Molecular cytogenetics studies in Reichardia tingetana: Physical mapping of heterochromatin,telomere repeats,and 5S and 45S rDNA by 4′, 6‐diamidino‐2‐phenylindole and fluorescence in situ hybridization

Abstract Molecular cytogenetics studies of A‐T‐rich regions, telomeres, and 5S and 45S rDNA sites on the chromosomes of Reichardia tingetana Roth (2n= 16; diploid) were done using 4′, 6‐diamidino‐2‐phenylindole (DAPI) and fluorescence in situ hybridization (FISH). The species were collected from three geographically isolated populations at Borg El Arab (salt marsh habitat), and Rashed and Shosha (sandy clay habitats) in Egypt. The three populations showed the chromosome number of all plants are diploid except for two tetraploid samples from Shosha. Plants from both Rashed and Shosha showed similarity in the distribution of six DAPI bands on six chromosomes, whereas those of Borg El Arab showed a distribution of 16 bands on 14 chromosomes. The FISH signals of the telomeres, and 5S and 45S rDNA, were at the telomeres of all chromosomes, two interstitial, and four terminal, respectively. The combination of DAPI and FISH showed colocalization of the DAPI bands with two 5S and two 45S rDNA loci. The increased number of DAPI bands in the cytotypes from the salt marsh habitat could indicate natural genetic adaptation through increasing the heterochromatin of A‐T‐rich regions.     

<Emphasis Type="Italic">Dunaliella salina</Emphasis> exopolysaccharides: a promising biostimulant for salt stress tolerance in tomato (<Emphasis Type="Italic">Solanum lycopersicum</Emphasis>)

Microalgal exopolysaccharides represent a potential sustainable alternative for the enhancement and protection of agricultural crops including management of both biotic and abiotic stress. In the present study, we investigated the potential of Dunaliella salina exopolysaccharides (PS) to attenuate the effect of salt stress on growth of Solanum lycopersicum, which was grown under different salinity levels (3 and 6 g L^?1 NaCl). The effects of PS treatment on plant growth, osmoprotectant molecules, protein content, and antioxidant enzymes activities of tomato plants under salt stress were analyzed. A metabolomics study showed that the exopolysaccharides released by D. salina contained sulfated moiety along with carbohydrates and uronic acids. The application of sulfated exopolysaccharides on tomato plants alleviated the salt stress and mitigated the decrease in length and dry weight of the plant’s shoot and root systems, as well as that of potassium (K⁺), and K⁺/Na⁺ ratio. Furthermore, the increase in proline, phenolic compounds, Na⁺, and antioxidant enzymes (CAT, POD, SOD) activities caused by salt stress were attenuated after the exopolysaccharide treatment. GC-MS metabolomics analysis showed that PS treatment allowed the activation and/or inhibition of various metabolic pathways involved in the plant’s tolerance to stress such as jasmonic acid-dependent pathways. This study shows the potential of microalgal exopolysaccharides for enhancing tomato tolerance to salt stress and highlights the possibility of their use as plant growth biostimulants under harsh environmental conditions.     

Iron requirement and siderophore production in Rhizobium ciceri during growth on an iron-deficient medium

Under conditions of iron limitation many rhizospheric bacteria produce siderophores, ferric iron-specific ligands, which may enhance plant growth by increasing the availability of iron near the roots. Thirty-five strains of Rhizobium ciceri, specific to chickpea (Cicer arietinum L.), were screened for their ability to grow on iron-deficient medium and to produce siderophores. Maximal growth of all strains previously depleted in iron was obtained in medium containing 5 to 10 m of ferric iron. When iron limitation was achieved by the addition of 2,2-bipyridyl or EDDHA [ethylene diamine di(o-hydroxyphenyl) acetic acid] to the medium, only two strains were able to scavenge iron and grow. Siderophore production by these two strains was detected by the Chrome Azurol S assay (CAS), a universal test for siderophores. No hydroxamate-type siderophores were detected in the supernatants of Rhizobium ciceri cultures. However, some strains secreted salicylic acid and 2,3-dihydroxybenzoic acid as phenolate-type siderophores. Addition of ferric iron to the culture medium increased growth yield significantly but depressed the production of siderophores. Although these compounds are produced in response to iron deficiency, nutritive components of the culture medium significantly affected their production. It seems that CuII, MoVI and MnII ions bound competitively with iron to siderophores, resulting in a 34 to 100% increase in production.     

