standby.
Example 2: identification of Strain SJ1
The physiological and biochemical characteristics of the strain SJ1 are detected according to the common bacteria System identification Manual.
When the strain SJ1 grows in LB culture medium, as shown in figure 1, bacterial colonies are yellow, have neat edges, smooth and moist surfaces and are opaque. The bacteria were observed under a microscope to be rod-shaped, without spores (FIG. 2). The physiological and biochemical characteristics of the strain: the strain has gram staining negative, amylase negative, contact enzyme positive and urease negative, and mannitol, starch, sucrose, maltose and glucose can be used. The growth is good at 20-35 ℃ after the resistance to 7% sodium chloride.
The bacterial strain SJ1 is subjected to extraction of genomic DNA, and then the DNA is subjected to PCR amplification by using universal primers 27F and 1492R of bacterial 16S rRNA genes. The PCR amplified product obtained was sequenced. BLAST analysis was performed on strain 16S rDNA sequences using NCBI nucleic acid database. The analysis results show that: the strain SJ1 belongs to Pantoea (Pantoea sp.) and has the homology of 99.1% -99.7% with the Pantoea agglomerans 16S rDNA sequence.
Referring to FIG. 3, phylogenetic analysis, strain SJ1 was found to cluster into a large branch with Pantoea agglomerans. Further in combination with the results of gram staining and other biochemical tests, it was shown that: the strain SJ1 is a pantoea agglomerans (Pantoea agglomerans), is named Pantoea agglomerans SJ and is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.29420. The 16S rDNA sequence of strain Pantoea agglomerans SJ is shown as SEQ ID NO. 1.
Example 3: determination of the phosphate-solubilizing ability and indoleacetic acid (IAA) production of Strain SJ1
1. Phosphate-solubilizing ability of strain SJ1
The bacterial strain SJ1 screened by the invention is inoculated into a test tube, shake-cultured for 24-30 hours by a shaking table, and then respectively inoculated into 100mL of inorganic phosphorus culture medium (10.0 g of glucose, 0.1g of ammonium sulfate, 0.2g of potassium chloride, 0.25g of magnesium sulfate heptahydrate, 5.0g of magnesium chloride hexahydrate, 5.0g of tricalcium phosphate, 1000mL of distilled water, pH 7.0, 115 ℃ C., sterilization for 30 min) containing tricalcium phosphate, and 100mL of organic phosphorus culture medium (10.0 g of glucose, 0.1g of ammonium sulfate, 0.2g of potassium chloride, 0.25g of magnesium sulfate heptahydrate, 5.0g of magnesium chloride hexahydrate, 5.0g of calcium phytate, 1000mL of distilled water, pH 7.0, 115 ℃ C., 30 min) containing tricalcium phosphate according to the inoculum size of 2 percent, and shake-cultured for 3 days at 28-30 ℃ C. Centrifuging the culture solution at 8000r/min, taking 20 mu L of supernatant, adding 1mL of molybdenum-antimony anti-chromogenic agent, fixing the volume to 10mL by distilled water, developing for 15-20 min, and measuring an OD value at 700 nm. A standard curve was prepared by gradient dilution of 5mg/L of the phosphorus standard solution to phosphorus contents of 0.00, 0.25, 0.50, 1.00, 1.25 and 1.50 mg/L. Adding molybdenum-antimony anti-color developing agent at room temperature, developing for 15-20 min, measuring the light absorption value and preparing a standard curve. The standard curve was determined to be y=0.5191x+0.0218 and the correlation coefficient R 2 = 0.9976.
According to the standard curve, the phosphorus content in the culture solution of the bacterial strain SJ1 containing tricalcium phosphate is 229.29mg/L, and the phosphorus content in the culture solution of the bacterial strain SJ1 containing calcium phytate is 272.10mg/L. The strain SJ1 has strong capability of dissolving calcium phosphate and calcium phytate, and can convert insoluble phosphorus into soluble phosphorus.
