Screening of indigenous phosphate⁃solubilizing bacteria from Liquidambar formosana Hance rhizosphere and its potential applications for improving plant growth

doi:10.3969/j.issn.1000-2006.201908022

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2020, Vol. 44 ›› Issue (3): 95-104.doi: 10.3969/j.issn.1000-2006.201908022

Screening of indigenous phosphate⁃solubilizing bacteria from Liquidambar formosana Hance rhizosphere and its potential applications for improving plant growth

SONG Juan¹(), XU Guofang², ZHAO Xing³, YAO Yao³, YANG Xuexiang³, TANG Ronglin³, CUI Jiawang¹, CHEN Fengmao¹(), REN Jiahong²

^1.Co -Innovation Center of the Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, China
^2.The Department of Biological Science and Technology, Changzhi University, Changzhi 046011, China
^3.Forest Farm of Jurong City, Jurong 212424, China

Received:2019-08-15 Revised:2020-01-18 Online:2020-05-30 Published:2020-06-11
Contact: CHEN Fengmao E-mail:Sjj818388@outlook.com;cfengmao@njfu.edu.cn

Abstract

Abstract: Objective

Isolate and identify efficient phosphate solubilizing bacteria(PSB) from Liquidambar formosana Hance rhizosphere soil and investigate the application in phosphorus release.

Method

the PSB strains were graded based on morphological and physiological-biochemical characteristics, microbial identification system (MIDI), BIOLOG analysis and 16S rRNA identification methods. The effect of these PSB strains (JX7，JX18，JX21 and WH10）on plant growth was evaluated by determining the minor and major mineral contents of L. formosana leaves using inductively-coupled plasma atomic emission spectrometry.

Result

Isolates JX7, JX18, WH10 and JX21 were closely related to Variovorax paradoxus, Bacillus cereus, B. mycoides and B.cereus, respectively. The JX21 showed the highest content of soluble phosphate was (9.35 ± 0.89) mg/L, followed by JX7 [（6.63±0.09) mg/L], WH10[ (6.16 ± 0.12 ) mg/L] and JX18 [(5.32 ± 0.07 ) mg/L], all the maximum phosphate was fermented after 72 h of incubation. Phosphate-solubilizing bacteria (JX7, WH10, JX18 and JX21) selected from L. formosana rhizosphere showed a significantly improvement in shoot height and ground diameter of L. formosana (P < 0.05). As well as, these selected strains promoted L. formosana in taking up mineral elements such as phosphorus, magnesium, calcium and zinc under greenhouse conditions.

Conclusion

The isolates showed good phosphate-solubilizing activity and had potential for use as biofertilizer agents. Further study is needed to evaluate beneficial traits to develop commercial applications.

Key words: Liquidambar formosana Hance, plant growth promoting bacteria, plant nutrients, phosphate-solubilizing bacteria, biofertilizer

CLC Number:

S763

SONG Juan, XU Guofang, ZHAO Xing, YAO Yao, YANG Xuexiang, TANG Ronglin, CUI Jiawang, CHEN Fengmao, REN Jiahong. Screening of indigenous phosphate⁃solubilizing bacteria from Liquidambar formosana Hance rhizosphere and its potential applications for improving plant growth[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2020, 44(3): 95-104.

