European Journal of Molecular Biotechnology

188-196

Authors: Eaman A. S. AL-Rubaee, Suha T. Abd, Nadia M. Kadim

The structural and optical properties of the titanium dioxide nanoparticles [TiO2 NPs] have been investigated using [UV-Vis] spectrophotometer and SEM. The produced nanoparticles show small and about sharp round peaks around 220 nm. The produced nanoparticles have a spherical shape with an average particle size ˂50 nm. The effect of titanium dioxide NPs was studied on the activity of Alkaline Phosphatase [ALP] in the saliva of 25 patients with gingivitis in comparison to 20 healthy subjects with the average age about 22–23 years for both groups. The results correlated with the observation that salivary alkaline phosphatase activity increase in patient with gingivitis in comparison to control group and salivary ALP activity inhibited by titanium dioxide nanoparticles.

210-227

Authors: Ignat Ignatov, Oleg Mosin

The isotopic composition, the temperature and the pH value of water were analyzed in experiments with prognosis of primary hydrosphere and hydrological conditions for origin of first organic forms in hot mineral water with HDO. For this aim we performed experiments with hot mineral water and seawater from Bulgaria and water with varying content of deuterium using IR- and DNES-spectroscopy. As model systems were used cactus juice of Echinopsis pachanoi and Mediterranean jellyfish Cotylorhiza tuberculata. The reactions of condensation–dehydration occurring in alkaline aqueous solutions at t = 65–95 0C and рН = 9–10, resulting in synthesis from unorganic molecules the larger organic molecules as polymers and short polipeptides, were discussed, as well as the possible mechanisms of the deuterium accumulation in form of HDO in hot water. It was shown that hot alkaline mineral water with temperature from +65 0C to +95 0C and the pH value from 9 to 11 is more suitable for the origination of life and living matter than other analyzed water samples. In hot mineral waters the local maximums in IR-spectra are more manifested compared to the local maximums obtained in IR-spectra of the same water at a lower temperature. The difference in the local maximums from +20 0C to +95 0C at each +5 0C according to the Student t-criterion makes up p < 0,05.

118-130

Authors: Firas Rashad Al-Samarai, Abdulkareem A. Al-Kazaz

The dramatic development of molecular genetics has laid the groundwork for genomics. It has introduced new generations of molecular markers for use in the genetic improvement of farm animals. These markers provide more accurate genetic information and better understanding of the animal genetic resources. Scientists, unfamiliar with the different molecular techniques tend to get lost as each has its own advantages and disadvantages. This review represents a trail to shade alight on the different types of molecular markers by introducing a brief summary on the development of genetic markers including both the classical genetic markers and more advanced DNA-based molecular markers. This review could be helpful to better understand the characteristics of different genetic markers and the genetic diversity of animal genetic resources.

156-173

Authors: Oleg Mosin, Ignat Ignatov

This paper deals with studying biosynthetic pathways of 2H-labeled purine ribonucleoside inosine excreted into liquid microbial culture (LC) by Gram-positive chemoheterotrophic bacterium Bacillus subtilis B-3157 while growing of this bacterium on heavy water (HW) medium with 2% (v/v) hydrolysate of deuterated biomass of the methylotrophic bacterium Brevibacterium methylicum B-5662 as a source of 2H-labeled growth substrates. Isolation of 2H-labeled inosine from LC was performed by adsorption/desorption on activated carbon with following extraction by 0,3 M ammonium–formate buffer (pH = 8,9), crystallization in 80% (v/v) EtOH, and ion exchange chromatography (IEC) on a column with AG50WX 4 cation exchange resin equilibrated with 0,3 M ammonium–formate buffer and 0,045 M NH4Cl. The investigation of deuterium incorporation into the inosine molecule by FAB method demonstrated incorporation of 5 deuterium atoms into the molecule (the total level of deuterium enrichment – 65,5 atom% 2H) with 3 deuterium atoms being included into the ribose and 2 deuterium atoms – into the hypoxanthine residue of the molecule. Three non-exchangeable deuterium atoms were incorporated into the ribose residue owing to the preservation in this bacterium the minor pathways of de novo glucose biosynthesis in 2H2O-medium. These non-exchangeable deuterium atoms in the ribose residue were originated from HMP shunt reactions, while two other deuterium atoms at C2, C8-positions in the hypoxanthine residue were synthesized from [2H]amino acids, primarily glutamine and glycine, that originated from deuterated hydrolysate. A glycoside proton at -N9-glycosidic bond could be replaced with deuterium via the reaction of СО2 elimination at the stage of ribulose-5-monophosphate formation from 3-keto-6-phosphogluconic acid with subsequent proton (deuteron) attachment at the С1-position of ribulose-5-monophosphate. Two other protons at C2(C3) and C4 positions in ribose residue could be replaced with deuterium via further enzimatic isomerization of ribulose-5-monophosphate into ribose-5-monophosphate.

56-62

Authors: Grigory A. Gerashchenkov, Gulnar R. Yasybaeva, Nataliya A. Rozhnova, Aleksey V. Chemeris

Apomixis is asexual seed reproduction without both meiosis and fertilization based on the complex developmental processes such as apomeiosis, parthenogenesis and specific endosperm development. This investigation is aimed at engineering of apomixis in Arabidopsis thaliana with sexual seed reproduction. The fragments of known genes of endosperm formation MEA, FIE, FIS2 and gene of apomeiosis DYAD (as control) were isolated using Q5 high fidelity DNA polymerase. These gene fragments of interest at the antisense orientation were fused with isolated constitutive and meiosis specific promoters of Arabidopsis at NcoI sites. The fused promoter-gene fragment modules were cloned in pCambia1301 at SalI cites. The engineered constructions will be used for the floral dip transformation of Arabidopsis and down regulation of these genes at engineering of apomixis.

