CN104396735A - Method for eliminating bacterial contamination in Momordica grosvenori tissue culture - Google Patents
Method for eliminating bacterial contamination in Momordica grosvenori tissue culture Download PDFInfo
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- CN104396735A CN104396735A CN201410488635.0A CN201410488635A CN104396735A CN 104396735 A CN104396735 A CN 104396735A CN 201410488635 A CN201410488635 A CN 201410488635A CN 104396735 A CN104396735 A CN 104396735A
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- luohanguo
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- germ contamination
- rejuvenation
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- 241001409321 Siraitia grosvenorii Species 0.000 title claims abstract description 60
- 238000011109 contamination Methods 0.000 title claims abstract description 40
- 238000000034 method Methods 0.000 title claims abstract description 35
- 235000011171 Thladiantha grosvenorii Nutrition 0.000 title claims abstract description 17
- 230000001580 bacterial effect Effects 0.000 title abstract 3
- 239000001963 growth medium Substances 0.000 claims abstract description 47
- 241000894006 Bacteria Species 0.000 claims abstract description 45
- 230000003716 rejuvenation Effects 0.000 claims abstract description 31
- 102000016943 Muramidase Human genes 0.000 claims abstract description 18
- 108010014251 Muramidase Proteins 0.000 claims abstract description 18
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 claims abstract description 18
- 239000004325 lysozyme Substances 0.000 claims abstract description 18
- 229960000274 lysozyme Drugs 0.000 claims abstract description 18
- 235000010335 lysozyme Nutrition 0.000 claims abstract description 18
- 229920001817 Agar Polymers 0.000 claims description 18
- 239000008272 agar Substances 0.000 claims description 18
- 239000011941 photocatalyst Substances 0.000 claims description 15
- 239000006870 ms-medium Substances 0.000 claims description 14
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 claims description 14
- 229930006000 Sucrose Natural products 0.000 claims description 13
- 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 claims description 13
- 239000005720 sucrose Substances 0.000 claims description 13
- NWBJYWHLCVSVIJ-UHFFFAOYSA-N N-benzyladenine Chemical compound N=1C=NC=2NC=NC=2C=1NCC1=CC=CC=C1 NWBJYWHLCVSVIJ-UHFFFAOYSA-N 0.000 claims description 12
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 11
- 229910052799 carbon Inorganic materials 0.000 claims description 11
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 10
- 238000012258 culturing Methods 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 10
- 238000005286 illumination Methods 0.000 claims description 8
- 238000011534 incubation Methods 0.000 claims description 8
- 229960002727 cefotaxime sodium Drugs 0.000 claims description 7
- AZZMGZXNTDTSME-JUZDKLSSSA-M cefotaxime sodium Chemical compound [Na+].N([C@@H]1C(N2C(=C(COC(C)=O)CS[C@@H]21)C([O-])=O)=O)C(=O)\C(=N/OC)C1=CSC(N)=N1 AZZMGZXNTDTSME-JUZDKLSSSA-M 0.000 claims description 7
- 229930027917 kanamycin Natural products 0.000 claims description 7
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- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 claims description 7
- 229930182823 kanamycin A Natural products 0.000 claims description 7
- 229960005322 streptomycin Drugs 0.000 claims description 7
- 239000004471 Glycine Substances 0.000 claims description 5
- JTEDVYBZBROSJT-UHFFFAOYSA-N indole-3-butyric acid Chemical compound C1=CC=C2C(CCCC(=O)O)=CNC2=C1 JTEDVYBZBROSJT-UHFFFAOYSA-N 0.000 claims description 5
- YGGXZTQSGNFKPJ-UHFFFAOYSA-N methyl 2-naphthalen-1-ylacetate Chemical compound C1=CC=C2C(CC(=O)OC)=CC=CC2=C1 YGGXZTQSGNFKPJ-UHFFFAOYSA-N 0.000 claims description 5
- IMQLKJBTEOYOSI-GPIVLXJGSA-N Inositol-hexakisphosphate Chemical compound OP(O)(=O)O[C@H]1[C@H](OP(O)(O)=O)[C@@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@@H]1OP(O)(O)=O IMQLKJBTEOYOSI-GPIVLXJGSA-N 0.000 claims description 4
- 229910019142 PO4 Inorganic materials 0.000 claims description 4
- IMQLKJBTEOYOSI-UHFFFAOYSA-N Phytic acid Natural products OP(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 IMQLKJBTEOYOSI-UHFFFAOYSA-N 0.000 claims description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 4
- 239000010452 phosphate Substances 0.000 claims description 4
- 235000002949 phytic acid Nutrition 0.000 claims description 4
- 229940068041 phytic acid Drugs 0.000 claims description 4
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- 238000004519 manufacturing process Methods 0.000 abstract description 3
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- 230000003197 catalytic effect Effects 0.000 abstract description 2
- 239000003242 anti bacterial agent Substances 0.000 abstract 1
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- 230000008030 elimination Effects 0.000 description 12
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- 241000196324 Embryophyta Species 0.000 description 3
- 150000004676 glycans Chemical class 0.000 description 3
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
