Comparative anatomy of Sirhookera (Orchidaceae) growing in Western Ghats of southern India

We compared the anatomical characteristics of vegetative organs, peduncle and mycorrhizal morphology of the two known species of Sirhookera (Epidendroideae, Orchidaceae) to identify anatomical markers for identification and the ecological adaptations of these species. The leaves are hypostomatic bearing tetracytic stomata and the walls of subsidiary cells are smooth in Sirhookera lanceolata and undulate in Sirhookera latifolia. On the adaxial and abaxial surfaces the leaves are covered by a thick cuticle. The hypodermis is dimorphic and present on both sides of the leaf; chlorenchyma is homogenous and the vascular bundles are collateral. The rhizome of Sirhookera possesses a single-layered epidermis, thick cuticle, thin-walled parenchymatous ground tissue containing starch grains and scattered collateral vascular bundles. A thick-walled sclerenchymatous band separates the cortex from the parenchymatous ground tissue comprising of banded cells in the peduncle. Starch grains are present in the ground tissue of the S. latifolia peduncle. The roots consist of the velamen, ∩-thickened exodermis, thin-walled cortex consisting of water-storage cells, O-thickened endodermis and a vascular cylinder with parenchymatous pith. Starch grains are present in the root cortical cells of S. lanceolata but absent in S. latifolia. Fungal pelotons that aids in nutrient acquisition were observed in the root cortical region of both species. The study revealed significant differences between the anatomical characteristics of the two species and that most of the anatomical features of Sirhookera relate to their ecological adaptations.     

TGFbeta intercepts nuclear glycogen synthase kinase 3beta to inhibit PDGF-induced DNA synthesis in mesangial cells

Here, we demonstrate a mechanism of TGFbeta-mediated inhibition of PDGF-induced DNA synthesis in mesangial cells. TGFbeta significantly inhibited nuclear Akt phosphorylation without any effect on PDGF-stimulated phosphorylation of PDGFR at PI 3 kinase binding site (Tyr-751). Remarkably, TGFbeta inhibited cyclin D1 and cyclin E expression with concomitant decrease in CDK2 activity induced by PDGF. More importantly, we demonstrate that TGFbeta significantly abolished Akt-mediated serine-9 phosphorylation of glycogen synthase kinase 3beta (GSK3beta), thus prevented its inactivation. Expression of inactive GSK3betaK85R mutant increased cyclin D1 expression and DNA synthesis similar to PDGF. These results provide the first evidence that TGFbeta intercepts Akt kinase activity in the nucleus to block inactivation of GSK3beta, leading to attenuation of PDGF-induced CDK2 activity and DNA synthesis.     

Nested and TaqMan® probe based quantitative PCR for the diagnosis of Ca. Phytoplasma in coconut palms

The root (wilt) disease caused by phytoplasma (Ca. Phytoplasma) is one of the major and destructive occurs in coconut gardens of Southern India. As this organism could not be cultured in vitro, the early detection in the palm is very much challenging. Hence, proper early diagnosis and inoculum assessment relay mostly on the molecular techniques namely nested and quantitative PCR (qPCR). So, the present study qPCR assay conjugated with TaqMan^® probe was developed which is a rapid, sensitive method to detect the phytoplasma. For the study, samples from different parts of infected coconut palms viz., spindle leaflets, roots and the insect vector—leaf hopper (Proutista moesta) were collected and assessed by targeting 16S rRNA gene. Further, nested PCR has been carried out using p1/p7 and fU5/rU3 primers and resulted in the amplification product size of 890 bp. From this amplified product, specifically a target of 69 bp from the 16S rRNA gene region has been detected through primers conjugated with Taqman probe in a step one instrument. The results indicated that the concentration of phytoplasma was more in spindle leaflets (8.9?×?10⁵ g of tissue) followed by roots (7.4?×?10⁵ g of tissue). Thus, a qPCR approach for detection and quantification of coconut phytoplasma was more advantageous than other PCR methods in terms of sensitivity and also reduced risk of cross contamination in the samples. Early diagnosis and quantification will pave way for the healthy coconut saplings selection and management under field conditions.