Figures/Tables 10

Table 1

Fig. 1

Fig. 2

Table 2

Table 3

Table 4

Table 5

Fig. 3

Fig. 4

Fig. 5

References 44

1	TAKAHASHI S，ANWAR M R.Wheat grain yield，phosphorus uptake and soil phosphorus fraction after 23 years of annual fertilizer application to an Andosol[J].Field Crop Res，2007，101(2)：160-171.DOI:10.1016/j.fcr.2006.11.003.
2	BUCHER M.Functional biology of plant phosphate uptake at root and mycorrhiza interfaces[J].New Phytol，2007，173(1)：11-26.DOI:10.1111/j.1469-8137.2006.01935.x.
3	MITTAL V，SINGH O，NAYYAR H，et al.Stimulatory effect of phosphate‍⁃‍solubilizing fungal strains (Aspergillusawamori and Penicilliumcitrinum) on the yield of chickpea (Cicerarietinum L.cv.GPF₂)[J].Soil Biol Biochem，2008，40(3)：718-727.DOI:10.1016/j.soilbio.2007.10.008.
4	WALPOLA B C, YOON M H. Isolation and characterization of phosphate solubilizing bacteria and their co⁃inoculation efficiency on tomato plant growth and phosphorous uptake[J]. Afr J Microbiol Res，2013, 7:266-275. DOI:10.5897/AJMR12.2282.
5	VIRUEL E，LUCCA M E，SIÑERIZ F.Plant growth promotion traits of phosphobacteria isolated from Puna，Argentina[J].Arch Microbiol，2011，193(7)：489-496.DOI:10.1007/s00203-011-0692-y.
6	ULÉN B，BECHMANN M，FÖLSTER J，et al.Agriculture as a phosphorus source for eutrophication in the north⁃west European countries，Norway，Sweden，United Kingdom and Ireland：a review[J].Soil Use Manag，2007，23(s1)：5-15.DOI:10.1111/j.1475-2743.2007.00115.x.
7	SINGH H，REDDY M S.Effect of inoculation with phosphate solubilizing fungus on growth and nutrient uptake of wheat and maize plants fertilized with rock phosphate in alkaline soils[J].Eur J Soil Biol，2011，47(1)：30-34.DOI:10.1016/j.ejsobi.2010.10.005.
8	DOBBELAERE S，CROONENBORGHS A，THYS A，et al.Effect of inoculation with wild type Azospirillumbrasilense and A.irakense strains on development and nitrogen uptake of spring wheat and grain maize[J].Biol Fertil Soils，2002，36(4)：284-297.DOI:10.1007/s00374-002-0534-9.
9	EGAMBERDIYEVA D.Plant‍⁃‍growth‍⁃‍promoting rhizobacteria isolated from a Calcisol in a semi⁃arid region of Uzbekistan：biochemical characterization and effectiveness[J].J Plant Nutr Soil Sci，2005，168(1)：94-99.DOI:10.1002/jpln.200321283.
10	REYES I，BERNIER L，ANTOUN H.Rock phosphate solubilization and colonization of maize rhizosphere by wild and genetically modified strains of Penicilliumrugulosum[J].Microb Ecol，2002，44(1)：39-48.DOI:10.1007/s00248-002-1001-8.
11	ZAIDI A，KHAN M，AMIL M.Interactive effect of rhizotrophic microorganisms on yield and nutrient uptake of chickpea (Cicerarietinum L.)[J].Eur J Agron，2003，19(1)：15-21.DOI:10.1016/s1161-0301(02)00015-1.
12	HINSINGER P，BRAVIN M，DEVAU N，et al.Soil⁃root⁃microbe interactions in the rhizosphere：a key to understanding and predicting nutrient bio availability to plants[J].RC Suelo Nutr Veg，2008，8(5)：39-47.DOI:10.4067/s0718-279120080004 00008.
13	SABER M S M, KABESH M O. Utilization of biofertilizers in field crop production. II. a comparison study on the effect of biofertilization or sulphur application on yield and nutrient uptake by lentil plants[J]. Eur J Soil Sci, 1990, 30: 415-422.
14	SUNDARA B，NATARAJAN V，HARI K.Influence of phosphorus solubilizing bacteria on the changes in soil available phosphorus and sugarcane and sugar yields[J].Field Crop Res，2002，77(1)：43-49.DOI:10.1016/s0378-4290(02)00048-5.
15	SHEN J，LI R，ZHANG F，et al.Crop yields，soil fertility and phosphorus fractions in response to long⁃term fertilization under the rice monoculture system on a calcareous soil[J].Field Crop Res，2004，86(2/3)：225-238.DOI:10.1016/j.fcr.2003.08.013.
16	NURAINI Y，ARFARITA N，SISWANTO B.Isolation and characteristic of nitrogen‍⁃‍fixing bacteria and phosphate‍⁃‍solubilizing bacteria from soil high in mercury in tailings and compost areas of artisanal gold mine[J].Agrivita J Agr Sci，2015，37(1)：0126-0537.DOI:10.17503/agrivita-2015-37-1-p001-007.
17	ILLMER P，SCHINNER F.Solubilization of inorganic calcium phosphates：solubilization mechanisms[J].Soil Biol Biochem，1995，27(3)：257-263.DOI:10.1016/0038-0717(94)00190-c.
18	BASHAN Y，KAMNEV A A，DE⁃‍BASHAN L E.A proposal for isolating and testing phosphate‍⁃‍solubilizing bacteria that enhance plant growth[J].Biology and Fertility of Soils，2013，49(1)：1-2.DOI:10.1007/s00374-012-0756-4.
19	KUNITSKY C, OSTERHOUT G, SASSER M. Identification of microorganisms using fatty acid methyl ester (FAME) analysis and the MIDI Sherlock Microbial Identification System［C］// MILLER M. Encyclopedia of rapid microbiological methods. River Grove, IL, USA:DAVIS Healthcare International Publishers: 2005: 1–17.
20	CHERKAOUI A，HIBBS J，EMONET S，et al.Comparison of two matrix‍⁃‍assisted laser desorption ionization‍⁃‍time of flight mass spectrometry methods with conventional phenotypic identification for routine identification of bacteria to the species level[J].J Clin Microbiol，2010，48(4)：1169-1175.DOI:10.1128/jcm.01881-09.
21	NATH D，MAURYA B R，MEENA V S.Documentation of five potassium⁃‍and phosphorus⁃‍solubilizing bacteria for their K and P‍⁃‍solubilization ability from various minerals[J].Biocatal Agric Biotechnol，2017，10：174-181.DOI:10.1016/j.bcab.2017. 03.007.
22	JACKSON M L，PRENTICE‍⁃‍HALL I N C，ENGLEWOOD CLIFFS N J. Soil chemical analysis[J].Soil Sci，1958，85(6)：339.DOI:10.1097/00010694-195806000-00014.
23	KRIEG N R,SNEATH P H A,STALEY J T, et al. Bergy’s manual of determinative bacteriology[J]. Jama⁃J Am Med Assoc,2000,15(170):408. DOI:10.1136/pgmj.15.170.408.
24	CHEN X，PIZZATTI C，BONALDI M，et al.Biological control of lettuce drop and host plant colonization by rhizospheric and endophytic streptomycetes[J].Front Microbiol，2016，7：714.DOI:10.3389/fmicb.2016.00714.
25	YU X，LIU X，ZHU T H，et al.Isolation and characterization of phosphate‍⁃‍solubilizing bacteria from walnut and their effect on growth and phosphorus mobilization[J].Biol Fertil Soils，2011，47(4)：437-446.DOI:10.1007/s00374-011-0548-2.
26	LI G E，WU X Q，YE J R，et al.Isolation and identification of phytate⁃degrading rhizobacteria with activity of improving growth of poplar and Masson pine[J].World J Microbiol Biotechnol，2013，29(11)：2181-2193.DOI:10.1007/s11274-013-1384-3.
27	SLABBINCK B，DE BAETS B，DAWYNDT P，et al.Towards large‍⁃‍scale FAME‍⁃‍based bacterial species identification using machine learning techniques[J].Syst Appl Microbiol，2009，32(3)：163-176.DOI:10.1016/j.syapm.2009.01.003.
28	DESPRÉS V R，NOWOISKY J F，KLOSE M，et al.Characterization of primary biogenic aerosol particles in urban，rural，and high⁃alpine air by DNA sequence and restriction fragment analysis of ribosomal RNA genes[J].Biogeosciences，2007，4(6)：1127-1141.DOI:10.5194/bg-4-1127-2007.
29	FYKSE E M，AARSKAUG T，THRANE I，et al.Legionella and non⁃Legionella bacteria in a biological treatment plant[J].Can J Microbiol，2013，59(2)：102-109.DOI:10.1139/cjm-2012-0166.
30	ALLEN R A，TOLLEY J O，JESAITIS A J.Preparation and properties of an improved photoaffinity ligand for the N⁃formyl peptide receptor[J].Biochim Et Biophys Acta BBA⁃Gen Subj，1986，882(3)：271-280.DOI:10.1016/0304-4165(86)90248-5.
31	SILINI⁃CHE H，SILINI A，GHOUL M，et al.Isolation and characterization of plant growth promoting traits of a rhizobacteria：Pantoeaagglomerans lma2[J].Pak J Biol Sci，2012，15(6)：267-276.DOI:10.3923/pjbs.2012.267.276.
32	HANIF M K，HAMEED S，IMRAN A，et al.Isolation and characterization of a Î²⁃propeller gene containing phosphobacterium Bacillus subtilis strain KPS⁃11 for growth promotion of potato (Solanumtuberosum L.)[J].Front Microbiol，2015，6：583.DOI:10.3389/fmicb.2015.00583.
33	LI X L，LUO L J，YANG J S，et al.Mechanisms for solubilization of various insoluble phosphates and activation of immobilized phosphates in different soils by an efficient and salinity⁃tolerant Aspergillus Niger strain An2[J].Appl Biochem Biotechnol，2015，175(5)：2755-2768.DOI:10.1007/s12010-014-1465-2.
34	BERCHEBRU L，RAMEIL P，GAUDIN J C，et al.Normalization of test and evaluation of biothreat detection systems：overcoming microbial air content fluctuations by using a standardized reagent bacterial mixture[J].J Microbiol Methods，2014，105：141-145.DOI:10.1016/j.mimet.2014.07.013.
35	RODRÍGUEZ H，FRAGA R，GONZALEZ T，et al.Genetics of phosphate solubilization and its potential applications for improving plant growth⁃promoting bacteria[J].Plant and Soil，2006，287(1/2)：15-21.DOI:10.1007/s11104-006-9056-9.
36	ZHAO K，PENTTINEN P，ZHANG X P，et al.Maize rhizosphere in Sichuan，China，hosts plant growth promoting Burkholderiacepacia with phosphate solubilizing and antifungal abilities[J].Microbiol Res，2014，169(1)：76-82.DOI:10.1016/j.micres.2013.07.003.
37	SMITH S E, READ D. Mycorrhizal symbiosis[M]. San Diego:USA:Academic Press, 1997.
38	BHATTACHARYYA D，GARLADINNE M，LEE Y H.Volatile indole produced by rhizobacterium Proteusvulgaris JBLS202 stimulates growth of Arabidopsisthaliana through auxin，cytokinin，and brassinosteroid pathways[J].J Plant Growth Regul，2015，34(1)：158-168.DOI:10.1007/s00344-014-9453-x.
39	PÉREZ⁃FLORES P L，VALENCIA⁃CANTERO E，ALTAMIRANO⁃HERNÁNDEZ J，et al.Bacillusmethylotrophicus M4⁃96 isolated from maize (Zeamays) rhizoplane increases growth and auxin content in Arabidopsisthaliana via emission of volatiles[J].Protoplasma，2017，254(6)：2201-2213.DOI:10.1007/s00709-017-1109-9.
40	CALVO P，WATTS D B，KLOEPPER J W，et al.Effect of microbial‍⁃‍based inoculants on nutrient concentrations and early root morphology of corn (Zeamays)[J].J Plant Nutr Soil Sci，2017，180(5)：636.DOI:10.1002/jpln.201770055.
41	RUDRESH D L，SHIVAPRAKASH M K，PRASAD R D.Effect of combined application of Rhizobium，phosphate solubilizing bacterium and Trichoderma spp.on growth，nutrient uptake and yield of chickpea (Ciceraritenium L.)[J].Appl Soil Ecol，2005，28(2)：139-146.DOI:10.1016/j.apsoil.2004.07.005.
42	GULL M，HAFEEZ F Y，SALEEM M，et al.Phosphorus uptake and growth promotion of chickpea by co‍⁃‍inoculation of mineral phosphate solubilising bacteria and a mixed rhizobial culture[J].Aust J Exp Agric, 2004，44(6)：623.DOI:10.1071/ea02218.
43	POONGUZHALI S，MADHAIYAN M，TONGMIN S. Isolation and identification of phosphate solubilizing bacteria from Chinese cabbage and their effect on growth and phosphorus utilization of plants[J]. J Microbiol biotechn,2008,18(4): 773-777. DOI: 10.1007/s12275-006-0149-1.
44	DE FREITAS J R，GERMIDA J J.A root tissue culture system to study winter wheat⁃rhizobacteria interactions[J].Appl Microbiol Biotechnol，1990，33(5)：589-595.DOI:10.1007/bf00172557.

