基于嗜铁素介导的吡咯伯克霍尔德氏菌JK-SH007促生作用机制研究

doi:10.3969/j.issn.1000-2006.201811058

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南京林业大学学报（自然科学版） ›› 2019, Vol. 43 ›› Issue (6): 165-172.doi: 10.3969/j.issn.1000-2006.201811058

基于嗜铁素介导的吡咯伯克霍尔德氏菌JK-SH007促生作用机制研究

闵莉静¹^,²(), 郭璐³, 叶建仁¹^,^*()

1.南京林业大学林学院,南方现代林业协同创新中心,江苏南京 210037
2.湖州师范学院生命科学学院,浙江省媒介生物学与病原控制重点实验室,浙江湖州 313000
3.湖州师范学院附属湖州市第一人民医院,浙江湖州 313000

收稿日期:2018-11-28 修回日期:2019-03-27 出版日期:2019-11-30 发布日期:2019-11-30
通讯作者: 叶建仁
基金资助:
国家自然科学基金项目(31270684);江苏农业自主创新项目(cx16-1005);2019年度湖州市第二批科技计划项目(2019Y203);湖州市自然科学基金项目(2015GY08);浙江省绿色农药2011协同创新中心开放基金项目(LSNY201820)

Mechanism of Burkholderia pyrrocinia JK-SH007 growth-promoting to plant via siderophore-mediation

MIN Lijing¹^,²(), GUO Lu³, YE Jianren¹^,^*()

1. College of Forestry, Nanjing Forestry University, Co-Innovation Center for the Sustainable Forestry in Southern China,Nanjing 210037, China
2. College of Life Science, Huzhou University, Huzhou 313000, China
3. Huzhou First People’s Hospital, Huzhou University, Huzhou 313000, China

Received:2018-11-28 Revised:2019-03-27 Online:2019-11-30 Published:2019-11-30
Contact: YE Jianren

摘要/Abstract

摘要：

【目的】分析吡咯伯克霍尔德菌 JK-SH007基于产嗜铁素的促生作用机制。【方法】通过萃取、过柱等操作,分离纯化,获得吡咯伯克霍尔德菌 JK-SH007所产嗜铁素(简称JK-SH007-嗜铁素)的纯品,通过HPLC和LC-MS对其结构进行鉴定,通过与病原菌共培养,测菌丝干质量的方法,分析其对3种杨树溃疡病病原真菌(金黄壳囊孢、拟茎点霉和七叶树壳梭孢)的抑制作用,以商品嗜铁素磺酸去铁胺(DFOB)作为对照,比较JK-SH007-细菌重悬液、1.4 mg/mL JK-SH007-嗜铁素溶液对番茄、黄瓜种子的萌发作用,并继续观察对黄瓜幼苗的促生作用和番茄挂果果品改善效果,评估JK-SH007-嗜铁素的直接与间接的促生作用。【结果】与对照相比,JK-SH007-嗜铁素显示出比DFOB更有效的抑制真菌生长的活性,3种病原真菌抑菌率分别为(89±0.9)%,(76±0.6)%和(90±0.7)%。同时,与JK-SH007-菌悬液相比,JK-SH007-嗜铁素溶液对番茄、黄瓜种子和幼苗活力的提高表现更好,而对于黄瓜幼苗的生长促进作用和番茄挂果的改善作用,其效果与菌悬液相近。【结论】嗜铁素为促生菌吡咯伯克霍尔德氏菌 JK-SH007的主要促生生防因子之一。

关键词: 吡咯伯克霍尔德氏菌, 嗜铁素, 抑菌, 促生

Abstract:

