Transgenic Amorphophallus konjac expressing synthesized acyl-homoserine lactonase (aiiA) gene exhibit enhanced resistance to soft rot disease.
Amorphophallus konjac is an important economic crop widely used in health products and biomaterials. However, this monocotyledonous plant's production is seriously restricted by soft rot disease. Some Bacillus thuringiensis strains generate an endocellular acyl homoserine lactonase (AiiA), which has inhibitory effect on soft rot pathogen through disrupting the signal molecules (N-acylhomoserine lactones, AHL) of their Quorum Sensing system. The aim of our study is to obtain transgenic A. konjac expressing AiiA protein and exhibiting resistance to soft rot. But till now, there is not any report about exogenous gene transformation in A. konjac. In this research, an Agrobacterium-mediated genetic transformation system was constructed. An aiiA gene was synthesized according to the codon usage in A. konjac. Embryogenic callus was infected with the A. tumefaciens strain EHA105 harboring the plant transformation plasmid pU1301 plus synthesized aiiA gene. After antibiotics screening, 34 plants were obtained. PCR analysis showed that positive amplified fragments were present in 21 out of these 34 lines. Southern blot analysis indicated that aiiA gene had integrated into the genome of A. konjac. Western blotting demonstrated that the target protein of interest was reactive with the antibody against AiiA. Further disease resistance detection revealed that all of the tested transgenic A. konjac lines exhibited high resistance to soft rot bacteria Erwinia carotovora subsp. Carotovora (Ecc) SCG1. The protocol is useful for the quality improvement of A. konjac through genetic transformation.
Ban
Huifang
H
State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, 430070 Wuhan, China.
Chai
Xinli
X
Lin
Yongjun
Y
Zhou
Ying
Y
Peng
Donghai
D
Zhou
Yi
Y
Zou
Yulan
Y
Yu
Ziniu
Z
Sun
Ming
M
D031056:Amorphophallus / Q000235:genetics* / Q000276:immunology / Q000382:microbiology*; D001413:Bacillus thuringiensis / Q000201:enzymology*; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D002265:Carboxylic Ester Hydrolases / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification; D018745:Genome, Plant / Q000235:genetics; D007113:Immunity, Innate / Q000276:immunology*; D008969:Molecular Sequence Data; D016973:Pectobacterium carotovorum / Q000502:physiology; D010935:Plant Diseases / Q000235:genetics / Q000276:immunology* / Q000382:microbiology*; D018515:Plant Leaves / Q000235:genetics / Q000382:microbiology; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction
D018744:DNA, Plant; D002265:Carboxylic Ester Hydrolases; C432003:N-acyl homoserine lactonase