Metrics

Comments

www.nldxb.njfu.edu.cn

Edited ＆ Published by Editorial Department of Nanjing Forestry University(Natural Sciences Edition)
Address： No.159 Longpan Road,Nanjing 210037 Jiangsu,P.R.China
E-mail ：xuebao@njfu.edu.cn , xuebao@njfu.com.cn
Telephone +86-25-85428247,85427076
Distributed by China International Book Trading Corporation P.O. Box 399, Beijing ,P.R. China

strain 菌株	D／mm (dissolved phosphorus circle diameter 溶磷圈直径)	d／mm (colony diameter 菌落直径)	D/d	ring type 圈类型
WH10	32.46 ± 0.60	14.34 ± 0.28	2.26 ± 0.003	grease ring溶脂圈
WH18	44.84 ± 0.40	36.34 ± 0.34	1.23 ± 0.002	grease ring溶脂圈
WH19	37.67 ± 0.80	18.62 ± 0.44	2.02 ± 0.005	grease ring溶脂圈
JX1	47.15 ± 0.30	34.70 ± 0.52	1.36 ± 0.012	grease ring溶脂圈
JX7	8.91 ± 0.50	3.50 ± 0.19	2.55 ± 0.005	hydrolysis ring水解圈
JX8	8.33 ± 0.70	4.02 ± 0.53	2.07 ± 0.101	hydrolysis ring水解圈
JX9	9.60 ± 0.34	4.22 ± 0.26	2.27 ± 0.060	hydrolysis ring水解圈
JX11	13.22 ± 0.26	4.66 ± 0.33	2.84 ± 0.146	hydrolysis ring水解圈
JX12	7.13 ± 0.43	5.10 ± 0.23	1.40 ± 0.021	hydrolysis ring水解圈
JX13	7.00 ± 0.54	3.38 ± 0.13	2.07 ± 0.080	hydrolysis ring水解圈
JX15	29.53 ± 1.38	15.10 ± 0.85	1.96 ± 0.019	grease ring溶脂圈
JX18	44.77 ± 1.64	29.08 ± 1.32	1.54 ± 0.014	grease ring溶脂圈
JX19	40.02 ± 1.14	29.97 ± 0.76	1.34 ± 0.005	grease ring溶脂圈
JX20	39.96 ± 0.83	27.04 ± 0.54	1.48 ± 0.002	grease ring溶脂圈
JX21	42.45 ± 0.53	25.48 ± 0.36	1.67 ± 0.004	grease ring溶脂圈