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- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 2
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- DQJCDTNMLBYVAY-ZXXIYAEKSA-N (2S,5R,10R,13R)-16-{[(2R,3S,4R,5R)-3-{[(2S,3R,4R,5S,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}-5-(ethylamino)-6-hydroxy-2-(hydroxymethyl)oxan-4-yl]oxy}-5-(4-aminobutyl)-10-carbamoyl-2,13-dimethyl-4,7,12,15-tetraoxo-3,6,11,14-tetraazaheptadecan-1-oic acid Chemical compound NCCCC[C@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)CC[C@H](C(N)=O)NC(=O)[C@@H](C)NC(=O)C(C)O[C@@H]1[C@@H](NCC)C(O)O[C@H](CO)[C@H]1O[C@H]1[C@H](NC(C)=O)[C@@H](O)[C@H](O)[C@@H](CO)O1 DQJCDTNMLBYVAY-ZXXIYAEKSA-N 0.000 description 1
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
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- 241000233866 Fungi Species 0.000 description 1
- 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 1
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- 102000002068 Glycopeptides Human genes 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 206010062717 Increased upper airway secretion Diseases 0.000 description 1
- 235000019082 Osmanthus Nutrition 0.000 description 1
- 241000333181 Osmanthus Species 0.000 description 1
- 241000505673 Scintilla Species 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 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 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
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- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
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- 238000009395 breeding Methods 0.000 description 1
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- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
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- 210000004027 cell Anatomy 0.000 description 1
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- -1 di(2-ethylhexyl)phosphate titanium oxide compound Chemical class 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- KYQODXQIAJFKPH-UHFFFAOYSA-N diazanium;2-[2-[bis(carboxymethyl)amino]ethyl-(carboxylatomethyl)amino]acetate Chemical compound [NH4+].[NH4+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O KYQODXQIAJFKPH-UHFFFAOYSA-N 0.000 description 1
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- 238000005516 engineering process Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 239000011790 ferrous sulphate Substances 0.000 description 1
- 235000003891 ferrous sulphate Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229930182478 glucoside Natural products 0.000 description 1
- 150000008131 glucosides Chemical class 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 229940064880 inositol 100 mg Drugs 0.000 description 1
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
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- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
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- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 229930189775 mogroside Natural products 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
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- 229960003512 nicotinic acid Drugs 0.000 description 1
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- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 239000004323 potassium nitrate Substances 0.000 description 1
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- 239000011734 sodium Substances 0.000 description 1
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- 239000011684 sodium molybdate Substances 0.000 description 1
- 235000015393 sodium molybdate Nutrition 0.000 description 1
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
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- 235000019190 thiamine hydrochloride Nutrition 0.000 description 1
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- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
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- 239000011686 zinc sulphate Substances 0.000 description 1
- 235000009529 zinc sulphate Nutrition 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for eliminating bacterial contamination in Momordica grosvenori tissue culture. The method comprises the following steps: 1, preparing an MS bacteria eliminating culture medium for Momordica grosvenori tissue culture; 2, inoculating multiple shoots contaminated by bacteria from Momordica grosvenori tissue culture into the MS bacteria eliminating culture medium, conducting bacteria eliminating culture to obtain sterile multiple shoots from tissue culture; 3, preparing an MS rejuvenation medium for Momordica grosvenori tissue culture; and 4, inoculating the obtained sterile multiple shoots from tissue culture into the MS rejuvenation culture medium to obtain healthy tissue culture seedling, wherein the MS bacteria eliminating culture medium is supplemented with antibiotics, non-light catalytic agent and lysozyme, and after bacteria eliminating culture, the influence of bacterial pollution on multiple seedlings can be eliminated in a short time. The invention can healthy seed source guarantee for germplasm preservation and industrialized production.
