Construction of EGFP-labeling system for visualizing the infection process of Xanthomonas axonopodis pv. citri in planta.
Xanthomonas axonopodis pv. citri (Xac) is the causal agent of citrus bacterial canker, an economically important disease to world citrus industry. To monitor the infection process of Xac in different citrus plants, the enhanced green florescent protein (EGFP) visualizing system was constructed to visualize the propagation and localization in planta. First, the wild-type Xac was isolated from the diseased leaves of susceptible 'Bingtang' sweet orange, and then the isolated Xac was labeled with EGFP by triparental mating. After PCR identification, the growth kinetics and pathogenicity of the transformants were analyzed in comparison with the wild-type Xac. The EGFP-labeled bacteria were inoculated by spraying on the surface and infiltration in the mesophyll of 'Bingtang' sweet orange leaves. The bacterial cell multiplication and diffusion processes were observed directly under confocal laser scanning microscope at different intervals after inoculation. The results indicated that the EGFP-labeled Xac releasing clear green fluorescence light under fluorescent microscope showed the infection process and had the same pathogenicity as the wild type to citrus. Consequently, the labeled Xac demonstrated the ability as an efficient tool to monitor the pathogen infection.
National Center for Citrus Improvement, Hunan Agricultural University, Changsha 410128, Hunan, China. llp31302@163.com
Li-Ping
LP
Liu
Zi-Niu
ZN
Deng
Jin-Wang
JW
Qu
Jia-Wen
JW
Yan
Vittoria
V
Catara
Da-Zhi
DZ
Li
Gui-You
GY
Long
Na
N
Li
D000704:Analysis of Variance; D032084:Citrus sinensis / Q000737:chemistry / Q000382:microbiology*; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010935:Plant Diseases / Q000382:microbiology*; D018515:Plant Leaves / Q000737:chemistry / Q000382:microbiology; D053532:Xanthomonas axonopodis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins