Interactive effects of carbon dioxide, temperature, and ultraviolet-B radiation on soybean (Glycine max L.) flower and pollen morphology, pollen production, germination, and tube lengths

Plant reproduction is highly vulnerable to global climate change components such as carbon dioxide concentration ([CO(2)]), temperature (T), and ultraviolet-B (UV-B) radiation. The objectives of this study were to determine the effects of season-long exposure to treatments of [CO(2)] at 360 (control) and 720 micromol mol(-1) (+CO(2)), temperature at 30/22 degrees C (control) and 38/30 degrees C (+T) and UV-B radiation 0 (control) and 10 kJ m(-2) d(-1) (+UV-B) on flower and pollen morphology, pollen production, germination, and tube lengths of six soybean genotypes (D 88-5320, D 90-9216, Stalwart III, PI 471938, DG 5630RR, and DP 4933RR) in sunlit, controlled environment chambers. The control treatment had 360 micromol mol(-1) [CO(2)] at 30/22 degrees C and 0 kJ UV-B. Plants grown either at +UV-B or +T, alone or in combination, produced smaller flowers with shorter standard petal and staminal column lengths. Flowers so produced had less pollen with poor pollen germination and shorter tube lengths. Pollen produced by the flowers of these plants appeared shrivelled without apertures and with disturbed exine ornamentation even at +CO(2) conditions. The damaging effects of +T and +UV-B were not ameliorated by +CO(2) conditions. Based on the total stress response index (TSRI), pooled individual component responses over all the treatments, the genotypes were classified as tolerant (DG 5630RR, D 88-5320: TSRI >-790), intermediate (D 90-9216, PI 471938: TSRI <-790 to >-1026), and sensitive (Stalwart III, DP 4933RR: TSRI <-1026). The differences in sensitivity identified among genotypes imply the options for selecting genotypes with tolerance to environmental stresses projected to occur in the future climates.     

An in vitro demonstration of peroxisome proliferation and increase in peroxisomal β-oxidation system mRNAs in cultured rat hepatocytes treated with ciprofibrate

Using the normal adult rat hepatocytes, plated on rat tail collagen-coated dishes and fed a chemically defined medium, we demonstrate here that ciprofibrate at 0.1 mM concentration, increases significantly the mRNA levels of fatty acyl-CoA oxidase, enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase bifunctional protein, and thiolase (the three enzymes of the β-oxidation system), and causes peroxisome proliferation. Increase in mRNA levels of these genes was evident within 1 h and was maximal 24 h after the addition of ciprofibrate. In hepatocytes cultured in the absence of ciprofibrate, the basal levels of these enzymes were low and further declined with time. Concomitant treatment of hepatocytes with cycloheximide did not inhibit or superinduce the mRNA levels, indicating that this induction may represent a primary (direct) effect of this compound on the expression of these genes and does not apparently involve short-lived repressor protein(s).     

Expansion of a unique region in the Marek's disease virus genome occurs concomitantly with attenuation but is not sufficient to cause attenuation

Pathogenic Marek's disease viruses (MDVs) have two head-to-tail copies of a 132-bp repeat. As MDV is serially passaged in cell culture, the virus becomes attenuated and the number of copies of the 132-bp repeat increases from 2 to often more than 20 copies. To determine the role of the repeats in attenuation, we used five overlapping cosmid clones that spanned the MDV genome to reconstitute infectious virus (rMd5). By mutating the appropriate cosmids, we generated clones of infectious MDVs that contained zero copies of the 132-bp repeats, rMd5(Delta132); nine copies of the 132-bp repeats, rMd5(9-132); and nine copies of the 132-bp repeats inserted in the reverse orientation, rMd5(rev9-132). After two passages in cell culture, wild-type Md5, rMd5, and rMd5(Delta132) were stable. However, rMd5(9-132) and rMd5(rev9-132) contained a population of viruses that contained from 3 to over 20 copies of the repeats. A major 1.8-kb mRNA, containing two copies of the 132-bp repeat, was present in wild-type Md5 and rMd5 but was not present in rMd5(Delta132), rMd5(9-132), rMd5(rev9-132), or an attenuated MDV. Instead, the RNAs transcribed from the 132-bp repeat region in rMd5(9-132) and rMd5(rev9-132) closely resembled the pattern of RNAs transcribed in attenuated MDVs. When inoculated into susceptible day-old chicks, all viruses produced various lesions. Thus, expansion of the number of copies of 132-bp repeats, which accompanies attenuation, is not sufficient in itself to attenuate pathogenic MDVs.     

Dynamic large branching hash tree based secure and efficient dynamic auditing protocol for cloud environment

Cluster Computing - Cloud computing model offers various platforms services and provides a scalable, on-demand service at any time-anywhere manner. However, in the outsourcing strategy, users no...     

