Boron promotes secretion of extracellular polysaccharides and indole-3-acetic acid by Rhizobium

doi:10.11686/cyxb2020220

Abstract

Abstract:

The aim of this study was to determine how boron promotes the production of extracellular polysaccharides and indole-3-acetic acid （IAA） by the Rhizobium strain gn5f. A label-free protein quantitative technique was used to analyze the effect of boron on the protein expression of Rhizobium strain gn5f. The results showed that 100 mg?L^-1 was the optimum boron concentration for inducing polysaccharide and IAA production by Rhizobium strain gn5f. A comparison between the control （no boron） and the 100 mg?L^-1boron treatment identified 54 differentially expressed proteins； 7 up-regulated proteins and 47 down-regulated proteins. Bioinformatics analysis showed that these differentially expressed proteins were related to energy production and conversion， fatty acid β-oxidation， gluconeogenesis， amino acid metabolism， and various metabolic processes. The proposed mechanism by which boron promotes the production of extracellular polysaccharides and IAA by Rhizobium gn5f is as follows： boron up-regulates NAD-dependent succinate-semialdehyde dehydrogenase， acetyl-coenzyme A synthetase， and succinate-semialdehyde dehydrogenase （NADP⁺）， which promote metabolic pathways related to the tricarboxylic acid cycle such as pyruvate metabolism， γ-aminobutyric acid （GABA） bypass metabolism， and glutamate metabolism. The energy and various carbohydrates （D-galactose and D-glucose residues， D-glucuronic acid） provided by these metabolic pathways promote the synthesis of extracellular polysaccharides. Finally， IAA is synthesized from tryptophan via indole-3-acetamide （IAM） or indole-3-pyruvate under the action of related enzymes， and boron contributes to this process by improving tryptophan synthesis.

Key words: boron, rhizobia, extracellular polysaccharides, IAA, proteomics

Yong-gang CHEN, Wen-juan KANG, Fang WU, Yun A, Shang-li SHI, Cui-mei ZHANG, Zi-li LI. Boron promotes secretion of extracellular polysaccharides and indole-3-acetic acid by Rhizobium[J]. Acta Prataculturae Sinica, 2021, 30(5): 42-51.

Figures/Tables 7

Fig.1 Effects of different boron concentrations on gn5f OD600

Fig.2 Growth curve of strain gn5f treated by boron before and after

Fig.3 Effect of boron on extracellular polysaccharides and IAA production of gn5f

Fig. 4 SDS-PAGE electrophoresis results

Fig.5 Functional enrichment analysis of boron-treated rhizobium gn5f differentially expressed protein

Fig.6 KEGG annotation statistics of gn5f differentially expressed protein of boron-treated rhizobium

Fig.7 Top 20 pathways with the most differentially expressed proteins from rhizobia gn5f

