CN102391972A - Cherry rhizosphere growth promotion pseudomonas sp. and application thereof - Google Patents
Cherry rhizosphere growth promotion pseudomonas sp. and application thereof Download PDFInfo
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Abstract
The invention discloses cherry rhizosphere growth promotion pseudomonas sp., the classification is named as pseudomonas sp., the cherry rhizosphere growth promotion pseudomonas sp. was collected in China General Microbiological Culture Collection Center on October 11, 2011, and the collection number is CGMCC No. 5319. The invention further discloses an application of the cherry rhizosphere growth promotion pseudomonas sp. in the promotion of growth of cherry. A bacterial agent prepared by a bacterial strain is inoculated into a cherry cutting seedling, and the result proves that the bacterial agent can obviously promote the growth of a root system of the cherry, improve the proportion of fine roots and increase the ground diameter of cherry shoots and the growth of seedling height. Compared with the contrast, the difference achieves the significant level. The cherry rhizosphere growth promotion pseudomonas sp. according to the invention is inoculated into fermented chicken manure for preparing a biological fertilizer, and the biological fertilizer is applied on the periphery of the root system of six-year-old large cherries, thereby being capable of significantly improving the vitality of the root system and activating phosphorus and potassium in the soil. The research result shows that the cherry rhizosphere growth promotion pseudomonas sp. according to the invention can obviously promote the growth and the development of the large cherries and improve a growth environment of the soil of the root system of the cherries.
Description
Technical field
The present invention relates to short living pseudomonas of a strain cherry rhizosphere and application thereof.
Background technology
Cherry (Cerasus pseudocerasus (Lindl.) G.Don) is that the Rosaceae (Rosaceae) cherry belongs to (Cerasus Mill.) plant; Be one of fruit maturation seeds the earliest in the deciduous fruit tree; Its fruit colour is bright-coloured, aromatic flavour, nutritious, and has certain value medical health care.The cherry fertilising is a link of can not ignore in the cherry management process.Fertilization Effect is except the influence that receives soil, the nutritive property of plant own and dose etc., and is also relevant with fertilizer variety etc.Present stage cherry fertilising is main with readily avaiable fertilizer; Though significantly improved cherry output, the readily avaiable fertilizer plant recovery of nutrient is low, causes soil compaction easily; The eutrophication of surface water and phreatic pollution also can cause the rapid decline of cherry fruit quality.
As everyone knows, mikrobe is present in occurring in nature widely, and is in close relations with soil fertility and crop micro-ecological environment, so many scholars connect mikrobe and agricultural, started the microbial agriculture engineering.The research and the utilisation technology of all kinds of probioticses constantly are in the news, and microbial fertilizer is exactly one of them.Microbial fertilizer can not only be improved the soil, and improves the ecotope of soil fauna, can also effectively improve the absorption of plant to soil nutrient.Therefore, seek new fertilizer, especially biomass fertilizers receives much concern with the research that substitutes or partly substitute chemical fertilizer.
The mikrobe of plant rhizosphere is many and enliven, and has constituted the distinctive micro-flora of rhizosphere.It is to survive in the plant rhizosphere scope that plant rhizosphere is urged endophytic bacteria (PGPR), can directly or indirectly promote plant-growth or have the bacteria beneficial of antagonistic action to be referred to as to pathogenic bacteria.The short endophytic bacteria of plant rhizosphere can be secreted the plant growth-promoting material, comprises plant hormone, VITAMINs, amino acid and other active organic molecule and verivate etc.; Also can improve the nutrient environment of plant rhizosphere, effective controlling plant disease etc.The monoid of the short endophytic bacteria of plant rhizosphere is various, and wherein existing useful population also has deleterious population.According to incompletely statistics, found to comprise that the short endophytic bacteria of more than 20 plant rhizospheres that belong to such as pseudomonas and bud pole bacterium has the report of disease prevention growth-promoting potential.At present, the screening of the short endophytic bacteria bacterial strain of relevant large cherry rhizosphere and the research of application do not appear in the newspapers.
Summary of the invention
To above-mentioned prior art, the invention provides short living pseudomonas of a strain cherry rhizosphere and application thereof.