2. IAA secretion amount measurement of Strain SJ1
Standard curves were prepared using analytically pure IAA, 0.2mg/mL IAA standard was diluted in gradient to 0, 5, 10, 15, 20, 25, 30mg/L concentrations and absorbance (OD 530) was determined using Salkowski chromogenic method. The standard curve is obtained as y=0.0275x+0.0131, and the correlation coefficient R 2 =0.9993.
Nitrogen culture medium (10.0 g of sucrose, 1.0g of ammonium sulfate, 2.0g of dipotassium hydrogen phosphate, 0.5g of magnesium sulfate heptahydrate, 0.1g of sodium chloride, 0.5g of yeast extract, pH 7.2 and 1000mL of distilled water) is subpackaged in test tubes, 4mL of each tube is sterilized, and 1mL of filtered and sterilized tryptophan is added to make the final concentration of tryptophan in the culture medium be 0.5mg/mL. Inoculating the target strain to the culture medium, culturing for 2d at 150r/min at the temperature of 28 ℃ in a shaking table, centrifuging the culture solution at 6000r/min for 10min, taking 1mL of supernatant, adding 50 mu L of 10mM/L orthophosphoric acid, adding 2mL of Sackowski's color reagent, fully mixing, developing color for 30min at the temperature of 25 ℃ in the dark, and measuring absorbance at 530 nm.
As shown by the standard curve, IAA content in the culture solution of the added strain SJ1 is 27.46mg/L.
Example 4: activation of strain SJ1 and preparation of bacterial suspension
A loop was picked from SJ1 glycerol tube strain with an inoculating loop, streaked onto LB solid medium by aseptic manipulation, and cultured at 28℃for 72h. Then selecting single colony of the SJ1 strain with vigorous growth, then marking the single colony on a new LB solid culture medium plate in three areas, and after culturing, selecting the single colony of the SJ1 strain to be inoculated in an LB liquid culture medium, and carrying out shaking culture for 20-30 h at 28 ℃ by a shaking table at 150-180 rpm.
Taking 100mL of culture solution containing the strain SJ1, centrifuging at 8000r/min to collect thalli, cleaning the thalli with sterile water, and then re-suspending thalli cells in 80-100 mL of sterile water to enable the cell number to reach more than 2X 10 8 CFU/mL, thus obtaining the strain SJ1 liquid microbial inoculum.
Example 5: strain SJ1 promotes growth of tea seedlings and improves tea polyphenol content
1. Potting experiments and biomass
Annual cuttage tea seedlings (medium tea 108, height about 11 cm) with the same growth size are selected and planted in flowerpots (height 15cm, diameter 25 cm). The nutrient soil for the growth of the tea seedlings is mainly a floating jade plate plant culture medium and is mixed with vermiculite and perlite according to the mass ratio of 2:1:1 are mixed evenly and then are subpackaged in flowerpots, and each flowerpot is subpackaged with 3kg. 2mL of the above bacterial suspension was applied near the root of each seedling as a treatment group (SJ 1), while the control group (CK) was applied with an equal volume of sterile water. Normally growing under natural conditions, watering at regular intervals, planting for 2 half months (about 75 days), and harvesting tea seedlings to measure plant height, fresh weight above ground, fresh weight below ground, dry weight above ground and dry weight below ground.
TABLE 1 Effect of Strain SJ1 on tea seed biomass
Note that: * Indicating significant differences between horizontal treatments with p <0.05
The growth status of the Control (CK) and treatment (SJ 1) potting is shown in fig. 4 and the final biomass measurements are shown in table 1.
As can be seen from table 1: after the strain is inoculated, the growth of tea seedlings is obviously enhanced, and compared with a control, the plant height, the overground fresh weight, the underground fresh weight, the overground dry weight and the underground dry weight are obviously improved by 10.94%,67.09%,35.40%,76.47% and 35.29%, and all reach obvious difference levels.