References 39

1	Ma L， Ma S Y， Chen G P， et al. Physiological responses of pea and nodule symbiosis to water stress. Acta Prataculturae Sinica， 2019， 28（9）： 96-109.
	马蕾，马绍英，陈贵平，等. 豌豆与根瘤共生对水分胁迫的生理响应. 草业学报， 2019， 28（9）： 96-109.
2	Ma X， Wang L L， Li W J， et al. Effects of different nitrogen levels on nitrogen fixation and seed production of alfalfa inoculated with rhizobia. Acta Prataculturae Sinica， 2013， 22（1）： 95-102.
	马霞，王丽丽，李卫军，等. 不同施氮水平下接种根瘤菌对苜蓿固氮效能及种子生产的影响. 草业学报， 2013， 22（1）： 95-102.
3	Afzal M， Khan S， Iqbal S， et al. Inoculation method affects colonization and activity of Burkholderia phytofirmans PsJN during phytoremediation of diesel-contaminated soil. International Biodeterioration & Biodegradation， 2013， 85： 331-336.
4	Li J F， Zhang S Q， Shi S L， et al. Position and quantity of endogensis rhizobia in alfalfa plant. Chinese Journal of Eco-Agriculture， 2009， 17（6）： 1200-1205.
	李剑峰，张淑卿，师尚礼，等. 苜蓿内生根瘤菌分布部位与数量变化动态. 中国生态农业学报， 2009， 17（6）： 1200-1205.
5	Li J F， Zhang S Q， Shi S L， et al. Infection and migration of marked rhizobia in alfalfa bud seedlings under the action of exogenous substance. Acta Agrestia Sinica， 2015， 23（6）： 1259-1264.
	李剑峰，张淑卿，师尚礼，等. 几种外源物质对内生根瘤菌侵染苜蓿芽苗并在植株体内运移的影响. 草地学报， 2015， 23（6）： 1259-1264.
6	Wu Y， Xi L Q. Dynamics of IAA produced by nitrogen fixation bacteria around oat root system. Journal of Anhui Agricultural Sciences， 2007， 35（15）： 4424-4425.
	吴瑛，席琳乔. 燕麦根际固氮菌分泌IAA的动态变化研究. 安徽农业科学， 2007， 35（15）： 4424-4425.
7	Dong R， Cao Y R. Research progress on the immune regulation of symbiotic nitrogen fixation between legumes and rhizobia. Biotechnology Bulletin， 2019， 35（10）： 25-33.
	董汝，曹扬荣. 豆科植物-根瘤菌共生固氮的免疫调控机制. 生物技术通报， 2019， 35（10）： 25-33.
8	Jordan J V， Anderson G R. Effect of boron on nitrogen fixation by azotobacter. Soil Science， 1950， 69（4）： 311-320.
9	Ei-hamdaoui A， Redondo N M， Rivilla R， et al. Effects of boron and calcium nutrition on the establishment of the Rhizobium leguminosarum-pea （Pisum sativum） symbiosis and nodule development under salt stress. Plant， Cell & Environment， 2003， 26（7）： 1003-1011.
10	Bonilla I. The aberrant cell walls of boron-deficient bean root nodules have no covalently bound hydroxyproline-proline-rich proteins. Plant Physiology， 1997， 115（4）： 1329-1340.
11	Miao Y Y， Zhou T， Shi S L， et al. Effect of born on migration and colonization by rhizobia and seeding growth in Medicago sativa. Acta Prataculturae Sinica， 2017， 26（4）： 120-133.
	苗阳阳，周彤，师尚礼，等. 硼对根瘤菌在紫花苜蓿体内运移和定殖及对幼苗生长的影响. 草业学报， 2017， 26（4）： 120-133.
12	Redondo N M， Rivilla R， Ei H A， et al. Research note： Boron deficiency affects early infection events in the pea-rhizobium symbiotic interaction. Functional Plant Biology， 2001， 28（8）： 819-823.
13	Song B Q， Pei X Y， Yan Z S， et al. Effects of boron on endogenous hormones of sugar beet. Chinese Agricultural Science Bulletin， 2014， 31（6）： 115-120.
	宋柏权，裴轩瑗，闫志山，等. 硼素对甜菜内源激素的影响研究. 中国农学通报， 2014， 31（6）： 115-120.
14	Yang Y， Gao K X， Wu Y， et al. Indole-3-acetic acid-mediated cross-kingdom signaling involved in plant-bacteria interactions. Biotechnology Bulletin， 2016， 32（8）： 14-21.
	杨扬，高克祥，吴岩，等. 吲哚乙酸跨界信号调节植物与细菌互作. 生物技术通报， 2016， 32（8）： 14-21.
15	Wu J， Li C J， Wang H， et al. Effect of boron deficiency on structure and nitrogenase activity of nodules in soybean plants. Acta Botanica Boreali-Occidentalia Sinica， 1999， 19（3）： 439-445.
	吴静，李春俭，王贺，等. 缺硼对大豆根瘤结构和功能的影响. 西北植物学报， 1999， 19（3）： 439-445.
16	Zhang B， Sun B S， Jin M， et al. Extraction and analysis of extracellular polymeric substances in membrane fouling in submerged MBR. Desalination， 2008， 227（1）： 286-294.
17	Zhao D Q， Yao T， Ma W B， et al. Effect of mixture of phosphate solubilizing bacteria and nodule bacteria on alfalfa seedling growth. Chinese Journal of Grassland， 2015， 37（5）： 57-61.
	赵冬青，姚拓，马文彬，等. 溶磷菌和根瘤菌混合菌剂对苜蓿苗期生长的影响. 中国草地学报， 2015， 37（5）： 57-61.
18	Zhang Y Y， Tang H， Guo Y H， et al. Establishment of two-dimensional gel electrophoresis technique for sheep serum. Xinjiang Agricultural Science， 2017， 54（1）： 190-196.
	张译元，唐红，郭延华，等. 绵羊血清蛋白双向凝胶电泳技术的建立. 新疆农业科学， 2017， 54（1）： 190-196.
19	Miao W H， Qu L L， Pang L， et al. Construction， expression， purification and evaluation of diagnostic effects of rubella virus compound epitope peptide. Chinese Journal Clinical Laboratory Science， 2015， 33（12）： 889-894.
	缪卫华，渠利利，庞露，等. 风疹病毒复合多表位肽的构建、表达、纯化及其诊断效能的初步评价. 临床检验杂志， 2015， 33（12）： 889-894.
20	Ma S L， Dong X， Xia X， et al. Impact of Streptococcus sanguis bacteriocins on the growth curve and intracellular free calcium concentration of Porphyromonas gingivalis. Journal of Oral Science Research， 2014， 30（2）： 108-111.
	马晟利，董雪，夏雪，等. 血链球菌细菌素对牙龈卟啉单胞菌生长曲线及细胞内钙离子浓度的影响. 口腔医学研究， 2014， 30（2）： 108-111.
21	Chai R J， Wang Y L. Effect of REE on growth of bacteria and fluorescence spectrum of DNA. Spectroscopy and Spectral Analysis， 2013， 33（7）： 1829-1833.
	柴瑞娟，王玉良. 镧和铈对两种细菌生长的影响及胞内DNA的荧光光谱分析. 光谱学与光谱分析， 2013， 33（7）： 1829-1833.
22	Montanari C， Kamdem S L S， Serrazanetti D I， et al. Synthesis of cyclopropane fatty acids in Lactobacillus helveticus and Lactobacillus sanfranciscensis and their cellular fatty acids changes following short term acid and cold stresses. Food Microbiology， 2010， 27（4）： 493-502.
23	Kabu M， Uyarlar C， Zarczynska K， et al. The role of boron in animal health. Journal of Elementology， 2015， 20（2）： 535-541.
24	Zhang W W. Calmodulin-like-protein in prokaryotes. Progress in Biochemistry and Biophysic， 1995， 22（2）： 98-102.
	张蔚文. 原核生物中的类钙调蛋白. 生物化学与生物物理进展， 1995， 22（2）： 98-102.
25	Redondo N M， Wilmot A R， Ei H A， et al. Relationship between boron and calcium in the N₂-fixing legume-rhizobia symbiosis. Plant， Cell and Environment， 2003， 26（11）： 1905-1915.
