CN101921718A - Dendrobium candidum endophyte with growth promotion function and use thereof - Google Patents
Dendrobium candidum endophyte with growth promotion function and use thereof Download PDFInfo
- Publication number
- CN101921718A CN101921718A CN 201010125409 CN201010125409A CN101921718A CN 101921718 A CN101921718 A CN 101921718A CN 201010125409 CN201010125409 CN 201010125409 CN 201010125409 A CN201010125409 A CN 201010125409A CN 101921718 A CN101921718 A CN 101921718A
- Authority
- CN
- China
- Prior art keywords
- cell
- sphingol single
- cctcc
- strain
- tissue culturing
- 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.)
- Granted
Links
- 230000012010 growth Effects 0.000 title claims abstract description 23
- 241000026010 Dendrobium candidum Species 0.000 title abstract description 10
- 239000007788 liquid Substances 0.000 claims abstract description 28
- 238000004321 preservation Methods 0.000 claims abstract description 11
- 241001135759 Sphingomonas sp. Species 0.000 claims abstract description 8
- 238000012258 culturing Methods 0.000 claims description 34
- 230000001580 bacterial effect Effects 0.000 claims description 7
- 241001076416 Dendrobium tosaense Species 0.000 claims description 4
- 230000001737 promoting effect Effects 0.000 abstract description 2
- 241000736131 Sphingomonas Species 0.000 abstract 2
- 210000001519 tissue Anatomy 0.000 description 43
- 241000196324 Embryophyta Species 0.000 description 30
- 241000894006 Bacteria Species 0.000 description 17
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 9
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 9
- 230000006872 improvement Effects 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 244000005700 microbiome Species 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 229920001817 Agar Polymers 0.000 description 5
- 239000008272 agar Substances 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 240000004638 Dendrobium nobile Species 0.000 description 4
- 241000233855 Orchidaceae Species 0.000 description 4
- 239000001888 Peptone Substances 0.000 description 4
- 108010080698 Peptones Proteins 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 235000019319 peptone Nutrition 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 241000234295 Musa Species 0.000 description 2
- 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 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 235000021015 bananas Nutrition 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- 238000005374 membrane filtration Methods 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 241001523681 Dendrobium Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000201295 Euphrasia Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 240000005373 Panax quinquefolius Species 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- 241001464945 Sphingopyxis macrogoltabida Species 0.000 description 1
- 230000036579 abiotic stress Effects 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000000702 anti-platelet effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 241000878392 bacterium sh1 Species 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 229940088623 biologically active substance Drugs 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- HEZMRBNCRSVIHU-UHFFFAOYSA-N calcium boric acid dihypochlorite Chemical class [Ca+2].Cl[O-].Cl[O-].OB(O)O HEZMRBNCRSVIHU-UHFFFAOYSA-N 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000012364 cultivation method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000003467 diminishing effect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000006353 environmental stress Effects 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- -1 polyoxyethylene Polymers 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000012882 sequential analysis Methods 0.000 description 1
- 239000002893 slag Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 230000024883 vasodilation Effects 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a sphingomonas sp. strain with preservation name of Sphingomonas (SH1), preserved in China Centre for Type Culture Collection (CCTCC) in Wuhan University, China, wherein the preservation date thereof is February 4, 2010 with preservation number thereof being CCTCC No: M 2010041. The invention also discloses a use of the sphingomonas sp. strain applied to promoting growth of dendrobium candidum, comprising the following specific uses: inoculating raw fermenting liquid or fermented liquid of the sphingomonas sp. strain CCTCC No: M 2010041 into a root of a dendrobium candidum seedling so as to promote the growth of the dendrobium candidum seedling.
Description
Technical field
The present invention relates to a kind of natural medicinal plant endogenetic bacteria---dendrobium candidum endophyte and uses thereof.
