Synchrotron-based FTIR microspectroscopy of chili resistance induced by Bacillus subtilis strain D604 against anthracnose disease

The aim of this study was to determine the resistance mechanisms of chili induced by the Bacillus subtilis strain D604 using synchrotron FTIR microspectroscopy (SR-FTIR). In this study, the strain D604 reduced anthracnose disease severity in chili plants by approximately 31.10%. The SR-FTIR spectral changes from the epidermis and mesophyll leaf tissue revealed higher integral areas for the C=O ester from lipids, lignin, or pectin (1770–1700?cm^?1) as well as polysaccharides (1200–900?cm^?1) in the treated samples of D606 and distilled water and then challenge inoculation with chili anthracnose pathogen, Colletotrichum acutatum. The secondary structure of the Amide I protein failed to convert from alpha helices (centered at 1650?cm^?1) to beta sheets (centered at 1600?cm^?1) in the mesophyll of samples not treated with D604. This study suggested that the strain D604 induced resistance against anthracnose pathogen in chili by inducing cellular changes related to defense compounds involved in plant defense mechanism.     

Cloning in Bacillus subtilis of temperature-dependent promoter fragments from Bacillus stearothermophilus and Bacillus subtilis

Abstract Using promoter-probe plasmids, more than 200 promoter-containing fragments from Bacillus stearothermophilus and Bacillus subtilis were cloned in B. subtilis . Among these, 15 promoter fragments were highly temperature-dependent in activity compared to the promoter sequence (TTGAAA for the −35 region, TATAAT for the −10 region) of the amylase gene, amyT , from B. stearothermophilus . Some fragments exhibited higher promoter activities at elevated temperature (48°C), others showed higher activities at lower temperature (30°C). Active promoter fragments at higher and lower temperatures were obtained mainly from the thermophile ( B. stearothermophilus ) and the mesophile ( B. subtilis ), respectively. A promoter fragment active at high temperature was sequenced, and the feature of the putative promoter region was discussed.     

Variability of protein and lipid composition of human subtantia nigra in aging: Fourier transform infrared microspectroscopy study

There is growing evidence that a variety of biochemical processes that underlie the most frequent neurodegenerative diseases may have much in common with those connected with natural aging. It was shown that they involve, among others, lipid peroxidation and/or generation of insoluble in water protein deposits (i.e. alpha-synuclein and/or beta amyloid). Therefore, it is likely that the analysis of changes in both lipid and protein composition may be interesting in the light of any potential pathologies occurring within the dopaminergic system during physiological aging. Thereby, this paper presents a methodology for the analysis of age-related changes in a lipid and protein composition within human subtantia nigra tissue by means of Fourier transform infrared microspectroscopy (FTIRM). Particularly, the changes in the lipid saturation, unsaturation as well as in the protein secondary structure were examined. The studies were carried out on samples from 35 individuals who died without any signs of neurologic dysfunctions. Our results show that the level of lipid saturation increases inside the subtantia nigra tissue with age, though the total content of lipid decreases with age of individuals. Moreover, the statistically significant decrease in the protein content within neuron bodies was observed. Interestingly, it is presented that the content of the anti-parallel beta sheets for neuron bodies decreases from seventh to eighth decades of life and subsequently markedly increases from eighth to ninth decades of life, whilst, as regards extraneuronal spaces, the opposite trends are reported i.e. increase from the seventh to eighth decades, and subsequent decrease in the ninth decade of life. These observations, though preliminary, shed the light on a putative contribution of various pathological lipid- and protein-related processes underlying senescence, suggesting a “biochemical link” between the aetiology of the most common neurodegenerative diseases and physiological aging.     

Evaluation of the biological stability of waste during landfill stabilization by thermogravimetric analysis and Fourier transform infrared spectroscopy

This study seeks to assess the biological stability of landfilled municipal solid waste (MSW) based on the changes in organic matter, as revealed by thermogravimetric analysis and Fourier transform infrared (FTIR) spectroscopy. Derivate thermogravimetry profiles (DTG) showed a reduction in peak intensity at 200-350 °C (DTG2), while an increase in peak intensity and a shift towards higher temperature at 400-600 °C (DTG3). The decrease in the peak intensity of the aliphatic methylene at 2920 and 2850 cm(-1), and the increase of aromatic substances and polysaccharide at 1640 cm(-1) in the FTIR spectra also confirm the changes. Well-fitted correlations of the peak intensity ratio (2920/1640) and peak area ratio (DTG2/DTG3) to C/N ratio were also established, confirming that the 2920/1640 and the DTG2/DTG3 ratios can be considered as reliable parameters for tracking the biological stability of MSW during landfill stabilization.     

