青枯雷尔氏菌温度相关致病基因yqhE功能研究

doi:10.16409/j.cnki.2095-039x.2023.03.003

中国生物防治学报 ›› 2023, Vol. 39 ›› Issue (1): 141-148.DOI: 10.16409/j.cnki.2095-039x.2023.03.003

青枯雷尔氏菌温度相关致病基因yqhE功能研究

周大祥^1,2, 杨俊年¹, 陈弘¹, 殷幼平², 熊书³

1. 重庆三峡学院生物与食品工程学院, 万州 404120;
2. 重庆大学生命科学学院/重庆市基因功能与调控重点实验室, 重庆 400300;
3. 重庆三峡医药高等专科学校基础医学部, 万州 404120

收稿日期:2022-07-15 出版日期:2023-02-08 发布日期:2023-02-21
通讯作者: 熊书,硕士,副教授,E-mail:24594294@qq.com
作者简介:周大祥,副教授,博士研究生,E-mail:dqzhou79@163.com
基金资助:
重庆市自然科学基金（cstc2016jcyjA0198）

Functional Analysis of Temperature Related Pathogenic Gene yqhE in Ralstonia solanacearum

ZHOU Daxiang^1,2, YANG Junnian¹, CHEN Hong¹, YIN Youping², XIONG Shu³

1. College of Biology and Food Engineering, Chongqing Three Gorges University, Wanzhou 404120, China;
2. College of Life Science, Chongqing University/Chongqing Key Lab of Genetic Function and Regulation, Chongqing 400030, China;
3. Department of Basic Medicine, Chongqing Three Gorges Medical College, Chongqing 404120, China

Received:2022-07-15 Online:2023-02-08 Published:2023-02-21

摘要/Abstract

摘要： 不同温度下青枯雷尔氏菌表达谱PPI网络分析发现，yqhE可能是青枯雷尔氏菌新的温度相关致病基因。克隆青枯雷尔氏菌CBM613的yqhE基因，结果发现该基因长度为831 bp，共编码276个氨基酸。基因敲除结果显示，28℃培养的yqhE突变株较野生型菌株致病力降低，病程延长，但并未彻底丧失致病力。20℃和28℃培养的yqhE突变株维生素C的生物合成皆被抑制；与28℃相比，20℃培养下野生型菌株维生素C的合成能力降低。突变株在20℃和28℃培养下甲基乙二醛（MG）和丙二醛（MDA）明显积累，其中20℃积累更多；20℃培养的野生型菌株MG和MDA比28℃积累更多。维生素C的生物合成被抑制导致青枯雷尔氏菌自由基清除能力减弱与氧化应激反应增强，MG和MDA积累产生细胞毒性，上述结果可能与28℃培养的突变株致病力减弱，20℃培养的野生型菌株致病力丧失有关。综上结果表明yqhE是青枯雷尔氏菌温度相关致病基因。

Abstract: The protein-protein interoperability (PPI) network analysis of the expression profiles of Ralstonia solanacearum at different temperatures showed that yqhE could be a new temperature related pathogenic gene of Ralstonia solanacearum. the gene yqhE was cloned from Ralstonia solanacearum strain CBM613. The results showed that gene yqhE was 831 bp in length, encoding 276 amino acids. Its function was further confirmed by gene knock-out. The results showed that the pathogenicity of the yqhE^－ mutant cultured at 28 ℃ was decreased and the course of disease was prolonged compared with the wild-type strain. Vitamin C biosynthesis of yqhE^－ mutant cultured at 20 ℃ and 28 ℃ were inhibited. The vitamin C biosynthesis ability of the wild-type strain cultured at 20 ℃ was reduced compared with 28 ℃. Methylglyoxal (MG) and malondialdehyde (MDA) of yqhE^－ mutant significantly accumulated at 20 ℃ and 28 ℃. MG and MDA of the wild-type strain cultured at 20 ℃ accumulated more than at 28 ℃. The inhibition of vitamin C biosynthesis resulted in the weakening of radical scavenging ability and the enhancement of oxidative stress reaction in Ralstonia solanacearum. The accumulation of MG and MDA produced cytotoxicity. The above results could be related to the weakening of pathogenicity in yqhE^－ mutant cultured at 28 ℃ and the loss of pathogenicity in wild-type strain cultured at 20 ℃. In conclusion, yqhE could be a temperature related pathogenic gene in Ralstonia solanacearum.

Key words: Ralstonia solanacearum, temperature related pathogenic gene, yqhE, gene knock-out, functional analysis

中图分类号:

S432.1

周大祥, 杨俊年, 陈弘, 殷幼平, 熊书. 青枯雷尔氏菌温度相关致病基因yqhE功能研究[J]. 中国生物防治学报, 2023, 39(1): 141-148.

ZHOU Daxiang, YANG Junnian, CHEN Hong, YIN Youping, XIONG Shu. Functional Analysis of Temperature Related Pathogenic Gene yqhE in Ralstonia solanacearum[J]. Chinese Journal of Biological Control, 2023, 39(1): 141-148.