strain 菌株	cell shape 菌体形态	Gram reaction 革兰氏染色	3% KOH	spores 芽孢	flagellum 鞭毛	aerobic condition 需氧情况	colony morphology 菌落形状
JX7	+	G-	+	-	-	+	colonies smaller, yellowish green, circular, slimy, moist 菌落较小，黄绿色，边缘整齐，光滑湿润
WH10	+	G+	-	+	+	+	larger colonies, creamy, circular, slimy, dry 菌落较大，乳白色，圆形,边缘整齐，光滑干燥
JX18	+	G+	-	+	+	+	larger colonies,whitish, circular, slimy, dry 菌落较大，白色圆形，边缘整齐，光滑干燥
JX21	+	G+	-	+	+	+	larger colonies, circular,the middle wrinkles, slimy, dry 菌落较大,圆形,边缘整齐，中间有皱褶，光滑干燥

project measureed 测定项目	JX7	WH10	JX18	JX21
BIOLOG result BIOLOG结果	Variovorax paradoxus 争论产碱菌	Bacillus mycoides 蕈状芽孢杆菌	Bacillus cereus 蜡状芽孢杆菌	Bacillus cereus 蜡状芽孢杆菌
oxidase test氧化酶	-	+	+	+
contact enzyme接触酶	+	+	+	+
lecithinase卵磷脂酶	-	+	+	+
gelatin hydrolysis 明胶水解	-	+	+	+
starch hydrolysis 淀粉水解	-	+	+	+
H₂S test产生H₂S	-	+	-	-
citrate utilization 柠檬酸盐利用	-	+	+	+
indole test 吲哚试验	+	-	+	+
methyl red test 甲基红试验	-	-	-	-
oges praskaure’s 伏普试验	+	+	+	+
nitrate reduction 硝酸盐还原	+	-	-	-

fatty acid 脂肪酸	JX18	JX21	WH10
13∶0 iso	6.72	6.77	6.08
14∶0 iso	3.10	3.40	3.11
14∶0	3.03	2.65	3.53
15∶0 iso	31.45	20.12	26.00
15∶0 anteiso	4.45	3.49	3.57
summed in feature 2 of 14∶0 3OH/16∶1 iso I	2.74	3.58	4.06
15∶1 w5c	no	4.30	3.51
16∶0 iso	5.25	5.13	5.10
summed in feature 3 of 16∶1 w6c/16∶1 w7c	8.21	8.95	7.61
16∶0	3.84	7.65	7.77
17∶1 iso w10c	3.94	2.38	2.98
17∶1 iso w5c	6.76	8.99	7.52
17∶0 iso	8.54	6.55	6.33
18∶0	2.18	6.40	no
summed in feature 8 of 18∶1 w7c	0.27	0.60	1.10
summed feature 2 of unknown 10.9525	2.74	3.58	4.06
summed feature 3 of 16∶1 w6c/16∶1 w7c	8.21	8.95	9.28
sim index	0.715	0.167	0.357

strain 菌株	specific name 种名	BIOLOG similarity		16SrDNA homology /% 16SrDNA同源性
strain 菌株	specific name 种名	Probability (PROB)/% BIOLOG可能性	(SIM) index value BIOLOG 相似性指数	16SrDNA homology /% 16SrDNA同源性
JX7	Variovorax paradoxus	99	0.648	97
WH10	Bacillus mycoides	83	0.636	95
JX18	Bacillus cereus	89	0.697	96
JX21	Bacillus cereus	94	0.720	97

Screening of indigenous phosphate⁃solubilizing bacteria from Liquidambar formosana Hance rhizosphere and its potential applications for improving plant growth

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 10

References 44

Related Articles 1

Recommended Articles

Metrics

Comments

Author

Reader

Reviewer