2. Tea polyphenol content determination
Collecting two leaves of one bud of tea seedling, deactivating enzyme at 105deg.C, and oven drying to constant weight. Grinding a dried sample in a mortar, weighing the sample, placing the sample in a centrifuge tube, adding the centrifuge tube into 70% methanol aqueous solution preheated at 70 ℃, fully stirring and wetting the sample uniformly by a glass rod, immediately transferring the sample into a water bath at 70 ℃, leaching for 10min, cooling the sample to room temperature after leaching, transferring the sample into a centrifugal machine for centrifugation at 8000rpm for 10min, transferring the supernatant into a 10mL volumetric flask, extracting residues once again by using 5mL of 70% methanol aqueous solution, repeating the above operation, and combining the extracting solutions to a constant volume of 10mL. After diluting the extract by 100 times, 1.0mL of the diluent is removed and placed in a graduated test tube, 5.0mL of Fu Lin Fen reagent is added and the mixture is shaken well. And (3) adding 4.0mL of 7.5% sodium carbonate solution into the reaction solution within 3 to 8 minutes, adding pure water to fix the volume to a scale, and shaking the solution uniformly. The mixture was left at room temperature for 60min. The absorbance at 765nm was measured.
The standard curve is prepared by adopting gallic acid, 1000 mug/mL of gallic acid standard solution is diluted to 10 mug/mL, 20 mug/mL, 30 mug/mL, 40 mug/mL and 50 mug/mL of gallic acid standard solution is subjected to gradient dilution, and the absorbance (OD 765) is measured by adopting Fu Lin Fenfa, so that the standard curve is y=0.0044x+0.0041, and the correlation coefficient R 2 =0.997.
Substituting the absorbance at 765nm measured by the sample into a regression equation, calculating according to dilution times, sample mass and the like, and drawing to obtain the graph 5. As can be seen from fig. 5: after inoculation of strain SJ1, the tea polyphenol content was significantly increased by 18.89% (p < 0.05) compared to the unvaccinated control group (CK).
Example 6: the bacterial strain SJ1 improves the phosphorus content of soil available phosphorus and tea seedlings
1. Soil pot culture
Annual cuttage tea seedlings (medium tea 108, about 11cm in height) with the same growth size are selected in the experiment, and planted in flowerpots (15 cm in height and 25cm in diameter). The soil to be tested is tea garden soil obtained from the county of Huoshan in Liu an of Anhui province, and 3kg of soil is packed in each pot. After transplanting the tea seedlings, 2mL of the above bacterial suspension (2X 10 8～9×108 CFU/mL) was applied near the root of each seedling as a treatment group (SJ 1), while the control group (CK) was applied with an equal volume of sterile water. The tea seedlings grow outdoors, are watered in proper amount regularly, are planted for more than 2 half months (about 80 to 85 days), the rhizosphere soil and the tea seedlings are collected, and the effective phosphorus content of the soil and the phosphorus content of roots and leaves of the tea seedlings are measured.
2. Soil available phosphorus determination
Determination of soil available phosphorus by ammonium fluoride-hydrochloric acid leaching method: weighing an air-dried sample passing through a 2mm sieve opening, placing the air-dried sample in a 200mL plastic bottle, adding an ammonium fluoride-hydrochloric acid leaching agent, shaking for 30min at 25 ℃, filtering with a phosphorus-free filter paper, sucking 10mL of filtrate in a 50mL volumetric flask, adding 10mL of boric acid solution, shaking uniformly, adding water to about 30mL, adding 2 drops of a dinitrophenol indicator, regulating the solution to be light yellow with sulfuric acid solution and ammonia water solution, adding 5mL of molybdenum-antimony anti-chromogenic agent, fixing the volume with pure water, standing for 30min at room temperature higher than 20 ℃, using a 1cm optical path cuvette at a wavelength of 700nm, taking a standard solution as a zero point, and performing colorimetric determination after zeroing. Substituting the measured value into a standard curve regression equation to calculate the phosphorus content in the sample.
The results are shown in FIG. 6. As can be seen from fig. 6: compared with the control group (CK) without inoculation, after the strain SJ1 is applied, the effective phosphorus content of the rhizosphere soil of the tea seedlings is obviously improved by 9.25 percent (p < 0.01).
