In rice (Oryza sativa L.), later flowering inferior spikelets (IS), which are located on proximal secondary branches, fill slowly and produce smaller and lighter grains than earlier flowering superior spikelets (SS). Many genes have been reported to be involved in poor grain filling of IS, however the underlying molecular mechanisms remain unclear. The present study determined that GF14f, a member of the 14-3-3 protein family, showed temporal and spatial differences in expression patterns between SS and IS. Using GF14f-RNAi plants, we observed that a reduction in GF14f expression in the endosperm resulted in a significant increase in both grain length and weight, which in turn improved grain yield. Furthermore, pull-down assays indicated that GF14f interacts with enzymes that are involved in sucrose breakdown, starch synthesis, tricarboxylic acid (TCA) cycle and glycolysis. At the same time, an increase in the activity of sucrose synthase (SuSase), adenosine diphosphate-glucose pyrophosphorylase (AGPase), and starch synthase (StSase) was observed in the GF14f-RNAi grains. Comprehensive analysis of the proteome and metabolite profiling revealed that the abundance of proteins related to the TCA cycle, and glycolysis increased in the GF14f-RNAi grains together with several carbohydrate intermediates. These results suggested that GF14f negatively affected grain development and filling, and the observed higher abundance of the GF14f protein in IS compared with SS may be responsible for poor IS grain filling. The study provides insights into the molecular mechanisms underlying poor grain filling of IS and suggests that GF14f could serve as a potential tool for improving rice grain filling.
We present a micropump with a simple planar design featuring compliant in-contact check valves in a single layer, which allows for a simple structure and easy system integration. The micropump, based on poly(dimethylsiloxane) (PDMS), primarily consists of a pneumatically driven thin membrane, a pump chamber, and two in-plane check valves. The pair of check valves is based on an in-contact flap–stopper configuration and is able to minimize leakage flow, greatly enhancing the reliability and performance of the micropump. Systematic experimental characterization of the micropump has been performed in terms of the frequency response of the pumping flow rate with respect to factors including device geometry (e.g. chamber height) and operating parameters (e.g. pneumatic driving pressure and backpressure). The results demonstrate that this micropump is capable of reliably generating a maximum flow rate of 41 μL min−1 and operating against a high backpressure of up to 25 kPa. In addition, a lumped-parameter theoretical model for the planar micropump is also developed for accurate analysis of the device behavior. These results demonstrate the capability of this micropump for diverse applications in lab-on-a-chip systems.
This paper presents label-free characterization of temperature-dependent biomolecular affinity binding on solid surfaces using a microcantilever-based device. The device consists of a Parylene cantilever one side of which is coated with a gold film and functionalized with molecules as an affinity receptor to a target analyte. The cantilever is located in a poly(dimethylsiloxane) (PDMS) microfluidic chamber that is integrated with a transparent indium tin oxide (ITO) resistive temperature sensor on the underlying substrate. The ITO sensor allows for real-time measurements of the chamber temperature, as well as unobstructed optical access for reflection-based optical detection of the cantilever deflection. To test the temperature-dependent binding between the target and receptor, the temperature of the chamber is maintained at a constant setpoint, while a solution of unlabeled analyte molecules is continuously infused through the chamber. The measured cantilever deflection is used to determine the target-receptor binding characteristics. We demonstrate label-free characterization of temperature-dependent binding kinetics of the platelet-derived growth factor (PDGF) protein with an aptamer receptor. Affinity binding properties including the association and dissociation rate constants as well as equilibrium dissociation constant are obtained, and shown to exhibit significant dependencies on temperature.
We present a lumped-parameter dynamic model for a planar poly(dimethylsiloxane) (PDMS) micropump using in-contact, virtually zero-leakage check valves to provide critical insights into the dynamic characteristics of the planar micropump operation. The model features a single-plane microfluidic chamber sandwiched between two check valves, which allows for reliable operation and easy system integration. Theoretic equations have been developed based on the coupled interaction of the fluid flow and the structural motion, combined with external pneumatic actuation directly applied. Furthermore, we have resolved the equations analytically and analyzed the dynamic performance of this micropump. Systematic analysis using this model studied comprehensive factors including the motion of the actuated diaphragm, the applied pressure, and pumping chamber height. Compared with experimental characterization data, this model shows good agreement with the experimental results, demonstrating its utility as a useful tool for analyzing and designing the planar micropump.
It is difficult for laser scanning confocal microscopy to obtain high- or ultra-high-resolution laser confocal images directly, which affects the deep mining and use of the embedded information in laser confocal images and forms a technical bottleneck in the in-depth exploration of the microscopic physiological and biochemical processes of plants. The super-resolution reconstruction model (SRGAN), which is based on a generative adversarial network and super-resolution reconstruction model (SRResNet), which is based on a residual network, was used to obtain single and secondary super-resolution reconstruction images of laser confocal images of the root cells of the hyperaccumulator Solanum nigrum. Using the peak signal-to-noise ratio (PSNR), structural similarity (SSIM) and mean opinion score (MOS), the models were evaluated by the image effects after reconstruction and were applied to the recognition of endocytic vesicles in Solanum nigrum root cells. The results showed that the single reconstruction and the secondary reconstruction of SRGAN and SRResNet improved the resolution of laser confocal images. PSNR, SSIM, and MOS were clearly improved, with a maximum PSNR of 47.690. The maximum increment of PSNR and SSIM of the secondary reconstruction images reached 21.7% and 2.8%, respectively, and the objective evaluation of the image quality was good. However, overall MOS was less than that of the single reconstruction, the perceptual quality was weakened, and the time cost was more than 130 times greater. The reconstruction effect of SRResNet was better than that of SRGAN. When SRGAN and SRResNet were used for the recognition of endocytic vesicles in Solanum nigrum root cells, the clarity of the reconstructed images was obviously improved, the boundary of the endocytic vesicles was clearer, and the number of identified endocytic vesicles increased from 6 to 9 and 10, respectively, and the mean fluorescence intensity was enhanced by 14.4% and 7.8%, respectively. Relevant research and achievements are of great significance for promoting the application of deep learning methods and image super-resolution reconstruction technology in laser confocal image studies.
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