Description
Technical field
The present invention relates to field of tissue culture, particularly relate to a kind of method eliminating germ contamination in Luohanguo cultivation.
Background technology
Momordica grosvenori is the medicine-food two-purpose rare traditional Chinese medicine that the Ministry of Public Health announces in the first batch, fruits nutrition is worth very high, containing abundant vitamin C and glucoside, fructose, glucose, protein, lipid etc., mogroside is a kind of natural sweetener of high sugariness low in calories, its sugariness can be rated as the first in the world, and low-heat, nontoxic, can be diabetes and bariatric patients etc. edible.Momordica grosvenori is sweet, sour, cool in nature, there is clearing heat and cooling blood, cough-relieving of promoting the production of body fluid, laxation toxin expelling, tender skin benefit effect such as face, moistening lung for removing phlegm, be the famous genunie medicinal materials in Guangxi.Guangxi Momordica grosvenori output accounts for more than 90% of the world, becomes the specialty industries that north, osmanthus is important, has obtained the protection of national geography famous special product.In recent years, increasing enterprise of China joins the ranks producing Momordica grosvenori, and Luohanguo With Plantlets of Tissue Culture market demand increases, and tissue cultures is the important means of Momordica grosvenori industrialized development, now achieves suitability for industrialized production.
MS medium is the minimal medium of most plants tissue-culturing quick-propagation.Its formula is as follows usually for MS tissue culture medium (TCM) of the prior art: macroelement: potassium nitrate 1900mg/L, ammonium nitrate 1650mg/L, calcium chloride 440mg/L, magnesium sulfate 370mg/L, potassium dihydrogen phosphate 170mg/L; Trace element: manganese sulphate 22.3mg/L, zinc sulphate 8.6mg/L, boric acid 6.3mg/L, copper sulphate 0.025mg/L, potassium iodide 0.83mg/L, cobalt chloride 0.025mg/L, sodium molybdate 0.25mg/L; Molysite: ferrous sulfate 27.8mg/L, b diammonium edta sodium 37.3mg/L; Organic principle: thiamine hydrochloride 0.1mg/L, glycine 2.0mg/L, inositol 100mg/L, nicotinic acid 0.5mg/L, puridoxine hydrochloride 0.5mg/L, sucrose 30000mg/L, agar 7000mg/L.
MS medium can meet nutrition and the physiological requirements of plant cell, and thus the scope of application is very wide, is the minimal medium of most plants tissue-culturing quick-propagation.Adopt in subculture and process of rooting culture in basic MS culture medium breeding process and can run into a large amount of germ contamination and produce vicious circle, time serious, media surface occurs that white or yellow water ooze sample material, make the plantlet in vitro poor growth of pollution, neither breed and also do not take root, even dead, test material cannot be preserved, and brings huge loss to follow-up test and seedling popularization.
The analog of the secondary metabolite that antibiotic is mainly produced by bacterium, mould or other microorganisms or Prof. Du Yucang, at low concentrations just can the synthesis of anti-bacteria cell wall, the synthesis of interferencing protein and suppress transcribing and copying of nucleic acid, have bacterium and well suppress and killing action.
Super nanometer scintilla di(2-ethylhexyl)phosphate titanium oxide compound without photocatalyst main component, not only all can produce strong catalytic degradation effect at no light condition but also under any condition, effectively can kill various bacteria, antibiotic rate is up to 99.99%, and the toxin that bacterium or fungi can be discharged decomposes and harmless treatment.
The main component of bacteria cell wall is peptide glycan, lysozyme is the alkaline enzyme of glutinous polysaccharide in a kind of energy hydrolytic bacteria, mainly through destroying the glycosidic bond in cell wall, the insoluble glutinous polysaccharide of bacteria cell wall is made to resolve into solubility glycopeptide, cell wall rupture content is caused to overflow and make bacterolysis, and lysozyme and phytic acid, polymeric phosphate, glycine etc. with the use of, have synergistic effect to lysozyme.