A Fourier transform infrared microspectroscopic imaging investigation into an animal model exhibiting glioblastoma multiforme

Glioblastoma multiforme (GBM) is a highly malignant human brain tumour for which no cure is available at present. Numerous clinical studies as well as animal experiments are under way with the goal being to understand tumour biology and develop potential therapeutic approaches. C6 cell glioma in the adult rat is a frequently used and well accepted animal model for the malignant human glial tumour. By combining standard analytical methods such as histology and immunohistochemistry with Fourier Transform Infrared (FTIR) microspectroscopic imaging and multivariate statistical approaches, we are developing a novel approach to tumour diagnosis which allows us to obtain information about the structure and composition of tumour tissues that could not be obtained easily with either method alone. We have used a “Stingray” FTIR imaging spectrometer to analyse and compare the compositions of coronal brain tissue sections of a tumour-bearing animal and those from a healthy animal. We have found that the tumour tissue has a characteristic chemical signature, which distinguishes it from tumour-free brain tissue. The physical-chemical differences, determined by image and spectral comparison are consistent with changes in total protein absorbance, phosphodiester absorbance and physical dispersive artefacts. The results indicate that FTIR imaging analysis could become a valuable analytic method in brain tumour research and possibly in the diagnosis of human brain tumours.     

Flagellin as a Biomarker for Bacillus subtilis Strains; Application to the DB9011 Strain and the Study of Interspecific Diversity in Amino-acid Sequences

Polymerase chain reaction (PCR) for the flagellin central domain coding region (FCD-PCR) was applied to the detection and discrimination of Bacillus subtilis DB9011, a strain with useful functions in agriculture. Cross-reactions were observed in 4 B. subtilis strains with similar flagellin genes (hag). Alignment of partial amino-acid sequences of flagellin and the results of PCR for the 16S/23S rRNA spacer in 11 B. subtilis strains suggested the presence of a group including strains with antifungal activity (DB9011 and others).     

枯草芽胞杆菌突变株ZC-7中性蛋白酶催化区域氨基酸突变导致其活力提高

目的：探讨枯草芽胞杆菌突变株ZC-7高产中性蛋白酶的原因。方法：用PCR方法分别扩增突变株ZC-7与出发菌株枯草芽胞杆菌AS1．398产中性蛋白酶的编码基因,测序比较二者基因的不同;在CPHmodels Server网站进行氨基酸序列分析,模拟突变前后中性蛋自酶的二级结构。结果：对比结果显示成熟肽中有5个氨基酸位点发生突变,其中3个位于酶的催化区域内;从预测的二级结构模型上可以看到突变位点所处区域的折叠结构发生细微变化。结论：先前研究中发现枯草芽胞杆菌AS1．398和突变株ZC-7发酵液中的酶蛋白含量基本相同,因此推测高产的原因不是酶量的增加,而是突变的氨基酸使酶与底物结合的部位更加适合催化水解反应,从而提高其比活力。     

Biomolecular investigation of human substantia nigra in Parkinson's disease by synchrotron radiation Fourier transform infrared microspectroscopy

Synchrotron radiation based-Fourier transform infrared microspectroscopy was used for preliminary investigation of the chemical composition and morphologies of the human substantia nigra of brain between normal and Parkinson's diseased tissues. The studies were carried out for thin tissue sections, focusing more particularly on nerve cell bodies, that are affected in Parkinson's disease (PD). The major spectral differences between normal (control) and PD tissues were identified at the following vibrational frequencies: 2930, 2850, 1655, 1380, 1236, 1173 and 1086 cm(-1). The infrared imaging of these biochemical markers show that for control cases the protein and nucleic acids functional groups (bands at: approximately 3300, approximately 3100, approximately 1655, approximately 1545, approximately 1240, approximately 1080 cm(-1)) are located mainly in the cell body. The spatial distribution of the band at 1740 cm(-1) (ester carbonyl stretching band) is quite dissimilar to the others, while it exhibits a minimal concentration in the cell body area. Contrarily, in PD samples, no clear evidence of variation of any of the vibrational fingerprint between cell body and the surrounding was noticed. Moreover, decrease of protein to lipid ratio as well as increase of amide I/amide II ratio were observed for PD case. The preliminary results strengthen the hypothesis that PD is a multietiological disorder. Moreover, the reported results clearly indicate that, in addition to a distinct visual observation, the diseased nerve cells exhibits change of their biochemical composition. It suggests that disturbances of normal functioning of SN neurons appear before their morphological atrophy.     

