CN102876598A - 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 PDF

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CN102876598A
CN102876598A CN2012103274413A CN201210327441A CN102876598A CN 102876598 A CN102876598 A CN 102876598A CN 2012103274413 A CN2012103274413 A CN 2012103274413A CN 201210327441 A CN201210327441 A CN 201210327441A CN 102876598 A CN102876598 A CN 102876598A
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pseudomonas fluorescens
growth
sweetgum
phosphate
chinese
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CN102876598B (en
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张明艳
陈凤毛
任嘉红
徐国芳
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Nanjing Forestry University
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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

A kind of Pseudomonas fluorescens and the application in promoting the echinopodospora jamaicensis growth thereof
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 Hamamelidaceae sweetgum subfamily Liquidambar, calls to be maple, black meal wood, sweet gum etc.Being the important indigenous tree of China, also is subtropical zone good fast-growing broad leaved and deciduous broad leaved seeds, and its wide adaptability, growth are typical deserted mountain vanguard tree seed rapidly.Safeguard the soil fertility successful, profitable on ecology, be one of first-selected seeds of artificial forest Tree Structure 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 large, and the forest land of plantation sweetgum often easily causes soil nutrient deficiency.Just affect 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 fast-growing sweetgum 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 widely cultivated 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, therefore phosphorus usually becomes the limiting factor of plant nutrition.Molten phosphorus microorganism is distributed widely in soil, 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 the 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.
Phosphate releasing 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 integration effect; Poplar bud plum, 2010).Phosphate-solubilizing bacteria can improve the phosphorus element supply 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 have no report.Therefore improving soil available phosphorus content just has very great 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 plant height effect phosphate-solubilizing pseudomonas fluorescens Pseudomonas fluorescens FXW-HS7A.Be preserved in Chinese Typical Representative 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, size is 0.43 * 0.64 μ m ~ 1.89 * 2.00 μ m approximately, without gemma, amphitrichous; Aerobic, oxydase reaction is positive, and catalase test is positive, and Starch Hydrolysis is negative, and M.R. and indole test are negative, and V.P. tests positive; Gelatine liquefication is positive, and the nitrate reductase negative produces ammonia and hydrogen sulfide production test negative, and the Citrate trianion growth test is 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 compare, 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 P 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 is in the situation that liquid shakes 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, this microbial inoculum can obviously promote growing of echinopodospora jamaicensis, and therefore, the present invention provides good strain resource for developing the echinopodospora jamaicensis microbial fertilizer special in the future.
Description of drawings
Pseudomonas fluorescens Pseudomonas fluorescens FXW-HS7A, be preserved in Chinese Typical Representative 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 that FXW-HS7A is to the short fruit of coming into force of sweetgum.
Embodiment
According to following embodiment, can make those skilled in the art understand better the present invention.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, take the phosphorus decomposing substratum that connects the blank seed liquor of equal volume as contrast (CK), each processes 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.
As can be drawn from Figure 1, 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.More than show, bacterial strain JW-JS1 all has stronger dissolving power to four kinds of Insoluble phosphates.
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), take the equivalent stroke-physiological saline solution as contrast, 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 rear 150 days growth situations and is seen Table 1, can find out, 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 respectively 15.09% and 8.97%, shortly comes into force fruit significantly.Fig. 2 is the photo 150 days time the behind the echinopodospora jamaicensis inoculation FXW-HS7A, can find out that postvaccinal echinopodospora jamaicensis is apparently higher than contrast.
Table 1 FXW-HS7A is on the impact of echinopodospora jamaicensis seedling growth
Figure DEST_PATH_GDA00002296897500041
Annotate: P<0.05, the not identical representative significant difference of same column different rows lowercase.
Figure IDA00002108209500011
Figure IDA00002108209500021

Claims (2)

1. Pseudomonas fluorescens FXW-HS7A, its Classification And Nomenclature is Pseudomonas fluorescens (Pseudomonas fluorescens), be preserved in Chinese Typical Representative culture collection center, culture presevation number is CCTCC M 2012260, and preservation day is on July 1st, 2012.
2. the application of Pseudomonas fluorescens claimed in claim 1 in promoting the echinopodospora jamaicensis growth.
CN 201210327441 2012-09-06 2012-09-06 Pseudomonas fluorescens and application of pseudomonas fluorescens for promoting growth of Chinese sweetgum Expired - Fee Related CN102876598B (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106811203A (en) * 2017-02-27 2017-06-09 成都康巴金生物技术有限公司 A kind of application of the preparation method and Pseudomonas fluorescens H4 of biological soil renovation agent
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
CN113817653A (en) * 2021-11-05 2021-12-21 昆明理工大学 Pseudomonas fluorescens BsEB-1 and application thereof

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
姚如斌等: "杨树解磷细菌蜡状芽孢杆菌和荧光假单胞菌发酵罐扩繁条件优化", 《林业科技开发》 *
李莎等: "接种乳黄粘盖牛肝菌和荧光假单胞菌对油松苗生长及猝倒病的影响", 《西北植物学报》 *
邱章忠: "杉木枫香混交林土壤化学性质和酶活性的研究", 《林业勘察设计(福建)》 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106811203A (en) * 2017-02-27 2017-06-09 成都康巴金生物技术有限公司 A kind of application of the preparation method and Pseudomonas fluorescens H4 of biological soil renovation agent
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
CN113817653A (en) * 2021-11-05 2021-12-21 昆明理工大学 Pseudomonas fluorescens BsEB-1 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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