Background technology
Herba Dendrobii belongs to the second largest genus in the orchid family (Orchidaceae) plant---Dendrobium (Dendrobium SW.), it is the superfine product in the stem of noble dendrobium, belong to aerial orchid section herbaceous plant, iron is green gains the name because of epidermis is for it, have nourishing Yin and clearing heat, the beneficial stomach that promotes the production of body fluid, moisten the lung and relieve the cough, make eye bright keep fit, effect such as easing pain and diminishing inflammation; Modern pharmacological research shows, effect such as that the stem of noble dendrobium also has is antitumor, anti-ageing, enhancing body immunizing power, vasodilation and anti-platelet aggregation.Because its great pharmaceutical use makes it become the whole nation and relates to pharmaceutical factory and the healthcare products processing enterprise raw material lifeblood of using stem of noble dendrobium class.But the growth conditions of Herba Dendrobii is very harsh, so natural production is very rare, more excessively excavates for a long time because of among the people, causes wild resource endangered.
China is annual according to statistics is that medicine, the healthcare products output value of raw material surpasses hundred million yuan of hundreds ofs with the stem of noble dendrobium, and quantity every year of the market requirement is all more than millions of kilograms, and in rising trend year by year.The Herba Dendrobii major part of Sheng Chaning is all from artificial culture in the market, and promptly the mode of training through group earlier expands numerous Herba Dendrobii, again through the acclimatization and transplants field.Yet the artificial culture Herba Dendrobii still exists breeding and seedling difficulty, transplant survival difficulty and poor growth Three Difficult Issues, and this is undoubtedly a serious obstruction to the development and use of Herba Dendrobii.
Endophyte of plant is to separate from the process plant tissue of surface sterilization or to obtain from the juice of inside plants, and apparent the going up of plant do not had the microorganism that harm does not promptly cause obvious illness.Endophyte of plant comprises reciprocal sharp altogether (mutualistic) and neutral (neutral) endosymbiontic microorganism.
At present from the various organs (root, stem, leaf, flower, fruit, seed) of a lot of plants, be separated to endophyte.They are existing many bibliographical informations aspect the intravital ubiquity of plant, abundant species diversity and functional diversity.Endophyte interacts with plant through secular, forms complicated relation between them.Have now found that endophyte has the effect of the following aspects to host plant:
(1) promotes the host plant growth, improve host's productivity, can strengthen the host to resistance sick, worm, thereby aspect agriculture production, biological control and the biological control significance arranged;
(2), strengthen the resistance of host plant to environment-stress.As resistance to arid, global warning, saline and alkaline, heavy metal and other toxic substance pollution etc.;
(3), endophyte produces new biologically active substance.Endophyte of plant is as the microorganism under a kind of special habitats, in plant materials, can produce rich and varied, have a multiple bioactive secondary metabolite, people have found from endophyte and have manyly had the important use that various bioactive novel substances show at pharmaceutical sector, aspect agriculture and be worth at present.
According to research both at home and abroad, in plant seedlings, introduce microorganism and will produce positive influence growth, the metabolism of seedling, can strengthen resistance to biological and the abiotic stress factor.Insert endophyte and also can carry out comprehensive adjustment, make it comparatively fast to adapt to extraneous complex environment plant strain growth and physiological metabolism.J Nowak is called " biotizafion " with this technology, such as Mycorrhizal (mycorrhization), bacteriumization (bacterization).Beneficial microorganism and little numerous seedling are carried out common cultivation, are the brand-new routes of producing high resistance high-quality seedling, and this extensive high-efficiency high-quality cultivation for Chinese medicinal materials has good application prospects.
At present, more about the endogenetic fungus research of orchid in the world, but the research of endogenetic bacteria is less, does not more have the Herba Dendrobii endogenetic bacteria to be used for the research report of short long candidum tissue culturing face.
Summary of the invention
The technical problem to be solved in the present invention provides a kind of Herba Dendrobii endogenetic bacteria---Sphingol single-cell strain, this bacterial strain is used for the seedlings of Dendrobium officinale promotes growth, helps to solve tissue cultured seedling poor growth in the present seedlings of Dendrobium officinale production process, the production cycle is long, cost is higher technical barrier.
In order to solve the problems of the technologies described above, the invention provides a kind of Sphingol single-cell strain, this Sphingol single-cell (Sphingomonas sp.) bacterial strain preservation name is called: Sphingol single-cell SH1, depositary institution: Chinese typical culture collection center, preservation address: Chinese Wuhan University, preservation date: on February 4th, 2010, preserving number: CCTCC NO:M2010041.
