CN102876598B - Pseudomonas fluorescens and application of pseudomonas fluorescens for promoting growth of Chinese sweetgum - Google Patents
Pseudomonas fluorescens and application of pseudomonas fluorescens for promoting growth of Chinese sweetgum Download PDFInfo
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- CN102876598B CN102876598B CN 201210327441 CN201210327441A CN102876598B CN 102876598 B CN102876598 B CN 102876598B CN 201210327441 CN201210327441 CN 201210327441 CN 201210327441 A CN201210327441 A CN 201210327441A CN 102876598 B CN102876598 B CN 102876598B
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Abstract
The invention discloses pseudomonas fluorescens FXW-HS7A with a classification name of the pseudomonas fluorescens. The pseudomonas fluorescens is preserved in China Typical Culture Collection Centre and has a preservation number of CCTCCM 2012260. The invention further discloses an application of the pseudomonas fluorescens for promoting growth of a Chinese sweetgum. In a condition of liquid shake-flask culture, bacterial strains disclosed by the invention have strong dissolving effects for insoluble phosphates such as tricalcium phosphate, iron phosphate, aluminium phosphate and hydroxyapatite; the bacterial stains are prepared into a fungicide, and the fungicide is inoculated in a Chinese sweetgum seedling; results indicate that the fungicide can obviously promote the growth of the Chinese sweetgum; and therefore, the pseudomonas fluorescens and the application of the pseudomonas fluorescens for promoting the growth of the Chinese sweetgum disclosed by the invention can provide an excellent bacterial strain resource for developing a special microbial fertilizer for the Chinese sweetgum in future.
Description
Technical field
The invention belongs to the microbial fertilizer technical field in the biological fertilizer field, be specifically related to a kind of echinopodospora jamaicensis rhizosphere high-efficient phosphate-solubilizing pseudomonas fluorescens and application thereof.
Background technology
Echinopodospora jamaicensis (Liquidambar formosana) is the tall and big deciduous tree of plum section sweetgum subfamily Liquidambar, calls to be maple, black meal wood, sweet gum etc.Being the important indigenous tree of China, also is that the good speed in subtropical zone is given birth to the broad leaved and deciduous broad leaved seeds, and its wide adaptability, growth are typical deserted mountain vanguard tree seed rapidly.Safeguard that the soil fertility effect is obvious, profitable on ecology, be one of first-selected seeds of artificial forest seeds structural adjustment, also be very potential afforestation seeds.View and admire, medicinal, all have vital role with aspects such as materials.In recent years, sweetgum enjoys attention in production, scientific research.
The echinopodospora jamaicensis growth is fast, and the water consumption amount of expending is big, and the forest land of plantation sweetgum often causes soil nutrient deficiency easily.Just influence the nutrient factor of sweetgum growth, available phosphorus content just is one of important factor of restriction sweetgum growth in the soil.Along with the huge of timber demand amount increased, the forest reserves progressively deficient, the plantation amount of the living sweetgum of speed obviously rises.At first only as ornamental tree species and scientific research forest, introduced a fine variety gradually afterwards and be green tree species, as the chief species of alleviating timber and Forest products demand anxiety, sweetgum is extensively planted now.Sweetgum has important economy and the ecological value.
Phosphorus is the required important nutrient of plant-growth, the phosphorus that plant can directly absorb seldom, so phosphorus usually becomes the limiting factor of plant nutrition.Molten phosphorus microorganism is distributed widely in soil, be that a class can reduce the micropopulation that phosphate fertilizer utilization efficiency and dissolving soil indissoluble phosphorus were fixed, improved to phosphate fertilizer, transforming soil indissoluble phosphorus, keep the soil phosphorus balance, improve phosphate fertilizer utilization efficiency, promote aspect such as plant growth to play an important role, by universally acknowledged be the phosphorus-activated biological control measure of soil (Bi Jiangtao, 2009) of Cheap highly effective environment-friendly type.
The release of phosphorus is main relevant with the factors such as decomposition of soil pH value, redox of soil reaction, microorganism secretion thing and organic substance, is result (Du Zhenyu, 2010 of multiple factor comprehensive action; Poplar bud plum, 2010).Phosphate-solubilizing bacteria can improve the plain supply of phosphorus of echinopodospora jamaicensis as the important microorganism of a class in the soil phosphorus circulation, accelerates the growth of echinopodospora jamaicensis.
At present, screening and the applied research of relevant echinopodospora jamaicensis rhizosphere high-efficient phosphate-solubilizing bacterial isolates do not appear in the newspapers.Therefore improving soil available phosphorus content just has very significant meaning, and becomes the key problem of numerous scholar's focuses.
Summary of the invention
Technical problem to be solved by this invention provides a kind of echinopodospora jamaicensis rhizosphere high-efficient phosphate-solubilizing pseudomonas fluorescens.
The technical problem that the present invention also will solve provides the application of above-mentioned echinopodospora jamaicensis rhizosphere high-efficient phosphate-solubilizing pseudomonas fluorescens.