【Objective】We aimed to explore the mechanism by which Burkholderia pyrrocinia JK-SH007, a gram-negative bacterium, promotes plant growth; we explored the effects of siderophores on pathogenic fungal activity and plant growth. 【Method】 A sample of JK-SH007-siderophores was isolated and purified using column chromatography, and the siderophore structure was identified using high-performance liquid chromatography and liquid chromatography-mass spectrometry. We cultivated three pathogens that cause poplar canker (Cytospora chrysosperma, Phomopsis macrospora and Fusicoccum aesculi) with siderophore samples to measure pathogen inhibition, and recorded the dry weight of mycelium at the end of the cultivation period. We compared the effects of a JK-SH007-siderophore solution, a JK-SH007 bacterial re-suspension, and a standard siderophore, Deferroamine B solution (DFOB) on plant growth (seed germination, cucumber seedling growth and tomato fruiting).【Result】Compared to DFOB, the JK-SH007-siderophore sample was more effective at inhibiting fungal growth. The pathogenic fungi were inhibited by (89±0.9)%, (76±0.6)% and (90±0.7)%, forC. chrysosperma, P. macrospora and F. aesculin, respectively. Compared to the JK-SH007 re-suspension, the JK-SH007-siderophore solution resulted in better seed germination and higher seedling vigor; however, there were no significant differences for cucumber seedling growth or tomato fruiting. 【Conclusion】We applied a pure siderophore sample, extracted and purified using chromatography, to plants, and compared this with a bacterial re-suspension sample and a standard commercial product. The pure siderophore sample strongly promoted plant growth and the application of a pure siderophore solution can achieve better anti-fungal effects than DFOB, a result which should inform future commercial development of JK-SH007 as a bio-fertilizer.

Key words: Burkholderia pyrrocinia, siderophore characterization, antifungi activity, plant growth promotion

中图分类号:

S763

闵莉静,郭璐,叶建仁. 基于嗜铁素介导的吡咯伯克霍尔德氏菌JK-SH007促生作用机制研究[J]. 南京林业大学学报（自然科学版）, 2019, 43(6): 165-172.

MIN Lijing, GUO Lu, YE Jianren. Mechanism of Burkholderia pyrrocinia JK-SH007 growth-promoting to plant via siderophore-mediation[J].Journal of Nanjing Forestry University (Natural Science Edition), 2019, 43(6): 165-172.DOI: 10.3969/j.issn.1000-2006.201811058.