The present invention also provides the purposes of above-mentioned Sphingol single-cell strain simultaneously: be used for the seedlings of Dendrobium officinale promotes growth.
Improvement as the purposes of Sphingol single-cell strain of the present invention: Sphingol single-cell CCTCC NO:M2010041 fermenation raw liquid or fermented liquid are inoculated in the base portion of candidum tissue culturing seedling, thereby promote the candidum tissue culturing seedling growth.
Dendrobium candidum endophyte of the present invention is the strain bacterium that the contriver is separated to from the Herba Dendrobii plant, shows that through traditional method evaluation and Molecular Identification it belongs to Sphingol single-cell, with its called after Sphingol single-cell SH1 (Sphingomonas sp.SH1).Be specially:
This dendrobium candidum endophyte separates the root from wild Herba Dendrobii plant, can obtain from wild Herba Dendrobii plant separation and Culture by the following method:
1), with the root of wild Herba Dendrobii after surface sterilization is handled, be cut into small pieces, be inoculated in special NA culture medium flat plate, cultivated 1～3 day down in 28～32 ℃;
This special NA substratum by basic NA substratum and the fresh Herba Dendrobii after grinding form, the composition of basic NA substratum consists of: peptone 10g, extractum carnis 3g, NaCl 5g, agar 15-20g and H
2O1000ml at the fresh Herba Dendrobii that adds on the above-mentioned basic NA substratum after 5g grinds, and regulates pH=7.4～7.6 with the NaOH of 1mol/L, must special NA substratum.
2), picking list bacterium colony is in the special NA culture medium flat plate line purifies and separates with above-mentioned step 1), places 28～32 ℃ to cultivate 1～3 day down;
3), repeat the cultivation of separating for several times purifying, until obtaining pure growth.
Dendrobium candidum endophyte of the present invention---Sphingol single-cell bacterium SH1 (Sphingomonas sp.SH1) has been deposited in Chinese representative microbial preservation center (CCTCC), preserving number: CCTCC NO:M 2010041 on February 4th, 2010.
Dendrobium candidum endophyte of the present invention---Sphingol single-cell SH1 (being designated hereinafter simply as Sphingol single-cell CCTCCNO:M 2010041) has following biological property:
1. colony morphology characteristic: well-grown on basic NA substratum, the bacterial colony feature: bacterium colony is rounded, and yellow is smooth, neat in edge, the bacterium colony umbo is carinate.
2. morphological features: spherical, diameter is 0.5～1.5 μ m, Gram-negative.
3. major physiological biochemical character: this bacterium can be on basic NA substratum, is that 20～40 ℃, pH value are well-grown in 6.0～8.0 the environment in growth temperature.
4. genetics feature: according to " common bacteria system identification handbook " (eastern elegant pearl, Cai Miaoying, Science Press, 2001), on the morphological specificity of Sphingol single-cell CCTCC NO:M 2010041 of the present invention, with domestic name to separate polyoxyethylene glycol Sphingol single-cell (Sphingomonas macrogoltabidus) the most approaching; 16S rDNA sequential analysis through this Sphingol single-cell CCTCC M2010041 shows that Sphingol single-cell CCTCC M 2010041 should belong to Sphingol single-cell and belong to.
Herba Dendrobii endogenetic bacteria SH1 of the present invention has growth promoting function to candidum tissue culturing seedling, can promote the long increase by 10%～25% of average root of tissue cultured seedling, increase the height 10%～30% of plant, increase aquatic foods kind and the dry weight 5%～20% of plant, can shorten 10～20 days production times of candidum tissue culturing seedling; Nontoxic to people and animals, environment is not polluted.Can in the production of candidum tissue culturing seedling, be applied, produce certain economic benefits.
When Herba Dendrobii endogenetic bacteria of the present invention is used to promote the Herba Dendrobii growth, only needing this Herba Dendrobii endogenetic bacteria---fermenation raw liquid or the fermented liquid of Sphingol single-cell CCTCC NO:M 2010041 are inoculated in the candidum tissue culturing seedling base portion, can obtain to promote the effect of candidum tissue culturing seedling growth.
