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将真养产碱菌JMP134质粒pJP4基因转移至本地土壤受体菌。

Gene transfer of Alcaligenes eutrophus JMP134 plasmid pJP4 to indigenous soil recipients.

作者信息

DiGiovanni G D, Neilson J W, Pepper I L, Sinclair N A

机构信息

Department of Soil, Water, and Environmental Science, University of Arizona, Tucson 85721, USA.

出版信息

Appl Environ Microbiol. 1996 Jul;62(7):2521-6. doi: 10.1128/aem.62.7.2521-2526.1996.

Abstract

This study evaluated the potential for gene transfer of a large catabolic plasmid from an introduced organism to indigenous soil recipients. The donor organism Alcaligenes eutrophus JMP134 contained the 80-kb plasmid pJP4, which contains genes that code for mercury resistance. Genes on this plasmid plus chromosomal genes also allow degradation of 2,4-dichloruphenoxyacetic acid (2,4-D). When JMP134 was inoculated into a nonsterile soil microcosm amended with 1,000 micrograms of 2,4-D g-1, significant (10(6) g of soil-1) populations of indigenous recipients or transconjugants arose. These transconjugants all contained an 80-kb plasmid similar in size to pJP4, and all degraded 2,4-D. In addition, all transconjugants were resistant to mercury and contained the tfdB gene of pJP4 as detected by PCR. No mercury-resistant, 2,4-D-degrading organisms with large plasmids or the tfdB gene were found in the 2,4-D-amended but uninoculated control microcosm. These data clearly show that the plasmid pJP4 was transferred to indigenous soil recipients. Even more striking is the fact that not only did the indigenous transconjugant population survive and proliferate but also enhanced rates of 2,4-D degradation occurred relative to microcosms in which no such gene transfer occurred. Overall, these data indicate that gene transfer from introduced organisms is an effective means of bioaugmentation and that survival of the introduced organism is not a prerequisite for biodegradation that utilizes introduced biodegradative genes.

摘要

本研究评估了一种大型分解代谢质粒从引入的生物体转移至本地土壤受体的可能性。供体生物真养产碱菌JMP134含有80 kb的质粒pJP4,该质粒含有编码汞抗性的基因。该质粒上的基因以及染色体基因还可使2,4-二氯苯氧乙酸(2,4-D)降解。当将JMP134接种到添加了1000微克2,4-D g-1的非无菌土壤微观环境中时,出现了数量可观(每克土壤10⁶个)的本地受体或转接合体。这些转接合体均含有一个大小与pJP4相似的80 kb质粒,并且都能降解2,4-D。此外,所有转接合体都对汞具有抗性,并且通过PCR检测发现含有pJP4的tfdB基因。在添加了2,4-D但未接种的对照微观环境中,未发现具有大质粒或tfdB基因的抗汞、降解2,4-D的生物体。这些数据清楚地表明质粒pJP4已转移至本地土壤受体。更引人注目的是,不仅本地转接合体群体得以存活和增殖,而且相对于未发生此类基因转移的微观环境,2,4-D的降解速率有所提高。总体而言,这些数据表明,从引入的生物体进行基因转移是生物强化的一种有效手段,并且引入生物体的存活并非利用引入的生物降解基因进行生物降解的先决条件。

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