     

Use of inert gas jets to measure the forces required for mechanical gene transfection

Background

Abnormal blood glucose (BG) concentrations have been associated with increased morbidity and mortality in both critically ill adults and infants. Furthermore, hypoglycaemia and glycaemic variability have both been independently linked to mortality in these patients. Continuous Glucose Monitoring (CGM) devices have the potential to improve detection and diagnosis of these glycaemic abnormalities. However, sensor noise is a trade-off of the high measurement rate and must be managed effectively if CGMs are going to be used to monitor, diagnose and potentially help treat glycaemic abnormalities.

Aim

To develop a tool that will aid clinicians in identifying unusual CGM behaviour and highlight CGM data that potentially need to be interpreted with care.

Methods

CGM data and BG measurements from 50 infants at risk of hypoglycaemia were used. Unusual CGM measurements were classified using a stochastic model based on the kernel density method and historical CGM measurements from the cohort. CGM traces were colour coded with very unusual measurements coloured red, highlighting areas to be interpreted with care. A 5-fold validation of the model was Monte Carlo simulated 25 times to ensure an adequate model fit.

Results

The stochastic model was generated using ~67,000 CGM measurements, spread across the glycaemic range ~2-10?mmol/L. A 5-fold validation showed a good model fit: the model 80% confidence interval (CI) captured 83% of clinical CGM data, the model 90% CI captured 91% of clinical CGM data, and the model 99% CI captured 99% of clinical CGM data. Three patient examples show the stochastic classification method in use with 1) A stable, low variability patient which shows no unusual CGM measurements, 2) A patient with a very sudden, short hypoglycaemic event (classified as unusual), and, 3) A patient with very high, potentially un-physiological, glycaemic variability after day 3 of monitoring (classified as very unusual).

Conclusions

This study has produced a stochastic model and classification method capable of highlighting unusual CGM behaviour. This method has the potential to classify important glycaemic events (e.g. hypoglycaemia) as true clinical events or sensor noise, and to help identify possible sensor degradation. Colour coded CGM traces convey the information quickly and efficiently, while remaining computationally light enough to be used retrospectively or in real-time.     

Raptor-rictor axis in TGFbeta-induced protein synthesis

Transforming growth factor-beta (TGFbeta) stimulates pathological renal cell hypertrophy for which increased protein synthesis is critical. The mechanism of TGFbeta-induced protein synthesis is not known, but PI 3 kinase-dependent Akt kinase activity is necessary. We investigated the contribution of downstream effectors of Akt in TGFbeta-stimulated protein synthesis. TGFbeta increased inactivating phosphorylation of Akt substrate tuberin in a PI 3 kinase/Akt dependent manner, resulting in activation of mTOR kinase. mTOR activity increased phosphorylation of S6 kinase and the translation repressor 4EBP-1, which were sensitive to inhibition of both PI 3 kinase and Akt. mTOR inhibitor rapamycin and a dominant negative mutant of mTOR suppressed TGFbeta-induced phosphorylation of S6 kinase and 4EBP-1. PI 3 kinase/Akt and mTOR regulated dissociation of 4EBP-1 from eIF4E to make the latter available for binding to eIF4G. mTOR and 4EBP-1 modulated TGFbeta-induced protein synthesis. mTOR is present in two multi protein complexes, mTORC1 and mTORC2. Raptor and rictor are part of mTORC1 and mTORC2, respectively. shRNA-mediated downregulation of raptor inhibited TGFbeta-stimulated mTOR kinase activity, resulting in inhibition of phosphorylation of S6 kinase and 4EBP-1. Raptor shRNA also prevented protein synthesis in response to TGFbeta. Downregulation of rictor inhibited serine 473 phosphorylation of Akt without any effect on phosphorylation of its substrate, tuberin. Furthermore, rictor shRNA increased phosphorylation of S6 kinase and 4EBP-1 in TGFbeta-independent manner, resulting in increased protein synthesis. Thus mTORC1 function is essential for TGFbeta-induced protein synthesis. Our data also provide novel evidence that rictor negatively regulates TORC1 activity to control basal protein synthesis, thus conferring tight control on cellular hypertrophy.     

