Abstract

One of the most studied mechanisms involved in bacterial evolution and diversification is conjugative transfer (CT) of plasmids. Plasmids able to transfer by CT often encode beneficial traits for bacterial survival under specific environmental conditions. Rhizobium etli CFN42 is a Gram-negative bacterium of agricultural relevance due to its symbiotic association with Phaseolus vulgaris through the formation of Nitrogen-fixing nodules. The genome of R. etli CFN42 consists of one chromosome and six large plasmids. Among these, pRet42a has been identified as a conjugative plasmid. The expression of the transfer genes is regulated by a quorum sensing (QS) system that includes a traI gene, which encodes an acyl-homoserine lactone (AHL) synthase and two transcriptional regulators (TraR and CinR). Recently, we have shown that pRet42a can perform CT on the root surface and inside nodules. The aim of this work was to determine the role of plant-related compounds in the CT of pRet42a. We found that bean root exudates or root and nodule extracts induce the CT of pRet42a in the plant rhizosphere. One possibility is that these compounds are used as nutrients, allowing the bacteria to increase their growth rate and reach the population density leading to the activation of the QS system in a shorter time. We tested if P. vulgaris compounds could substitute the bacterial AHL synthesized by TraI, to activate the conjugation machinery. The results showed that the transfer of pRet42a in the presence of the plant is dependent on the bacterial QS system, which cannot be substituted by plant compounds. Additionally, individual compounds of the plant exudates were evaluated; among these, some increased and others decreased the CT. With these results, we suggest that the plant could participate at different levels to modulate the CT, and that some compounds could be activating genes in the conjugation machinery.

Highlights

  • Plasmid conjugative transfer (CT) is a widely distributed mechanism that allows the distribution of genetic material across microorganisms

  • We showed the capacity of plasmid pRet42a of R. etli CFN42 (i.e derivative CFNX559) to be transferred at high frequency to other bacteria on the surface of the roots and inside the nodules generated during the interaction between this strain and P. vulgaris [32]

  • Tubes were inoculated with CFNX559 as a donor and CFN2001 as a recipient strain, and the CT frequencies were evaluated in the medium of the tubes, and on the root surface

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Summary

Introduction

Plasmid conjugative transfer (CT) is a widely distributed mechanism that allows the distribution of genetic material across microorganisms. Role of plant compounds in conjugative transfer of pRet42a grants PICT2016-0210 and PIP 2014-0420 to GTT. LABV received a Fellowship 384814 from Consejo Nacional de Ciencia y Tecnologıa. There was no additional external funding received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

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