3. The bacterial strain SJ1 improves the phosphorus content of the leaf part and the root part of the tea seedling
Accurately weighing uniformly-mixed and ground dry plant samples (leaf and root samples of tea seedlings) respectively, placing the dry plant samples in a digestion tube, adding concentrated sulfuric acid, slightly and uniformly shaking, standing overnight, covering a neck-bending small funnel, firstly digesting at low temperature on the digestion furnace until white smoke disappears, taking down the digestion tube, slightly cooling, and dropwise adding 30% H 2O2 about 0.5mL; continuously and lightly shaking the digestion tube, continuously heating and slightly boiling for about 5min, taking down and slightly cooling, repeatedly adding a few drops of H 2O2, and then digestion, repeating the steps for a plurality of times (about 2-3 times), wherein each time of adding H 2O2 is gradually reduced, and continuously digesting for 5-15 min after the digestion is colorless or clear, so as to remove the residual H 2O2. The digestion tube is removed, cooled, the neck funnel is rinsed with a small amount of water, and the rinsing liquid flows into the digestion tube. The digestion solution was transferred completely intact to a 100mL volumetric flask and fixed in volume. Filtering with quantitative filter paper to a triangular flask, sucking a certain amount of filtrate into a 50mL volumetric flask, adding water to about 30mL, adding 2 drops of dinitrophenol indicator, regulating the solution to be slightly yellow with sodium hydroxide solution or sulfuric acid solution, then adding 5mL of molybdenum-antimony color-developing inhibitor, and fixing the volume. Placing at 20deg.C or above for 30min for color development, and measuring and recording absorbance at 700 nm.
The results are shown in fig. 7 and 8. As can be seen from fig. 7 and 8: compared with the unvaccinated control group (CK), after the strain SJ1 is applied, the total phosphorus content of leaves and roots of the tea seedlings is obviously improved by 5.10% and 18.60%, respectively. The application of the strain SJ1 can promote the absorption and utilization of the tea seedlings to the phosphorus nutrition.
In conclusion, the strain SJ1 has the characteristics of phosphate dissolution, indoleacetic acid production and the like, can remarkably promote the growth of tea seedlings, increase the content of tea polyphenol in the tea, and increase the phosphorus content in rhizosphere soil, root and leaf parts of the tea seedlings, and has important effects of improving the utilization of phosphorus nutrient substances by the tea seedlings, promoting the growth of the tea seedlings and improving the quality of the tea.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, and alternatives falling within the spirit and principles of the invention.
Claims (10)
1. The endophyte is a strain SJ1, is classified and named as Pantoea agglomerans Pantoea agglomerans, and is preserved in China general microbiological culture collection center (CGMCC) No.29420.
2. Endophyte promoting tea seedling growth and increasing tea polyphenol content according to claim 1, wherein the 16S rDNA sequence of strain SJ1 is shown in SEQ ID No. 1.
3. Endophyte promoting tea seedling growth and increasing tea polyphenol content according to claim 1, wherein strain SJ1 is used for dissolving insoluble inorganic phosphorus, organic phosphorus, and producing indoleacetic acid.
4. Endophyte promoting tea seedling growth and increased tea polyphenol content as claimed in claim 3 wherein the poorly soluble inorganic and organic phosphorus comprises tricalcium phosphate and calcium phytate.
5. Endophyte promoting tea seedling growth and increasing tea polyphenol content according to claim 1, wherein strain SJ1 is used to promote tea seedling growth and increase tea polyphenol content of tea leaves.
6. Use of an endophyte according to any one of claims 1 to 5 to promote the growth of tea seedlings and increase the content of tea polyphenols in the preparation of a microbial fertilizer.
7. A microbial inoculum characterized in that it is prepared by using endophytic bacteria for promoting growth of tea seedlings and increasing tea polyphenol content according to any one of claims 1 to 6.
8. The microbial inoculant according to claim 7, wherein the microbial inoculant is a biological liquid microbial inoculant and the effective viable count of the strain SJ1 is greater than 2 hundred million/ml.
9. Use of a microbial agent according to claim 7 or 8 for promoting the growth of tea seedlings and/or increasing the tea polyphenol content of tea leaves.
10. Use of the microbial inoculum according to claim 7 or 8 for increasing phosphorus content in soil and promoting absorption and utilization of phosphorus nutrition by tea seedlings.
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