参考文献

[1] 周大祥, 龙泉洲, 殷幼平, 等. 青枯雷尔氏菌Ⅲ型分泌系统hrcN基因的功能分析[J]. 中国生物防治学报, 2021, 37 (6):1353-1362.
[2] Wei Z, Huang J F, Hu J, et al. Altering transplantation time to avoid periods of high temperature can efficiently reduce bacterial wilt disease incidence with tomato[J]. PLoS ONE, 2015, 10(10):e0139313.
[3] Jacobs J M, Babujee L, Meng F, et al. The in planta transcriptome of Ralstonia solanacearum:conserved physiological and virulence strategies during bacterial wilt of tomato[J]. mBio, 2012, 3(4):e00114-12.
[4] Zhou D X, Li GL, Sun H L, et al. In silico studies reveal RSc1154 and RhlE as temperature-related pathogenic proteins of Ralstonia solanacearum[J]. FEMS Microbiology Letters, 2019, 366(15):1-8.
[5] Yum D Y, Lee B Y, Pan J G. Identification of the yqhE and yafB genes encoding two 2,5-diketo-D-gluconate reductases in Escherichia coli[J]. Applied and Environmental Microbiology, 1999, 65(8):3341-3346.
[6] Habrych M, Rodriguez S, Stewart J D. Purification and identification of an Escherichia coli beta-keto ester reductase as 2,5-diketo-D-gluconate reductase YqhE[J]. Biotechnology Progress, 2002, 18(2):257-261.
[7] Thornalley P J, Langborg A, Minhas H S. Formation of glyoxal, methylglyoxal and 3-deoxyglucosone in the glycation of proteins by glucose[J]. Biochemical Journal, 1999, 344(1):109-116.
[8] Rondeau P, Bourdon E. The glycation of albumin:Structural and functional impacts[J]. Biochimie, 2011, 93(4):645-658.
[9] Ko J, Kim I, Yoo S et al. Conversion of methylglyoxal to acetol by Escherichia coli aldo-Keto reductases[J]. Journal of Microbiology, 2005,187(16):5782-5789.
[10] Lee C, Kim I, Park C. Glyoxal detoxification in Escherichia coli K-12 by NADPH dependent aldo-keto reductases[J]. Journal of Microbiology, 2013, 51(4):527-530.
[11] Khurana S, Powers D B, Anderson S, et al. Crystal structure of 2,5-diketo-D-gluconic acid reductase A complexed with NADPH at 2.1-A resolution[J]. Proceedings of the National Academy of Sciences of the United States of America, 1998, 95(12):6768-6773.
[12] Khurana S, Sanli G, Powers D B, et al. Molecular modeling of substrate binding in wild-type and mutant Corynebacteria 2,5-diketo-D-gluconate reductases[J]. Proteins Structure Function and Bioinformatics, 2000, 39(1):68-75.
[13] Zelić B, Pavlović N, Delić V, et al. Optimization of pH and temperature in the process of bioconversion of glucose to 2,5-diketo-D-gluconic acid[J]. Chemical and Biochemical Engineering Quaterly, 2002, 16(1):7-11.
[14] Kaswurm V, Pacher C, Kulbe K D, et al. 2,5-Diketo-gluconic acid reductase from Corynebacterium glutamicum:Characterization of stability, catalytic properties and inhibition mechanism for use in vitamin C synthesis[J]. Process Biochemistry, 2012, 47(1):2012-2019.
[15] Togo T. Cell membrane disruption stimulates cAMP and Ca²⁺signaling to potentiate cell membrane resealing in neighboring cells[J]. Biology Open, 2017, 6(12):1814-1819.
[16] He L Y, Sequeira L, Kelman A. Characteristics of strains of Pseudomonas solanacearum[J]. Plant Disease, 1983, 67(2):1357-1361.
[17] Meng F, Yao J, Allen C. A MotN mutant of Ralstonia solanacearum is hypermotile and has reduced virulence[J]. Journal of Bacteriology, 2011, 193(10):2477-2486.
[18] Miller E N, Jarboe L R, Yomano L P, et al. Silencing of NADPH-dependent oxidoreductase genes (yqhD and dkgA) in furfural-resistant ethanologenic Escherichia coli[J]. Applied and Environmental Microbiology, 2009, 75(13):4315-4323.
[19] Chen M, Nan Chen N, Wang J W, et al. Involvement of a FAD-Linked Oxidase RSc0454 for Expression of the Type III Secretion System and Pathogenicity in Ralstonia solanacearum[J]. Molecular Plant-microbe Interactions, 2021, 34(11):1228-1235.
[20] Tang M X, Bouchez O, Cruveiller S, et al. Modulation of Quorum Sensing as an Adaptation to Nodule Cell Infection during Experimental Evolution of Legume Symbionts[J]. Mbio, 2020, 11(1):e03129-19.
[21] Qi P P, Huang M L, Hu X H, et al. A Ralstonia solanacearum effector targets TGA transcription factors to subvert salicylic acid signaling[J]. The Plant Cell, 2022, 34(5):1666-1683.
[22] Gao L L, Hu Y D, Liu J C, et al. Stepwise metabolic engineering of Gluconobacter oxydans WSH-003 for the direct production of 2-keto-L-gulonic acid from D-sorbitol[J]. Metabolic Engineering, 2014, 24(5):30-37.
[23] Kaswurm V, Van Hecke W, Kulbe K D, et al. Engineering of a bi-enzymatic reaction for efficient production of the ascorbic acid precursor 2-keto-L-gulonic acid[J]. Biochemical Engineering Journal, 2013(3), 79:104-111.
[24] Sanli G, Banta S, Aanderson S, et al. Structural alteration of cofactor specificity in Corynebacterium 2,5-diketo-D-gluconic acid reductase[J]. Protein Science, 2004, 13(2):504-512.

