Augmenting salt tolerance in rice by regulating uptake and tissue specific accumulation of Na+ - through Ca2+ -induced alteration of biochemical events.

The protective effect of Ca2+ against NaCl toxicity was investigated in two rice varieties with contrasting for salt tolerance to understand the mechanistic details of the antagonism to address adverse effects of salinity on agriculture. The study primarily examined the influence of Ca2+ on expression/activity of the effectors and regulators involved in Na+ translocation. Calcium reduced uptake of Na+ concomitant with higher tissue K+ /Na+ in seedlings, comparatively more in salt-tolerant Nona Bokra than in salt-sensitive IR-64, together with a significant increase in root PM H+ ATPase in the former, but not in the latter. Increased antagonism in Nona Bokra could be the result of Ca2+ signalling-mediated phosphorylation of PM H+ ATPase in roots caused by a significant Ca2+ -dependent increase in expression of OsCIPK24, which did not occur in IR-64. Furthermore, significant Ca2+ -mediated NaCl-induced increase in transcription of 14-3-3 protein in Nona Bokra, but not in IR-64, might also lead to a greater protective effect of Ca2+ in the former, as 14-3-3 protein is essential for activating PM H+ ATPase. Thus, efficient functioning of PM H+ ATPase could be key in determining resistance of plants to salinity, implying that identification of the Ca2+ -dependent kinase phosphorylating the PM H+ ATPase threonine residue and manipulation of its expression, together with expression of 14-3-3 proteins could be an important strategy to improve salt tolerance of crops and their cultivation in salt-affected lands.

1-MCP treatment enhanced expression of genes controlling endosperm cell division and starch biosynthesis for improvement of grain filling in a dense-panicle rice cultivar.

High ethylene production in dense-panicle rice cultivars impacts grain filling. 1-MCP (ethylene action inhibitor) treatment increased assimilates partitioning, cell number and size and expression of starch synthesizing enzyme genes of developing caryopses mostly in the basal spikelets of panicle at early post-anthesis stage. The gain in cell number was less compared to the increase of size. High ethylene production in spikelets matched with greater expression of ethylene receptor and signal transducer genes. Genes encoding cell cycle regulators CDK, CYC and CKI expressed poorly on 9 DAA. 1-MCP treatment enhanced their expression; the increase of expression was higher for CDKs and lower for CKIs in basal compared to apical spikelets. Greater expression of CDKB2:1 might have lifted cytokinesis of nascent peripheral cells of endosperm, while promotion of CDKAs, CYCD2:2 and inhibition of CYCB2:2 expression contributed to endoreduplication of central cells increasing cell size and DNA ploidy level. It is concluded that the process of endoreduplication, which begins at mid-grain filling stage, is crucially linked with the final caryopsis size of rice grain. The enhanced endosperm growth brought about by repressed ethylene action during the first few days after anthesis seems to be associated with the overall increased cell cycle activity and sink strength.

Computational prediction and experimental validation of a novel miRNA in Suaeda maritima, a halophyte.

The lack of available transcriptome data for plants of no economic or agronomic importance limits the identification of miRNAs in many species. Considering the possible similarity of the transcriptome between related species, the present study used expressed sequence tags (ESTs) of Suaeda salsa and Suaeda glauca to identify conserved miRNAs, which were validated in a halophyte, Suaeda maritima, with the aim of identifying salt-responsive miRNAs from naturally salt-tolerant plants, information on which is limited. In this study, computational analysis predicted three miRNA sequences by mapping non-redundant miRNA sequences from miRBase 16.0 on 1534 ESTs of S. salsa and S. glauca. The expression of one could be validated in S. maritima, and was named sma-miR1867. This miRNA was downregulated in response to NaCl treatment. It was predicted to target ferredoxin-thioredoxin reductase (FTR), cell division control protein 6 (CDC6), and ubiquitin-protein ligase (UPL) in S. salsa and/or S. glauca. However, only UPL could be amplified in S. maritima, and RT-qPCR showed that it was upregulated in response to NaCl treatment. These results indicate that, in halophytes, FTR and CDC6 may promote carbon metabolism and cell division, respectively, in the presence of salt, while UPL may regulate the abundance of proteins that are important for salt tolerance in halophytes. Thus, sma-miR1867 could be an essential component of salt resistance in halophytes.

Real time PCR expression analysis of gene encoding p5cs enzyme and proline metabolism under NaCI salinity in rice.