In order to solve the problems of the technologies described above, the technical solution used in the present invention is as follows:
On April 23rd, 2011, to check the echinopodospora jamaicensis rhizosphere soil of park, the Lingshan from the Huangshan, Anhui Province, screening has obtained a plant height and has imitated phosphorus decomposing Pseudomonas fluorescens Pseudomonas fluorescens FXW-HS7A.Be preserved in Chinese typical culture collection center, address: China. Wuhan. Wuhan University, postcode 430072, culture presevation number: CCTCC M 2012260, preservation day is on July 1st, 2012.
The main biological property of CCTCC M 2012260 bacterial strains: on the beef-protein medium flat board, bacterium colony is less, and is pale yellow, circle; The thalline direct rod shape, Gram-negative, about 0.43 * 0.64 μ m of size ~ 1.89 * 2.00 μ m, no gemma, amphitrichous; Aerobic, the oxydase reaction positive, the catalase test positive, starch hydrolysis feminine gender, M.R. and indole test feminine gender, V.P. tests positive; The gelatine liquefication positive, the nitrate reductase test is negative, produces ammonia and hydrogen sulfide production test feminine gender, the Citrate trianion growth test positive.
CCTCC M 2012260 bacterial strain 16SrDNA gene orders are seen shown in the SEQ ID No.1.Sequence in survey 16SrDNA sequence and the GenBank database is carried out BLAST comparison, and the result shows: bacterial strain CCTCC M2012260 and Rhodopseudomonas (Pseudomonas) homology is all very high, with the similarity of P.fluorescens be 99%.Combining form, physiological and biochemical property and 16SrDNA sequential analysis are accredited as Pseudomonas fluorescens (Pseudomonas fluorescens).
CCTCC M 2012260 bacterial strains can change insoluble inorganic phosphorus in the soil into available state phosphorus and absorb for echinopodospora jamaicensis, promote the echinopodospora jamaicensis growth.
Beneficial effect: echinopodospora jamaicensis rhizosphere high-efficient phosphate-solubilizing pseudomonas fluorescens of the present invention shakes at liquid under the situation of training, to insoluble phosphate tricalcium phosphate (Ca
3(PO
4)
2), tertiary iron phosphate (FePO
44H
2O), aluminum phosphate (AlPO
4) and hydroxyapatite (Ca
10(PO
4)
6(OH)
2) have stronger solute effect, to compare with contrast (CK), difference is extremely remarkable; Echinopodospora jamaicensis rhizosphere high-efficient phosphate-solubilizing pseudomonas fluorescens of the present invention is made microbial inoculum inoculation echinopodospora jamaicensis seedling.The result shows that this microbial inoculum can obviously promote growing of echinopodospora jamaicensis, and therefore, the present invention provides good strain resource for developing echinopodospora jamaicensis special microorganism fertilizer in the future.
Description of drawings
Pseudomonas fluorescens Pseudomonas fluorescens FXW-HS7A, be preserved in Chinese typical culture collection center, address: China. Wuhan. Wuhan University, postcode 430072, culture presevation number: CCTCC M 2012260, preservation day is on July 1st, 2012.
Fig. 1 is that the FXW-HS7A bacterial strain is to the dissolving power of insoluble phosphate tricalcium phosphate, tertiary iron phosphate, aluminum phosphate and hydroxyapatite.
Fig. 2 is the short fruit of coming into force of the sweetgum of FXW-HS7A.
Embodiment
According to following embodiment, can make those skilled in the art understand the present invention better.Embodiment is described only to be used for explanation the present invention, and should also can not limit the present invention described in detail in claims.
The phosphorus decomposing test of embodiment 1:FXW-HS7A bacterial strain laboratory.
Phosphorus decomposing culture medium A: glucose 10g, Ca
3(PO
4)
25g, MgCl
25g, KCl 0.2g, MgSO
47H
2O 0.25g, (NH
4)
2SO
40.1g, distilled water 1000mL, pH 7.0.
Phosphorus decomposing substratum B: with hydroxyapatite (Ca
10(PO
4)
6(OH)
2) replace the Ca in the phosphorus decomposing culture medium A
3(PO
4)
2, other composition is identical with content.
Phosphorus decomposing culture medium C: with tertiary iron phosphate (FePO
44H
2O) Ca in the replacement phosphorus decomposing culture medium A
3(PO
4)
2, other composition is identical with content.
Phosphorus decomposing substratum D: with aluminum phosphate (AlPO
4) replace the Ca in the phosphorus decomposing culture medium A
3(PO
4)
2, other composition is identical with content.
