As metabolic centers, plant organelles participate in maintenance, defense, and signaling. MSH1 is a plant-specific protein involved in organellar genome stability in mitochondria and plastids. Plastid depletion of MSH1 causes heritable, non-genetic changes in development and DNA methylation. We investigated the msh1 phenotype using hemi-complementation mutants and transgene-null segregants from RNAi suppression lines to sub-compartmentalize MSH1 effects. We show that MSH1 expression is spatially regulated, specifically localizing to plastids within the epidermis and vascular parenchyma. The protein binds DNA and localizes to plastid and mitochondrial nucleoids, but fractionation and protein-protein interactions data indicate that MSH1 also associates with the thylakoid membrane. Plastid MSH1 depletion results in variegation, abiotic stress tolerance, variable growth rate, and delayed maturity. Depletion from mitochondria results in 7%-10% of plants altered in leaf morphology, heat tolerance, and mitochondrial genome stability. MSH1 does not localize within the nucleus directly, but plastid depletion produces non-genetic changes in flowering time, maturation, and growth rate that are heritable independent of MSH1. MSH1 depletion alters non-photoactive redox behavior in plastids and a sub-set of mitochondrially altered lines. Ectopic expression produces deleterious effects, underlining its strict expression control. Unraveling the complexity of the MSH1 effect offers insight into triggers of plant-specific, transgenerational adaptation behaviors.
Bioactive compounds were investigated in eight cultivars of broccoli grown under North Indian conditions over 2 years. The cultivars showed significant differences in phenols, antioxidant activity and other antioxidant constituents including ascorbic acid, β‐carotene, α‐tocopherol and chlorophyll. Free phenolics ranged from 19.60 to 41.40 mg/100 g fresh weight and on an average constituted 73% of total extractable phenolics. Cultivars Punjab broccoli and Packman had the highest ascorbic acid content, whereas green sprouting broccoli had the highest β‐carotene content. Total antioxidant activity as determined by ferric reducing antioxidant power ranged from 2.05 to 3.56 µmol Trolox/g fresh weight. Free radical scavenging activity as estimated by 2, 2‐diphenyl‐1‐picrylhydrazyl, ranged from 57 to 74%. There was strong positive correlation between free phenolics and antioxidant activity. Results indicate that both cultivars and year affect the bioactive composition in broccoli. PRACTICAL APPLICATIONS Broccoli is a potential healthy vegetable with enhanced antioxidant activity because of its high phenolic content. It also comprises a mixture of other antioxidants including ascorbic acid, carotenoids and α‐tocopherol which are known to prevent the onset of chronic diseases. Among broccoli cultivars, Aishwarya, Packman and Punjab broccoli seem to be promising ones, to be used in breeding programs to improve the quality of broccoli under Indian conditions.
To apply motor proteins as natural nanomolecular machines to transporting systems in nanotechnology, complete temporal control over ON/OFF switching of the motility is necessary. We have studied the photoresponsive inhibition properties of azobenzene-tethered peptides for regulation of kinesin-microtubule motility. Although a compound containing a peptide having an amino acid sequence derived from the kinesin's C-terminus (a known inhibitor of kinesin's motor domain) and also featuring a terminal azobenzene unit exhibited an inhibition effect, the phototunability of this behavior upon irradiation with UV or visible light was only moderate. Unexpectedly, newly synthesized peptides featuring the reverse sequence of amino acids of the C-terminus of kinesin exhibited excellent photoresponsive inhibition. In particular, azobenzene-CONH-IPKAIQASHGR completely stopped and started the motility of kinesin-microtubules in its trans- and cis-rich states, respectively, obtained after irradiation with visible and UV light, respectively. A gliding motility system containing this photoresponsive inhibitor allowed in situ control of the motion of microtubules on a kinesin-coated glass substrate. It is expected that the present results on the photoresponsive nanomotor system open up new opportunities to design nanotransportation systems.