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参考文献 35

[1]	COMPANT S, NOWAK J, COENYE T, et al. Diversity and occurrence of Burkholderia spp. in the natural environment[J]. Fems Microbiology Reviews, 2010, 32(4):607-626. DOI: 10.1111/j.1574-6976.2008.00113.x. doi: 10.1111/j.1574-6976.2008.00113.x
[2]	叶建仁, 任嘉红, 李浩, 等. 洋葱伯克霍尔德氏菌及其在林木病害防治中的应用[J]. 南京林业大学学报(自然科学版), 2013, 37(4):149-155. DOI: 10.3969/j.issn.1000-2006.2013.04.028.
	YE J R, REN J H, LI H, et al. Application and its prospect analysis for Burkholderia cepacia in forest disease control [J]. Journal of Nanjing Forestry University (Natural Sciences Edition), 2013, 37(4):149-155.
[3]	刘丹丹, 李敏, 刘润进. 我国植物根围促生细菌研究进展[J]. 生态学杂志, 2016, 35(3):815-824. DOI: 10.13292/j.1000-4890.201603.033.
	LIU D D, LI M, LIU R J. Rent advances in the study of plant growth-promoting rhizobacteria in China[J]. Chinese Journal of Ecology, 2016, 35(3):815-824.
[4]	SCHMIDT S, BLOM J F, PERNTHALER J, et al. Production of the antifungal compound pyrrolnitrin is quorum sensing-regulated in members of the Burkholderia cepacia complex[J]. Environmental Microbiology, 2009, 11(6):1422-1437. DOI: 10.1111/j.1462-2920.2009.01870.x. doi: 10.1111/emi.2009.11.issue-6
[5]	LI G X, WU X Q, YR J R. Biosafety and colonization of Burkholderia multivorans WS-FJ9 and its growth-promoting effects on poplars[J]. Appl Microbiol Biotechnol, 2013, 97(24):10489-10498. DOI: 10.1007/s00253-013-5276-0. doi: 10.1007/s00253-013-5276-0
[6]	COENYE T, VANDAMME P, LIPUMA J J, et al. Updated version of the Burkholderia cepacia complex experimental strain panel[J]. Journal of Clinical Microbiology, 2003, 41(6):2797-2798. DOI: 10.1128/JCM.41.6.2797-2798.2003. doi: 10.1128/JCM.41.6.2797-2798.2003
[7]	林超, 余贤美, 王春妮, 等. 土壤嗜铁细菌C19的筛选、分子鉴定及其对炭疽菌和镰刀菌的拮抗作用[J]. 热带作物学报, 2009, 30(1):94-98. DOI: 10.3969/j.issn.1000-2561.2009.01.019.
	LIN C, YU X M, WANG C N, et al. Isolation of sderophore producing bacteria C19 from the soil and analysis of its Antagonism[J]. Chinese Journal of Tropical Crops, 2009, 30(1):94-98.
[8]	林超, 郑服丛, 贺春萍, 等. 嗜铁细菌C19对黄瓜种子发芽的影响及对芒果炭疽病生防效果初步评价[J]. 中国农学通报, 2009, 25(1):232-235.
	LIN C, ZHENG F C, HE C P, et al. The effect of siderophore-producing bacteria C19 on cucumber seed germination and biocontrol effects on mango anthracnose[J]. Chinese Agricultural Science Bulletin, 2009, 25(1):232-235.
[9]	REN J H, YE J R, LIU H, et al. Isolation and characterization of a new Burkholderia pyrrocinia strain JK-SH007 as a potential biocontrol agent[J]. World Journal of Microbiology & Biotechnology, 2011, 27(9):2203-2215. DOI: 10.1007/s11274-011-0686-6. doi: 10.1007/s11274-011-0686-6
[10]	任嘉红, 班虎栋, 叶建仁, 等. 吡咯伯克霍尔德氏菌JK-SH007的发酵条件及其对杨树溃疡病的防治效果[J]. 中国生物防治学报, 2010, 28(4):300-306. DOI: 10.3724/SP.J.1142.2010.40486.
	REN J H, BAN H D, YE J R. et al. Fermentation conditions of antagonistic strain Burkholderia pyrrocinia JK-SH007 and its control effect on poplar canker disease [J]. Chinese Journal of Biological Control, 2010, 28(4):300-306.
[11]	任嘉红, 王艳芳, 叶建仁. 杨树腐烂病拮抗细菌的筛选及其定殖研究[J]. 西部林业科学, 2009, 38(2):6-11. DOI: 10.3969/j.issn.1672-8246.2009.02.002.
	REN J H, WANG Y F, YE J R. Screening and colonizationof popular antagonistic bacteria against Valsa sordida [J]. Journal of West China Forestry Science, 2009, 38(2):6-11.
[12]	陈飞飞, 黄金思, 王翕韫, 等. 吡咯伯克霍尔德氏菌JK-SH007多聚半乳糖醛酸酶基因的克隆及表达分析[J]. 南京林业大学学报(自然科学版), 2018, 42(4):127-133. DOI: 10.1111/j.1365-2672.2004.02262.x.
	CHEN F F, HUANG J S, WANG X Y, et al. Cloning and expression analysis of polygalacturonase gene (BpPG) from Burkholderia pyrrocinia JK-SH007 [J]. Journal of Nanjing Forestry University (Natural Sciences Edition), 2018, 42(4):127-133.
[13]	任嘉红, 李浩, 刘辉, 等. 吡咯伯克霍尔德氏菌JK-SH007对杨树根际微生物数量及功能多样性的影响[J]. 林业科学, 2016, 52(5):126-133. DOI: 10.11707/j.1001-7488.201605015.
	REN J H, LI H, LIU H, et al. Influence of Burkholderia pyrrocinia JK-SH007 on the microbial population and functional diversity of microbial communities in the rhizosphere soil of poplar [J]. Scientia Silvaesinicae, 2016, 52(5):126-133.
[14]	窦承阳, 王焱, 张岳峰, 等. 3种促生有益微生物在上海梨树上的应用[J]. 南京林业大学学报(自然科学版), 2017, 41(4):186-190. DOI: 10.3969/j.issn.1000-2006.201512029.
	DOU C Y, WANG Y, ZHANG Y F, et al. An application of three somatotrophic bacteria on pear in Shanghai.[J]. Journal of Nanjing Forestry University (Natural Sciences Edition), 2017, 41(4):186-190.
[15]	李浩, 任嘉红, 叶建仁, 等. 吡咯伯克霍尔德氏菌JK-SH007分批发酵动力学模型的构建[J]. 南京林业大学学报(自然科学版), 2016, 40(2):173-178. DOI: 10.3969/j.issn.1000-2006.2016.02.029.
	LI H, REN J H, YE J R, et al. Construction of batch fermentation kinetics model of Burkholderia pyrrocinia strain JK-H007 [J]. Journal of Nanjing Forestry University (Natural Sciences Edition), 2016, 40(2):173-178.
[16]	任嘉红, 吴小芹, 刘辉, 等. 吡咯伯克霍尔德氏菌JK-SH007抗菌蛋白的分离纯化[J]. 微生物学通报, 2010, 37(6):872-880. DOI: 10.13344/j.microbiol.china.2010.06.018.