Summary of the invention
The invention provides a kind of method eliminating germ contamination in Luohanguo cultivation.Momordica grosvenori go add antibiotic in bacterium culture medium, remove without photocatalyst, lysozyme Luohanguo cultivate in germ contamination, impact on tufted seedling can be polluted, for preserving seed and factorial praluction provide provenance to ensure by eradicate bacteria at short notice.Active carbon is added in rejuvenation culture medium, utilize the strong suction-operated of active carbon, adsorbable residual antibiotic and other hormone, to alleviate the impact of antibiotic use on Multiple Buds, return taking root and growing the dark surrounds built under approximate natural growthing condition of culture, make that Multiple Buds stem segment length is high, stem stalk is sturdy, mounted blade, leaf look dark green.
Technical scheme provided by the invention is:
Eliminate a method for germ contamination in Luohanguo cultivation, comprise the following steps:
Step one: the MS required for the cultivation of preparation Luohanguo removes bacterium culture medium;
Step 2: the Momordica grosvenori group training Multiple Buds by germ contamination is inoculated into MS and goes in bacterium culture medium, through past bacterium cultivation, obtains aseptic group training Multiple Buds;
Step 3: the MS rejuvenation culture medium required for the cultivation of preparation Luohanguo;
Step 4: the aseptic group training Multiple Buds obtained is inoculated in MS rejuvenation culture medium, cultivates through rejuvenation, obtain healthy tissue-cultured seedling;
Described MS goes also to be added with without photocatalyst and lysozyme in bacterium culture medium, is added with active carbon in MS rejuvenation culture medium.
Preferably, in the method for germ contamination in described elimination Luohanguo cultivation, described MS goes bacterium culture medium to be the agar adding the 6-benzylaminopurine of 0.5mg/L, the kanamycin of 20-120mg/L, the Cefotaxime Sodium of 2-20mg/L and the streptomycin of 5-50mg/L, the sucrose of 30g/L and 4.5g/L in basic MS medium, and its pH value is 5.8.
Preferably, in the method for germ contamination in described elimination Luohanguo cultivation, described MS goes bacterium culture medium to be the agar adding the 6-benzylaminopurine of 0.5mg/L, the kanamycin of 90mg/L, the Cefotaxime Sodium of 10mg/L and the streptomycin of 25mg/L, the sucrose of 30g/L and 4.5g/L in basic MS medium, and its pH value is 5.8.
Preferably, described elimination Luohanguo cultivate in germ contamination method in, described MS go also to be added with in bacterium culture medium in phytic acid that mass fraction is 0.01-0.05%, polymeric phosphate, glycine any one or a few.
Preferably, in the method for germ contamination in described elimination Luohanguo cultivation, described MS rejuvenation culture medium is the agar adding the 6-benzylaminopurine of 0.05mg/L, the methyl α-naphthyl acetate of 0.01mg/L, the indolebutyric acid of 0.01mg/L, the sucrose of 30g/L and 4.5g/L in basic MS medium, and its pH value is 5.8.
Preferably, described elimination Luohanguo cultivate in germ contamination method in, described in go bacterium to cultivate and in rejuvenation cultivation, the temperature of culturing room is 24-27 DEG C, and intensity of illumination is 2000lux, and light application time is 10-12 hour/day, and incubation time is 30 days.
Preferably, in the method for germ contamination in described elimination Luohanguo cultivation, described is add before described agar adds without photocatalyst and lysozyme.
Preferably, in the method for germ contamination in described elimination Luohanguo cultivation, the concentration of described active carbon is 1.0g/L.
Preferably, in the method for germ contamination in described elimination Luohanguo cultivation, the described mass fraction without photocatalyst is 0.5-1.0%.
Preferably, in the method for germ contamination in described elimination Luohanguo cultivation, the mass fraction of described lysozyme is 0.03-0.07%.
The invention provides a kind of method eliminating germ contamination in Luohanguo cultivation.Momordica grosvenori go add antibiotic in bacterium culture medium, remove without photocatalyst, lysozyme Luohanguo cultivate in germ contamination, impact on tufted seedling can be polluted, for preserving seed and factorial praluction provide provenance to ensure by eradicate bacteria at short notice.Active carbon is added in rejuvenation culture medium, utilize the strong suction-operated of active carbon, adsorbable residual antibiotic and other hormone, to alleviate the impact of antibiotic use on Multiple Buds, return taking root and growing the dark surrounds built under approximate natural growthing condition of culture, make that Multiple Buds stem segment length is high, stem stalk is sturdy, mounted blade, leaf look dark green.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, can implement according to this with reference to specification word to make those skilled in the art.