The aim of this study is to investigate in vitro antioxidant activities and the phytochemical screening endophytes. Seven different endophytic fungi were isolated from different parts of the plant and their extracts subjected to know antioxidant properties and phytochemical screening. Phytochemical analysis revealed the presence of tannins, flavonoids, steroids, alkaloids, phenols and proteins from different solvents extracts of different endophytes. The antioxidant activity was evaluated by six separated methods: scavenging of free radical DPPH, FRAP, TBA, superoxide radical, FTC and iron methods. All seven different endophytes yielded almost all phytochemicals in methanol extracts which were tested. The endophytes A. niger, Penicillium sp. and Trichoderma sp. have shown potential in vitro antioxidant activities. Further work is needful to isolate the exact compound which is responsible for antioxidant activity and biophysical characterization will be done in the future.
Regulated transportation of nanoscale objects with a high degree of spatiotemporal precision is a prerequisite for the development of targeted molecular delivery. In vitro integration of the kinesin-microtubule motor system with synthetic molecules offers opportunities to develop controllable molecular shuttles for lab-on-a-chip applications. We attempted a combination of the kinesin-microtubule motor system with push-pull type azobenzene tethered inhibitory peptides (azo-peptides) through which reversible, spatiotemporal control over the kinesin motor activity was achieved locally by a single, visible wavelength. The fast thermal relaxation of the cis-isomers of azo-peptides offered us quick and complete resetting of the trans-state in the dark, circumventing the requirement of two distinct wavelengths for two-way switching of kinesin-driven microtubule motility. Herein, we report the manipulation of selected, single microtubule movement while keeping other microtubules at complete rest. The photoresponsive inhibitors discussed herein would help in realizing complex bionanodevices.
Artificial control of bio-nanomachines should have a major impact on the development of controllable transport systems for specific cargo transport on chips. Precise spatiotemporal control and local regulation of the bio-motor activity will, however, be necessary if we are to accomplish such a goal. In this study, we exploited the photoswitching properties of azobenzene-based high-energy molecules and inhibitors to control a single kinesin-driven microtubule that has potential to work as a nanocarrier for molecular cargos. In particular, we could influence the local concentration and dispersion of the microtubules at any desired position and time by irradiating a local area of the motility system at one wavelength, while irradiating the entire area at another wavelength, to enrich either cis or trans isomers of photoswitches in the selected region. Furthermore, various regulations (e.g., transporting, bending, breaking) of single microtubules were possible while almost arresting ambient microtubules-all without the need for any surface patterning.
An experimental study on heat transfer enhancement in an automobile radiator using Al2O3/water-ethylene glycol (EG) nanofluids is carried out. Heat transfer enhancement studies can help in the design of lighter and more compact radiators for the same given load, which in turn can improve the fuel economy of the automobile. A closed loop experimental setup is designed using a commercial automobile radiator for the study. The effect of adding EG to water on the overall heat conductance (UA) is studied using two mixtures of water-EG proportions, 90:10 and 80:20 (by volume). They showed a reduction in UA by 20% and 25% respectively. Experiments have also been done using Al2O3/water-EG nanofluids. The nanofluid was prepared using an 80:20 mixture and 0.1% (vol.) of Al2O3 nanoparticles. The addition of nanoparticles enhanced the heat transfer performance by 37 %. All the experiments have been conducted at a constant coolant flow rate and coolant inlet temperatures varying from 40 o C to 70 o C. The results showed that the heat transfer performance of the radiator reduced with the addition of EG and increased with the addition of nanoparticles to the water-EG mixture.
Recently we demonstrated the photoregulation of the activity of kinesin-1 using an azobenzene-tethered peptide (azo-peptide: Azo-Ile-Pro-Lys-Ala-Ile-Gln-Ala-Ser-His-Gly-Arg-OH). To understand the mechanism behind this photoswitchable inhibition, here we studied the structure-property relationships of a range of azo-peptides through systematic variations in the structures of the peptide and azobenzene units. The vital peptide sequence for kinesin inhibition-mediated through electrostatic, hydrophobic and C-Hπ interactions-was the same as that for the self-inhibition of kinesin. We also identified substituents on the azobenzene capable of enhancing the photoswitchability of inhibition. As a result, we developed a new inhibitor featuring a relatively short peptide unit (-Arg-Ile-Pro-Lys-Ala-Ile-Arg-OH) and an azobenzene unit bearing a para-OMe group. In the trans form of its azobenzene unit, this finely tuned inhibitor stopped the kinesin-driven gliding motility of microtubules completely at a relatively low concentration, yet allowed gliding motility with a relatively high velocity in the cis form obtained after UV irradiation.
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