RESUMEN
Although droplet microfluidics has been studied for the past two decades, its applications are still limited due to the low productivity of microdroplets resulting from the low integration of planar microchannel structures. In this study, a microfluidic system implementing inverse colloidal crystals (ICCs), a spongious matrix with regularly and densely formed three-dimensional (3D) interconnected micropores, was developed to significantly increase the throughput of microdroplet generation. A new bottom-up microfabrication technique was developed to seamlessly integrate the ICCs into planar microchannels by accumulating non-crosslinked spherical PMMA microparticles as sacrificial porogens in a selective area of a mold and later dissolving them. We have demonstrated that the densely arranged micropores on the spongious ICC of the microchannel function as massively parallel micronozzles, enabling droplet formation on the order of >10 kHz. Droplet size could be adjusted by flow conditions, fluid properties, and micropore size, and biopolymer particles composed of polysaccharides and proteins were produced. By further parallelization of the unit structures, droplet formation on the order of >100 kHz was achieved. The presented approach is an upgrade of the existing droplet microfluidics concept, not only in terms of its high throughput, but also in terms of ease of fabrication and operation.
RESUMEN
Salicylic acid (SA), an essential secondary messenger for plant defence responses, plays a role in maintaining a balance (trade-off) between plant growth and resistance induction, but the detailed mechanism has not been explored. Because the SA mimic benzothiadiazole (BTH) is a more stable inducer of plant defence than SA after exogenous application, we analysed expression profiles of defence genes after BTH treatment to better understand SA-mediated immune induction. Transcript levels of the salicylic acid glucosyltransferase (SAGT) gene were significantly lower in BTH-treated Nicotiana tabacum (Nt) plants than in SA-treated Nt control plants, suggesting that SAGT may play an important role in SA-related host defence responses. Treatment with BTH followed by SA suppressed SAGT transcription, indicating that the inhibitory effect of BTH is not reversible. In addition, in BTH-treated Nt and Nicotiana benthamiana (Nb) plants, an early high accumulation of SA and SA 2-O-ß-d-glucoside was only transient compared to the control. This observation agreed well with the finding that SAGT-overexpressing (OE) Nb lines contained less SA and jasmonic acid (JA) than in the Nb plants. When inoculated with a virus, the OE Nb plants showed more severe symptoms and accumulated higher levels of virus, while resistance increased in SAGT-silenced (IR) Nb plants. In addition, the IR plants restricted bacterial spread to the inoculated leaves. After the BTH treatment, OE Nb plants were slightly larger than the Nb plants. These results together indicate that SAGT has a pivotal role in the balance between plant growth and SA/JA-mediated defence for optimum plant fitness.
