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Glutamine (Gln) is a critical intermediate in nitrogen metabolism in all organisms. Here, a whole cell biosensor (GlnLux) for Gln was constructed by transforming a bacterial Gln auxotroph with a constitutive lux reporter. The biosensor was optimized for sensitivity, linearity, efficiency, specificity and robustness to permit detection of Gln in vitro and in vivo. The optimized GlnLux biosensor achieved nanomolar sensitivity with Gln standards. Extracts from only 1 mg of maize (Zea mays L.) leaf tissue were sufficient for Gln detection by GlnLux. Measurements of Gln in leaf extracts by GlnLux correlated with quantification by high performance liquid chromatography (Spearman r = 0.95). GlnLux permitted indirect in planta imaging of Gln using a CCD camera, enabling identification of plants that had been fertilized with nitrogen. Imaging using GlnLux also resolved predicted spatial differences in leaf Gln concentration. In a second application, it was demonstrated that GlnLux embedded into agar permits non-destructive screening of co-inoculated bacterial colonies for biological nitrogen fixation (BNF). GlnLux agar was able to distinguish a Bradyrhizobium japonicum wild type strain (nif+) from a mutant strain defective in nitrogenase (nif-) following ≥8 h of co-incubation. The technology was used to screen a bacterial endophyte diversity library cultured from Zea mays (L.) seeds for biological nitrogen fixation. / OMAFRA

Sustainable vegetable production is one of the most active areas of vegetable research and of concern to all producers. Everyone, both producers and consumers, are concerned with sustainability. Brussels sprouts and sweet basil are high value commodities, but increasing global concerns about water availability, insect-pest problems, and costly fertilizer inputs severely impact the growth and production of these crops. Low tunnels covered with spun-bonded fabric can improve production of vegetables and herbs in Virginia and the U.S. This study investigated the performance of Brussels sprouts and basil grown under low tunnels (LTs), and their relationship with water use efficiency, nitrogen use efficiency, and the level of protection against insect injury. Low tunnels increased yield, number of sprouts, and water use efficiency of Brussels sprout production. In addition, LTs decreased irrigation requirements, irrigation events, leaf feeding injury, and insect populations in comparison to open field. Similarly, LTs increased summer production of sweet basil as measured by fresh weight and biomass. In addition, plant N uptake was greater under the LTs; however, the increase in nitrogen use efficiency was inconsistent. / Master of Science in Life Sciences / Brussels sprouts and sweet basil are economically important cash crops on the East Coast. Brussels sprouts is a Cole crop and an important source of dietary fiber, vitamins (A, C & K), calcium, iron, manganese and antioxidants. Similarly, sweet basil is a member of the mint family and important high-value herb in the U.S. and the world. It is mainly grown for culinary purposes as a dried and fresh spice in the U.S. However, demand for these commodities is increasing. Low tunnels (LTs) covered with spunbonded fabric can be a practical management tool to increase yield. Results from this study indicate that LTs increase yield of Brussels sprouts and basil, water use efficiency and total nitrogen uptake, while reducing insect pest infestation. Therefore, LTs can be a useful tool to improve sustainability of Brussels sprouts and basil production.

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