Reversible binding of zinc in Plasmodium falciparum Sir2: Structure and activity of the apoenzyme

Reversible zinc chelation via thiol groups of cysteines leading to modulation of activity in redox regulated proteins forms a basis for switching on–off of various biochemical processes. Silent information regulator 2 (Sir2), a NAD⁺ dependent deacetylase, contains a non-catalytic zinc ion coordinated by thiol groups of cysteines. Using Plasmodium falciparum Sir2 (PfSir2), we have examined the effect of zinc removal on the structure and activity of this enzyme. Our studies show that the enzyme with high affinity for zinc exhibits partial collapse of structure upon removal of the metal ion. Zinc reconstitution of apo PfSir2 led to recovery of both structure and activity highlighting the reversibility of the process.     

Indirect regeneration of Withania somnifera and comparative analysis of withanolides in in vitro and greenhouse grown plants

The present study reports an efficient protocol for indirect shoot organogenesis and plantlets regeneration of Withania somnifera (L.) Dunal. Leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of 6-benzylaminopurine (BAP) and indole-3-acetic acid (IAA). The highest callus induction rate (89.5 %) and shoot regeneration rate (92 %) were obtained when 2 mg dm⁻³ BAP was combined with 0.5 mg dm⁻³ IAA. Three major withanolides (withaferine A, 12-deoxywithastramonolide and withanolide A) were investigated in different plant organs from in vitro and greenhouse grown plants. Leaves contained higher contents of withanolides and phenolics than roots or stems, whereas roots contained the highest contents of flavonoids and polysacharides. In vitro grown plants contained greater contents of phenolics, flavonoids and polysaccharides while lower contents of withanolides than greenhouse grown plants.     

Application of RAPD markers for characterization of γ-ray-induced rose mutants and assessment of genetic diversity

Six parent and their 12 gamma ray-induced somatic flower colour mutants of garden rose were characterized to discriminate the mutants from their respective parents and understanding the genetic diversity using Random amplification of polymorphic DNA (RAPD) markers. Out of 20 primers screened, 14 primers yielded completely identical fragments patterns. The other 7 primers gave highly polymorphic banding patterns among the radiomutants. All the cultivars were identified by using only 7 primers. Moreover, individual mutants were also distinguished by unique RAPD marker bands. Based on the presence or absence of the 48 polymorphic bands, the genetic variations within and among the 18 cultivars were measured. Genetic distance between all 18 cultivars varied from 0.40 to 0.91, as revealed by Jaccard’s coefficient matrix. A dendrogram was constructed based on the similarity matrix using the Neighbor Joining Tree method showed three main clusters. The present RAPD analysis can be used not only for estimating genetic diversity present in gamma ray-induced mutants but also for correct identification of mutant/new varieties for their legal protection under plant variety rights.     

Arsenic affects mineral nutrients in grains of various Indian rice (Oryza sativa L.) genotypes grown on arsenic-contaminated soils of West Bengal

The exposure of paddy fields to arsenic (As) through groundwater irrigation is a serious concern that may not only lead to As accumulation to unacceptable levels but also interfere with mineral nutrients in rice grains. In the present field study, profiling of the mineral nutrients (iron (Fe), phosphorous, zinc, and selenium (Se)) was done in various rice genotypes with respect to As accumulation. A significant genotypic variation was observed in elemental retention on root Fe plaque and their accumulation in various plant parts including grains, specific As uptake (29–167 mg kg^?1 dw), as well as As transfer factor (4–45%). Grains retained the least level of As (0.7–3%) with inorganic As species being the dominant forms, while organic As species, viz., dimethylarsinic acid and monomethylarsonic acid, were non-detectable. In all tested varieties, the level of Se was low (0.05–0.12 mg kg^?1 dw), whereas that of As was high (0.4–1.68 mg kg^?1 dw), considering their safe/recommended daily intake limits, which may not warrant their human consumption. Hence, their utilization may increase the risk of arsenicosis, when grown in As-contaminated areas.     