Regulation of proline accumulation in seedlings of rice (Oryza sativa L. cv. Lunishree) was investigated. The increasing concentration of NaCl from 85 mM to 425 mM NaCl progressively increased proline content in rice. The maximum increase in proline content was recorded at 425 mM NaCl concentration as compared to control and other concentrations of NaCl. The highest significant activity of proline synthesizing enzymes, delta1-Pyrrolline-5-carboxylate synthetase, delta1-Pyrrolline-5-carboxylate reductase and Ornithine-δ- aminotransferase with lowest activity of proline hydrolysis enzymes;Proline dehydrogenase was also recorded at 425 mM NaCl salinity over control and other concentrations of NaCI with insignificant increase in the activity of delta1-Pyrrolline-5-carboxylate synthetase and Ornithine-δ-aminotransferase at 85 mM NaCI over control. It was found that the transcript of gene encoded with p5cs is up regulated about 1.35 folds under salinity stress. This gene synthesis an osmo protectant to help the plant resist the change in osmotic imbalances. Externally addition of MnCl2 at 300 mg/220 ml 1/2 strength Hoagland solution, having 1% NaCI, was also seen to increase 893.9% proline content of this variety as compared to control.

Compact panicle architecture is detrimental for growth as well as sucrose synthase activity of developing rice kernels.

The increase of spikelet number in the panicles of modern super rice has made the architecture compact, as the extra spikelets are accommodated mostly on secondary branches than on primary branches. However, the grain yield did not improve because of poor grain filling, which was more visible in the basal spikelets than apical spikelets. The objective of this study was to examine the effect of the compactness and positional difference of spikelets in the panicle on grain filling by comparing the activity and genetic expression of starch synthesising enzymes in the developing kernels of lax-(Upahar and CR3856-45-11-2-7-2-5 (CR-45)) and compact-(Mahalaxmi and CR3856-29-14-2-1-1-1 (CR-29)) panicle cultivars. Upahar and Mahalaxmi are genetically related, whereas CR-45 and CR-29 are recombinant inbred lines. The grain carbohydrate concentration and activity of sucrose synthase (SUS) enzyme were estimated during the active period of grain filling. Further, expression of isoforms of SUS, ADP glucose pyrophosphorylase (APL and APS for large and small units respectively) and starch synthase (SS and GBSS for soluble and granule bound starch synthases respectively) were also assayed through PCR studies. The genotype approach used revealed grain SUS activity and starch concentration high and sugar concentration low in the lax- compared with compact-panicle cultivars and in the apical spikelets compared with basal ones. The margin of variation between apical and basal spikelets was higher in the compact- than the lax-panicle cultivars. Genetic expression of most of the isoforms of the enzymes was higher in the lax- than the compact-panicle cultivars as seen in RT-PCR studies. A quantitative appraisal of transcript levels of isoforms in the qRT-PCR identified greater expression of SUS3 in the basal spikelets of Upahar than that in Mahalaxmi and in CR-45 over CR-29, most prominently during the active period of grain filling. We conclude that proximal location as well as increased density of spikelets on panicles affected SUS3 expression in the basal spikelets. The metabolic dominance of a spikelet in rice panicle is dependent on the expression of the genes for different isoforms of starch synthesising enzymes, but the expression of SUS3 could be more specific than the others. SUS3 expression is most active during grain filling of the lax-panicle cultivars, but its dominance is reduced significantly in the kernels of the compact-panicle cultivars.

Analysis of proline metabolic enzymes in Oryza sativa under NaCl stress.

The regulation of proline accumulation in seedlings of rice (Oryza sativa L. cv. Badami) was investigated. The increasing concentration of NaCl from 0.5 to 2.5% progressively increased the proline content in rice. Proline accumulation caused by NaCl was related to protein proteolysis, an increase in OAT, P5CS, PSCR activity, a decrease in PDH activity. The maximum increase in proline content was recorded at 2.5% NaCl concentration as compared to control and other concentrations of NaCl. The highest significant activity of proline synthesizing enzymes, Delta1-Pyrrolline-5-carboxylate synthetase, Delta1-Pyrrolline-5-carboxylate reductase and Ornithine-delta-aminotransferase with a lowest activity of proline hydrolysis enzymes; Proline dehydrogenase were also recorded at 2.5% salinity over control and other concentrations of NaCl with a in-significant increase in the activity of Delta1-Pyrrolline-5-carboxylate synthetase and Ornithine-delta-aminotransferase at 0.5 % concentration of NaCl over control. Externally the addition of 300mg MnCl2, 220 ml(-1) 1/2 strength Hoagland solution, having 1% NaCl, was seen to increase a 893.9% in proline content of this variety as compared to control.

