Protein and cDNA sequences of Bowman-Birk protease inhibitors from the cowpea (Vigna unguiculata Walp.)

The protein and gene sequences of the cowpea Bowman-Birk type trypsin inhibitor which confers enhanced insect resistance to transgenic tobacco plants, and of cowpea trypsin/chymotrypsin inhibitors are presented. There are regions of high conservation and high divergence within the 5 leader, mature protein and 3 non-coding regions of the Bowman-Birk inhibitors and in the genes which encode them in different members of this family within the Leguminosae. The practical implications of this finding for studies on the evolution of plants and the utilization of these genes for enhancing insect resistance is discussed.     

Automatic whole broth multi-fermentor sampling

Summary An efficient, aseptic method of obtaining whole broth fermentation samples was developed based on a piston-valve, a local sample loop, and an ability to drive the entire sample volume with sterile air through a sample line and into a remote tube. The configuration delivers 10-ml samples 10 m away with about 4 ml of broth wasted in the sampling process. An autosampler was enhanced and programmed to control acquisition into chilled tubes. The autosampler-based system represents a convenient way to provide frequent samples to profile intracellular and extracellular components for yeast and bacterial fermentations. A configuration to provide sampling from six fermentors with a multi-rack autosampler will be presented.     

Endogenous cytokinins in vernalized winter wheat grains

Summary The soybean callus assay was used to study the effect of vernalization on the endogenous cytokinin levels of the grains of winter wheat (Triticum vulgare L.), cv. Ferto and Fema. Vernalization of the grains was carried out at 3° C in the dark for 0, 9, 18, 27, 36 and 45 days. Cytokinin activities could be detected in both the aqueous and butanol extracts of vernalized grains of both cultivars. It is suggested that flowering of winter wheat may involve changes in the levels of endogenous cytokinins during vernalization.     

Evaluation of Antibacterial Activity of Lavandulapedunculata subsp. atlantica (Braun‐Blanq.) Romo Essential Oil and Selected Terpenoids against Resistant Bacteria Strains–Structure–Activity Relationships

The genus Lavandula is known for its different uses in traditional medicine. This study is interested in the chemical composition of Lavandulapedunculata subsp.atlantica (Braun‐Blanq. ) Romo as well as evaluating its antibacterial potential against multi‐resistant strains. The analysis of Lavandulaatlantica essential oil (LAEO) allows the identification of 47 components representing 93.6 % of all identified. The main constituent of LAEO was camphor (50.4 %), followed by fenchone (14.1 %) and camphene (5.6 %). The antibacterial assays revealed that LAEO was active against all the studied bacteria. A preliminary study of the relationship between certain terpenoids and antibacterial activity was also carried out in order to note the compound(s) that are responsible for LAEO's antibacterial activity. This study showed that the activity of the essential oil may be due to the presence of certain minor compounds such as carvone, considering the presence of the synergistic effect between the essential oil.     

An improved method for micropropagation and regeneration of date palm (Phoenix dactylifera L.)

We developed a novel large-scale micropropagation pathway for date palm (Phoenix dactylifera L.) based on organogenesis. We obtained organogenic stems from shoot tip explants of the Moroccan date palm cultivar Najda, and investigated shoot proliferation from these organogenic stems in vitro on various media; Beauchesne medium (BM) and Murashige and Skoog medium (MS) at full-strength, half-strength, and one-third-strength, containing various concentrations (0, 0.25, 0.5, and 1 mg/L) of 2-naphthoxyacetic acid (NOAA) and kinetin. The optimal medium during the multiplication phase was half-strength Murashige and Skoog medium (MS/2) supplemented with 0.5 mg/L NOAA and 0.5 mg/L kinetin (23.5 morphologically superior shoots per explant, with low vitrification rates). For the shoot elongation phase, shoots were transferred to the same proliferation medium, or to MS or MS/2 media without plant growth regulators (PGRs). Shoots elongated rapidly and showed a high rate of root formation on media supplemented with PGRs. For example, on MS/2 medium containing 1 mg/L NOAA and 1 mg/L kinetin, the average shoot length was 15.1 cm, the average number of roots per shoot was 6.2, and their average length was 3.4 cm. On PGR-free media, shoots were shorter with wider and greener leaves, and had fewer roots. The plantlets were transferred to a greenhouse for acclimation. The survival rate after 2 months was related to the medium used during the elongation phase; >90 % of shoots that were cultured on PGR-free media survived, while there was a poor survival rate of shoots that had been cultured on media containing PGRs.     

By Regulating Mitochondrial Ca2+-Uptake UCP2 Modulates Intracellular Ca2+

Introduction

The possible role of UCP2 in modulating mitochondrial Ca²⁺-uptake (mCa²⁺-uptake) via the mitochondrial calcium uniporter (MCU) is highly controversial.

Methods

Thus, we analyzed mCa²⁺-uptake in isolated cardiac mitochondria, MCU single-channel activity in cardiac mitoplasts, dual Ca²⁺-transients from mitochondrial ((Ca²⁺)m) and intracellular compartment ((Ca²⁺)c) in the whole-cell configuration in cardiomyocytes of wild-type (WT) and UCP2^-/- mice.

Results

Isolated mitochondria showed a Ru360 sensitive mCa²⁺-uptake, which was significantly decreased in UCP2^-/- (229.4±30.8 FU vs. 146.3±23.4 FU, P<0.05). Single-channel registrations confirmed a Ru360 sensitive voltage-gated Ca²⁺-channel in mitoplasts, i.e. mCa1, showing a reduced single-channel activity in UCP2^-/- (Po,total: 0.34±0.05% vs. 0.07±0.01%, P<0.05). In UCP2^-/- cardiomyocytes (Ca²⁺)m was decreased (0.050±0.009 FU vs. 0.021±0.005 FU, P<0.05) while (Ca²⁺)c was unchanged (0.032±0.002 FU vs. 0.028±0.004 FU, P>0.05) and transsarcolemmal Ca²⁺-influx was inhibited suggesting a possible compensatory mechanism. Additionally, we observed an inhibitory effect of ATP on mCa²⁺-uptake in WT mitoplasts and (Ca²⁺)m of cardiomyocytes leading to an increase of (Ca²⁺)c while no ATP dependent effect was observed in UCP2^-/-.

Conclusion

Our results indicate regulatory effects of UCP2 on mCa²⁺-uptake. Furthermore, we propose, that previously described inhibitory effects on MCU by ATP may be mediated via UCP2 resulting in changes of excitation contraction coupling.