Embodiment
The isolation cultivation method of embodiment 1, Sphingol single-cell CCTCC NO:M 2010041, carry out following steps successively:
1, gathers the root sample of the Herba Dendrobii plant of wild health in the Tonglu, Zhejiang, fully clean 25KHz ultrasonication 5 minutes with tap water;
2, under aseptic condition, to step 1 gained for the examination material with aseptic water washing 2 times, then successively with the alcohol of volumetric concentration 75% and saturated Eusol to carrying out surface sterilization for the examination material, thereby kill for trying the material surface microorganism;
3, in gnotobasis, with step 2 gained for the examination material with aseptic water washing 3 times, get the aseptic water washing liquid coating that last 1 flushing of 100 μ l produced and be inoculated in the basic NA substratum, 30 ℃ cultivate 24h after, observe and have or not bacterium colony to produce.Verify in view of the above whether this sterilization method all kills for examination material surface microorganism.
If observe conclusion be: do not have bacterium colony to produce, then carry out following step 4; If observe conclusion be: have bacterium colony to produce, then return step 2.
4, the root sample that will handle well is cut into 0.5～1cm segment, inserts in the special NA culture medium flat plate, is positioned over 30 ℃ and cultivates 1～3 day;
This special NA substratum is made up of basic NA substratum and fresh Herba Dendrobii abrasive slag; The composition of basis NA substratum consists of: peptone 10g, extractum carnis 3g, NaCl 5g, agar 18g and H
2O1000ml at the fresh Herba Dendrobii that adds on the above-mentioned basic NA substratum after 5g grinds, and regulates pH=7.4～7.6 with the NaOH of 1mol/L, must special NA substratum.
5, the bacterial classification that grows on the special NA culture medium flat plate of picking above-mentioned steps 4 gained places 30 ℃ to cultivate picking list bacterium colony 1～3 day in special NA culture medium flat plate line purifies and separates;
Repeat aforesaid operations, till obtaining single bacterium colony.
6, single bacterium colony of step 5 picking gained being connected to special NA medium slant is stored in 4 ℃ of refrigerators.
A kind of single bacterium colony of gained is a dendrobium candidum endophyte---Sphingol single-cell SH1 (Sphingomonas sp.SH1), be deposited in Chinese representative microbial preservation center (CCTCC) on February 4th, 2010, preservation address: Chinese Wuhan University, preservation date: on February 4th, 2010, preserving number: CCTCC NO:M 2010041.Its sequence is shown in SEQID NO:1.
The cultivation producing method for seed of embodiment 2, Sphingol single-cell CCTCC NO:M 2010041, carry out following steps successively:
1, Sphingol single-cell CCTCC NO:M 2010041 is transferred to basic NA medium slant, cultivated 2 days for 30 ℃; Get the inclined-plane seed;
Basis NA medium slant prescription is: peptone 10g, extractum carnis 3g, NaCl 5g, agar 18g and H
2O1000ml, pH=7.4～7.6.
2, with transfering loop picking 0.5g step 1 gained slant strains, be inoculated in the 250ml triangular flask that 100ml LB liquid nutrient medium is housed, cultivate 30h with the 200r/m speed oscillation, get Sphingol single-cell CCTCC NO:M2010041 fermenation raw liquid in 28 ℃;
LB liquid culture based formulas is: peptone 10g, yeast extract paste 10g, NaCl 5g and H
2O1000ml, pH=7.4～7.6.
The promotion experiment of 2010041 pairs of candidum tissue culturing seedling growths of embodiment 3, Sphingol single-cell CCTCC NO:M:
Following experiment all with in the candidum tissue culturing substratum of improvement the candidum tissue culturing seedling cultivated as experimental subjects.
The prescription of candidum tissue culturing substratum of improvement is: employings 1/2MS is a minimum medium, adds bananas juice 10%/L, agar 7g/L, and sucrose 20g/L is 5.8 with the NaOH adjustment pH value of 1mol/L.That is: in the 1/2MS of every L, add 0.1L bananas juice, 7g agar and 20g sucrose, and to adjust the pH value be 5.8.