Choline acetyltransferase activity of spinal cord cell cultures increased by co-culture with muscle and by muscle-conditioned medium

Activity of the enzyme choline acetyltransferase (CAT), which mediates the synthesis of the neurotransmitter, acetylcholine, was increased up to 20- fold in spinal cord (SC) cells grown in culture with muscle cells for 2 wk. This increase was directly related to the duration of co-culture as well as to the cell density of both the SC and muscle involved and was not affected by the presence of the acetylcholine receptor blocking agent, α-bungarotoxin. Glutamic acid decarboxylase (GAD) activity was often markedly decreased in SC-muscle cultures while the activities of acetylcholinesterase and several other enzymes were little changed. Increased CAT activity was also observed when SC cultures were maintained in medium which had been conditioned by muscle cells or by undifferentiated cells from embryonic muscle. Muscle-conditioned medium (CM) did not affect the activities of SC cell GAD or acetylcholinesterase. Dilution or concentration of the CM directly affected its ability to increase SC CAT activity , as did the duration and timing of exposure of the SC cells to the CM. The medium could be conditioned by muscle cells in the presence or absence of serum, and remained effective after dialysis or heating to 58 degrees C. Membrane filtration data were consistent with the conclusion that the active material(s) in CM had a molecular weight in excess of 50,000 daltons. We conclude that large molecular weight material that is released by muscle cells is capable of producing a specific increase in CAT activity of SC cells.     

Influence of Exogenous Hormones on Somatic Embryogenesis in Hevea brasiliensis

Generating somatic embryos from the inner teguments of Heveaseeds is difficult. Like other ligneous plants, the rubber-treeis generally considered to be recalcitrant with regard to somaticembryogenesis. In this paper we show that when conventionallyused amounts of both 3, 4-dichlorophenoxyacetic acid (3, 4-D)and benzylaminopurine (BAP) are reduced by 2–4 times andwhen the length of exposure to these hormones is also reduced,the embryogenic capacity of calli increases without affectingcallogenesis. The germination of several embryos and plantletswas obtained. We conclude that an excess of auxin and cytokininin the medium, even of a weak auxin such as 3, 4-D, and theprolonged exposure to these hormones suppress the embryogeniccapacity of the callus. Hevea brasiliensis, rubber-tree, somatic embryogenesis, hormones     

Comparative detection,density, and reproductive performance of Kirtland's warbler in jack and red pine

The formerly endangered Kirtland's warbler (Setophaga kirtlandii) is among a growing number of conservation-reliant species that depend on active management to avoid reverting to endangered status. Because the Kirtland's warbler is a habitat specialist of young, even-aged jack pine (Pinus banksiana), managers of the recovery effort stressed creating new jack pine stands and monitoring numbers of singing males through an annual census using single visits to individual stands. Kirtland's warbler will occupy and breed in red pine (P. resinosa), but red pine has not been surveyed for Kirtland's warblers in the annual population census. Furthermore, the current monitoring approach cannot determine their species detection probability or individual detection probability, which is essential to evaluate both red pine use and the accuracy of the census. From 2016–2018 we estimated density and detection probabilities in jack pine and red pine stands through repeated visits to a limited number of stands rather than single visits to many stands. Estimates of species detection probability indicated that ≥1 male Kirtland's warbler would be detected on most sites when any were present, but individual detection probabilities were less and varied by stand type, indicating that single visits to sites would underestimate numbers and that accurate estimation of detection probability was important for estimation of density in different stand types. We offer quantitative estimates of detection probabilities for determination of Kirtland's warbler population size in jack pine versus red pine stands in the same areas and breeding seasons. Managers of Kirtland's warblers should incorporate detection probabilities into population surveys to achieve more accurate estimates of population size.     

Vibrio cholerae laboratory infection of the adult house fly Musca domestica

The present study was designed to test the hypothesis that house flies may be capable of specifically harbouring ingested Vibrio cholerae in their digestive tracts. Flies were continuously fed green fluorescent protein (GFP)‐labelled, non‐O1/non‐O139 environmental strains of V. cholerae. Bacterial burdens were quantitatively measured using plate counts and localization was directly observed using confocal microscopy. Vibrio cholerae were present in the fly alimentary canal after just 4 h, and reached a plateau of ～10⁷ colony‐forming units (CFU)/fly after 5 days in those flies most tolerant of the pathogen. However, individual flies were resistant to the pathogen: one or more flies were found to carry < 180 V. cholerae CFU at each time‐point examined. In flies carrying V. cholerae, the pathogen was predominantly localized to the midgut rather than the rectal space or crop. The proportion of house flies carrying V. cholerae in the midgut was dose‐dependent: the continuous ingestion of a concentrated, freshly prepared dose of V. cholerae increased the likelihood that fluorescent cells would be observed. However, V. cholerae may be a transient inhabitant of the house fly. This work represents the first demonstration that V. cholerae can inhabit the house fly midgut, and provides a platform for future studies of host, pathogen and environmental mediators of the successful colonization of this disease vector.     

