Adaptive optical third-harmonic generation microscopy for in vivo imaging of tissues.

Third-harmonic generation microscopy is a powerful label-free nonlinear imaging technique, providing essential information about structural characteristics of cells and tissues without requiring external labelling agents. In this work, we integrated a recently developed compact adaptive optics module into a third-harmonic generation microscope, to measure and correct for optical aberrations in complex tissues. Taking advantage of the high sensitivity of the third-harmonic generation process to material interfaces and thin membranes, along with the 1,300-nm excitation wavelength used here, our adaptive optical third-harmonic generation microscope enabled high-resolution in vivo imaging within highly scattering biological model systems. Examples include imaging of myelinated axons and vascular structures within the mouse spinal cord and deep cortical layers of the mouse brain, along with imaging of key anatomical features in the roots of the model plant Brachypodium distachyon. In all instances, aberration correction led to enhancements in image quality.

Adaptive optical third-harmonic generation microscopy for in vivo imaging of tissues.

Third-harmonic generation microscopy is a powerful label-free nonlinear imaging technique, providing essential information about structural characteristics of cells and tissues without requiring external labelling agents. In this work, we integrated a recently developed compact adaptive optics module into a third-harmonic generation microscope, to measure and correct for optical aberrations in complex tissues. Taking advantage of the high sensitivity of the third-harmonic generation process to material interfaces and thin membranes, along with the 1,300-nm excitation wavelength used here, our adaptive optical third-harmonic generation microscope enabled high-resolution in vivo imaging within highly scattering biological model systems. Examples include imaging of myelinated axons and vascular structures within the mouse spinal cord and deep cortical layers of the mouse brain, along with imaging of key anatomical features in the roots of the model plant Brachypodium distachyon. In all instances, aberration correction led to significant enhancements in image quality.

Plant host domestication and soil nutrient availability determine positive plant microbial response across the Solanum genus.

Domestication of crops has changed how crops shape their associated microbial communities compared with their progenitors. However, studies testing how crop domestication-driven differences in rhizosphere microbial communities affect plant health are limited mostly to specific symbiont pairings. By conducting a soil manipulation greenhouse study, we examined plant growth and yield in response to differences in microbial communities and nutrient availability across a variety of wild, landrace, and commercially available 'Modern' potatoes. Coupled with this, we conducted 16S and internal transcribed spacer (ITS) amplicon sequencing to examine plant host- and soil treatment-driven differences in microbial community composition on potato plant roots. We found that the plant response to microbes (PRM) was context dependent. In low nutrient conditions, landraces responded positively to the presence of live soil microbial inocula. Conversely, modern potato varieties responded positively only in high nutrient conditions. Amplicon sequencing found differences in bacterial communities due to environmental and temporal factors. However, potato clade (e.g. Andigenum, Chiletanum, Solanum berthaultii, and 'Modern') alone did not lead to differences in microbial communities that accounted for PRM differences. Differences in PRM between landraces and modern potatoes, and the correlation of PRM to microbial diversity, suggest that domestication and subsequent breeding have altered the S. tuberosum response to rhizosphere microbiomes between Andigenum, Chiletanum, and North American potato varieties.

16S rRNA analysis of diversity of manure microbial community in dairy farm environment.

Dairy farms generate a considerable amount of manure, which is applied in cropland as fertilizer. While the use of manure as fertilizer reduces the application of chemical fertilizers, the main concern with regards to manure application is microbial pollution. Manure is a reservoir of a broad range of microbial populations, including pathogens, which have potential to cause contamination and pose risks to public and animal health. Despite the widespread use of manure fertilizer, the change in microbial diversity of manure under various treatment processes is still not well-understood. We hypothesize that the microbial population of animal waste changes with manure handling used in a farm environment. Consequential microbial risk caused by animal manure may depend on manure handling. In this study, a reconnaissance effort for sampling dairy manure in California Central Valley followed by 16S rRNA analysis of content and diversity was undertaken to understand the microbiome of manure after various handling processes. The microbial community analysis of manure revealed that the population in liquid manure differs from that in solid manure. For instance, the bacteria of genus Sulfuriomonas were unique in liquid samples, while the bacteria of genus Thermos were observed only in solid samples. Bacteria of genus Clostridium were present in both solid and liquid samples. The population among liquid samples was comparable, as was the population among solid samples. These findings suggest that the mode of manure application (i.e., liquid versus solid) could have a potential impact on the microbiome of cropland receiving manure as fertilizers.