Endophytic colonization and in planta nitrogen fixation by a diazotrophic Serratia sp. in rice

Nitrogen fixing endophytic Serratia sp. was isolated from rice and characterized. Re-colonization ability of Serratia sp. in the rice seedlings as endophyte was studied under laboratory condition. For detecting the re-colonization potential in the rice seedlings, Serratia sp. was marked with reporter genes (egfp and Kmr) using transposon mutagenesis. The conjugants were screened for re-colonization ability and presence of nif genes using PCR. Further, the influence of flavonoids and growth hormones on the endophytic colonization and in planta nitrogen fixation of Serratia was also investigated. The flavonoids, quercetin (3 microg/ml) and diadzein (2 microg/ml) significantly increased the re-colonization ability of the endophytic Serratia, whereas the growth hormones like IAA and NAA (5 microg/ml) reduced the endophytic colonization ability of Serratia sp. Similarly, the in planta nitrogen fixation by Serratia sp. in rice was significantly increased due to flavonoids. The inoculation of endophytic diazotrophs increased the plant biomass and biochemical constituents.     

Effect of Extremely Low Power Time-Varying Electromagnetic Field on Germination and Other Characteristics in Foxtail Millet (Setaria italica) Seeds

The ability of extremely low, time-varying electromagnetic field (EMF) to improve germination efficacy was studied in Foxtail millet (Setaria italica) seeds using response surface methodology. An optimal factorial central composite design was chosen to optimize the EMF with three critical factors, viz. frequency, intensity, and duration. The adequacy of the model and fitness was evaluated by analysis of variance and regression coefficients. This model suggested that the factors, frequency, and intensity had a significant impact on germination. Optimal conditions for germination were observed to be 10 Hz frequency, 30,007 nT intensity, and 30-min duration with an observed germination percentage of 93.0, and a predicted germination percentage of 92.92. Magneto-priming was found to increase the germination efficacy (15.66%), shoot length (27.78%), total seedling length (20.30%), seedling dry mass (26.49%), and water uptake (34.48% at 80 min) showing significant output when compared with the control and positive controls. Remarkable improvements were observed in germination parameters such as vigor index-1 (39.14%), vigor index-2 (46.28%), speed of germination (27.52%), and emergence index (12.50%). Magneto-priming was found to reduce the levels of germination-specific enzymes, viz. α-amylase, protease, and dehydrogenase, while it enhanced the levels of antioxidant enzymes, viz. catalase (114.63%) and superoxide dismutase (19.62%), triggering fast germination and early vigor of seedlings. This study clearly showed that EMF priming significantly improved the germination effect and other characteristics of Foxtail millet seeds. Bioelectromagnetics. © 2020 Bioelectromagnetics Society     

Suppression of root rot of mung bean (Vigna radiata L.) by Streptomyces sp. is associated with induction of peroxidase and polyphenol oxidase

Five strains of Streptomyces sp. were evaluated in vitro for their ability of inhibiting the mycelial growth of Macrophomina phaseolina, the causal agent of root rot of mung bean (Vigna radiata L.). Among the Streptomyces sp. strains tested, PDK showed the maximum in vitro inhibition of mycelial growth of M. phaseolina and recorded an inhibition zone of 21?mm. The strains CBE, MDU, SA and ANR recorded inhibition zones of 18, 16, 13 and 11?mm, respectively. These Streptomyces sp. strains were tested for their growth-promoting efficiency on mung bean seedlings. Among them, CBE and PDK recorded the maximum increase in shoot length, root length and seedling vigour compared with control, followed by MDU. Three Streptomyces sp. strains (CBE, MDU and PDK) that showed higher levels of inhibition of growth of M. phaseolina in dual culture assay and plant growth-promoting activity were tested for their biocontrol activity against root rot under greenhouse and field conditions. Seed treatment or soil application with powder formulation of Streptomyces sp. strains CBE, MDU and PDK was effective in controlling root rot disease; but, combined application through seed and soil increased the efficacy in both the greenhouse and field trials. Among the treatments, seed treatment plus soil application with powder formulation of Streptomyces sp. strain CBE proved to be most effective, which reduced the root rot incidence from 26.8% (with non-bacterised seeds) to 4.0% in Trial I and from 32.0 to 4.9% in Trial II. The above treatment recorded the highest yield in both the field trials, and the yield increase was 78 and 74% over control in Trial I and Trial II, respectively. Isozyme analysis of the Streptomyces sp.-treated plants indicates that seed treatment plus soil application strongly induce the activities of peroxidase (PO-1 and PO-2) and polyphenol oxidase (PPO-2 and PPO-3) in mung bean. Among the three strains tested, Streptomyces sp. strain MDU- treated plants showed higher levels of activities of PO and PPO. Based on the above findings, it can be concluded that both the direct inhibition of pathogen and induced resistance might be involved in the control of root rot of mung bean by Streptomyces sp.