Glutamate-354 of the CP43 polypeptide interacts with the oxygen-evolving Mn4Ca cluster of photosystem II: a preliminary characterization of the Glu354Gln mutant

In the recent X-ray crystallographic structural models of photosystem II, Glu354 of the CP43 polypeptide is assigned as a ligand of the O2-evolving Mn4Ca cluster. In this communication, a preliminary characterization of the CP43-Glu354Gln mutant of the cyanobacterium Synechocystis sp. PCC 6803 is presented. The steady-state rate of O2 evolution in the mutant cells is only approximately 20% compared with the wild-type, but the kinetics of O2 release are essentially unchanged and the O2-flash yields show normal period-four oscillations, albeit with lower overall intensity. Purified PSII particles exhibit an essentially normal S2 state multiline electron paramagnetic resonance (EPR) signal, but exhibit a substantially altered S2-minus-S1 Fourier transform infrared (FTIR) difference spectrum. The intensities of the mutant EPR and FTIR difference spectra (above 75% compared with wild-type) are much greater than the O2 signals and suggest that CP43-Glu354Gln PSII reaction centres are heterogeneous, with a minority fraction able to evolve O2 with normal O2 release kinetics and a majority fraction unable to advance beyond the S2 or S3 states. The S2-minus-S1 FTIR difference spectrum of CP43-Glu354Gln PSII particles is altered in both the symmetric and asymmetric carboxylate stretching regions, implying either that CP43-Glu354 is exquisitely sensitive to the increased charge that develops on the Mn4Ca cluster during the S1-->S2 transition or that the CP43-Glu354Gln mutation changes the distribution of Mn(III) and Mn(IV) oxidation states within the Mn4Ca cluster in the S1 and/or S2 states.     

Stable isotope ratios of carbon and nitrogen in pollen grains in order to characterize plant functional groups and photosynthetic pathway types

Measurements of delta(13)C, delta(15)N and C : N ratios on modern pollen grains from temperate plants, including whole grains as well as extracted sporopollenin, were analysed in order to characterize physiological plant types at the pollen level and to determine the variation of these parameters in modern pollen grains of the same climatic area. Measurements are presented for 95 batches of whole modern pollen from 58 temperate species and on the stable fraction of modern pollen grains, chemically extracted sporopollenin, for two modern species. Fourier transform infrared (FTIR) and cross-polarization and magic-angle spinning (CP/MAS) sporopollenin spectra were conducted in parallel. C(3) and C(4) plants can be separated by delta(13)C measurements based on pollen. Probabilistic assignments to plant functional groups (herbaceous, deciduous woody, evergreen woody) of C(3) plants by the means of a discriminant analysis can be made for C : N ratios and for delta(13)C. The results are related to other studies on sporopollenin in order to use this method in future work on fossil samples. Stable isotope measurements on pollen allow improved pollen diagrams, including forms that cannot be differentiated at species level, increasing the accuracy and resolution of plant physiological type distribution in quaternary and older fossil sediments.     

Characterization of Bacillus subtilis strains in Thua nao, a traditional fermented soybean food in northern Thailand

AIMS: To clarify the diversity of Bacillus subtilis strains in Thua nao that produce high concentrations of products useful in food manufacturing and in health-promoting compounds. METHOD AND RESULTS: Production of amylase, protease, subtilisin NAT (nattokinase), and gamma-polyglutamic acid (PGA) by the Bacillus subtilis strains in Thua nao was measured. Productivity of protease NAT by these strains tended to be higher than by Japanese commercial natto-producing strains. Molecular diversity of isolated strains was analysed via randomly amplified polymorphic DNA-PCR fingerprinting. The strains were divided into 19 types, including a type with the same pattern as a Japanese natto-producing strain. CONCLUSION: B. subtilis strains that could be a resource for effective production of protease, amylase, subtilisin NAT, or PGA were evident in Thua nao produced in various regions in northern Thailand. SIGNIFICANCE AND IMPACT OF THE STUDY: This study clearly demonstrated the value of Thua nao as a potential resource of food-processing enzymes and health-promoting compounds.     

