Investigations on the antioxidative defence responses to NaCl stress in a mangrove, Bruguiera parviflora: Differential regulations of isoforms of some antioxidative enzymes

Two-month-old healthy seedlings of a true mangrove, Bruguiera parviflora, raised from propagules in normal nursery conditions were subjected to varying concentrations of NaCl for 45 d under hydroponic culture conditions to investigate the defence potentials of antioxidative enzymes against NaCl stress imposed oxidative stress. Changes in the activities of the antioxidative enzymes catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (POX), glutathione reductase (GR) and superoxide dismutase (SOD) were assayed in leaves to monitor the temporal regulation. Among the oxidative stress triggered chemicals, the level of H₂O₂ was significantly increased while total ascorbate and total glutathione content decreased. The ratio of reduced to oxidized glutathiones, however, increased due to decreased levels of oxidized glutathione in the leaf tissue. Among the five antioxidative enzymes monitored, the APX, POX, GR and SOD specific activities were significantly enhanced at high concentration (400 mM NaCl), while the catalase activities declined, suggesting both up and downregulations of antioxidative enzymes occurred due to NaCl imposed osmotic and ionic stress. Analysis of the stress induced alterations in the isoforms of CAT, APX, POX, GR and SOD revealed differential regulations of the isoforms of these enzymes. In B. parviflora one isoform of each of Mn-SOD and Cu/Zn-SOD while three isoforms of Fe-SOD were observed by activity staining gel. Of these, only Mn-SOD and Fe-SOD2 content was preferentially elevated by NaCl treatment, whereas isoforms of Cu/Zn-SOD, Fe-SOD1 and Fe-SOD3 remained unchanged. Similarly, out of the six isoforms of POX, the POX-1,-2,-3 and -6 were enhanced due to salt stress but the levels of POX-4 and -5 remained same as in control plants suggesting preferential upregulation of selective POX isoforms. Activity staining gel revealed only one prominent band of APX and this band increased with increased salt concentration. Similarly, two isoforms of GR (GR1 and GR2) were visualized on activity staining gel and both these isoforms increased upon salt stress. In this mangrove four CAT-isoforms were identified, among which the prominent CAT-2 isoform level was maximally reduced again suggesting differential downregulation of CAT isoforms by NaCl stress. The results presented in this communication are the first report on the resolutions of isoforms APX, POX and GR out of five antioxidative enzymes studied in the leaf tissue of a true mangrove. The differential changes in the levels of the isoforms due to NaCl stress may be useful as markers for recognizing salt tolerance in mangroves. Further, detailed analysis of the isoforms of these antioxidative enzymes is required for using the various isoforms as salt stress markers. Our results indicate that the overproduction of H₂O₂ by NaCl treatment functions as a signal of salt stress and causes upregulation of APX, POX, GR and deactivations of CAT in B. parviflora. The concentrations of malondialdehyde, a product of lipid peroxidation and lipoxygenase activity remained unchanged in leaves treated with different concentrations of NaCl, which again suggests that the elevated levels of the antioxidant enzymes protect the plants against the activated oxygen species thus avoiding lipid peroxidation during salt stress.     

A Fragrant Solution to Soil Remediation

The ability of scented geraniums (Pelargonium sp. ‘Frensham’) plants to tolerate, uptake, and accumulate lead was assessed compared with two well-established metal accumulators, Indian mustard (Brassica juncea) and sunflower (Helianthus annuus), under greenhouse conditions. The efficiency of the photosynthetic apparatus and the number and size of active photosynthetic reaction centers (expressed as chlorophyll a fluorescence ratios of variable fluorescence to maximal fluorescence [Fv/Fm] and variable fluorescence to unquenchable portion of fluorescence [Fv/Fo], respectively) were affected to varying degrees at all metal concentrations in all the plants tested. Lead exposure did not significantly affect the efficiency of photosystem II activity or the number and size of the photosynthetic reaction centers in scented geraniums, but the ratios decreased significantly in Indian mustard and sunflower plants following lead exposure. In addition to tolerating high levels of lead, the scented geraniums accumulated in excess of 3000 mg of lead per kg DW of shoot and above 60,000 mg of lead per kg DW of root tissue. Additionally, scented geraniums exposed to a mixture of metals (lead + cadmium + nickel) had the ability to uptake in excess of 4.72% Pb + 0.44% Cd + 0.52% Ni per kg of root DW, as well as 0.17% Pb + 0.07% Cd + 0.14% Ni per kg of shoot DW within 14 d, indicated the potential for existence of more than one functional tolerance and metal uptake mechanism(s). The cellular localization of lead was assessed using transmission electron microscopy coupled with an X-ray microanalyzer. Lead accumulation was observed in the apoplasm and in the cytoplasm, vacuoles, and as distinct globules (potentially as lead-lignin or lead-phosphate complexes) on the cell membrane and cell wall. We conclude that the ability of scented geraniums to tolerate high lead accumulation in its biomass is due, in part, to limiting damage to photosynthetic apparatus and metal detoxification by formation of metal complexes.     

