The Effects of luxR_19 on the Synthesis of Phenzaine-1-carboxamide in Pseudomonas chlororaphis HT66

doi:10.13560/j.cnki.biotech.bull.1985.2015.08.024

Abstract

Abstract: In order to study the potential effects of regulator gene luxR_19 on the synthesis of antibacterial substance phenazine-1-carboxamide（PCN）and physiological-biochemical characteristics in Pseudomonas chlororaphis HT66, strains of gene luxR_19-knockout, -complementation, -overexpression and -point-mutation were constructed, and then the growth curve, fermentation production of PCN, swimming and swarming activities were tested at the target strains. The results showed that the knockout of luxR_19 lowered the production of PCN and the over-expression of gene luxR_19 increased the double production of PCN compared to the wild type;luxR_19 gene affected the expression of several genes relating to the regulation or synthesis of phenazines. Besides, the knockout of luxR_19 had no effects on the growth and swimming activity, however, promoted the swarming activity of HT66. The production of PCN in point-mutation strain of luxR_19（G85A）was similar to that in wild type. The above results indicated that luxR_19 was the positive regulator of PCN production, and could affect the swarming activity of HT66. Besides, the point-mutation at 254^th site in luxR_19 did not affect the production of PCN.

Key words: Pseudomonas chlororaphis, HT66, phenazine-1-carboxamide, transcriptional regulator of LuxR family

Chen Yawen, Peng Huasong, Wang Wei, Zhang Xuehong. The Effects of luxR_19 on the Synthesis of Phenzaine-1-carboxamide in Pseudomonas chlororaphis HT66[J]. Biotechnology Bulletin, 2015, 31(8): 166-173.

References

[1] 张锋华, 许煜泉, 张雪洪.农用杀菌剂吩嗪-1-羧酸的生物合成与基因调控[J].农药研究与应用, 2006（5）：4-7.
[2] 张云, 胡洪波, 彭华松, 等.产PCA基因工程菌M18G反复补料分批培养研究[J].工业微生物, 2007：16-20.
[3] Han SH, Lee SJ, Moon JH, et al. GacS-dependent production of 2R, 3R-butanediol by Pseudomonas chlororaphis O6 is a major determinant for eliciting systemic resistance against Erwinia carotovora but not against Pseudomonas syringae pv. tabaci in tobacco[J]. Mol Plant-Microbe Interact, 2006, 19（8）：924-930.
[4] Chin-A-Woeng TF, Bloemberg GV, van der Bij AJ, et al. Biocontrol by phenazine-1-carboxamide-producing Pseudomonas chlororaphis PCL1391 of tomato root rot caused by Fusarium oxysporum f. sp. radicis-lycopersici[J]. Mol Plant-Microbe Interact, 1998, 11（11）：1069-1077.
[5] Liu H, He Y, Jiang H, et al. Characterization of a phenazine-producing strain Pseudomonas chlororaphis GP72 with broad-spectrum antifungal activity from green pepper rhizosphere[J]. Curr Microbiol, 2007, 54（4）：302-306.
[6] Shen X, Hu H, Peng H, et al. Comparative genomic analysis of four representative plant growth-promoting rhizobacteria in Pseudomonas[J]. BMC Genomics, 2013, 14（1）：271.
[7] Loper JE, Hassan KA, Mavrodi DV, et al. Comparative genomics of plant-associated Pseudomonas spp.：insights into diversity and inheritance of traits involved in multitrophic interactions[J]. PLoS Genet, 2012, 8（7）：e1002784.
[8] Park J, Oh S, Anderson A, et al. Production of the antifungal compounds phenazine and pyrrolnitrin from Pseudomonas chlororaphis O6 is differentially regulated by glucose[J]. Lett Appl Microbiol, 2011, 52（5）：532-537.
[9] Maddula V, Pierson E, Pierson L. Altering the ratio of phenazines in Pseudomonas chlororaphis（aureofaciens）strain 30-84：effects on biofilm formation and pathogen inhibition[J]. J Bacteriol, 2008, 190（8）：2759-2766.
[10] Huang L, Chen MM, Wang W, et al. Enhanced production of 2-hydroxyphenazine in Pseudomonas chlororaphis GP72[J]. Appl Microbiol Biotechnol, 2011, 89（1）：169-177.
[11] 张平原, 彭华松, 张雪洪.高产吩嗪-1-甲酰胺的绿针假单胞菌的诱变与基因工程育种[J].上海交通大学学报：农业科学版, 2015, 33（2）：90-94.
[12] He W, Lei J, Jiu Y, et al. The LuxR family members gdmri and gdmrii are positive regulators of geldanamycin biosynthesis in Streptomyces hygroscopicus 17997[J]. Arch Microbiol, 2008, 189（5）：501-510.
[13] Whistler CA, Pierson III LS. Repression of phenazine antibiotic production in Pseudomonas aureofaciens strain 30-84 by RpeA[J]. J Bacteriol, 2003, 185（13）：3718-3725.
[14] Fernández-piñar R, Cámara M, Soriano MI, et al. Ppor, an orphan luxr-family protein of Pseudomonas putida KT2440, modulates competitive fitness and surface motility independently of n-acylhomoserine lactones[J]. Environmental Microbiology Reports, 2011, 3（1）：79-85.