FXW-HS7A inoculation NA substratum (extractum carnis 3g with activation, peptone 10g, sodium-chlor 5g, distilled water 1000mL, pH 7.2 ~ 7.4) in, 28 ℃ of shaking culture 18 ~ 24h are as seed liquor, with seed liquor by 1%(v/v) inoculum size is inoculated into respectively 50mL phosphorus decomposing culture medium A is housed, B, C, in the 100mL triangular flask of D, be contrast (CK) with the phosphorus decomposing substratum that connects the blank seed liquor of equal volume, each handles three repetitions, 28 ℃, after 180r/min cultivates 3d, 4 ℃ of fermented liquids, the centrifugal 10min of 10000r/min, molybdenum antimony resistance colorimetric method is measured titanium pigment content in the fermented liquid, the results are shown in Figure 1.
From Fig. 1, can draw, respectively with insoluble phosphate tricalcium phosphate (Ca
3(PO
4)
2), tertiary iron phosphate (FePO
44H
2O), aluminum phosphate (AlPO
4) and hydroxyapatite (Ca
10(PO
4)
6(OH)
2) be unique phosphorus source, behind the cultivation 3d, titanium pigment content all is higher than contrast in the FXW-HS7A fermented liquid.Show that more than bacterial strain JW-JS1 all has stronger dissolving power to four kinds of insoluble phosphorus sources.
The test of embodiment 2:FXW-HS7A greenhouse pot culture.
With after the bacterial strain FXW-HS7A activation, be inoculated in a small amount of thalline of transfering loop picking 50mLNA substratum (extractum carnis 3g, peptone 10g are housed, sodium-chlor 5g, distilled water 1000mL, pH 7.2 ~ 7.4) the 100mL triangular flask in, 29 ℃, 200r/min shaking culture 48h.(4 ℃, behind 6000r/min) the centrifugal 5min, stroke-physiological saline solution rinse thalline 3 times, stroke-physiological saline solution is regulated bacteria suspension (10 to fermented liquid
8Cfu/ml) make Inoculant.Inoculation echinopodospora jamaicensis (seedling age 1 year) is contrast with the equivalent stroke-physiological saline solution, and inoculum size is seedling 5mL/ strain.10 repetitions of every processing place the greenhouse unified management, and illumination is 12h/ days, waters in good time.
Echinopodospora jamaicensis is inoculated back 150 days growth situations and is seen Table 1, and as can be seen, inoculation FXW-HS7A can significantly promote the growth of echinopodospora jamaicensis seedling.Compare with contrast (CK), the rate of increase of the processing of inoculation FXW-HS7A on height of seedling and stem are thick reached 15.09% and 8.97% respectively, shortly comes into force fruit significantly.Fig. 2 for echinopodospora jamaicensis inoculation FXW-HS7A after photo 150 days the time, postvaccinal echinopodospora jamaicensis is apparently higher than contrast as can be seen.
The influence of the echinopodospora jamaicensis seedling growth of table 1 FXW-HS7A
Annotate: P＜0.05, same column different rows lowercase representative significant difference inequality.
Claims (2)
1. a Pseudomonas fluorescens (Pseudomonas fluorescens) FXW-HS7A, its called after Pseudomonas fluorescens (Pseudomonas fluorescens) of classifying, be preserved in Chinese typical culture collection center, culture presevation number is CCTCC NO:M2012260, and preservation day is on July 1st, 2012.
2. the application of the described Pseudomonas fluorescens of claim 1 (Pseudomonas fluorescens) FXW-HS7A in promoting the growth of echinopodospora jamaicensis seedling.
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| CN106811203B (en) * | 2017-02-27 | 2019-12-24 | 成都康巴金生物技术有限公司 | Preparation method of biological soil remediation agent and application of pseudomonas fluorescens H4 |
| CN110699303A (en) * | 2019-11-19 | 2020-01-17 | 河南工业大学 | High-efficiency phosphate-solubilizing pseudomonas as well as microbial agent and application thereof |
| CN113817653B (en) * | 2021-11-05 | 2023-04-25 | 昆明理工大学 | Pseudomonas fluorescens BsEB-1 and application thereof |
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Application publication date: 20130116 Assignee: Yangzhou little apple gardening Co.,Ltd. Assignor: Nanjing Forestry University Contract record no.: 2018320000233 Denomination of invention: Pseudomonas fluorescens and application of pseudomonas fluorescens for promoting growth of Chinese sweetgum Granted publication date: 20130918 License type: Common License Record date: 20181024 Application publication date: 20130116 Assignee: Beijing Huamei Wanxiang Technology Co.,Ltd. Assignor: Nanjing Forestry University Contract record no.: 2018320000235 Denomination of invention: Pseudomonas fluorescens and application of pseudomonas fluorescens for promoting growth of Chinese sweetgum Granted publication date: 20130918 License type: Common License Record date: 20181024 |
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Application publication date: 20130116 Assignee: Jiangsu Jing ancient environment construction Limited by Share Ltd. Assignor: Nanjing Forestry University Contract record no.: 2018320000363 Denomination of invention: Pseudomonas fluorescens and application of pseudomonas fluorescens for promoting growth of Chinese sweetgum Granted publication date: 20130918 License type: Common License Record date: 20181129 |
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