Ectopic expression of pigeonpea cold and drought regulatory protein (CcCDR) in yeast and tobacco affords multiple abiotic stress tolerance

Pigeonpea (Cajanus cajan L.) cold and drought regulatory protein encoding gene (CcCDR) has been introduced into yeast and tobacco for its functional validation. In yeast, expression of CcCDR imparted marked tolerance against abiotic stresses exerted by PEG and NaCl. Transgenic tobacco lines, expressing CcCDR under the control of CaMV35S and rd29A promoters, when subjected to mannitol, NaCl and cold (4 °C) stress, developed into healthy plants with profuse root system, increased biomass, root length and chlorophyll content in contrast to the weak-stunted wild-type plants. Transgenic plants also showed increased levels of proline, reducing sugars and endogenous abscisic acid (ABA) content. Exogenous ABA treatment resulted in increased hypersensitivity and decreased stomatal aperture size of transgenic plants compared to wild type. Localization studies confirmed that CcCDR could enter the nucleus as revealed by intense fluorescence, indicating its plausible interaction with various nuclear proteins. The overall results amply demonstrate the intrinsic effect of CcCDR in bestowing multiple abiotic stress tolerance at cellular and whole plant levels. Accordingly, the multipotent CcCDR seems promising as a prime candidate gene to fortify crop plants with abiotic stress tolerance.     

Studies on the production and application of cellulase from Trichoderma reesei QM-9414

High yielding mutant strain, Trichoderma reesei QM-9414, was employed for the cellulase enzyme production. Enzyme production conditions (pH, inoculum age and concentration, and organic supplements) were optimized. The ability of partially purified enzyme to hydrolyze various regionally abundant lignocellulosic raw materials was studied. Enzymatic hydrolysis conditions (temperature, pH, enzyme and substrate concentrations) were optimized. Temperature 50v°C, pH 4.5, enzyme concentration 40 FPU/g substrate and substrate concentration 2.5% were found to be optimum for the maximum yields of sugars. #-glucosidase supplementation was found to increase both the sugar yield and hydrolysis rate, and shorten the reaction time significantly.     

Identification of the potential regions of Epap-1 that interacts with V3 loop of HIV-1 gp120

Early pregnancy associated protein-1 (Epap-1), a 90 kDa glycoprotein present in first trimester placental tissue, inhibits HIV-1 entry through interaction with HIV-1 gp120 at V3 and C5 regions. In the present study, we have identified the specific 32 mer region of Epap-1 that can interact with V3 loop. This was achieved by docking between Epap-1 molecular model and gp120 and studying the interaction of peptides with gp120 in vitro. Out of four peptides analyzed, two peptides (P-2 and P-3) showed significant interaction with V3 domain (N = 8; N = 7) of gp120. In the studies conducted using soluble gp120 and virus, peptide P-2 has shown conserved interaction at V3 loop regions recognized by 257D and F425 antibodies and higher anti-viral activity. Also, P-2 inhibited cell fusion mediated dye transfer between gp120 expressing HL2/3 and CD4 expressing Sup T1 cells suggesting its inhibition of viral entry, which is further confirmed by its action on HIV infection mediated by Tat activated beta gal expression in TZM-bl cells. Further optimization of P-2 peptide showed that the anti-viral activity and gp120 interaction residues lie in the N-terminal region of the peptide. These results together suggest that P-2 inhibits viral entry through specific interaction at V3 loop region.     

Design and synthesis of 2-substituted benzoxazoles as novel PTP1B inhibitors

A series of benzoxazole compounds containing oxamic acid were synthesized and screened for the PTP1B inhibition. Compound 31d showed best biochemical potency (K_i) of 6.7 μM. Structure–activity relationship were explained with the help of molecular modeling approach.     

Modeling and analysis of soybean (Glycine max. L) Cu/Zn,Mn and Fe superoxide dismutases

Superoxide dismutase (SOD, EC 1.15.1.1) is an important metal-containing antioxidant enzyme that provides the first line of defense against toxic superoxide radicals by catalyzing their dismutation to oxygen and hydrogen peroxide. SOD is classified into four metalloprotein isoforms, namely, Cu/Zn SOD, Mn SOD, Ni SOD and Fe SOD. The structural models of soybean SOD isoforms have not yet been solved. In this study, we describe structural models for soybean Cu/Zn SOD, Mn SOD and Fe SOD and provide insights into the molecular function of this metal-binding enzyme in improving tolerance to oxidative stress in plants.     

Engineering dynamics of growth factors and other therapeutic ligands

Peptide growth factors and other receptor-binding cytokine ligands are of interest in contemporary molecular health care approaches in applications such as wound healing, tissue regeneration, and gene therapy. Development of effective technologies based on operation of these regulatory molecules requires an ability to deliver the ligands to target cells in a reliable and well-characterizable manner. Quantitative information concerning the fate of peptide ligands within tissues is necessary for adequate interpretation of experimental observations at the tissue level and for truly rational engineering design of ligand-based therapies. To address this need, we are undertaking efforts to elucidate effects of key molecular and cellular parameters on temporal and spatial distribution of cytokines in cell population and cell/matrix systems. In this article we summarize some of our recent findings on dynamics of growth factor depletion by cellular endocytic trafficking, growth factor transport through cellular matrices, and growth factor production and release by autocrine cell systems. (c) 1996 John Wiley & Sons, Inc.