The present invention realizes through following technical scheme:
One strain cherry rhizosphere is urged living pseudomonas, and its classification called after pseudomonas pseudomonas sp. is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on October 11st, 2011, and deposit number is CGMCC No.5319.
The colony characteristics of the short living pseudomonas of said cherry rhizosphere is: rounded opaque after cultivating 24h on the LB flat board, and smooth surface; The somatic cells size is the μ m of (0.5-1.0) μ m * (1.5-2.5); Do not produce gemma, G
-, the straight or little knee shape of cell.
The physiological and biochemical property of the short living pseudomonas of said cherry rhizosphere is: gelatin liquification test is negative, and the starch hydrolysis experiment is negative, and indole test is negative, and methyl red test is negative; Fu-Pu tests negative, and nitrate reduction test is positive, and Citrate trianion utilization test is positive, and hydrogen sulfide production test is negative; It is positive that glucose produces acid, glucose aerogenesis feminine gender, and mobility test is positive, and fructose test is negative; Cane sugar test is negative, and the N.F,USP MANNITOL test is negative, and catalase test is positive, and Nagler's reaction is negative; The urase examination is negative, and the ornithine decarboxylase test is negative, and the lysine decarboxylase test is negative.
The optimal culture conditions of the short living pseudomonas of said cherry rhizosphere is: LB substratum: peptone 10g, yeast extract paste 5g, NaCl10g, zero(ppm) water 1000ml.Temperature: 37 ℃, pH 7.0.
The short living pseudomonas of said cherry rhizosphere can be used to prepare microbial fertilizer.During application, cultivate the short living pseudomonas of cherry rhizosphere, get bacteria suspension, bacteria suspension is as liquid fertilizer, and the concentration of bacteria suspension is preferably 1.0 * 10
8Cfu/ml.Or: cultivate short the livings pseudomonas of cherry rhizosphere, must bacteria suspension, then with bacterial suspension inoculation in fertilizer, process the cherry biological fertilizer, the concentration of bacteria suspension is preferably 5.0 * 10
8The cfu/ milliliter, viable count is preferably 0.5 * 10 in the cherry fertilizer
8Cfu/g.
Said fertilizer is the chicken manure that becomes thoroughly decomposed.
The present invention also provides a kind of microbial fertilizer, obtains through following preparation method: cultivate the short living pseudomonas of cherry rhizosphere, get bacteria suspension, bacteria suspension is as liquid fertilizer.Or: cultivate short the livings pseudomonas of cherry rhizosphere, must bacteria suspension, then with bacterial suspension inoculation in fertilizer, process the cherry biological fertilizer.
Show that after deliberation the bacterial suspension inoculation of short the livings pseudomonas preparation of cherry rhizosphere of the present invention can obviously promote the cherry root growth in the cherry cuttage seeding, improve radicula (＜2mm) ratio, the growth of increase cherry seedling leading thread and height of seedling.Compare with contrast, difference reaches utmost point level of signification.The short living pseudomonas of cherry rhizosphere of the present invention is inoculated into processes bio-feritlizer in the fermentation poultry dung, be applied to around the big cherry root of life in 6 years, can significantly improve the root system vigor, phosphorus in the activating soil and potassium.Above result of study shows; The short living pseudomonas of cherry rhizosphere of the present invention can obviously promote large cherry to grow; Improve the growing environment of cherry rhizosphere soil, the present invention provides good strain resource for developing in the future the special-purpose microbial fertilizer of large cherry, will be widely used.
Description of drawings
One strain cherry rhizosphere is urged living pseudomonas; Bacterial strain is YT3; This strain classification called after pseudomonas pseudomonas sp. is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation on October 11st, 2011, and deposit number is CGMCC No.5319.
Fig. 1 is the cherry root system comparison diagram of YT3 treatment group and control group.
Embodiment
Below in conjunction with embodiment the present invention is further described.
