Automated detection of Karnal bunt teliospores

Karnal bunt is a fungal disease which infects wheat and, when present in wheat crops, yields it unsatisfactory for human consumption. Due to the fact that Karnal bunt (KB) is difficult to detect in the field, samples are taken to laboratories where technicians use microscopes and methodically search for KB teliospores. AlliedSignal Federal Manufacturing and Technologies (FM and T), working with the Kansas Department of Agriculture, created a system which utilizes pattern recognition, feature extraction, and neural networks to prototype an automated detection system for identifying KB teliospores. System hardware consists of a biological compound microscope, motorized stage, CCD camera, frame grabber, and a PC. Integration of the system hardware with custom software comprises the machine vision system. Fundamental processing steps involve capturing an image from the slide, while concurrently processing the previous image. Features extracted from the acquired imagery are then processed by a neural network classifier which has been trained to recognize spore-like objects. Images with spore-like objects are reviewed by trained technicians. Benefits of this system include: (1) reduction of the overall cycle-time; (2) utilization of technicians for intelligent decision making (vs. manual searching); (3) a regulatory standard which is quantifiable and repeatable; (4) guaranteed 100% coverage of the cover slip; and (5) significantly enhanced detection accuracy

Programmable Ligand Detection System in Plants through a Synthetic Signal Transduction Pathway

There is an unmet need to monitor human and natural environments for substances that are intentionally or unintentionally introduced. A long-sought goal is to adapt plants to sense and respond to specific substances for use as environmental monitors. Computationally re-designed periplasmic binding proteins (PBPs) provide a means to design highly sensitive and specific ligand sensing capabilities in receptors. Input from these proteins can be linked to gene expression through histidine kinase (HK) mediated signaling. Components of HK signaling systems are evolutionarily conserved between bacteria and plants. We previously reported that in response to cytokinin-mediated HK activation in plants, the bacterial response regulator PhoB translocates to the nucleus and activates transcription. Also, we previously described a plant visual response system, the de-greening circuit, a threshold sensitive reporter system that produces a visual response which is remotely detectable and quantifiable.We describe assembly and function of a complete synthetic signal transduction pathway in plants that links input from computationally re-designed PBPs to a visual response. To sense extracellular ligands, we targeted the computational re-designed PBPs to the apoplast. PBPs bind the ligand and develop affinity for the extracellular domain of a chemotactic protein, Trg. We experimentally developed Trg fusions proteins, which bind the ligand-PBP complex, and activate intracellular PhoR, the HK cognate of PhoB. We then adapted Trg-PhoR fusions for function in plants showing that in the presence of an external ligand PhoB translocates to the nucleus and activates transcription. We linked this input to the de-greening circuit creating a detector plant.Our system is modular and PBPs can theoretically be designed to bind most small molecules. Hence our system, with improvements, may allow plants to serve as a simple and inexpensive means to monitor human surroundings for substances such as pollutants, explosives, or chemical agents