Synthesis and QSAR studies on hypotensive 1-[3-(4-substituted phenylthio) propyl]-4-(substituted phenyl) piperazines

A series of 1-[3-(4-substituted phenylthio) propyl]-4-(substituted phenyl) piperazines has been synthesized and evaluated for hypotensive activity. The QSAR studies indicate that resonance and hydrophobic parameters of the aryl substituents are important for hypotensive activity. The similar role of resonance parameter in describing the variance of 5-HT(2A) receptor binding affinities of these compounds suggests a possible role of 5-HT(2A) receptors in mediating the hypotensive action of title compounds.     

Beyond host-pathogen interactions: microbial defense strategy in the host environment

Many extracellular pathogenic bacteria colonize human or animal bodies through evasion of the host immune system, a process called host-pathogen interaction. What happens when other intruders try to invade the same host and try to establish themselves in the same niche is largely unknown. In one well-studied case, Pseudomonas aeruginosa is known to secrete the protein azurin as a weapon against such invaders as cancers, parasites and viruses. The production of such weapons by pathogenic bacteria could provide important insights into how a pathogen responds in the post-colonization state to impede other intruders for its own survival. Moreover, these molecules might find use in the pharmaceutical industry as next-generation therapeutics.     

Spatio-temporal analysis of development of basal roots of common bean (Phaseolus vulgaris L.)

Temporal development of roots is key to the understanding of root system architecture of plants which influences nutrient uptake, anchorage and plant competition. Using time lapse imaging we analyzed developmental patterns of length, growth angle, depth and curvature of Phaseolus basal roots from emergence till 48 h in two genotypes, {"type":"entrez-nucleotide","attrs":{"text":"B98311","term_id":"3000390","term_text":"B98311"}}B98311 and TLP19 with contrasting growth angles. In both genotypes all basal roots appeared almost simultaneously, but their growth rates varied which accounted for differences in root length. The growth angles of the basal roots fluctuated rapidly during initial development due to oscillatory root growth causing local bends. Beyond 24 h, as the root curvature stabilized, so did the growth angle. Therefore growth angle of basal roots is not a very reliable quantity for characterizing root architecture, especially during early seedling development. Comparatively, tip depth is a more robust measure of vertical distribution of the basal roots even during early seedling development.Key words: basal root, kinematics, root architecture, root growth, spatiotemporal analysis, root imagingVertical and horizontal placements of the roots in the soil influence plant performance through acquisition of below ground resources like water and nutrients, plant anchorage and intra- and inter-plant competition.^1–4 Therefore the architecture of the root system plays important roles in regulating plant growth and yield, especially under abiotic stresses.⁵ As a seedling grows to become a mature plant, the root architecture develops continuously in response to various cues e.g., genotypic, environmental, hormonal, etc. Therefore studies of root architecture of plants of different ages are important for understanding the influence of these cues in regulating plant growth.The root scaffold of a plant is comprised of different types of roots with different functions. A mature common bean (Phaseolus vulgaris L.) plant has root system consisting of primary, adventitious, lateral and basal roots. Among these, the basal roots are typically the earliest emerging secondary roots from the hypocotyl⁶ forming a major part of the mature root system. We have recently demonstrated important differences in architectural traits of the basal roots of common bean in the early seedling stage between two contrasting class of genotypes and how auxin-ethylene interplay regulates these traits.⁷ While this study of basal roots at a fixed time allows assessment and comparison of root development up to that point of time, investigation of the temporal events of emergence and growth of the basal roots is important and complementary to the understanding of their architectural traits. Therefore in the present study, we examined the detailed developmental patterns of basal roots through time lapse imaging in two genotypes.We chose two bean genotypes with contrasting basal root growth angles (BRGA) relative to the gravity—{"type":"entrez-nucleotide","attrs":{"text":"B98311","term_id":"3000390","term_text":"B98311"}}B98311 producing basal roots of smaller BRGA (41.7° ± 14°) and TLP19 having roots of larger BRGA (56.4° ± 18°).⁸ The germinated seedling with 2–3 cm radical was transferred to the blue germination paper (Anchor Paper Co., St. Paul, MN), which was suspended in nutrient solution⁷ inside a growth chamber (ACMAS Technocracy Limited, Delhi, India) maintained at 25 ± 1°C. Time lapse photography was carried out for 48 h at 30 min intervals using Nikon D200 digital camera fitted with a macro lens to obtain high resolution digital images of the roots. Imaging started from the visibility of the protrusions of emerging basal root along the root-shoot interface. A computer program was developed in Matlab^® 7.8 (Mathworks, Natick) to analyze the images semi-automatically. From every image the computer program identified the basal roots using contrast of color between the roots (mostly white) and the germination paper (blue). Root midlines were determined following the methodology of Miller et al.⁹ and smoothed using the method of overlapping polynomials. Length of the midline is root length. The angle between gravity and the line connecting the root tip to the base is BRGA.⁷ The vertical distance of the root tip from the base of the lowest emerging root along the gravity vector is tip depth. From the midline, root curvature was also determined using the equation κ=x′y″−y′x″(x′2+y′2)3/2,(1) where [x(x), y(s)] is coordinate of any point along the root midline, s is normalized distance along the midline, and the primes denote derivatives with respect to s. Here positive curvature signifies bending upward and vice versa.     

