Journal of Cell and Molecular Research

Number 2(2), 2010

56-60
Hemocyanin subunits composition as a molecular marker for identification of Intra-specific variations of Gammarus

Authors: Saber Zahri, Gadir Nouri-Ganbalani and Shahram Agahi

Abstract

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Crustacean hemocyanin represents a unique case of molecular heterogeneity among oxygen-carrying proteins. Two populations of Gammarus lacustris and six populations of Gammarus komareki were identified and isolated from aquatic habitats of Ardabil (northwest of Iran). The populations were nominated based on the name of the isolated region. The Principal component analysis of morphological absolute attributes showed that the populations were relatively dissociated. The Meshkin population of G. komareki was completely different from the other populations. The analysis of relative attributes did not show any dissociation. The Hemolymph profile analysis using native PAGE revealed 5 to 7 hemocyanin subunits for four populations of G. komareki and 8 subunits for two populations of G. lacustris. The Detection of copper ions using bathocuproin sulfonate on the native PAGE surface confirmed the hemocyanin bands. Using SDS PAGE, 1-3 and 1 protein bands were separated for hemolymph samples from G. lacustris and G. komareki, respectively. Molecular weights of the bands were 75 - 80 kDa. Comparison of our results with the previous published data confirms the species specificity of hemocyanin subunit composition. These results imply that the ecological variation may affect the morphological traits, but the hemocyanin profiles are useful as a molecular marker for the taxonomic studies and identifying very closely-related species of Gammarus.

61-66
Irrigation effects on pds and bch genes expression of the Iranian Saffron

Authors: Nasrin Moshtaghi, Robab Ghahremanzadeh and Seyyed Hasan Marashi

Abstract

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Number of views: 377

Saffron (Crocus sativus L.) is an indigenous and the most valuable and marginal plant in Iran. However, limited knowledge exists on its molecular biology. The importance of this plant is due to the color, flavor and medicinal properties of its red stigmas. Saffron stigmas contain a high amount of carotenoids such as crocetin and crocin. Two genes, bch and pds, have essential role in carotenoid production. In this study, the effect of four different irrigation regimes was evaluated on the expression of bch and pds genes. Semi-quantitative RT-PCR showed no significant difference in the expression levels of genes of interest related to the internal standard (18S rRNA). Results of Real-Time PCR assays showed that the expression of bch and pds genes were affected by irrigation treatments as their expression decreased in irrigated plants in comparison to non irrigated ones, exept for one irrigation treatment (one irrigation in September) where the pds gene expression showed higher level. However, the expression profile of the genes was almost the same in all treatments. The comparison between results of two techniques indicated that the Real-Time PCR is more accurate for determination of the level of transcript in the Iranian saffron. It was interesting that by decreasing of irrigation, the expression level of these two genes increased indicating that abiotic stress and drought can affect on the gene expression relating to the saffron color.

67-73
Association of growth trait and Leptin gene polymorphism in Kermani sheep

Authors: Moslem Shojaei, Mohammadreza Mohammad Abadi, Masood Asadi Fozi, Omid Dayani, Amin Khezri and Masoumeh Akhondi

Abstract

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Number of views: 352

Identification of genes affecting energy balance, milk yield and feed intake is an interesting area of researches in animal breeding. Leptin gene polymorphism is associated with key economic affair. Considering rich resources for animals, in our country, accomplishing a few assays to identify a gene that controls her traits with molecular genetics, and identifying the candidate genes in sheep breeds using DNA test can greatly help to her breeding progress. For analyzing Leptin gene polymorphism and its association with growth traits in Kermani sheep, blood samples of 120 sheep of both genes rearing at breeding centre of Shahre Babak were taken. In addition growth traits were measured. PCR was performed to amplify 275 bp fragments of exon 3 from Leptin gene. Then Single Strand Conformation Polymorphism (SSCP) of PCR product was performed and Leptin band patterns (genotypes) were obtained using acrylamid gel and silver staining. For Leptin gene 10 genotypes including A/A, C/C, A/B, A/C, A/B/C, A/B/E, A/B/F, A/C/F, A/B/D/E and A/B/C/F were obtained. The results of this study showed that the growth traits are significantly affected by the genotypes. Accordingly, A/B/E, A/C, A/B/C/F and A/B/C/F genotypes had higher body weight at 3, 6, 9 and 12 months of ages respectively. The animals with A/B, A/B, A/B/F and A/B/D/E genotypes had the smallest body weight at 3, 6, 9 and 12 months of ages respectively. It is suggested that polymorphism in Leptin gene loci can be used as a selective criterion to improve growth traits in Kermani sheep.

