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.
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.
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.
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.
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