The screening of the short living pseudomonas of embodiment 1 cherry rhizosphere
(1) screening process:
Through the gradient dilution method, from the cherry rhizosphere soil sample of Tai, Shandong Province, isolate 73 strain isolates, from the rhizosphere soil sample in Nanma cherry garden, Yiyuan County, Zibo City, Shandong Province, isolate 373 strain isolates.Through three zoning collimation methods the bacterium that is separated to is carried out purifying, and judge whether purifying of bacterial strain, the bacterium behind the purifying is numbered through microscopy, picking list bacterium colony, it is subsequent use to be transferred on the LB inclined-plane preservation.Filter out 25 strains and protect green effect bacterial strain preferably through protecting green method, in conjunction with radish cotyledon weightening finish method, finishing screen is selected the bacterium that 1 strain has the good short fruit of coming into force, and is designated as YT3.
(2) colony characteristics and thalli morphology
Rounded opaque after cultivating 24h on the LB flat board, smooth surface.The somatic cells size is the μ m of (0.5-1.0) μ m * (1.5-2.5).Do not produce gemma, G
-, the straight or little knee shape of cell.
(3) physiological and biochemical property
Gelatin liquification test is negative, and the starch hydrolysis experiment is negative, and indole test is negative, and methyl red test is negative; Fu-Pu tests negative, and nitrate reduction test is positive, and Citrate trianion utilization test is positive, and hydrogen sulfide production test is negative; It is positive that glucose produces acid, glucose aerogenesis feminine gender, and mobility test is positive, and fructose test is negative; Cane sugar test is negative, and the N.F,USP MANNITOL test is negative, and catalase test is positive, and Nagler's reaction is negative; The urase examination is negative, and the ornithine decarboxylase test is negative, and the lysine decarboxylase test is negative.
(4) 16S rDNA sequential analysis
The 16SrDNA gene order is seen shown in the nucleotides sequence tabulation.With the sequence alignment in survey 16SrDNA sequence and the GenBank DB, the result shows: YT3 and Pseudomonas putida are in together in the minimum branch, and the similarity of its 16S rDNA sequence and Pseudomonasputida (D84020) reaches 97.16%.In conjunction with colonial morphology; Physiological and biochemical property and 16S rDNA sequential analysis; Be accredited as pseudomonas Pseudomonas sp., be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation on October 11st, 2011, deposit number is CGMCC No.5319.
Embodiment 2 pot experiments
With the short living subtilis YT3 activation on the LB inclined-plane of the cherry rhizosphere of preserving, picking one articulating is gone in the beef-protein medium, and in 37 ℃, shaking culture is 2 days under the 180rpm condition, and transferring the bacterium liquid hold-up is 2.5 * 10
8Cfu/ml (suspension A), subsequent use.The field planting cherry seedling is stained with root in suspension A after.The suspension A for preparing is transferred bacterial concentration to 1.0 * 10
8Cfu/ml (suspension B) gets 50ml suspension B and evenly waters around cherry shoot root portion.The cherry seedling is stained with root in sterilized water after, the sterilized water of pouring equivalent is contrast.Normal management, fresh weight, the capillary root that stops to measure the cherry root system in the back in cherry seedling then is heavy, height of seedling and leading thread.Result such as table 1 are with shown in Figure 1.
From table, can find out, total root that YT3 handles cherry is heavy, radicula is heavy (＜2mm), height of seedling and leading thread compare according to having improved 18.33%, 66.51%, 15.28% and 18.84% respectively.Difference all reaches level of signification.More than show: bacterial strain YT3 grows to cherry, and especially the growth to the capillary root has obvious facilitation.
Table 1
Embodiment 3 field tests
The short living pseudomonas YT3 of large cherry rhizosphere is in the reciprocation type vibrator, and in 37 ℃, shaking culture is 2 days under the 180rpm condition, and transferring the bacterium liquid hold-up is 5.0 * 10
8Cfu/ml is inoculated in the chicken manure that becomes thoroughly decomposed, and processes cherry biological fertilizer (0.5 * 10
8Cfu/g).Every tree is used cherry biological fertilizer 8.5kg, is contrast with the chicken manure that becomes thoroughly decomposed of equivalent.Fertilization mode adopts four lateral sulcus to execute, and ditch length and width and height are respectively 60,30 and 20cm, with backfill again behind cherry biological fertilizer and the abundant mixing of soil that digs out.12 trees of every processing are provided with guard rows.