	REN J H, WU X Q, L H, et al. Isolation and purification of antifungal protein from Burkholderia pyrrocinia JK-SH007 [J]. Microbiology China, 2010, 37(6):872-880.
[17]	NEILANDS J B. Microbial iron compounds[J]. Annual Review of Biochemistry, 1981, 50(1):715-31. DOI: 10.1146/annurev.bi.50.070181.003435. doi: 10.1146/annurev.bi.50.070181.003435
[18]	ARNOW L E. Colorimetric determination of the components of 3,4-dihydroxyphenylalaninetrosine mixtures[J]. Circulation Journal Official Journal of the Japanese Circulation Society, 1937, 50(1):531-537. DOI: 10.1146/annurev.bi.50.070181.003435.
[19]	SHENKER M, OLIVER I, HELMANN M, et al. Utilization by tomatoes of iron mediated by a siderophore produced by Rhizopus arrhizus[J]. Journal of Plant Nutrition, 1992, 15(10):2173-2182. DOI: 10.1080/01904169209364466. doi: 10.1080/01904169209364466
[20]	SCHWYN B, NEILANDS J B. Universal chemical assay for the detection and determination of siderophores[J]. Analytical Biochemistry, 1987, 160(1):47-56. DOI: 10.1016/0003-2697(87)90612-9. doi: 10.1016/0003-2697(87)90612-9
[21]	LIU X, YANG G M, GUAN D X, et al. Catecholate-siderophore produced by As-resistant bacterium effectively dissolved FeAsO₄ and promoted Pteris vittata growth[J]. Environmental Pollution, 2015(206):376-381. DOI: 10.1016/j.envpol.2015.07.034.
[22]	BERNER I, GREINER M, METZGER J, et al. Identification of enterobactin and linear dihydroxybenzoylserine compounds by HPLC and ion spray mass spectrometry (LC/MS and MS/MS)[J]. Biology of Metals, 1991, 4(2):113. DOI: 10.1007/BF01135388. doi: 10.1007/BF01135388
[23]	ABDUL-BAKI A A, ANDERSON J D. Vigor determination in soybean seet by multiple criteria[J]. Crop Science, 1973, 13:630-633.DOI: 10.2135/cropsci1973.0011183×001300060013X. doi: 10.2135/cropsci1973.0011183X001300060013x
[24]	AlI S, CHARLES T C, GLICK B R. Amelioration of high salinity stress damage by plant growth-promoting bacterial endophytes that contain ACC deaminase[J]. Plant Physiology & Biochemistry, 2014(80):160-167. DOI: 10.1016/j.plaphy.2014.04.003.
[25]	郑国华, 王金昌, 王小红. 植物根际细菌的促生机制[J]. 江西科学, 2012, 30(4):454-458. DOI: 10.3969/j.issn.1001-3679.2012.04.011.
	ZHENG G H, WANG J C, WANG X H. The deveopment of research on high efficiency phosphate solubilizing bacteria. Jiangxi Science, 2012, 30(4):454-458.
[26]	FREYBERG D P, ABUDARI K, RAYMOND K N. Coordination chemistry of microbial iron transport compounds. 17. Preparation and structural characterization of tris(N-methylthiobenzohydroxamato) obalt(III), -chromium(III), -iron(III), and-manganese(III)[J]. Inorganic Chemistry, 1979, 11(8):3037-3043. DOI: 10.1002/chin.198008084.
[27]	DAVE B P, ANSHUMAN K, HAJELA P. Siderophores of halophilic archaea and their chemical characterization[J]. Indian Journal of Experimental Biology, 2006, 44(4):340-344. DOI: 10.1016/S1051-0443(99)70113-2.
[28]	SCHIPPERS B, BAKKER A W, BAKKER P A H M. Interactions of deleterious and beneficial rhizosphere microorganisms and the effect of cropping practices[J]. Annual Review of Phytopathology, 1987, 25(1):339-358. DOI: 10.1146/annurev.py.25.090187.002011. doi: 10.1146/annurev.py.25.090187.002011
[29]	VOISARD C, KEEL C, HAAS D, et al. Cyanide production by Pseudomonas fluorescens helps suppress black root rot of tobacco under gnotobiotic conditions[J]. Embo Journal, 1989, 8(2):351-358. DOI: 10.1002/j.1460-2075.1989.tb03384.x. doi: 10.1002/embj.1989.8.issue-2
[30]	PAL K K, TILAK K V, SAXENA A K, et al. Suppression of maize root diseases caused by Macrophomina phaseolina, Fusarium moniliforme and Fusarium graminearum by plant growth promoting rhizobacteria[J]. Microbiological Research, 2001, 156(3):209-223. DOI: 10.1078/0944-5013-00103. doi: 10.1078/0944-5013-00103
[31]	YU X, AI C, XIN L, et al. The siderophore-producing bacterium, Bacillus subtilis CAS15, has a biocontrol effect on Fusarium wilt and promotes the growth of pepper[J]. European Journal of Soil Biology, 2011, 47(2):138-145. DOI: 10.1016/j.ejsobi.2010.11.001. doi: 10.1016/j.ejsobi.2010.11.001
[32]	MCLOUGHLIN T J, QUINN J P, BETTERMANN A, et al. Pseudomonas cepacia suppression of sunflower wilt fungus and role of antifungal compounds in controlling the disease[J]. Appl Environ Microbiol, 1992, 58(5):1760-1763. DOI: 10.1007/BF00713942. doi: 10.1128/aem.58.5.1760-1763.1992
[33]	FOLSCHWEILLER N, SCHALK I J, CELIA H, et al. The pyoverdin receptor FpvA, a TonB-dependent receptor involved in iron uptake by Pseudomonas aeruginosa[J]. Membrane Biochemistry, 2000, 17(3):123-133. DOI: 10.1080/09687680050197356.
[34]	AHMED E, HOLMSTROM S J M. Siderophores in environmental research: roles and applications[J]. Microbial Biotechnology, 2014, 7(3):196-208. DOI: 10.1111/1751-7915.12117. doi: 10.1111/1751-7915.12117
[35]	KLOEPPER J W, LEONG J, TEINTZE M, et al. Enhanced plant growth by siderophores produced by plant growth-promoting rhizobacteria[J]. Nature, 1980, 286(5776):885-886. DOI: 10.1038/286885a0. doi: 10.1038/286885a0

