Construction and application of luminescent strain of Pectobacterium carotovorum harboring luxCDABE operon
Pectobacterium carotovorum is one of the top ten plant bacterial pathogens in the world. In order to facilitate the observation of the pathogen infection process at the early stage and in vivo quantification of bacterial growth without the tissue extraction, we generated a P. carotovorum strain expr...
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| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2021-10-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2021.02.221 |
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| Summary: | Pectobacterium carotovorum is one of the top ten plant bacterial pathogens in the world. In order to facilitate the observation of the pathogen infection process at the early stage and in vivo quantification of bacterial growth without the tissue extraction, we generated a P. carotovorum strain expressing the luxCDABE operon, which encodes a luciferase and enzymes that produce its substrate, thus bacteria that express this operon emit light spontaneously. The luxCDABE operon was cloned into the vector pBBR1MCS2, and the resulting pBBR1MCS2-luxCDABE plasmid was transformed into P. carotovorum subsp. carotovorum LMG2404, namely LMG2404-LUX. Compared with LMG2404, LMG2404-LUX had no significant differences in growth rate, biofilm formation and motility. Luminescent signal intensities of LMG2404-LUX were linearly correlated with the bacterial concentrations and not significantly reduced after successive subculturing for ten times. In addition, those signals were stable at the temperature and pH suitable for bacterial growth. When inoculating plants, LMG2404-LUX and LMG2404 had similar pathogenicity, and the signal intensity of LMG2404-LUX was positively correlated with its infection and proliferation. Collectively, P. carotovorum tagged with luxCDABE could greatly facilitate rapid bacterial quantification and in vivo observation of bacterial infection in host tissues, which provides a new tool for the research on P. carotovorum. |
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| ISSN: | 1008-9209 2097-5155 |