Differentiation of the American and Oriental Maydeae Accessions Using the Maize Simple Sequence Repeat (SSR) Markers

The present study evaluated the utility of maize microsatellite markers in genotype differentiation and analysis of genetic diversity among the Oriental Maydeae accessions (Coix aquatica and Chionachne koenigii) vis-à-vis the American Maydeae (maize, teosintes and maize-teosinte derivatives). Analysis of 27 microsatellite loci distributed throughout the maize genome led to the detection of 89 SSR alleles at average of 3.30 per locus. Polymorphism information content (PIC) values and observed genetic distances showed the existence of large variability among the various accessions. The maize SSR markers, particularly nc013, phi059, bnlg1523 and umc1014, were highly effective in differentiating the Oriental Maydeae accessions from the American Maydeae. Cluster analysis and Principal Component Analysis based on the SSR data clearly indicated significant genetic divergence between the Oriental Maydeae and the American Maydeae accessions, as well as between Coix aquatica and Chionachne koenigii. Molecular analyses would be valuable in gaining a better understanding of genera belonging to the Oriental Maydeae, the lesser known wild relatives of maize.     

Estimation of distribution function in bivariate competing risk models

We consider lifetime data involving pairs of study individuals with more than one possible cause of failure for each individual. Non-parametric estimation of cause-specific distribution functions is considered under independent censoring. Properties of the estimators are discussed and an illustration of their application is given.     

Deglycosylated milin unfolds via inactive monomeric intermediates

The effect of deglycosylation on the physiological and functional organization of milin was studied under different denaturizing conditions. Trifluoromethanesulfonic acid mediated deglycosylation resulted in insoluble milin, which was found to be soluble only in 1.5 M GuHCl with native-like folded structure. Kinetic stability, proteolytic activity, and dimeric association were lost in deglycosylated milin. Urea-induced unfolding revealed two inactive, highly stable equilibrium intermediates at pH 7.0 and pH 2.0. These intermediates were stable between 5.5–6.5 and 5.0–6.0 M total chaotropes (urea + 1.5 M GuHCl) at pH 7.0 and pH 2.0, respectively. GuHCl-induced unfolding was cooperative and noncoincidental with a broad transition range (2.0–5.0 M) at pH 7.0 and pH 2.0. Equilibrium unfolding of deglycosylated milin by urea and GuHCl substantiates the involvement of various inactive monomeric intermediates. This study provides a way to understand the role of glycosylation in the unfolding mechanism, stability, and functional activity of the serine protease milin.     

Simian immunodeficiency virus-induced mucosal interleukin-17 deficiency promotes Salmonella dissemination from the gut

Salmonella typhimurium causes a localized enteric infection in immunocompetent individuals, whereas HIV-infected individuals develop a life-threatening bacteremia. Here we show that simian immunodeficiency virus (SIV) infection results in depletion of T helper type 17 (TH17) cells in the ileal mucosa of rhesus macaques, thereby impairing mucosal barrier functions to S. typhimurium dissemination. In SIV-negative macaques, the gene expression profile induced by S. typhimurium in ligated ileal loops was dominated by TH17 responses, including the expression of interleukin-17 (IL-17) and IL-22. TH17 cells were markedly depleted in SIV-infected rhesus macaques, resulting in blunted TH17 responses to S. typhimurium infection and increased bacterial dissemination. IL-17 receptor-deficient mice showed increased systemic dissemination of S. typhimurium from the gut, suggesting that IL-17 deficiency causes defects in mucosal barrier function. We conclude that SIV infection impairs the IL-17 axis, an arm of the mucosal immune response preventing systemic microbial dissemination from the gastrointestinal tract.     