Phase behaviour and crystallinity of plant cuticular waxes studied by Fourier transform infrared spectroscopy

The phase behaviour of cuticular waxes from leaves of Hedera helix L. and Juglans regia L. was studied by Fourier transform infrared spectroscopy. For this purpose reconstituted waxes, isolated cuticular membranes, dewaxed polymer matrix membranes and whole leaves were studied in the horizontal attenuated total reflection and transmission modes. Melting curves of cuticular waxes were derived from temperature-dependent changes in the absorption maximum of the symmetric stretching mode of CH₂ groups (ν_s, at approx. 2856–2848 cm⁻¹). With increasing temperature absorption band doublets due to CH₂ scissoring (δ_sciss) and rocking (δ_rock) movements (at approx. 1473–1471 and 730–720 cm⁻¹, respectively) indicative of an orthorhombic arrangement of alkyl chains merged into a single peak. The area ratio of the peaks at approx. 720 and 730 cm⁻¹ was used as a measure for aliphatic crystallinity of plant cuticular waxes at a given temperature. The investigations of reconstituted cuticular waxes and those still embedded in isolated cuticles or in situ on the leaf produced comparable results. The findings are discussed in terms of the properties of the cuticular transport barrier. Received: 21 March 1997 / Accepted: 25 April 1997     

Green synthesis and characterization of Carica papaya leaf extract coated silver nanoparticles through X-ray diffraction,electron microscopy and evaluation of bactericidal properties

The evolution of nanotechnology and the production of nanomedicine from various sources had proven to be of intense value in the field of biomedicine. The smaller size of nanoparticles is gaining importance in research for the treatment of various diseases. Moreover the production of nanoparticles is eco-friendly and cost effective. In the present study silver nanoparticles were synthesized from Carica papaya leaf extract (CPL) and characterized for their size and shape using scanning electron microscopy and transmission electron microscopy, respectively. Fourier transform infrared spectroscopy (FTIR), Energy dispersive X-ray spectroscopy (EDS/EDX) and X-ray diffraction spectroscopy (XRD) were conducted to determine the concentration of metal ions, the shape of molecules. The bactericidal activity was evaluated using Luria Bertani broth cultures and the minimum inhibition concentration (MIC) and minimum bactericidal concentration (MBC) were estimated using turbidimetry. The data analysis showed size of 50–250 nm spherical shaped nanoparticles. The turbidimetry analysis showed MIC and MBC was >25 μg/mL against both Gram positive and Gram negative bacteria in Luria Bertani broth cultures. In summary the synthesized silver nanoparticles from CPL showed acceptable size and shape of nanoparticles and effective bactericidal activity.     

Hydrogen-isotope composition of leaf water in C3 and C4 plants: its relationship to the hydrogen-isotope composition of dry matter

The natural abundance hydrogen-isotope composition of leaf water ( ) and leaf organic matter ( _D ^org ) was measured in leaves of C₃ and C₄ dicotyledons and monocotyledons. The value of leaf water showed a marked diurnal variation, greatest enrichment being observed about midday. However, this variation was greater in the more slowly transpiring C₄ plants than in C₃ plants under comparable environmental conditions. A model based on analogies with a constant feed pan of evaporating water was developed and the difference between C₃ and C₄ plants expressed in terms of either differences in kinetic enrichment or different leaf morphology. Microclimatic and morphological features of the leaves which may be associated with this factor are discussed. There was no daily excursion in the _D ^org value in leaves of either C₃ or C₄ plants. When _D ^org values were referenced to the mean values during the period of active photosynthesis, the discrimination against deuterium during photosynthetic metabolism (D) was greater in C₃ plants (-117 to -121) than in C₄ plants (-86 to -109).These results show that the different water use strategies of C₃ and C₄ plants are responsible for the measured difference in deuterium-isotope composition of leaf water. However, it is unlikely that these physical processes account fully for the differences in hydrogen-isotope composition of the products of C₃ and C₄ photosynthetic metabolism.Symbols Hydrogen-isotope composition of leaf water - _D ^org hydrogen-isotope composition of leaf organic matter