A new glycosidic flavonoid from Jwarhar mahakashay (antipyretic) Ayurvedic preparation.

The aqueous extract of Jwarhar mahakashay Ayurvedic preparation (from the roots of Hemidesmus indicus R. Br., Rubia cordifolia L., Cissampelos pareira L.; fruits of Terminalia chebula Retz., Emblica officinalis Gaertn., Terminalia bellirica Roxb., Vitis vinifera L., Grewia asiatica L., Salvadora persica L. and granules of Saccharum officinarum L.) has been used as a traditional antipyretic. Experimental studies confirmed its antipyretic-analgesic effect with very low ulcerogenicity and toxicity. Flavonoids, glycosides and tannins were later found to be present in the extract. Detailed chemical investigations were undertaken after hydrolysis of extract using spectroscopic and chromatography methods to determine its active chemical constituent. UV-Visible spectroscopy showed absorbance maxima at 220 and 276 nm, while fourier transform infra-red investigations indicated an end carboxylic O-H structure at 2940 cm(-1) suggesting the presence of glycoside-linked flavonoids. Thin layer chromatography and high performance liquid chromatography also confirmed the possibility of at least one major and two minor compounds in this abstract. Detailed examination using gas chromatography-mass spectrometry led to the identification of the principal component as 2-(1-oxopropyl)-benzoic acid, which is quite similar to the active compound found in the standard drug Aspirin (2-acetyl-oxybenzoic acid).

Seasonal appearance of Chlorophyceae phytoplankton bloom by river discharge off Paradeep at Orissa Coast in the Bay of Bengal.

Characteristics of the monsoonal bloom of phytoplankton at Orissa Coast in the Bay of Bengal were studied through bimonthly observation from April 2001 to December 2002. Three photosynthetic pigments chlorophyll-a (Chl a), chlorophyll-b (Chl b) and carotenoid (Car) were analyzed by absorption spectroscopic method. The seasonal variation of Chl a included phytoplankton bloom in the coastal area during monsoon period. The water column integrated Chl a reached to 68 mg m(-2) at the station-1(St1), and amounted to 20 mg m(-2) at 30 km off the river mouth during August 2001. In contrast the same amount was found at 15 km off the Mahanadi river mouth during August 2002. Salinity during this period varied from 5 psu at the St1 to 27 psu at the edge of the bloom area. The total amount of river discharge in the monsoon period calculated from daily river discharge data reported by Water Resources Department in India was 84 x 10(9) m(3) during 2001 and 20 x 10(9) m(3) during 2002. Both nitrate and phosphate concentrations showed negative quadratic relationship with salinity throughout the observation period. Extrapolated nitrate and phosphate concentration discharge from the Mahanadi river were 10.8 and 4 microg-at l(-1), respectively. Microscopic identification revealed dominance of fluvial Chlorophyceae and diatoms during the monsoon period showing influence of the freshwater discharge.

Salt tolerance in aquatic macrophytes: possible involvement of the antioxidative enzymes.

Possible involvement of the antioxidative enzymes in salt tolerance was investigated in three submerged aquatic macrophytes, viz. Hydrilla verticillata, Najas indica and Najas gramenia, which differed in their tolerance to salinity. NaCl in light resulted in significant decrease in the chlorophyll a/b ratio in H. verticillata, which was significantly neutralized by the presence of DABCO (diazabicyclo[2.2.2]octane), sodium benzoate or mannitol along with the NaCl in the exposure medium indicating induction of oxidative stress upon salt treatment. All the three test species showed enhanced activity of superoxide dismutase (SOD; EC 1.15.1.1) and peroxidases (EC 1.11.1.7) upon exposure to either NaCl or SWS (seawater salinity); the increases in their activity in response to NaCl were positively correlated with the salt tolerance of the plants. Only N. gramenia showed considerable increase in the activity of catalase (EC 1.11.1.6), while only H. verticillata exhibited significant increase in the activity of ascorbate peroxidase (EC 1.11.1.11) in response to the salt treatment. A higher constitutive level of the antioxidative enzymes in the salt-tolerant N. gramenia than in the salt-sensitive H. verticillata coupled with significant increase in the activity of these enzymes, particularly of catalase and SOD, in the former upon salinity treatment suggested their active involvement in salt tolerance. The oxidative stress induced upon the salt treatment also indicated a possible involvement of the antioxidative enzymes in the process. The ion interaction study revealed significant modifying effect of Ca(2+), and also of Mg(2+) and K(+), on the changes induced by Na(+) in the activity of catalase and SOD suggesting that relative build-up of these ions inside cells could be of much importance in regulation of synthesis of antioxidative enzymes.