26	Zhang S Q， Li J F， Shi S L， et al. Quantity and ecological dominance of endogenesis rhizobia in bud seedling and seeds of alfalfa. Chinese Journal of Grassland， 2009， 31（5）： 90-95.
27	Martin S， Dominic T N， Patrick C. Influence of boric acid on energy metabolism and stress tolerance of Candida albicans. Journal of Trace Elements in Medicine and Biology， 2018， 49： 140-145.
28	Qin G， Zong Y， Chen Q， et al. Inhibitory effect of boron against Botrytis cinerea on table grapes and its possible mechanisms of action. International Journal of Food Microbiology， 2010， 138（1/2）： 145-150.
29	Mustafa K， Cangir U U， Katarzyna Z， et al. The role of boron in animal health. Journal of Elementology， 2015， 20（2）： 535-541.
30	Hu H， Zhang R B， Qin J M， et al. Enzymatic characterization of acetyl CoA synthetase and its application in biosynthesis of mevalonate. Chinese Journal of Applied and Environmental Biology， 2016， 22（3）： 357-362.
	胡红，张汝兵，秦杰明，等. 乙酰辅酶A合成酶的酶学性质及其在甲羟戊酸合成中的应用. 应用与环境生物学报， 2016， 22（3）： 357-362.
31	Wang L H， Yang Z W， Zhu L， et al. Cloning and prokaryotic expression of sweet sorghum succinic semialdehyde dehydrogenase SbSSADH. Biotechnology Bulletin， 2015， 31（7）： 83-90.
	王龙海，杨泽伟，朱莉，等. 甜高粱琥珀酸半醛脱氢酶SbSSADH基因克隆及原核表达. 生物技术通报， 2015， 31（7）： 83-90.
32	Sànchez M， Alvarez M A， Balana R， et al. Properties and functions of two succinic-semialdehyde dehydrogenases from Pseudomonas putida. Biochim Biophys Acta， 1988， 953（3）： 249-257.
33	Peter W， Hermann H. Substrate stereochemistry of the enoyl-CoA hydratase reaction. European Journal of Biochemistry， 1975， 54（1）： 247-252.
34	Xiao B C， Rong X H， Qin L Y， et al. Theoretical study on the chemical mechanism of enoyl-CoA hydratase and the form of inhibitor binding. Journal of Molecular Modeling， 2014， 20（9）： 2411-2422.
35	Zhang D， Liu Z X， Choi C S， et al. Mitochondrial dysfunction due to long-chain acyl-CoA dehydrogenase deficiency causes hepatic steatosis and hepatic insulin resistance. Proceedings of the National Academy of Sciences， 2007， 104（43）： 17075-17080.
36	Tang D J， He Y Q， Feng J X， et al. Xanthomonas campestris pv. campestris possesses a single gluconeogenic pathway that is required for virulence. Journal of Bacteriology， 2005， 187（17）： 6231-6237.
37	Wang W W， Guan D W， Ma M C， et al. Genetic diversity and phylogeny of soybean rhizobia isolated from Northeast China. Soybean Science， 2013， 32（4）： 433-437.
	王卫卫，关大伟，马鸣超，等. 东北地区大豆根瘤菌遗传多样性与系统发育研究. 大豆科学， 2013， 32（4）： 433-437.
38	Wang X F. The proteomics research of tap root inflation in sugar beet （Beta vulgaris L.） by iTRAQ. Huhhot： Inner Mongolia Agriculture University， 2016.
	王雪峰. 基于iTRAQ技术对甜菜块根膨大蛋白质组学的研究. 呼和浩特：内蒙古农业大学， 2016.
39	Janczarek M. Environmental signals and regulatory pathways that influence exopolysaccharide production in rhizobia. International Journal of Molecular Sciences， 2011， 12（12）： 7898-7933.