Embodiment 1:
Eliminate a method for germ contamination in Luohanguo cultivation, comprise the following steps:
Step one: the MS required for the cultivation of preparation Luohanguo removes bacterium culture medium, described MS goes bacterium culture medium to be the streptomycin, the polymeric phosphate of 0.05%, the agar without photocatalyst, the lysozyme of 0.03%, the sucrose of 30g/L and 4.5g/L of 0.5% that add the 6-benzylaminopurine of 0.5mg/L, the kanamycin of 20mg/L, the Cefotaxime Sodium of 2mg/L and 5mg/L in basic MS medium, its pH value is 5.8, the temperature of Qu Jun culturing room is 24 DEG C, intensity of illumination is 2000lux, light application time is 10 hours/day, and incubation time is 30 days;
In the method for germ contamination in described elimination Luohanguo cultivation, described is add before described agar adds without photocatalyst and lysozyme.
Step 2: the Momordica grosvenori group training Multiple Buds by germ contamination is inoculated into MS and goes in bacterium culture medium, through past bacterium cultivation, obtains aseptic group training Multiple Buds;
Step 3: the MS rejuvenation culture medium required for the cultivation of preparation Luohanguo, described MS rejuvenation culture medium is the agar adding the 6-benzylaminopurine of 0.05mg/L, the methyl α-naphthyl acetate of 0.01mg/L, the indolebutyric acid of 0.01mg/L, the active carbon of 1.0g/L, the sucrose of 30g/L and 4.5g/L in basic MS medium, its pH value is 5.8, the temperature of rejuvenation culturing room is 24 DEG C, intensity of illumination is 2000lux, light application time is 10 hours/day, and incubation time is 30 days;
Step 4: the aseptic group training Multiple Buds obtained is inoculated in MS rejuvenation culture medium, cultivates through rejuvenation, obtain healthy tissue-cultured seedling.
Embodiment 2:
Eliminate a method for germ contamination in Luohanguo cultivation, comprise the following steps:
Step one: the MS required for the cultivation of preparation Luohanguo removes bacterium culture medium, described MS goes bacterium culture medium to be the streptomycin, the phytic acid of 0.01%, the agar without photocatalyst, the lysozyme of 0.07%, the sucrose of 30g/L and 4.5g/L of 1.0% that add the 6-benzylaminopurine of 0.5mg/L, the kanamycin of 120mg/L, the Cefotaxime Sodium of 20mg/L and 50mg/L in basic MS medium, its pH value is 5.8, the temperature of Qu Jun culturing room is 27 DEG C, intensity of illumination is 2000lux, light application time is 12 hours/day, and incubation time is 30 days;
In the method for germ contamination in described elimination Luohanguo cultivation, described is add before described agar adds without photocatalyst and lysozyme.
Step 2: the Momordica grosvenori group training Multiple Buds by germ contamination is inoculated into MS and goes in bacterium culture medium, through past bacterium cultivation, obtains aseptic group training Multiple Buds;
Step 3: the MS rejuvenation culture medium required for the cultivation of preparation Luohanguo, described MS rejuvenation culture medium is the agar adding the 6-benzylaminopurine of 0.05mg/L, the methyl α-naphthyl acetate of 0.01mg/L, the indolebutyric acid of 0.01mg/L, the active carbon of 1.0g/L, the sucrose of 30g/L and 4.5g/L in basic MS medium, its pH value is 5.8, the temperature of rejuvenation culturing room is 27 DEG C, intensity of illumination is 2000lux, light application time is 12 hours/day, and incubation time is 30 days;
Step 4: the aseptic group training Multiple Buds obtained is inoculated in MS rejuvenation culture medium, cultivates through rejuvenation, obtain healthy tissue-cultured seedling.