Use after 2 years rapid available phosphorus, available potassium and the root system vigor of measuring soil, the result sees table 2, can find out, uses the bio-fertilizer that short the livings pseudomonas YT3 of large cherry rhizosphere processes and can promote rapid available phosphorus and quick-acting potassium content in the soil, raising root system vigor.Compare with contrast (CK), the rhizosphere fertile rapid available phosphorus of handling of short living pseudomonas biological and the raising rate of available potassium have reached 22.13% and 13.28% respectively, and activation is obvious.
Table 2
Claims (9)
1. a strain cherry rhizosphere is urged living pseudomonas; It is characterized in that: this strain classification called after pseudomonas pseudomonas sp.; Be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on October 11st, 2011, deposit number is CGMCC No.5319.
2. the application of the short living pseudomonas of the described cherry rhizosphere of claim 1 in the preparation microbial fertilizer.
3. application according to claim 2 is characterized in that: during application, cultivate the short living pseudomonas of cherry rhizosphere, get bacteria suspension, bacteria suspension is as liquid fertilizer.
4. application according to claim 3 is characterized in that: the concentration of said bacteria suspension is 1.0 * 10
8Cfu/ml.
5. application according to claim 2 is characterized in that: during application, cultivate short the livings pseudomonas of cherry rhizosphere, must bacteria suspension, then with bacterial suspension inoculation in fertilizer, process the cherry biological fertilizer.
6. application according to claim 5 is characterized in that: the concentration of said bacteria suspension is 5.0 * 10
8Cfu/ml, viable count is 0.5 * 10 in the cherry biological fertilizer
8Cfu/g.
7. application according to claim 5 is characterized in that: said fertilizer is the chicken manure that becomes thoroughly decomposed.
8. microbial fertilizer is characterized in that: obtain through following preparation method: cultivate short the livings pseudomonas of the described cherry rhizosphere of claim 1, must bacteria suspension, and bacteria suspension is as liquid fertilizer.
9. microbial fertilizer is characterized in that: obtain through following preparation method: cultivate short the livings pseudomonas of the described cherry rhizosphere of claim 1, must bacteria suspension, then with bacterial suspension inoculation in fertilizer, process the cherry biological fertilizer.
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|---|---|---|---|
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Cited By (5)
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| CN106119148A (en) * | 2016-06-07 | 2016-11-16 | 江苏联海生物科技有限公司 | One pseudomonas and application thereof |
| CN109402004A (en) * | 2018-11-07 | 2019-03-01 | 山东农业大学 | One plant of Jian Shi Aeromonas and its application in raising stress resistance of plant |
| CN113943190A (en) * | 2021-12-09 | 2022-01-18 | 陕西师范大学 | Radix scrophulariae bud water-retention root-promotion probiotic powder |
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- 2011-11-23 CN CN201110376284A patent/CN102391972B/en not_active Expired - Fee Related
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102742495A (en) * | 2012-07-31 | 2012-10-24 | 山东省林业科学研究院 | Method for utilizing non-woven fabric container to perform sweet cherry dwarfing rootstock twig cuttage seedling culture and matrix combination |
| CN102742495B (en) * | 2012-07-31 | 2013-11-06 | 山东省林业科学研究院 | Method for utilizing non-woven fabric container to perform sweet cherry dwarfing rootstock twig cuttage seedling culture and matrix combination |
| CN102942420A (en) * | 2012-11-30 | 2013-02-27 | 山东省林业科学研究院 | Special microbial agent and bio-organic fertilizer for sweet cherry and preparation methods of special microbial agent |
| CN102942420B (en) * | 2012-11-30 | 2014-03-05 | 山东省林业科学研究院 | Special microbial agent and bio-organic fertilizer for sweet cherry and preparation methods of special microbial agent |
| CN106119148A (en) * | 2016-06-07 | 2016-11-16 | 江苏联海生物科技有限公司 | One pseudomonas and application thereof |
| CN106119148B (en) * | 2016-06-07 | 2019-10-01 | 江苏联海生物科技有限公司 | One pseudomonas and its application |
| CN109402004A (en) * | 2018-11-07 | 2019-03-01 | 山东农业大学 | One plant of Jian Shi Aeromonas and its application in raising stress resistance of plant |
| CN113943190A (en) * | 2021-12-09 | 2022-01-18 | 陕西师范大学 | Radix scrophulariae bud water-retention root-promotion probiotic powder |
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