80-87

Authors: Da´ad Ali Hussain

One of these major genes is growth hormone and growth hormone receptor (GH, GHR) genes which are related to production and reproduction traits in livestock. Associations were analysed between the bovine growth hormone (GH) as well as growth hormone receptor (GHR) combined with their genotypes (GH/AluI& GHR/AluI). There is extensive literature on the genetic polymorphism of GH and GHR in cattle, but perusal of literature has indicated paucity of information on these two genes in buffalo. This study aimed to evaluate the genetic polymorphism within growth hormone and its receptor genes in Iraqi buffalo using PCR-RFLP technique. Genomic DNA extracted from 100 healthy buffaloes and amplified using primers that were designed from the cattle GH and GHR gene sequences. All tested buffalo DNA amplified fragments at 428bp for GH 5th exon. The digestion with AluI endonuclease gave three types of genotypes. The Distribution of the three genotypes and allele frequency was calculated according to Hardy-Weinberg equation, were LL=94 (94.09%), LV=6 (5.82%) and VV=0 (0.09%), so that most of buffaloes was homozygous (LL) due to the presence of the restriction site at position 52^53 (AG^CT) and less heterozygous was for the valine allele as compared with leucine allele homozygous. The amplified fragments of GHR gene obtained from all tested buffalo DNA at 342-bp were digested with AluI endonuclease. The result showed that all tested buffaloes are genotyped as GG for GHR gene, where amplified fragments were digested into two digested fragments at 241and 101-bp due to the presence of AluI restriction site at position (AG^CT).

103-114

Authors: Sajedeh Saeedfar, Marzieh Negahban, Mohammad Mahmoodi Soorestani

Plot trials were carried out in a research field in Tehran (Iran) to determine the effect of drought stress on the essential oil content and some of the plant biochemical characteristics of Anise Hyssop (Agastache foeniculum [Pursh] Kuntze), which is a valuable medicinal plant. Drought stress was conducted at different levels including: well-water (100% FC), mild drought stress (85% FC), moderate drought stress (70% FC), severe drought stress (55% FC), 100% FC (vegetative stage) 85% FC (reproductive stage), 100% FC (vegetative stage) 70% FC (reproductive stage), and 85% FC (vegetative stage) 100% FC (reproductive stage). The experiment was arranged as a RCBD with three replications. The output results showed that, water deficit stress significantly (P≤0.05) increased activities of antioxidant enzymes (Superoxide Dismutase, Catalase, Glutathione Peroxidase) as well as Essential Oil yield and Abscisic Acid content. Lipid and protein oxidation (malondialdehyde and dityrosine contents) also increased significantly under severe water deficit stress. According to the results, severe drought conduction (55% FC) is the optimum level of soil moisture to plant Anise Hyssop under water deficit stress.

4-11

Authors: Rabah Chadli, Aman Bouzid, Khadidja Bouzid, Hamida Nader

This research concerns the study of antibacterial properties of different aqueous extracts of the bark of the pomegranate (Punica granatum L.). Three bacterial strains were used in this test: Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella. Very interesting bactericidal properties of aqueous extracts of the bark of the pomegranate were found on bacteria. The inhibition zones have a very large diameter up to 20 mm and the MIC and MBC are low, of the order of 0.78 mg/ml. This work has shown antibacterial activity against three bacteria may contribute to the fight against infectious diseases, and possibly offer the possibility of using pomegranate peel pharmaceutical and food industries.

27-36

Authors: Ali Saeed Atiyah AL-Janabi

In this study gene expression for DGAT1 gene was analyzed. Diacylglycerol acyltransferases (DGATs) catalyze the final step of the triacylglycerol (TAG) biosynthesis of the Kennedy pathway. Two major gene families have been shown to encode DGATs, DGAT1 (type-1) and DGAT2 (type-2). Gene expression were analyzed for 10 Olive cultivars (Olea europaea L.) (Khaderi, Qaysi, Manzenillo, Baashiqi, Arabqween, Nabali, Labeeb, Dahkan, Shami and Sorani). Different plant organs as plant materials (mature leaves, mesocarp and seeds for drups) used for analysis. Two methods for analysis gene expression were used, first method was called semi – quantitative and second method was called relative – quantitative, used in relative method (Real time PCR and Actine gene as Housekeeping gene). On the other hand chemical analysis was used on fruits like moisture % and oil % of dry and fresh weigh. The results revealed the following: DGAT1 gene expression in leaves, mesocarp and seeds by two methods (semi- quantitative and relative quantity) were the convergent results and clear, also if this results compared with chemical analysis shows that the best cultivars were Arabqween, Khaderi, Qaysi and Labeeb. The cultivars Shami and Khaderi then were contain in fruits desirable qualities of olive oil, low moisture and high oil percentages ratios. While Nabali, Manzanello, and Sorani cultivars middle desirable quantity, and Baashiqi and Dahkan cultivars had undesirable because of low oil quantity and high moisture in contain fruits. Some cultivars have low intensity in semi- quantitative and little fold in relative quantity but it have high oil in contain fruits that may be indicate that these cultivars were complete gene expression and begin to accumulation and save oil in tissue. Therefore particular emphasis was given to the temporal regulation of olive DGATs during drupe development. In olive fruit, TAGs are formed and stored in both the mesocarp and the seed .Two drupe compartments that have different physiological functions and roles and also display difference in the mode of TAG accumulation. DGATI share an overlapping expression pattern after 28 WAF, suggesting that they probably function at those stages. However, following maximal mRNA levels at 22 WAF, DGAT1 transcription declined substantially.