|Table of Contents|

Overexpression of Pectinase Gene in Endophytic Bacillus Strains and Its Effect on Colonization(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

2013 05
Research Field:
Publishing date:


Overexpression of Pectinase Gene in Endophytic Bacillus Strains and Its Effect on Colonization
FAN Xiaojing YANG Ruixian QIU Sixin HU Fangping
(1College of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou 350002, China) (2Key Laboratory for Biopesticide and Chemical Biology, Ministry of Education, Fujian Agriculture and Forestry University, Fuzhou 350002, China) (3Department of Environmental Engineering and Chemistry, Luoyang Institute of Science and Technology, Luoyang 471023, China) (4Crop research institute, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China)
endophytic Bacillus pectinase gene Real-time PCR overexpression colonization
Q948.12+2.3 : Q78

In order to investigate the correlation between pectinase gene of endogenous Bacillus and colonization in plants, the full length genes of pectinase from two endophytic Bacillus strains were cloned; the overexpressing vectors of pectinase genes were constructed by inserting the full length genes of pectinase into shuttle vector pGFP4412, then transformed into the endophytic Bacillus strains and overexpressed. Pectinase enzymatic assays showed that the diameters of hydrolyzed circles caused by the wild type strains were only 50% of the overexpressing mutants on LB agar plates. The real-time quantitative PCR results showed that the relative mRNA transcription levels of pectinase gene in mutant bacteria were 82 and 85 times as much as the level in the control strains (BS2-gfp and TB2-gfp), respectively. The colonizing bacteria amounts of pectinase mutants and the control strains in the tissues of cabbage within 28 d indicated a significant difference between mutants and control strains in roots, stems and leaves in the first 1-3 d after inoculation (P < 0.05). The amount of mutant bacteria in roots was notably higher than that of control strains in the 7th day (P < 0.05). But for the other statistic data, no significant difference was detected. The results of continuous isolations during 28 d showed a remarkably higher population density in mutants than the control strains at the beginning, and a gradually decreasing difference between the two. This study presented preliminary evidence that pectinase gene plays a role in endophytic Bacillus bacteria colonization in plant at the initial stage. Fig 7, Tab 2, Ref 23