[1]	Zhi-min WEI, Bin SUN, Cheng FANG, Zi-wen DAI, Man-qiang LIU, Jia-guo JIAO, Feng HU, Hui-xin LI, Li XU. Co-inoculation with rhizobia and azotobacter affects the growth of Vicia villosa [J]. Acta Prataculturae Sinica, 2021, 30(5): 94-102.
[2]	Ning ZHANG, Yun-xin CAO, Wei XU, Zhi-hui CAHNG. Effects of biosolids on the growth and auxin metabolism of Poa pratensis under drought stress [J]. Acta Prataculturae Sinica, 2021, 30(3): 167-176.
[3]	XING Yi-mei, DONG Li, ZHAN Li-feng, CAI Hua, YANG Sheng-qiu, SUN Na. Effect of mixed inoculation of Glomus mosseae and Sinorhizobium melilotion alkali resistance of alfalfa [J]. Acta Prataculturae Sinica, 2020, 29(9): 136-145.
[4]	YAN Jing, XIA Li, SHENG Xia-fang, HE Lin-yan. Isolation of heavy metal-tolerant Sinorhizobium meliloti and the effect on copper uptake of alfalfa, perennial ryegrass and Sorghum bicolor plants grown on copper-contaminated soil [J]. Acta Prataculturae Sinica, 2019, 28(2): 102-111.
[5]	ZHENG Chun-feng, LIU Chun-zeng, LI Ben-yin, Lü Yu-hu, PAN Zi-liang, CAO Wei-dong. Effects of boron foliar spray on grain-setting characteristics of Chinese milk vetch (Astragalus sinicus) [J]. Acta Prataculturae Sinica, 2019, 28(11): 192-199.
[6]	ZHAO Tao, MA Chun-hui, WANG Dong, JING Yong-yuan, XI Lin-qiao. Correlation between rhizobia distribution and the physical/chemical properties in soil of winter wheat intercropped with sweet clover [J]. Acta Prataculturae Sinica, 2018, 27(4): 45-55.
[7]	SHU Jian-Hong, WANG Pu-Chang, LI Xian-Gang, WANG Xiao-Li, LI Xiao-Dong. Isolation and screening of inorganic phosphate-solubilizing bacteria and their effect on the growth of Bromus cartharticus [J]. Acta Prataculturae Sinica, 2017, 26(5): 173-180.
[8]	MIAO Yang-Yang, ZHOU Tong, SHI Shang-Li, KANG Wen-Juan, ZHANG Yun-Ting. Effect of boron on migration and colonization by rhizobia and seedling growth in Medicago sativa [J]. Acta Prataculturae Sinica, 2017, 26(4): 120-133.
[9]	ZENG Qing-Fei, WANG Qian, LU Rui-Xia, LIU Zheng-Shu, WU Jia-Hai, WANG Xiao-Li. Identification of soybean growth-promoting rhizobacteria and their effects on the growth and quality of Glycine max and Lotus corniculatus [J]. Acta Prataculturae Sinica, 2017, 26(1): 99-111.
[10]	WANG Xue-Cui, MA Xiao-Tong, HAN Mei, CAO Wei-Dong, ZHANG Hong-Liang, BAI Jin-Shun, ZENG Nao-Hua, GAO Song-Juan, ZHOU Guo-Peng, WANG Yan-Qiu. Screening of rhizobia of common vetch (Vicia sativa) in Qinghai, and assessment of symbiont salt tolerance [J]. Acta Prataculturae Sinica, 2016, 25(8): 145-153.
[11]	CHEN Jing, HAN Gui-Qing, SHAN Chen, ZHAN Hai-Ling, LI Ji-Kai, LIU Hui-Ying, ZHANG Yue-Xue. Proteomic methods for removing high-abundance proteins in alfalfa leaf [J]. Acta Prataculturae Sinica, 2015, 24(7): 131-138.
[12]	LEI Xue-jun,WU Yi,HUANG Xin,CAI Jian-bo,LI Zhuo,WU Lin-shi. Phosphate dissolving capability and the affect of phosphate-dissolving bacteria in the rhizosphere on Sorghum bicolor [J]. Acta Prataculturae Sinica, 2014, 23(6): 274-278.
[13]	PAN Ming-hong,LING Yao,JING Wen,MA Hong-ping,PENG Yan. Genetic diversity and phylogeny of rhizobia isolated from white clover in Sichuan Province [J]. Acta Prataculturae Sinica, 2014, 23(5): 143-152.
[14]	XUE Liang, LIU Jian-feng, SHI Sheng-qing, WEI Yuan, CHANG Er-mei, GAO Ming, JIANG Ze-ping. A review on the progress of proteomic study on plant responses to heavy metal stress [J]. Acta Prataculturae Sinica, 2013, 22(4): 300-311.
[15]	ZHANG Ying, ZHU Ying, YAO Tuo,QI Juan, RONG Liang-yan. Interactions of four PGPRs isolated from pasture rhizosphere [J]. Acta Prataculturae Sinica, 2013, 22(1): 29-37.