处理 treatment	发芽率/% germination index		胚根长/cm radicle length		幼苗活力指数/cm SVI
处理 treatment	黄瓜 cucumber	番茄 tomato	黄瓜 cucumber	番茄 tomato	黄瓜 cucumber	番茄 tomato
无菌水(对照)	90.0 c	86.7 c	4.1 c	6.8 c	3.6 d	5.9 c
JK-SH007-重悬液	96.7 b	90.7 b	4.3 c	6.9 c	4.2 c	6.3 b
JK-SH007-嗜铁素	100 a	100 a	6.2 a	7.8 a	6.2 a	7.8 a
DFOB	100 a	87.5 c	5.6 b	7.2 b	5.6 b	6.3 b

处理 treatments	株高/cm total plant height	根长/cm root length	总叶绿素含量/ (mg·L^-1) total chlorophyll content	湿质量/g wet weight
处理 treatments	株高/cm total plant height	根长/cm root length	总叶绿素含量/ (mg·L^-1) total chlorophyll content	茎 stem	根 root	茎 stem	根 root	挂果数 No. of fruits	果质量/g fruit weight	果径/cm fruit diameter	糖含量/ (^oBx) degrees Brix
DFOB	55.31 b	5.24 c	2.95 a	93.42 b	4.22 a	8.42 c	0.63 c	16 c	70.52 d	5.19 b	3.88 c
JK-SH007-嗜铁素	62.78 a	6.16 b	2.67 b	124.51 a	3.82 c	9.45 a	0.89 a	18 b	84.53 c	5.33 a	4.24 ab
JK-SH007-重悬液	53.82 b	6.24 a	2.74 b	86.72 c	3.94 b	8.84 b	0.78 b	22 a	88.78 a	5.40 a	4.25 a
无菌水(对照)	43.18 c	4.34 d	1.74 c	83.46 d	2.93 d	7.62 d	0.56 d	17 bc	85.37 b	5.35 a	4.12 b

基于嗜铁素介导的吡咯伯克霍尔德氏菌JK-SH007促生作用机制研究

Mechanism of Burkholderia pyrrocinia JK-SH007 growth-promoting to plant via siderophore-mediation

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