The effect of mesenchymal stem cells as co-culture in in vitro nuclear maturation of ovine oocytes

This study compared the effects of ovine mesenchymal stem cells (MSCs) and ovine oviductal epithelial cells (OECs) as feeder cells in cell free culture systems (HEPES-modified tissue culture medium, TCM199) supplemented with polyvinyl alcohol (PVA) or fetal calf serum (FCS) on in vitro oocyte maturation and subsequent embryo development (IVM/IVC). Cumulus-oocyte complexes (COCs) were harvested from ovine ovaries and subjected to IVM in the above-mentioned culture media. After culture for 24 h, nuclear maturation of the oocytes was evaluated by 4, 6-diamino- 2-phenylindole (DAPI) staining. After fertilization the presumptive zygotes were cultured under identical culture conditions and embryo development was evaluated. The percentage of oocytes at nuclear maturation (metaphase II) cultured in the MSC group was higher than for the IVM medium + PVA group (P<0.05), while between MSCs, OECs and IVM medium + FCS it was non-significant. The rates (%) of cleavage and the percentage of total blastocysts in MSCs and the IVM medium + FCS group were higher than for OECs and the IVM medium + PVA group (P<0.05). These rates were non-significant between MSCs and the IVM medium + FCS group or between OECs and the IVM medium + PVA group. The percentage of hatched blastocysts (%) was significantly increased in MSCs and the IVM medium + FCS group when compared to OECs and the IVM medium + PVA group (P<0.05). In conclusion, the effects of mesenchymal stem cells as co-culture on oocyte maturation and the successive embryo development in vitro are similar to those in the medium supplemented with FCS. This study suggests that co-culturing with mesenchymal stem cells may be a promising alternative to FCS-medium

EFFECT OF MEDIA AND INDOLE BUTYRIC ACID (IBA) CONCENTRATIONS ON ROOTING OF RUSSIAN OLIVE (ELAEAGNUS ANGUSTIFOLIA L.) SEMI-HARD WOOD CUTTINGS

ABSTRACT This study was conducted with the main purpose of determination of the most suitable IBA concentrations and media on rooting of Russian olive (Elaeagnus angustifolia L.) semi hardwood cuttings. A two factorial experiment was laid out a completely randomized design (CRD) with three replications and each replication consisted of 12 cuttings. Treatments consist of five concentrations of IBA (0, 1000, 2000, 3000 and 4000 mg L -1 ) and two rooting media (sand and cocopeat + perlite 1:1 by volume). Results showed that significant difference of the most the experimental treatments. The highest rooting percentage (97.22%), root number (7.15) and root length (12.67 cm) were obtained in cuttings were treated with 2000, 4000 and 1000 mg L -1 IBA in cocopeat + perlite rooting medium, respectively. There was no significant difference in most traits between 2000 and 4000 mg L -1 IBA except on rooting percentage. Finally our results revealed that 2000 mg L -1 IBA in cocopeat + perlite rooting medium was the suitable treatment for propagation by semi-hard wood cutting

Evaluation of Zinc solubilization potential by different strains of Fluorescent Pseudomonads

Zinc solubilizing ability of Pseudomonas fluorescent was evaluated using zinc oxide, zinc carbonate and zinc sulphide in both plate and broth media assays. Forty bacterial strains and 0.1% of each chemical source in six replications were used. Colony and halo diameters were measured after incubating the plates for 48h in incubator. Zn solubilizing ability of 40 mentioned strains in three replications was studied with ZnO and ZnCO3 solutions in broth assay. The soluble zinc and pH were measured after five days. The results showed, only 8 of 40 strains could form clearing zone in plate assay. Halo diameter, ratio of the halo diameter to the colony diameter and area respectively for zinc oxide and zinc carbonate were as following, respectively: 0.60- 1.32 cm, 1.20-2.64 and 0.95-2.60 cm2, 0.13-1.70 cm, 0.27-2.99 and 0.31-4.10 cm2. There were no halos observed in zinc sulphide. The concentration of soluble Zn for ZnO was 28-625 mg/l and pH was shifted from 7.0-7.2 to 3.90-6.50 and for ZnCO3 was 247-753 mg/l and pH was shifted from 7.0-7.2 to 3.5-6.3 after 5 days of inoculation in 28°C