Experiment 1, Sphingol single-cell CCTCC NO:M 2010041 fermenation raw liquids test one to the candidum tissue culturing seedling promotes growth:
Utilize Sphingol single-cell CCTCC NO:M 2010041 fermenation raw liquids of the foregoing description 2 gained, fermenation raw liquid is diluted, make its bacterial concentration be adjusted into 10 with sterilized water
3CFU/ml is inoculated in this dilution secondary fermentation stoste in the candidum tissue culturing substratum of improvement and has cultivated 15 days 25 strain candidum tissue culturing seedling base portions, as experimental group; Be specially: the dilution secondary fermentation stoste that adds 50 μ l at every strain candidum tissue culturing seedling base portion.
The sterilized water of equal volume is replaced above-mentioned dilution secondary fermentation stoste, and all the other contents are identical, as the blank group.
Above-mentioned experimental group and blank group are all cultivated under the intensity of illumination condition of 25 ℃ of constant temperature, whole day 1000lx.
Cultivation results is as follows:
After 60 days as seen, the average upgrowth situation of the candidum tissue culturing seedling of experimental group obviously is better than the blank group, tissue cultured seedling average root length has increased by 20.5% than blank group in the experimental group, plant height has increased by 17.8% than blank group, and tissue cultured seedling plant fresh weight and dry weight have increased by 16.3% and 15.9% than blank group respectively.
Experiment 2, Sphingol single-cell CCTCC NO:M 2010041 fermenation raw liquids test two to the candidum tissue culturing seedling promotes growth:
With the dilution after bacterial concentration by 10
3CFU/ml makes 10 into
1CFU/ml is to have cultivated 30 days candidum tissue culturing seedling as experimental subjects in the candidum tissue culturing substratum of improvement; All the other equivalent experiments 1.
Cultivation results is as follows:
After 60 days as seen, the average upgrowth situation of the candidum tissue culturing seedling of experimental group all obviously is better than the blank group, tissue cultured seedling average root length has increased by 11.3% than blank group in the experimental group, plant height has increased by 14.7% than blank group, and tissue cultured seedling plant fresh weight and dry weight have increased by 10.2% and 9.7% than blank group respectively.
Experiment 3, Sphingol single-cell CCTCC NO:M 2010041 fermented liquids test one to the candidum tissue culturing seedling promotes growth:
Is the membrane filtration of 0.45 μ m with Sphingol single-cell CCTCC NO:M 2010041 fermenation raw liquids of the foregoing description 2 gained with the aperture, must not have fermented liquid.Get this no fermented liquid and be inoculated in the candidum tissue culturing substratum of improvement and cultivated 15 days 25 strain candidum tissue culturing seedlings, as experimental group; Be specially: the no fermented liquid that adds 50 μ l at every strain candidum tissue culturing seedling base portion; After 15 days, add the no fermented liquid of 50 μ l again at every strain candidum tissue culturing seedling base portion.
The sterilized water of equal volume is replaced above-mentioned no fermented liquid, and all the other contents are identical, as the blank group.
Above-mentioned experimental group and blank group are all cultivated under the condition with experiment 1, and cultivation results is as follows:
After 60 days as seen, the average upgrowth situation of the candidum tissue culturing seedling of experimental group is better than the blank group, tissue cultured seedling average root length has increased by 7.8% than blank group in the experimental group, plant height has increased by 7.3% than blank group, and tissue cultured seedling plant fresh weight and dry weight have increased by 6.6% and 6.3% than blank group respectively.
Experiment 4, Sphingol single-cell CCTCC NO:M 2010041 fermented liquids test two to the candidum tissue culturing seedling promotes growth:
Is the membrane filtration of 0.45 μ m with Sphingol single-cell CCTCC NO:M 2010041 fermenation raw liquids of the foregoing description 2 gained with the aperture, must not have fermented liquid.Get this no fermented liquid and be inoculated in the candidum tissue culturing substratum of improvement and cultivated 30 days 25 strain candidum tissue culturing seedlings, as experimental group; Be specially: the no fermented liquid that adds 100 μ l at every strain candidum tissue culturing seedling base portion.
