[국내논문]Interaction of Pseudostellaria heterophylla with Quorum Sensing and Quorum Quenching Bacteria Mediated by Root Exudates in a Consecutive Monoculture System원문보기
Zhang, Liaoyuan
(Department of Chemical Engineering, Konkuk University)
,
Guo, Zewang
(Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, College of Life Sciences, Gutian Edible Fungi Research Institute, Fujian Agriculture and Forestry University)
,
Gao, Huifang
(Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, College of Life Sciences, Gutian Edible Fungi Research Institute, Fujian Agriculture and Forestry University)
,
Peng, Xiaoqian
(Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, College of Life Sciences, Gutian Edible Fungi Research Institute, Fujian Agriculture and Forestry University)
,
Li, Yongyu
(College of Horticulture, Fujian Agriculture and Forestry University)
,
Sun, Shujing
(Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, College of Life Sciences, Gutian Edible Fungi Research Institute, Fujian Agriculture and Forestry University)
,
Lee, Jung-Kul
(Department of Chemical Engineering, Konkuk University)
,
Lin, Wenxiong
(Key Laboratory of Biopesticide and Chemical Biology, Ministry of Educa)
Many plant-pathogenic bacteria are dependent on quorum sensing (QS) to evoke disease. In this study, the population of QS and quorum quenching (QQ) bacteria was analyzed in a consecutive monoculture system of Pseudostellaria heterophylla. The isolated QS strains were identified as Serratia marcescen...
Many plant-pathogenic bacteria are dependent on quorum sensing (QS) to evoke disease. In this study, the population of QS and quorum quenching (QQ) bacteria was analyzed in a consecutive monoculture system of Pseudostellaria heterophylla. The isolated QS strains were identified as Serratia marcescens with SwrIR-type QS system and exhibited a significant increase over the years of monoculture. Only one QQ strain was isolated from newly planted soil sample and was identified as Bacillus thuringiensis, which secreted lactonase to degrade QS signal molecules. Inoculation of S. marcescens to P. heterophylla root could rapidly cause wilt disease, which was alleviated by B. thuringiensis. Furthermore, the expression of lactonase encoded by the aiiA gene in S. marcescens resulted in reduction of its pathogenicity, implying that the toxic effect of S. marcescens on the seedlings was QS-regulated. Meanwhile, excess lactonase in S. marcescens led to reduction in antibacterial substances, exoenzymes, and swarming motility, which might contribute to pathogensis on the seedlings. Root exudates and root tuber extracts of P. heterophylla significantly promoted the growth of S. marcescens, whereas a slight increase of B. thuringiensis was observed in both samples. These results demonstrated that QS-regulated behaviors in S. marcescens mediated by root exudates played an important role in replanting diseases of P. heterophylla.
Many plant-pathogenic bacteria are dependent on quorum sensing (QS) to evoke disease. In this study, the population of QS and quorum quenching (QQ) bacteria was analyzed in a consecutive monoculture system of Pseudostellaria heterophylla. The isolated QS strains were identified as Serratia marcescens with SwrIR-type QS system and exhibited a significant increase over the years of monoculture. Only one QQ strain was isolated from newly planted soil sample and was identified as Bacillus thuringiensis, which secreted lactonase to degrade QS signal molecules. Inoculation of S. marcescens to P. heterophylla root could rapidly cause wilt disease, which was alleviated by B. thuringiensis. Furthermore, the expression of lactonase encoded by the aiiA gene in S. marcescens resulted in reduction of its pathogenicity, implying that the toxic effect of S. marcescens on the seedlings was QS-regulated. Meanwhile, excess lactonase in S. marcescens led to reduction in antibacterial substances, exoenzymes, and swarming motility, which might contribute to pathogensis on the seedlings. Root exudates and root tuber extracts of P. heterophylla significantly promoted the growth of S. marcescens, whereas a slight increase of B. thuringiensis was observed in both samples. These results demonstrated that QS-regulated behaviors in S. marcescens mediated by root exudates played an important role in replanting diseases of P. heterophylla.
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제안 방법
heterophylla were collected from both newly planted and replanted fields at the harvesting time, and the adjacent uncultivated field soil sample was used as a control. The soil samples of three conditions (uncultivated soil, newly planted soil, and replanted soil) were randomly chosen and analyzed. Five grams of three fresh samples was resuspended in 50 ml of sterile 0.
heterophylla were tested in pots by exogenous addition of diluted cultures (inoculated 2 cm away from the seedling root). The assessment of B. thuringiensis against S. marcescens was performed by adding the diluted cultures of different proportions (S. marcescens:B. thuringiensis, 2:3, 1:1, 7:3, 4:1, and 9:1), which were inoculated 2 cm away from the seedling root. Equal amounts of LB medium and E.