Chemical and pharmacological evaluation of different ayurvedic preparations of iron.

Ayurvedic preparations of metallic iron commonly categorised as different 'putas' of 'Louha Bhasma' was chemically analysed and pharmacologically investigated in iron deficiency anemia. Atomic absorption spectral (AAS) study of different putas of Louha Bhasma revealed the presence of various proportions of important metals along with varied concentration of iron in it. The effect of a representative puta viz. 50 puta of Louha Bhasma in the management of agar gel diet and phlebotomy induced iron deficiency anemia in animal model was found to be statistically highly significant (P < 0.001) in comparison to the control and standard drug Fefol treated groups.

Value of Mantoux Test in diagnosis of tuberculosis in chronic illness.

Total 240 cases of 2-12 years age group who had contact of tuberculosis indirectly or directly and had chronic illness were screened for tuberculosis. Out of 240 cases, 180 cases were diagnosed as tuberculosis. Special emphasis was given on Mantoux Test (M.T.), though the battery of tests were done to diagnose tuberculosis. Out of 180 cases of Tuberculosis, M.T. was Positive in 97 cases (40.41%) and repeat M.T. was done after 3 weeks and was converted to Positive in 20 cases (8.33%). Thus only M.T. help in diagnosis of Tuberculosis in almost 50% of the cases.

PIP: Childhood tuberculosis (TB) in India affects more than 40% of individuals under the age of 15 years. Diagnosis of childhood TB is difficult due to the various clinical presentations and the cost of diagnosis kits. This article reports on the value of the Mantoux test in the diagnosis of TB in children. 240 children between the ages of 2 and 12 years were studied; 180 were diagnosed as having TB. These children had a history of chronic illness and direct or indirect contact with TB. All patients were tested for TB using tests such as Mantoux, Roentgenogram, E.S.R., and complete hemogram. Nutritional status was determined using criteria established by the Indian Academy of Pediatrics. 97 (40.41%) of the 180 children with TB had a positive diagnosis with the Mantoux test. The 4-8 year age group was found to have the greatest number of cases in this study. 60/180 (33%) positive TB cases were reported for this group. It was noted that the higher the socio-economic status, the lower the incidence of TB.

Epidemiology of Japanese encephalitis.

Seven hundred and sixty-two cases of Japanese Encephalitis (JE) were studied during the last 5 years (1985-1989) in relation to age, sex, religion, nutritional status, living habits, exposure to domestic animals and mosquitos, clinical profile, seasonal variation and mortality pattern. The maximum occurrence was in 1987-1988 and it showed a preponderance in males (51-82%). The disease is progressively decreasing in Muslims (3-7%) and gradually increasing in tribes (25-60%). Children in the age-group of 6-7 years (19-25%) were maximally affected and the disease was rare in infancy. The common features were coma, convulsions, neck rigidity and fever (88-97%). Gastrointestinal manifestations were rare (3.6%) but were associated with the highest mortality. About 80-95% had exposure to domestic animals directly or indirectly and 95% of the patients were not using mosquito nets. The CSF protein and sugar content were normal, with or without slight leucocytosis while the lymphocyte count was variable. The CSF and blood picture had no significant relation with clinical presentation and prognosis.

Effect of methyl mercuric chloride treatment on haematological characteristics and erythrocyte morphology of Swiss mice.

Effects of methyl mercuric chloride (MMC) on the blood parameters of Swiss mice (Mus musculus) were studied. The mice received an initial dose of MMC (24 mg kg(-1) body wt) as intraperitoneal injection followed by a second similar dose on the 14th day of the first dose administration. Significant (p < or = 0.001) decreases in haemoglobin content, red blood cell (RBC) count and haematocrit value were observed in the MMC injected mice when compared to the control mice. The effect of the second dose was severe, after which no significant recovery in the values of these parameters was observed. The result also showed a high degree of mercury accumulation in the blood of the MMC exposed mice. Interesting features were marked in the erythrocyte morphology of the exposed mice. An initial shrinkage followed by swelling of the cells was observed after each injection. The outline of the exposed cells was irregular with beak like or small finger like projections. Rupturing and disintegration of the erythrocyte membrane, leading to erythrolysis, were also noticed.

Effect of herbal compound on maturity onset diabetes.

The clinical study conducted to evaluate the ant diabetes effect of a herbal compound drug consisting two herbs is presented here.