The sterilized water of equal volume is replaced above-mentioned no fermented liquid, and all the other contents are identical, as the blank group.
Above-mentioned experimental group and blank group are all cultivated under the condition with experiment 1, and cultivation results is as follows:
After 60 days as seen, the average upgrowth situation of the candidum tissue culturing seedling of experimental group is better than the blank group, the long average root length of tissue cultured seedling average root has increased by 4.5% than blank group in the experimental group, plant height has increased by 5.3% than blank group, and tissue cultured seedling plant fresh weight and dry weight have increased by 6.1% and 5.3% than blank group respectively in the experimental group.
At last, it is also to be noted that what more than enumerate only is several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.
Sequence table
SEQ?ID?NO：1
agagtttgat?cctggctcag?aacgaacgct?ggcggcatgc?ctaacacatg?caagtcgaac 60
gaaggcttcg?gccttagtgg?cgcacgggtg?cgtaacgcgt?gggaatctgc?ccttaggttc 120
ggaataacag?ctggaaacgg?ctgctaatac?cggatgatat?cgcgagatca?aagatttatc 180
gcctgaggat?gagcccgcgt?tggattaggt?agttggtggg?gtaaaggcct?accaagccga 240
cgatccatag?ctggtctgag?aggatgatca?gccacactgg?gactgagaca?cggcccagac 300
tcctacggga?ggcagcagtg?gggaatattg?gacaatgggc?gaaagcctga?tccagcaatg 360
ccgcgtgagt?gatgaaggcc?ctagggttgt?aaagctcttt?tacccgggaa?gataatgact 420
gtaccgggag?aataagcccc?ggctaactcc?gtgccagcag?ccgcggtaat?acggaggggg 480
ctagcgttgt?tcggaattac?tgggcgtaaa?gcgcacgtag?gcggctttgt?aagtcagagg 540
tgaaagcctg?gagctcaact?ccagaactgc?ctttgagact?gcatcgcttg?aatccaggag 600
aggtcagtgg?aattccgagt?gtagaggtga?aattcgtaga?tattcggaag?aacaccagtg 660
gcgaaggcgg?ctgactggac?tggtattgac?gctgaggtgc?gaaagcgtgg?ggagcaaaca 720
ggattagata?ccctggtagt?ccacgccgta?aacgatgata?actagctgtc?cgggcacttg 780
gtgcttgggt?ggcgcagcta?acgcattaag?ttatccgcct?ggggagtacg?gccgcaaggt 840
taaaactcaa?aggaattgac?gggggcctgc?acaagcggtg?gagcatgtgg?tttaattcga 900
agcaacgcgc?agaaccttac?cagcgtttga?catggtagga?cgacttccag?agatggattt 960
cttcccttcg?gggacctaca?cacaggtgct?gcatggctgt?cgtcagctcg?tgtcgtgaga 1020
tgttgggtta?agtcccgcaa?cgagcgcaac?cctcgccttt?agttaccatc?atttggttgg 1080
gtactctaaa?ggaaccgccg?gtgataagcc?ggaggaaggt?ggggatgacg?tcaagtcctc 1140
atggccctta?cgcgctgggc?tacacacgtg?ctacaatggc?aactacagtg?ggcagcgacc 1200
ctgcgagggc?gagctaatcc?ccaaaagttg?tctcagttcg?gattgttctc?tgcaactcga 1260
gagcatgaag?gcggaatcgc?tagtaatcgc?ggatcagcat?gccgcggtga?atacgttccc 1320
aggccttgta?cacaccgccc?gtcacaccat?gggagttgga?ttcacccgaa?ggcgttgcgc 1380
caacctagca?ataggaagca?ggcgaccacg?gtgggttcag?cgactggggt?gaagtcgtaa 1440
caaggtagcc 1450
Claims (3)
1. Sphingol single-cell strain, it is characterized in that: this Sphingol single-cell (Sphingomonas sp.) bacterial strain preservation name is called: Sphingol single-cell SH1, depositary institution: Chinese typical culture collection center, preservation address: Chinese Wuhan University, preservation date: on February 4th, 2010, preserving number: CCTCC NO:M 2010041.
2. the purposes of Sphingol single-cell strain as claimed in claim 1 is characterized in that: be used for the seedlings of Dendrobium officinale promotes growth.
3. the purposes of Sphingol single-cell strain according to claim 2 is characterized in that: Sphingol single-cell CCTCC NO:M 2010041 fermenation raw liquids or fermented liquid are inoculated in the base portion of candidum tissue culturing seedling, thereby promote the candidum tissue culturing seedling growth.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101254098A CN101921718B (en) | 2010-03-16 | 2010-03-16 | Dendrobium candidum endophyte with growth promotion function and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101254098A CN101921718B (en) | 2010-03-16 | 2010-03-16 | Dendrobium candidum endophyte with growth promotion function and use thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101921718A true CN101921718A (en) | 2010-12-22 |
| CN101921718B CN101921718B (en) | 2011-12-07 |
Family
ID=43336936
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010101254098A Expired - Fee Related CN101921718B (en) | 2010-03-16 | 2010-03-16 | Dendrobium candidum endophyte with growth promotion function and use thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101921718B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104830707A (en) * | 2015-01-09 | 2015-08-12 | 江苏大学 | Endogenous sphingosine monas sourced from pennisetum purpureum and application thereof |
| CN105112309A (en) * | 2015-09-25 | 2015-12-02 | 江苏农林职业技术学院 | Umbelopsis nana and application thereof in promoting growth of Dendrobium officinale |
| CN105112308A (en) * | 2015-09-25 | 2015-12-02 | 江苏农林职业技术学院 | Gibberella moniliformis and application of gibberella moniliformis to growth promotion and drought resistance for dendrobium officinale |
| CN105132293A (en) * | 2015-09-25 | 2015-12-09 | 江苏农林职业技术学院 | Alternaria tenuissima and application of alternaria tenuissima to growth promotion and drought resistance of dendrobium officinale |
| CN105219656A (en) * | 2015-09-25 | 2016-01-06 | 江苏农林职业技术学院 | A kind of Fusarium oxysporum and the application in promotion Growth of Dendrobium candidum thereof |
| CN112493252A (en) * | 2020-12-29 | 2021-03-16 | 浙江大学 | Sphingomonas cucurbitae and application of fermentation product thereof in preventing and treating rice bacterial diseases |
-
2010
- 2010-03-16 CN CN2010101254098A patent/CN101921718B/en not_active Expired - Fee Related
Non-Patent Citations (3)
| Title |
|---|
| 《中国新药杂志》 20080505 廖晨阳等 石斛内生菌的筛选和培养优化 第17卷, 第9期 2 * |
| 《中草药》 20020625 高徽徽 三种内生真菌对铁皮石斛、金线莲生长影响的研究 第33卷, 第6期 2 * |
| 《信阳师范学院学报：自然科学版》 20090430 陈玉栋等 铁皮石斛内生真菌的分离与鉴定 第22卷, 第2期 2 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104830707A (en) * | 2015-01-09 | 2015-08-12 | 江苏大学 | Endogenous sphingosine monas sourced from pennisetum purpureum and application thereof |
| CN104830707B (en) * | 2015-01-09 | 2018-01-16 | 江苏大学 | A kind of interior raw Sphingol single-cell for coming from napier grass and application thereof |
| CN105112309A (en) * | 2015-09-25 | 2015-12-02 | 江苏农林职业技术学院 | Umbelopsis nana and application thereof in promoting growth of Dendrobium officinale |
| CN105112308A (en) * | 2015-09-25 | 2015-12-02 | 江苏农林职业技术学院 | Gibberella moniliformis and application of gibberella moniliformis to growth promotion and drought resistance for dendrobium officinale |
| CN105132293A (en) * | 2015-09-25 | 2015-12-09 | 江苏农林职业技术学院 | Alternaria tenuissima and application of alternaria tenuissima to growth promotion and drought resistance of dendrobium officinale |
| CN105219656A (en) * | 2015-09-25 | 2016-01-06 | 江苏农林职业技术学院 | A kind of Fusarium oxysporum and the application in promotion Growth of Dendrobium candidum thereof |
| CN105112308B (en) * | 2015-09-25 | 2018-03-09 | 江苏农林职业技术学院 | An a kind of beading are red mould and its in dendrobium candidum growth-promoting and the application in drought resisting |