1. Screening of quorum sensing (QS) and quorum quenching (QQ) bacteria in uncultivated soil, newly planted soil, and replant soil of P. heterophylla using the biosensor strain C. violaceum CV026, where 100 colonies of each condition were randomly selected from the plates.
thuringiensis was determined from the optical density at 600 nm using a spectrophotometer (UV-1800, Mapada). All the experiments were performed in parallel triplicate tests.
heterophylla. In this study, we attempted to analyze the population of QS and QQ bacteria in the consecutive monoculture system of P. heterophylla and its relationship with replanting diseases. Screening of QS and QQ bacteria showed that 32 QS and one QQ bacteria were isolated from three samples of uncultivated, newly planted, and replanted soils.
대상 데이터
In previous studies, the QS system in the genus Serratia showed diversity, and QS luxIR homologs, including SmaIR, SpnIR, SplIR, SprIR, and SwrIR from the genus Serratia have been reported [32]. To confirm the genes in the QS system of the 32 isolates, 10 pairs of primers were designed and used to amplify the QS luxIR homologs using the genomic DNA of the 32 isolates. The sequencing results showed that these isolated strains only contained the SwrIR genes (GenBank Accession No.
성능/효과
heterophylla, which resulted in increased replanting disease incidence [38, 42]. These findings implied that replanting diseases of P. heterophylla might result from its interaction with soil pathogenic organisms mediated by root exudates from the consecutive monoculture system of P. heterophylla.
heterophylla with different years of monoculture was analyzed using the reporter strain Chromobacterium violaceum CV026. Thirty-two QS strains and one QQ strain were isolated and identified as Serratia marcescens and Bacillus thuringiensis, respectively. Subsequently, the SwrIR genes of the QS system in S.
Based on their physiological and biochemical characteristics as well as the 16S rDNA sequences, 32 QS bacteria and one QQ bacteria were identified as S. marcescens and B. thuringiensis respectively. The genes of QS system in the isolated S.
thuringiensis respectively. The genes of QS system in the isolated S. marcescens were amplified by PCR with 10 pairs of primers designed according to the reported gene sequences of the QS system (SpnIR, SplIR, SprIR, SmaIR, and SwrIR) from the genus Serratia (Table 1). The gene encoding lactonase in B.
1A). Among these isolates, 5, 12, and 15 colonies were obtained from uncultivated soil, newly planted soil, and replanted soil, respectively, which indicated that the number of QS bacteria increased significantly over the years of monoculture under consecutive monoculture conditions. However, only one QQ bacteria, isolated from newly planted soil, exhibited the ability of degrading AHL (Fig.
16S rDNA sequencing showed that the QQ isolate was a strain of Bacillus. Sequence alignment by the BLAST program revealed a consistently high identity (more than 98% identity) with species of B. thuringiensis.
thuringiensis under the same condition. These results demonstrated that the root exudates and root tuber extract of P. heterophylla provided adequate nutrients for the growth of S. marcescens, and led to rapid pathogen proliferation during the consecutive monoculture process of P. heterophylla.
According to the above results, the isolated QS and QQ strains were identified as S. marcescens and B. thuringiensis, respectively. In previous studies, the QS system in the genus Serratia showed diversity, and QS luxIR homologs, including SmaIR, SpnIR, SplIR, SprIR, and SwrIR from the genus Serratia have been reported [32].
To confirm the genes in the QS system of the 32 isolates, 10 pairs of primers were designed and used to amplify the QS luxIR homologs using the genomic DNA of the 32 isolates. The sequencing results showed that these isolated strains only contained the SwrIR genes (GenBank Accession No. EKF66792.1 and AAO38761.1) of the QS system (Fig. S1), which have been reported in S. marcescens MG1. The isolated QQ strain was identified as B.
S1). Sequence alignment indicated that the amplified fragment from the isolated QQ strain shared 99% identity with the reported aiiA gene of B. thuringiensis. Based on the findings above, the isolated QS strains should belong to S.
No disease symptoms could be observed during the entire experiment period using the treatment of the mixed strains at the ratio of 2:3. These results showed that exogenous addition of QQ strain B. thuringiensis could efficiently alleviate plant infection by the QS strain S. marcescens, whereas the imbalances between these two strains could result in increase of replanting disease incidence in consecutive monoculture.
The above results showed that the pathogenicity of S. marcescens on the seedlings of P. heterophylla could be alleviated by B. thuringiensis. However, it is not clear whether alleviation of the S.
4B). These results showed that S. marcescens pathogenicity on the P. heterophylla seedlings was regulated by the bacterial QS system.
4). The results demonstrated that S. marcescens was dependent on the QS system to evoke disease. The SwrIR genes of the QS system in S.
5). The results showed the wild S. marcescens exhibited antibacterial activity against B. thuringiensis and E. coli, which was defective in S. marcescens with overexpressed aiiA. Obvious reductions in exoenzyme production and swarming motility by S.
In conclusion, we analyzed and identified 32 QS strains of S. marcescens and one QQ strain of B. thuringiensis in the consecutive monoculture system of P. heterophylla. The QS strain of S.
heterophylla might provide a suitable environment and adequate nutrient for its proliferation. As shown in Fig. 2, root exudates and root tuber extracts of P. heterophylla could efficiently promote the growth of S. marcescens, whereas a slight increase for the growth of B. thuringiensis was observed. In addition, S.
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