Structural basis for regulation of the Crk signaling protein by a proline switch

Proline switches, controlled by cis-trans isomerization, have emerged as a particularly effective regulatory mechanism in a wide range of biological processes. Here we report the structures of both the cis and trans conformers of a proline switch in the Crk signaling protein. Proline isomerization toggles Crk between two conformations: an autoinhibitory conformation, stabilized by the intramolecular association of two tandem SH3 domains in the cis form, and an uninhibited, activated conformation promoted by the trans form. In addition to acting as a structural switch, the heterogeneous proline recruits cyclophilin A, which accelerates the interconversion rate between the isomers, thereby regulating the kinetics of Crk activation. The data provide atomic insight into the mechanisms that underpin the functionality of this binary switch and elucidate its remarkable efficiency. The results also reveal new SH3 binding surfaces, highlighting the binding versatility and expanding the noncanonical ligand repertoire of this important signaling domain.     

Effects of Hydroponic Solution EC,Substrates, PPF and Nutrient Scheduling on Growth and Photosynthetic Competence During Acclimatization of Micropropagated <Emphasis Type="Italic">Spathiphyllum</Emphasis> plantlets

In vitro regenerated shoots of Spathiphyllum from bioreactor were hydroponically cultured for 30 days. The response of plant growth and photosynthesis to different substrates, photosynthetic photon flux (PPF), nutrient scheduling and electrical conductivity (EC) of hydroponic solution were studied. The best plant growth response was observed in perlite based substrates with moderate PFF (70–100μmol m⁻² s⁻¹). Highest fresh weight, dry weight, shoot length, root length, root number and photosynthetic characteristics (chlorophyll, carotenoids and Fv/Fm) was observed in continuous immersion system. Plant growth responses, photosynthetic rate, stomatal conductance and transpiration rate were also found to be affected by EC levels. The optimum EC of a balanced nutrient solution was recorded as 1.2 dS m⁻¹. Photosynthetic activity was also characterized in terms of photochemical efficiency using measurements of chlorophyll fluorescence. Fv/Fm (it is a measure of the intrinsic or maximum efficiency of PSII i.e. the quantum efficiency if all PSII centers were open) also decreased significantly in plants grown under higher EC level; a decrease in this parameter indicates down regulation of photosynthesis or photoinhibition. Antioxidant defense enzymes such as catalase (CAT), ascorbate peroxidase (APX), peroxidase (POD), glutathione reductase (GR) and monodehydroascorbate reductase (MDHAR) significantly elevated in the leaves and roots of plantlets at higher EC levels. This increase could reflect a defense response to the cellular damage provoked by higher EC levels in the nutrient solution.