| CN105132293B (en) * | 2015-09-25 | 2018-03-09 | 江苏农林职业技术学院 | A kind of Alternaria tenuissima and its application in dendrobium candidum growth-promoting, drought resisting |
| CN105219656B (en) * | 2015-09-25 | 2018-03-30 | 江苏农林职业技术学院 | A kind of Fusarium oxysporum and its application in Growth of Dendrobium candidum is promoted |
| CN112493252A (en) * | 2020-12-29 | 2021-03-16 | 浙江大学 | Sphingomonas cucurbitae and application of fermentation product thereof in preventing and treating rice bacterial diseases |
| CN112493252B (en) * | 2020-12-29 | 2021-09-21 | 浙江大学 | Sphingomonas cucurbitae and application of fermentation product thereof in preventing and treating rice bacterial diseases |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101921718B (en) | 2011-12-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102976801B (en) | Method for producing functional microorganism organic fertilizer by using food residue | |
| CN103330258B (en) | Cordyceps militaris health-care beverage prepared by liquid submerged fermentation and preparation method thereof | |
| CN102613518B (en) | Jerusalem artichoke pickle produced by direct-vat-set lactobacillus brevis leavening agent, and process of same | |
| CN101921718B (en) | Dendrobium candidum endophyte with growth promotion function and use thereof | |
| CN102703342B (en) | Bacillus velezensis ZJ20 strain and liquid preparations thereof | |
| CN107475134B (en) | One plant of Fusarium solani and its application in prevention and treatment dendrobium candidum southern blight | |
| CN108401794A (en) | A kind of armillaria mellea accreting with Rhizoma Gastrodiae liquid spawn production method and cultigen special culture media | |
| CN101955902B (en) | New brevibacillus brevis strain and application thereof | |
| CN104988082A (en) | Debaryomyces hansenii strain and applications thereof | |
| CN112725228A (en) | Virginia streptomycete capable of preventing and treating peach brown rot and application thereof | |
| CN106434490A (en) | Ginseng bacterium TY15-2 with effects of disease prevention and growth promotion and application thereof | |
| CN106071524B (en) | A kind of asparagus fermented beverage preparation method with nutrition health-care functions | |
| CN108913622B (en) | Bacillus megaterium BM22 and preparation and application of spore powder thereof | |
| CN106978359B (en) | One plant of Fusarium solani and its application in the anti-root rot of dendrobium candidum | |
| CN109645490A (en) | Application of the lactobacillus plantarum CQPC02 in the food or drug of preparation prevention diabetes | |
| CN103004465A (en) | Coprinus comatus strain and preparation method | |
| CN103060234A (en) | Biocontrol strain KMXU28 for controlling paris rhizome root rot and biocontrol agent thereof | |
| CN101928688A (en) | Abamectin pesticide residue degrading bacteria and bactericide thereof | |
| CN108841752B (en) | Bacillus megaterium BM22 and application of spore liquid preparation thereof in preventing and treating cyclamen persicum radices | |
| CN101855994B (en) | Production method of anoectochilus symbiosis seedling | |
| CN111607621A (en) | Yeast capable of producing rose fragrance and application of yeast in Lingwu jujube enzyme | |
| CN102986537A (en) | Tricholoma lobayense strain KJH-3 and preparation method thereof | |
| CN102986536A (en) | Flammulina velutipes strain and preparation method | |
| CN107353060A (en) | Mushroom culture composition, agalloch eaglewood mushroom and agalloch eaglewood mushroom cultural method | |
| CN105385611B (en) | A kind of plum surprise saccharomycete and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20101222 Assignee: Deqing Muge Ecological Agriculture Co., Ltd. Assignor: Zhejiang Sci-Tech University Contract record no.: 2017330000019 Denomination of invention: Dendrobium candidum endophyte with growth promotion function and use thereof Granted publication date: 20111207 License type: Common License Record date: 20170328 |
|
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20111207 Termination date: 20210316 |