Towards identifying host cell-type specific response patterns to bacterial endosymbiosis

Publikation: Bog/antologi/afhandling/rapportPh.d.-afhandling

  • Srdjan Gavrilovic, Danmark
The establishment of Symbiotic Nitrogen Fixation (SNF) is a complex process. It requires highly sophisticated signal exchanges between host plant and bacteria in order to fine-tune the molecular mechanisms necessary for optimal performance of the symbiosis, which ultimately determines the evolutionary success of the interaction. Precise spatial and temporal coordination of the
expression of numerous genes is necessary to develop and maintain such a complex interaction. Many gene expression studies have been conducted to gain insight into bacterial root endosymbiosis. From a historical point of view, available techniques have relied heavily on whole organ analyses that disregard specificities of individual cell types. To address this issue we aimed to develop a technology for comparative global analysis of mature mRNA and small RNA
populations at the cell type specific level in the model plant Lotus japonicus.
A powerful approach referred to here as Defined Expression and RNA Affinity co-Purification (DERAP) was developed to study gene expression and small RNA populations in the host roots during early phases of signal exchange at the cell-type level. As a basis for DERAP analysis of the L. japonicus root during symbiosis establishment, we generated tools to express molecular elements of the cellular RNA binding machinery in a cell-type defined manner. To this end, a set of promoters that allows cell type specific expression in all major root cell types of the responsive zone, namely epidermis with elongating root hairs, inner cortex, endodermis, phloem and xylem, were characterized in L. japonicus. In combination with tagged forms of a Ribosomal surface
Protein (RP) and the viral small RNA binding protein P19, these promoters were introduced into L. japonicus ecotype Gifu, and whole plant transformants were regenerated. These will form a basis for isolating transcriptionally active mRNA fractions associated with ribosomes and 21 nt long small RNAs from targeted cell populations.
OriginalsprogEngelsk
ForlagAarhus University, Faculty of Science and Technology
Antal sider113
Rekvirerende organGraduate School of Science and Technology
StatusUdgivet - 4 dec. 2012

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