Embodiment 3:
Eliminate a method for germ contamination in Luohanguo cultivation, comprise the following steps:
Step one: the MS required for the cultivation of preparation Luohanguo removes bacterium culture medium, described MS goes bacterium culture medium to be the streptomycin, the glycine of 0.03%, the agar without photocatalyst, the lysozyme of 0.05%, the sucrose of 30g/L and 4.5g/L of 0.75% that add the 6-benzylaminopurine of 0.5mg/L, the kanamycin of 70mg/L, the Cefotaxime Sodium of 11mg/L and 27mg/L in basic MS medium, its pH value is 5.8, the temperature of Qu Jun culturing room is 25 DEG C, intensity of illumination is 2000lux, light application time is 11 hours/day, and incubation time is 30 days;
In the method for germ contamination in described elimination Luohanguo cultivation, described is add before described agar adds without photocatalyst and lysozyme.
Step 2: the Momordica grosvenori group training Multiple Buds by germ contamination is inoculated into MS and goes in bacterium culture medium, through past bacterium cultivation, obtains aseptic group training Multiple Buds;
Step 3: the MS rejuvenation culture medium required for the cultivation of preparation Luohanguo, described MS rejuvenation culture medium is the agar adding the 6-benzylaminopurine of 0.05mg/L, the methyl α-naphthyl acetate of 0.01mg/L, the indolebutyric acid of 0.01mg/L, the active carbon of 1.0g/L, the sucrose of 30g/L and 4.5g/L in basic MS medium, its pH value is 5.8, the temperature of rejuvenation culturing room is 25 DEG C, intensity of illumination is 2000lux, light application time is 11 hours/day, and incubation time is 30 days;
Step 4: the aseptic group training Multiple Buds obtained is inoculated in MS rejuvenation culture medium, cultivates through rejuvenation, obtain healthy tissue-cultured seedling.
Although embodiment of the present invention are open as above, but it is not restricted to listed in specification and embodiment utilization, it can be applied to various applicable the field of the invention completely, for those skilled in the art, can easily realize other amendment, therefore do not deviating under the universal that claim and equivalency range limit, the present invention is not limited to specific details and illustrates here and the embodiment described.
Claims (10)
1. eliminate a method for germ contamination in Luohanguo cultivation, comprise the following steps:
Step one: the MS required for the cultivation of preparation Luohanguo removes bacterium culture medium;
Step 2: the Momordica grosvenori group training Multiple Buds by germ contamination is inoculated into MS and goes in bacterium culture medium, through past bacterium cultivation, obtains aseptic group training Multiple Buds;
Step 3: the MS rejuvenation culture medium required for the cultivation of preparation Luohanguo;
Step 4: the aseptic group training Multiple Buds obtained is inoculated in MS rejuvenation culture medium, cultivates through rejuvenation, obtain healthy tissue-cultured seedling;
It is characterized in that, described MS goes also to be added with without photocatalyst and lysozyme in bacterium culture medium, is added with active carbon in MS rejuvenation culture medium.
2. a kind of method eliminating germ contamination in Luohanguo cultivation as claimed in claim 1, it is characterized in that, described MS goes bacterium culture medium to be the agar adding the 6-benzylaminopurine of 0.5mg/L, the kanamycin of 20-120mg/L, the Cefotaxime Sodium of 2-20mg/L and the streptomycin of 5-50mg/L, the sucrose of 30g/L and 4.5g/L in basic MS medium, and its pH value is 5.8.
3. a kind of method eliminating germ contamination in Luohanguo cultivation as claimed in claim 1, it is characterized in that, described MS goes bacterium culture medium to be the agar adding the 6-benzylaminopurine of 0.5mg/L, the kanamycin of 90mg/L, the Cefotaxime Sodium of 10mg/L and the streptomycin of 25mg/L, the sucrose of 30g/L and 4.5g/L in basic MS medium, and its pH value is 5.8.
4. a kind of method eliminating germ contamination in Luohanguo cultivation as claimed in claim 1, it is characterized in that, described MS go also to be added with in bacterium culture medium in phytic acid that mass fraction is 0.01-0.05%, polymeric phosphate, glycine any one or a few.
5. a kind of method eliminating germ contamination in Luohanguo cultivation as claimed in claim 1, it is characterized in that, described MS rejuvenation culture medium is the agar adding the 6-benzylaminopurine of 0.05mg/L, the methyl α-naphthyl acetate of 0.01mg/L, the indolebutyric acid of 0.01mg/L, the sucrose of 30g/L and 4.5g/L in basic MS medium, and its pH value is 5.8.