1 Surette MA, Sturz AV, Lada RR, Nowak J. Bacterial endophytes in processing carrots (Daucus carota L. var. sativus): their localization, population density, biodiversity and their effects on plant growth [J]. Plant Soil, 2003, 253 (2): 381-390 2 Ryan RP, Germaine K, Franks A, Ryan DJ, Dowling DN. Bacterial endophytes: recent developments and applications [J]. FEMS Microbiol Lett, 2008, 278 (1): 1-9 3 Yang J, Kloepper JW, Ryu CM. Rhizosphere bacteria help plants tolerate abiotic stress [J]. Trends Plant Sci, 2008, 14 (1): 1-4 4 Chen XH, Koumoutsi A, Scholz R, Borriss R. More than anticipated-production of antibiotics and other secondary metabolites by Bacillus amyloliquefaciens FZB42 [J]. J Mol Microb Biotech, 2009, 16 (1-2): 14-24 5 Niu D, Liu HX, Jiang CH, Wang YP, Wang QY, Jin HL, Guo JH. The plant growth-promoting Rhizobacterium Bacillus cereus AR156 induces systemic resistance in Arabidopsis thaliana by simultaneously activating salicylate- and jasmonate/ethylene-dependent signaling pathways [J]. Mol Plant Microbe Interact, 2011, 24 (5): 533-542 6 Lugtenberg BJJ, Dekkers L, Bloemberg GV. Molecular determinants of rhizosphere colonization by Pseudomonas [J]. Annu Rev Phytopathol, 2001, 39: 461-490 7 Ji XL, Lu GB, Gai YP, Zheng CC, Mu ZM. Biological control against bacterial wilt and colonization of mulberry by an endophytic Bacillus subtilis strain [J]. FEMS Microbiol Ecol, 2008, 65 (3): 565-573 8 Iniguez AL, Dong Y, Carter HD, Ahmer BM, Stone JM, Triplett EW. Regulation of enteric endophytic bacterial colonization by plant defenses [J]. Mol Plant Microbe Interact, 2005, 18 (2): 169-178 9 Miché L, Battistoni F, Gemmer S, Belghazi M, Reinhold-Hurek B. Upregulation of jasmonate-inducible defense proteins and differential colonization of roots of Oryza sativa cultivars with the endophyte Azoarcus sp. [J]. Mol Plant Microbe Interact, 2006, 19 (5): 502-511 10 Fuentes-Ramírez LE, Caballero-Mellado J, Sepúlveda J, Martínez-Romero E. Colonization of sugarcane by Acetobacter diazotrophicus is inhibited by high N-fertilization [J]. FEMS Microbiol Ecol, 1999, 29 (2): 117-128 11 Yaryura PM, Leon M, Correa OS, Kerber NL, Pucheu NL, García AF. Assessment of the role of chemotaxis and biofilm formation as requirements for colonization of roots and seeds of soybean Plants by Bacillus amyloliquefaciens BNM339 [J]. Curr Microbiol, 2008, 56 (6): 625-632 810 应用与环境生物学报 Chin J Appl Environ Biol http://www.cibj.com/果胶酶基因在内生芽孢杆菌中的过表达及其对定殖的影响 5期 12 Iniguez AL, Dong Y, Carter HD, Ahmer BM, Stone JM, Triplett EW. Regulation of enteric endophytic bacterial colonization by plant defenses [J]. Mol Plant Microbe Interact, 2005, 18 (2): 169-178 13 Cooley MB, Miller WG, Mandrell RE. Colonization of Arabidopsis thaliana with Salmonella enterica and enterohemorrhagic Escherichia coli O157:H7 and competition by Enterobacter asburiae [J]. Appl Environ Microbiol, 2003, 69 (8): 4915-4926 14 Kovtunovych G, Lar O, Kamalova S, Kordyum V, Kleiner D, Kozyrovska N. Correlation between pectate lyase activity and ability of diazotrophic Klebsiella oxytoca VN 13 to penetrate into plant tissues [J]. Plant Soil, 1999, 215 (1): 1-6 15 Compant S, Reiter B, Sessitsch A, Nowak J, Clement C, Barka EA. Endophytic colonization of Vitis vinifera L. by plant growth-promoting bacterium Burkholderia sp. strain PsJN [J]. Appl Environ Microbiol, 2005, 71 (4): 1685-1693 16 Yasbin RE, Wilson GA, Young FE. Transformation and transfection in lysogenic strains of Bacillus subtilis: Evidence for selection induction of prophage in competent cells [J]. J Bacteriol, 1975, 121: 296-304 17 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2?ΔΔCt method [J]. Methods, 2001, 25 (4): 402-408 18 范晓静, 邱思鑫, 吴小平, 洪永聪, 蔡学清, 胡方平. 绿色荧光蛋白基因标记内生枯草芽孢杆菌[J]. 应用与环境生物学报, 2007, 13 (4): 530-534 [Fan XJ, ? Qiu SX? ,Wu XP, Hong YC, Cai XQ, Hu FP. Endophytic Bacillus subtilis strain BS-2 labeled with green fluorescent protein gene [J]. Chin J Appl Environ Biol, 2007, 13 (4): 530-534] 19 Rosenblueth M, Martinez-Romero E. Bacterial endophytes and their interactions with hosts [J]. Mol Plant Microbe Interact, 2006, 19 (8): 827-837 20 Reinhold-Hurek B, Hurek T. Living inside plants: bacterial endophytes [J]. Curr Opin Plant Biol, 2011, 14 (4): 435-443 21 Van’t Slot KAE, Knogge W. A dual role for microbial pathogen-derived effector proteins in plant disease and resistance [J]. Crit Rev Plant Sci, 2002, 21 (3): 229-271 22 Dudeja SS, Giri R, Saini R, Suneja-Madan P, Kothe E. Interaction of endophytic microbes with legumes [J]. J Basic Microbiol, 2012, 52 (3): 248-260 23 Monteiro RA, Balsanelli E, Wassem R, Marin AM, Brusamarello-Santos LCC, Schmidt MA, Tadra-Sfeir MZ, Pankievicz VCS, Cruz LM, Chubatsu LS, Pedrosa FO, Souza EM. Herbaspirillum-plant interactions: microscopical, histological and molecular aspects [J]. Plant Soil, 2012, 356 (1-2): 175-196


Last Update: 2013-10-28