Development of new cytoplasmic-genetic male-sterile lines in pigeonpea from crosses betweenCajanus cajan (L. Millsp. andC. scarabaeoides (L.) Thouars

Exploitation of hybrid vigour is quite possible in cross-pollinated crops. However, pigeonpea is a grain legume crop with a moderate level of cross-pollination (20-70%), which is mainly aided by insect pollinators. As a first step, hybrids based on genetic male sterility (GMS) were developed in pigeonpea, but the hybrid seed production technique is not farmer-friendly, because in the hybrid seed production plot 50% of the population, which are male-fertile in the female rows, have to be eliminated in time before contamination. This requires skilled labour and is a time-consuming process, which increases the cost of hybrid seed production. Therefore, the objective of this study was to develop cytoplasmic-genetic male-sterile (CGMS) lines in pigeonpea through wide hybridization, which would be very suitable for hybrid seed production. Two CGMS lines, viz. CORG 990052 A and CORG 990047, were developed by interspecific hybridization of Cajanus cajan and C. scarabaeoides. Restorers were identified and three CGMS-based pigeonpea hybrids were developed. The hybrid COPH 3 is found to be promising in Tamil Nadu State, India.     

Purification and characterization of a novel protease from the latex of Pedilanthus tithymaloids

A novel protease was purified to homogeneity from the latex of Pedilanthus tithymaloids by a simple purification procedure involving ammonium sulfate precipitation and cation-exchange chromatography. The molecular weight of the protease was estimated to be approximately 63.1 kDa and the extinction coefficient (epsilon(1%)(280nm)) was 28.4. The enzyme hydrolyzes denatured natural substrates like casein, azoalbumin and azocasein with a high specific activity but little activity towards synthetic substrates. The pH and temperature optima were pH 8.0-9.5 and 65-70 degrees C, respectively. The proteolytic activity of the enzyme was inhibited by different protease-specific inhibitors (e.g., thiol, serine, metallo, etc.) up to a certain extent but not completely by any class of inhibitors. The enzyme was relatively stable towards pH change, temperature, denaturants and organic solvents. The enzyme consists of five disulfide bridges compared to three observed in most plant cysteine proteases. Overall, the striking features of this protease are its high molecular weight, high cysteine content and only partial inhibition of activity by different classes of protease inhibitors contrary to known proteases from other plant sources. The enzyme is named as pedilanthin as per the protease nomenclature.     

Endosperm Protein Quality and Kernel Modification in the Quality Protein Maize Inbred Lines

Twenty-two selected quality protein maize (QPM) lines, including 13 lines developed in India (DMRQPM series) and nine lines released by CIMMYT, Mexico (CML series), were evaluated for their endosperm protein content and quality, besides kernel modification in terms of vitreousness. Endosperm protein contents in 13QPMlines were on par or better than that of the normal maize ‘checks’ (Trishulata and Parkash). The QPM endosperm proteins showed significantly higher % tryptophan as well as EF-1α (a multifunctional protein with a positive and highly significant correlation with lysine content in the endosperm) contents, in comparison with the normal maize genotypes. Evaluation of kernel modification revealed considerable scope for accumulation of endosperm modifiers in some of the QPM lines. Positive and highly significant correlation was revealed between tryptophan and EF-1α contents in the endosperm proteins, whereas the correlations between the quality parameters with kernel modification in the QPM genotypes were found to be non-significant. The study led to the identification of some promising QPM lines, such as DMRQPM-37, DMRQPM-44, CML176, CML142 and CML149, which could be effectively deployed in the QPM breeding programmes.     

Ozone as a selective disinfectant for nonaseptic fungal cultivation on corn-processing wastewater

Treatment of wet corn-milling wastewater with filamentous fungi was investigated as a means of obtaining fungal biomass as an additional byproduct. Competitive bacterial growth is a common problem during this nonaseptic treatment process. Selective disinfection with ozone was evaluated for eliminating bacterial populations during fungal cultivation. Three laboratory-scale continuous flow aerated reactors were operated under nonaseptic conditions at 38 degrees C, hydraulic retention time of 8h and pH of 4. The bacterial population was reduced by one log with respect to the control when ozone was dosed at a concentration above 47+/-2mg/L. An ozone dosage of about 57mg/L was found to be most effective in improving both fungal biomass production and soluble chemical oxygen demand (SCOD) removal (up to 90%). Fungal biomass concentration increased from c. 1.45g/L (control) to c. 1.75g/L at a 57-mg/L ozone dosage. Higher and lower dosages of ozone resulted in poorer fungal growth and lower SCOD removal.