6. as claimed in claim 1 a kind of eliminate Luohanguo cultivate in the method for germ contamination, it is characterized in that, described in go bacterium to cultivate and in rejuvenation cultivation, the temperature of culturing room is 24-27 DEG C, intensity of illumination is 2000lux, and light application time is 10-12 hour/day, and incubation time is 30 days.
7. a kind of method eliminating germ contamination in Luohanguo cultivation as claimed in claim 1, is characterized in that, described is add before described agar adds without photocatalyst and lysozyme.
8. a kind of method eliminating germ contamination in Luohanguo cultivation as claimed in claim 1, it is characterized in that, the concentration of described active carbon is 1.0g/L.
9. a kind of method eliminating germ contamination in Luohanguo cultivation as claimed in claim 1, it is characterized in that, the described mass fraction without photocatalyst is 0.5-1.0%.
10. a kind of method eliminating germ contamination in Luohanguo cultivation as claimed in claim 1, it is characterized in that, the mass fraction of described lysozyme is 0.03-0.07%.
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105613299A (en) * | 2016-02-05 | 2016-06-01 | 王少荣 | Method for eliminating bacterial contamination of chrysanthemum tissue cultured seedlings |
| CN105660402A (en) * | 2016-02-05 | 2016-06-15 | 王少荣 | Method for saving Jiyichao chrysanthemum tissue culture seedling from mold contamination |
| CN105684905A (en) * | 2016-02-05 | 2016-06-22 | 王少荣 | Method for remedying bacterial contamination of tissue culture seedlings of blueberry |
| CN106035092A (en) * | 2016-07-18 | 2016-10-26 | 中国科学院新疆生态与地理研究所 | Method for recovering and rejuvenating moss in culture medium polluted by fungus and mold |
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| CN107278899A (en) * | 2017-07-28 | 2017-10-24 | 韦荣昌 | Promote the method for Momordica grosvenori UGT6 gene expressions |
| CN108522273A (en) * | 2018-02-05 | 2018-09-14 | 孝感市鲁建农业开发有限公司 | A kind of Siraitia grosvenorii implantation methods rich in various trace elements |
| CN109757381A (en) * | 2019-03-20 | 2019-05-17 | 丽江海贝瑞生物科技有限公司 | A kind of bletilla striata tissue culture and rapid propagation method |
| CN115918540A (en) * | 2022-12-26 | 2023-04-07 | 广西壮族自治区药用植物园 | Method for eliminating bacterial contamination in artemisia annua tissue culture |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1631105A (en) * | 2004-12-31 | 2005-06-29 | 中国科学院武汉植物园 | Clonal propagation method of Momordica grosvenori germchit |
| CN103125385A (en) * | 2013-01-28 | 2013-06-05 | 浙江红叶园艺有限公司 | Cutting propagation method of strengthening antipollution factor zelkova schneideriana |
-
2014
- 2014-09-22 CN CN201410488635.0A patent/CN104396735B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1631105A (en) * | 2004-12-31 | 2005-06-29 | 中国科学院武汉植物园 | Clonal propagation method of Momordica grosvenori germchit |
| CN103125385A (en) * | 2013-01-28 | 2013-06-05 | 浙江红叶园艺有限公司 | Cutting propagation method of strengthening antipollution factor zelkova schneideriana |
Non-Patent Citations (4)
| Title |
|---|
| CHANG WEL等: "Bactericidal Activity of TiO2 Photocatalyst In Aqueous Media: Toward a Solar-Assisted Water Disinfection System", 《ENVIRON. SCI.TECHNOL.》 * |
| 刘志良: "不同浓度空气触媒对小区室内空气负离子和空气污染的影响研究", 《黑龙江生态工程职业学院学报》 * |
| 张鹏等: "动物源溶菌酶研究进展", 《中国生物工程杂志》 * |
| 潘丽梅等: "无籽罗汉果组培无菌系的构建", 《中国农学通报》 * |
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Application publication date: 20150311 Assignee: Cenxi Xulong Agricultural Technology Co.,Ltd. Assignor: GUANGXI BOTANICAL GARDEN OF MEDICINAL PLANTS Contract record no.: X2023980045868 Denomination of invention: A Method of Eliminating Bacterial Contamination in Tissue Culture of Siraitia grosvenorii Granted publication date: 20160817 License type: Common License Record date: 20231106 |