青枯雷尔氏菌温度相关致病基因yqhE功能研究

Functional Analysis of Temperature Related Pathogenic Gene yqhE in Ralstonia solanacearum

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

联系我们

[1]	郑雪芳, 陈燕萍, 肖荣凤, 陈梅春, 陈峥, 刘欣, 王阶平, 刘波. 福建青枯雷尔氏菌的遗传多样性及其生防放线菌的筛选[J]. 中国生物防治学报, 2022, 38(5): 1269-1279.
[2]	周大祥, 龙泉洲, 殷幼平, 熊书. 青枯雷尔氏菌Ⅲ型分泌系统hrcN基因的功能分析[J]. 中国生物防治学报, 2021, 37(6): 1353-1362.
[3]	陈莹莹, 王亚楠, 吕斌娜, 谭晓东, 李世东, 孙漫红, 马桂珍. 粉红螺旋聚孢霉67-1短链脱氢酶基因CrSdr功能研究[J]. 中国生物防治学报, 2021, 37(4): 749-760.
[4]	冯印印, 李斌, 杨洋, 何佶弦, 安航, 闫芳芳, 安德荣. 烟草青枯病菌拮抗细菌的筛选、鉴定和抑菌机制研究[J]. 中国生物防治学报, 2021, 37(2): 331-339.
[5]	汪章勋, 徐秀珍, 冯健雨, 周权, 黄勃. MrXrn1基因参与调控罗伯茨绿僵菌的产孢和致病[J]. 中国生物防治学报, 2020, 36(5): 721-728.
[6]	李信申, 陈建, 肖运萍, 黄瑞荣, 魏林根, 华菊玲. 中生菌素和生石灰对芝麻青枯病的联合防控效果[J]. 中国生物防治学报, 2020, 36(3): 472-478.
[7]	李昕玥, 王丽荣, 李梅, 吴蓓蕾, 蒋细良. 哈茨木霉C2H2型转录因子Tha09974功能研究[J]. 中国生物防治学报, 2019, 35(3): 407-415.
[8]	马胜龙, 凡肖, 姚俊敏, 吴华川, 束长龙, 商铭达, 张韶芮, 杨郑豪, 关雄, 黄天培. 苏云金芽胞杆菌XL6 BTXL6_11095基因对生物被膜相关表型的调控[J]. 中国生物防治学报, 2019, 35(1): 53-62.
[9]	郑雪芳, 刘波, 朱育菁, 葛慈斌, 刘国红. 番茄青枯病生防芽胞杆菌的筛选与鉴定[J]. 中国生物防治学报, 2016, 32(5): 657-665.
[10]	徐显皓, 陶立彬, 马正, 俞晓平. 淀粉酶产色链霉菌1628中toyF基因的克隆与丰加霉素合成相关性[J]. 中国生物防治学报, 2016, 32(4): 511-517.
[11]	刘艳, 葛蓓孛, 刘炳花, 赵文珺, 张克诚. 武夷菌素产生菌CK?15遗传操作系统的优化[J]. 中国生物防治学报, 2016, 32(4): 524-530.
[12]	刘增亮, 李梅, 张玉杰, 张林, 杨晓燕, 蒋细良. 哈茨木霉Thga1基因敲除突变株对几种植物病原真菌的离体拮抗作用[J]. , 2014, 30(3): 370-375.
[13]	刘国红1,2 , 刘波2 , 林乃铨1 . 青枯雷尔氏菌生防芽胞杆菌的快速筛选及其鉴定[J]. , 2013, 29(3): 473-480.
[14]	张晔1,2 , 孙漫红1 , 李世东1 , 罗明2 . 链孢粘帚霉HL-1-1内切葡聚糖酶基因克隆及功能分析[J]. , 2013, 29(1): 74-82.
[15]	张彦1,2 , 车建美1 , 刘波1 , 唐乐尘2 . 蜡样芽孢杆菌ANTI-8098A在番茄内的定殖和对青枯病的防治研究[J]. , 2011, 27(2): 221-227.