Evidence for enhanced dissipation of chlorpyrifos in an agricultural soil inoculated with Serratia rubidaea strain ABS 10.

The insecticide 14C-chlorpyrifos was found mineralized in a Tunisian soil with repeated exposure to it. From this soil, a bacterial strain was isolated that was able to grow in a minimal salt medium (MSM) supplemented with 25 mg L-1 of chlorpyrifos. It was characterized as Serratia rubidaea strain ABS 10 using morphological and biochemical analyses, as well as 16S rRNA sequencing. In a liquid culture, the S. rubidaea strain ABS 10 was able to dissipate chlorpyrifos almost entirely within 48 h of incubation. Although the S. rubidaea strain ABS 10 was able to grow in an MSM supplemented with chlorpyrifos and dissipate it in a liquid culture, it was not able to mineralize 14C-chlorpyrifos. Therefore, it can be concluded that the dissipation capability of this bacteria might be attributed to its capacity to adsorb CHL. It can also be ascribed to other reasons such as the formation of biogenic non-extractable residues. In both non-sterile and sterile soil inoculated with S. rubidaea strain ABS 10, chlorpyrifos was more rapidly dissipated than in controls with DT50 of 1.38 and 1.05 days, respectively.

Evaluation of protein extraction methods for Vitis vinifera leaf and root proteome analysis by two-dimensional electrophoresis.

An efficient protein extraction method is crucial to ensure successful separation by two-dimensional electrophoresis (2-DE) for recalcitrant plant species, in particular for grapevine (Vitis vinifera L.). Trichloroacetic acid-acetone (TCA-acetone) and phenol extraction methods were evaluated for proteome analysis of leaves and roots from the Tunisian cultivar 'Razegui'. The phenol-based protocol proved to give a higher protein yield, a greater spot resolution, and a minimal streaking on 2-DE gels for both leaf and root tissues compared with the TCA-based protocol. Furthermore, the highest numbers of detected proteins on 2-DE gels were observed using the phenol extraction from leaves and roots as compared with TCA-acetone extraction.

Preservation of endangered Tunisian grapevine cultivars using embryogenic cultures

The preservation of embryogenic lines derived from several endangered local grapevine cultivars was studied. Embryogenic calluses were obtained from immature anthers of eight cultivars, sampled on both fruity-cuttings and field grown vines. Anthers at the 'separated flower' stage, derived from fruity-cuttings, resulted in an increased induction of somatic embryogenesis, compared to those derived from the field. Pro-embryogenic calluses were induced on Chée and Pool (1987) basal medium, supplemented with 9 micron M of 2,4-dichlorophenoxyacetic acid (2,4-D) and 11.35 micron M of thidiazuron (TDZ) under dark conditions. Different anther zones (filament, abaxial, adaxial, lateral zones and entire anthers) were involved in somatic embryogenesis induction. The percentages of granular and yellowish pro-embryogenic calluses ranged between 15.6 percent and 34.8 percent in 'Kahli Kerkennah' and 'Muscat Raf-raf' cultivars, respectively. Although, morphological diversifications of pro-embryogenic calluses (several necrosis and spontaneous maturation) were observed on the induction mediumafter 5 subcultures. The reduction of 2,4-D and TDZ levels to 4.52 micron M and 2.89 micron M respectively, induced granular and yellowish embryogenic material. Thus, Chée and Pool (1987) (CP) enriched with 4.52 micron M of 2,4-D and 2.89 micron M of TDZ revealed to be the most appropriate for long-term maintenance. In fact, all the cultivars presented high and regular embryo maturation rates after 12, 24, 36 and 48 months of cultivation on this medium, under light conditions. After 4 years, they still exhibit high germination and regeneration abilities. Germination of somatic embryos was achieved on Murashige and Skoog (1962) basal-medium, with rates ranging from 69 percent to 96 percent. Only 5 percent of somatic embryos were concerned by morphological variations. The regenerated plantlets presented a normal phenotype under controlled greenhouse conditions, compared to mother plants.