Occurrence of aflatoxin B1 and ochratoxin A in Lebanese cultivated wheat

An extensive survey of filamentous fungi isolated from wheat grown and consumed in Lebanon and their capacity to produce aflatoxin B₁ (AFB₁) and ochratoxin A (OTA) was conducted to assess fungi potential for producing these toxins in wheat. From the 468 samples of wheat kernel, collected at preharvest stage from different locations during 2008 and 2009 cultivation seasons, 3,260 fungi strains were isolated with 49.4% belonging to Penicillium spp. and 31.2% belonging to Aspergillus spp. Penicillium spp. was detected on wheat samples with a high amount of P. verrucosum (37.0%). Among the different Aspergillus spp. isolated, A. niger aggregate was predominant and constituted 37.3%. whereas the isolation rate of A. flavus and A. ochraceus was 32.2 and 25.6%, respectively. The ability to produce OTA and AFB₁ by isolates belonging to Aspergillus spp. and Penicillium spp. was analyzed by high performance liquid chromatography with fluorescence detector (HPLC-FLD). It was found that 57.0% of Penicillium spp. and 80% of A. ochraceus isolates tested produced OTA, respectively, at maximum concentrations of 53 and 65 μg/g CYA. As for the aflatoxinogenic ability, 45.3% of A. flavus produced AFB₁, with maximum concentration of 40 μg/g CYA. A total of 156 wheat samples were analyzed for the levels of OTA and AFB₁ by HPLC-FLD. The results showed that 23.7% were contaminated with OTA, at a concentration higher than 3 μg/kg and 35.2% of these samples were contaminated with AFB₁ at concentration higher than 2 μg/kg. The risks originating from toxin levels in wheat produced in Lebanon should be monitored to prevent their harmful effects on public health.     

Catabolism of Tryptophan by Trypanosoma evansi

ABSTRACT. Trypanosoma brucei gambiense , which causes human African trypanosomiasis, catabolizes the aromatic amino acid tryptophan via an initial aminotransferase catalyzed reaction to form several indole end products, which have been suggested to contribute to the pathogenesis of trypanosomiasis. To determine if this same pathway exists in T. evansi , the closely related trypanosome pathogen of domestic animals, tryptophan catabolism was examined in vitro and in vivo. As is the case with human African trypanosomes, T. evansi catabolized tryptophan to form indole-3-pyruvic acid and smaller amounts of indole-3-acetic acid and indole-3-lactic acid. Large concentrations of indole-3-pyruvic acid are excreted in urine of trypanosome-infected mice. However, indole-3-ethanol could not be detected in incubates of T. evansi or T. b. gambiense , even though the latter species had previously been reported to form this neutral metabolite. A new, previously unreported tryptophan metabolite was isolated and partially characterized from incubates of T. evansi and T. b. gambiense. Although the functional significance of tryptophan catabolism to trypanosomatids remains obscure, the pathway is quantitatively significant in all species examined thus far.     

Glutathione S-transferase omega in the lung and sputum supernatants of COPD patients

Background

The major contribution to oxidant related lung damage in COPD is from the oxidant/antioxidant imbalance and possibly impaired antioxidant defence. Glutathione (GSH) is one of the most important antioxidants in human lung and lung secretions, but the mechanisms participating in its homeostasis are partly unclear. Glutathione-S-transferase omega (GSTO) is a recently characterized cysteine containing enzyme with the capability to bind and release GSH in vitro. GSTO has not been investigated in human lung or lung diseases.

Methods

GSTO1-1 was investigated by immunohistochemistry and Western blot analysis in 72 lung tissue specimens and 40 sputum specimens from non-smokers, smokers and COPD, in bronchoalveolar lavage fluid and in plasma from healthy non-smokers and smokers. It was also examined in human monocytes and bronchial epithelial cells and their culture mediums in vitro.

Results

GSTO1-1 was mainly expressed in alveolar macrophages, but it was also found in airway and alveolar epithelium and in extracellular fluids including sputum supernatants, bronchoalveolar lavage fluid, plasma and cell culture mediums. The levels of GSTO1-1 were significantly lower in the sputum supernatants (p = 0.023) and lung homogenates (p = 0.003) of COPD patients than in non-smokers.

Conclusion

GSTO1-1 is abundant in the alveolar macrophages, but it is also present in extracellular fluids and in airway secretions, the levels being decreased in COPD. The clinical significance of GSTO1-1 and its role in regulating GSH homeostasis in airway secretions, however, needs further investigations.