Conjugative transfer of preferential utilization of aromatic compounds from Pseudomonas putida CSV86
Conjugative transfer of preferential utilization of aromatic compounds from Pseudomonas putida CSV86
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DOI：
10.1007/s10532-007-9117-7
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发表时间：
2008-02-01
期刊：
影响因子：
3.6
通讯作者：
Phale, Prashant S.
中科院分区：
文献类型：
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作者：
Basu, Aditya;Phale, Prashant S.
恶臭假单胞菌 CSV86 优先利用萘 (Nap)、水杨酸盐 (Sal)、苯甲醇 (Balc) 和甲基萘 (MN)，而不是葡萄糖。甲基萘通过环羟基化以及侧链羟基化途径代谢。虽然降解性能稳定，但在固化剂存在下，获得 Nap(-)Sal(-)MN(-)Balc(-) 表型的频率增加至 11%。该特性通过接合转移至嗜麦芽寡养单胞菌 CSV89，每个供体细胞的频率为 7 x 10(-8)。转接合子是Nap(+)Sal(+)MN(+)Balc(+)，并通过环和侧链羟基化途径代谢MN。转接合子还显示出相对于葡萄糖优先利用芳香族化合物，这表明优先降解特性的转移。当转接合子在葡萄糖或 2YT 上生长时，转移的特性完全丧失。从 CSV86 和转接合子中检测和分离质粒 DNA 的尝试未成功。通过使用 1,2-二羟基萘双加氧酶和儿茶酚 2,3-双加氧酶 (C23O) 特异性引物的 PCR 以及使用总 DNA 作为模板和 C23O PCR 片段作为探针的 DNA-DNA 杂交，证实了降解基因的转移及其随后从转接合子中的丢失。这些结果表明可能的共轭元素参与以下过程：（i）芳香族化合物的代谢，（ii）MN的环和侧链羟基化途径，以及（iii）芳香族化合物相对于葡萄糖的优先利用。
Pseudomonas putida CSV86 utilizes naphthalene (Nap), salicylate (Sal), benzyl alcohol (Balc), and methylnaphthalene (MN) preferentially over glucose. Methylnaphthalene is metabolized by ring-hydroxylation as well as side-chain hydroxylation pathway. Although the degradation property was found to be stable, the frequency of obtaining Nap(-)Sal(-)MN(-)Balc(-) phenotype increased to 11% in the presence of curing agents. This property was transferred by conjugation to Stenotrophomonas maltophilia CSV89 with a frequency of 7 x 10(-8) per donor cells. Transconjugants were Nap(+)Sal(+)MN(+)Balc(+) and metabolized MN by ring- as well as side-chain hydroxylation pathway. Transconjugants also showed the preferential utilization of aromatic compounds over glucose indicating transfer of the preferential degradation property. The transferred properties were lost completely when transconjugants were grown on glucose or 2YT. Attempts to detect and isolate plasmid DNA from CSV86 and transconjugants were unsuccessful. Transfer of degradation genes and its subsequent loss from the transconjugants was confirmed by PCR using primers specific for 1,2-dihydroxynaphthalene dioxygenase and catechol 2,3-dioxygenase (C23O) as well as by DNA-DNA hybridizations using total DNA as template and C23O PCR fragment as a probe. These results indicate the involvement of a probable conjugative element in the: (i) metabolism of aromatic compounds, (ii) ring- and side-chain hydroxylation pathways for MN, and (iii) preferential utilization of aromatics over glucose.