Bacteria colonize plant roots and engage in reciprocal interactions with their hosts. However, the contribution of individual taxa or groups of bacteria to plant nutrition and fitness is not well characterized due to a lack of in\uffc2\uffa0situ evidence of bacterial activity.
To address this knowledge gap, we developed an analytical approach that combines the identification and localization of individual bacteria on root surfaces via gold\uffe2\uff80\uff90based in\uffc2\uffa0situ hybridization with correlative NanoSIMS imaging of incorporated stable isotopes, indicative of metabolic activity.
We incubated Kosakonia strain DS\uffe2\uff80\uff901\uffe2\uff80\uff90associated, gnotobiotically grown rice plants with 15N\uffe2\uff80\uff93N2 gas to detect in\uffc2\uffa0situ N2 fixation activity. Bacterial cells along the rhizoplane showed\uffc2\uffa0heterogeneous patterns of 15N enrichment, ranging from the natural isotope abundance levels up to 12.07 at% 15N (average and median of 3.36 and 2.85 at% 15N, respectively, n\uffe2\uff80\uff89=\uffe2\uff80\uff89697 cells).
The presented correlative optical and chemical imaging analysis is applicable to a broad range of studies investigating plant\uffe2\uff80\uff93microbe interactions. For example, it enables verification of the in\uffc2\uffa0situ metabolic activity of host\uffe2\uff80\uff90associated commercialized strains or plant growth\uffe2\uff80\uff90promoting bacteria, thereby disentangling their role in plant nutrition. Such data facilitate the design of plant\uffe2\uff80\uff93microbe combinations for improvement of crop management.
Bacteria colonize plant roots and engage in reciprocal interactions with their hosts. However, the contribution of individual taxa or groups of bacteria to plant nutrition and fitness is not well characterized due to a lack of in\uffc2\uffa0situ evidence of bacterial activity.
To address this knowledge gap, we developed an analytical approach that combines the identification and localization of individual bacteria on root surfaces via gold\uffe2\uff80\uff90based in\uffc2\uffa0situ hybridization with correlative NanoSIMS imaging of incorporated stable isotopes, indicative of metabolic activity.
We incubated Kosakonia strain DS\uffe2\uff80\uff901\uffe2\uff80\uff90associated, gnotobiotically grown rice plants with 15N\uffe2\uff80\uff93N2 gas to detect in\uffc2\uffa0situ N2 fixation activity. Bacterial cells along the rhizoplane showed\uffc2\uffa0heterogeneous patterns of 15N enrichment, ranging from the natural isotope abundance levels up to 12.07 at% 15N (average and median of 3.36 and 2.85 at% 15N, respectively, n\uffe2\uff80\uff89=\uffe2\uff80\uff89697 cells).
The presented correlative optical and chemical imaging analysis is applicable to a broad range of studies investigating plant\uffe2\uff80\uff93microbe interactions. For example, it enables verification of the in\uffc2\uffa0situ metabolic activity of host\uffe2\uff80\uff90associated commercialized strains or plant growth\uffe2\uff80\uff90promoting bacteria, thereby disentangling their role in plant nutrition. Such data facilitate the design of plant\uffe2\uff80\uff93microbe combinations for improvement of crop management.