74-80
Proline, sodium and potassium concentration changes in gamma rays and NaCl treated potato calli

Authors: Behrokh Shojaie, Ali Akbar Ehsanpour* and Mohammad Reza Abdi

Abstract

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In this study, the effects of gamma rays on changes of proline, sodium and potassium in potato calli (Solanum tuberosoum L.) cultivar "White Desiree" were investigated. After irradiation of calli by 60 and 80 Gy of gamma radiation, irradiated and non irradiated calli were transferred to a callus production medium containing 0, 30, 60, 90, 120 and 150 mM NaCl. After 26 days post treatment amounts of proline, Na+ and K+ were measured. The results showed that generally in each of gamma ray treatment, amounts of proline at 120 and 150 mM NaCl were significantly more than 0, 30, 60 and 90 mM salt. A significant difference observed only at 150 mM NaCl between irradiated and non irradiated calli. This result could be related to the proline role in osmotic adjustment at high concentration of salt. The amounts of sodium increased by increasing salt the concentration but the amount of potassium decreased. These results showed that the sodium influx inhibited the potassium uptake, and may be because of strong inhibitory effect of sodium on the potassium uptake system or efflux potassium from the cells. The Na+/K+ ratio decreased at 150 mM by 60 and 80 Gy of gamma radiation.

81-85
Conservation of Colutea gifana, a rare and potential ornamental species, using in vitro method

Authors: Mahnaz Kiani, Homa Zarghami, Ali Tehranifar and Farshid Memariani

Abstract

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Number of views: 356

In vitro methods provide a variety of tools to supplement traditional methods for collection, propagation and preservation of endangered plant species. In this study, an efficient protocol was developed for in vitro propagation of Colutea gifana, a rare and endangered plant species with limited reproductive capacity that grows in a narrow area of Iran. Single node explants were used for a series of experiments to select the appropriate disinfection method and growth regulators for establishment, proliferation and rooting stages. Explants showed the highest establishment percent after treatment with 2% Sodium hypochlorite (NaOCl) for 15 min, cultured in MS medium containing 2.2 µM 6-benzylaminopurine (BA) and 1 µM indole-3-butyric acid (IBA). In proliferation stage, 8.8 µM of BA was more effective cytokinin than Kinetin (Kin) and Thidiazuron (TDZ) for growth induction of axillary shoots. In vitro rooting of proliferated shoots was induced in half- strength MS medium in all concentrations of both tested auxins i.e. IBA and α-naphthalene acetic acid (NAA). Eighty percent of the plantlets were successfully acclimatized to ex vitro conditions, showed normal development. These plants were used to replenish declining populations in the collection sites and conserve C. gifana from extinction.

86-92
In vitro selection for NaCl tolerance in Thymus vulgaris L.

Authors: Akram Zia, Farkhonde Rezanejad and Abbas Safarnejad

Abstract

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Number of views: 318

Thyme (Thymus vulgaris L.) is an aromatic and medicinal plant which is very important for the herbal industry. In this research, in vitro selection for NaCl tolerance was investigated in this plant. Hypocotyl and shoot explants (apical meristem and cotyledonary leaves) of sterilized seedlings were cultured in MS medium supplemented with different combinations of 2, 4-dichlorophenoxyacetic acid (2, 4-D), napthaleneacetic acid (NAA), kinetin (KIN) and N6-benzyladenine (BA) for callus induction. Results showed that the maximum frequency of callus induction was obtained on MS medium containing 1:0.5 ratio of 2, 4-D:Kin (mgl-1), while optimal callus induction with the best quality and regeneration potential was achieved in 1:1 ratio of NAA:BA (mgl-1). The calli grown in 1:1 ratio of NAA:BA (mgl-1) were transferred to NaCl supplemented medium at 50, 75 and 100 mM concentrations. Fresh and dry weights, percentage of necrosis and regeneration of calli were determined after 4 weeks. There were significant differences between fresh and dry weights of calli in different concentrations of NaCl. The highest and the lowest of fresh and dry weights of calli from hypocotyl explants were observed in 0 (control) and 100 mM NaCl concentrations, respectively. But fresh weight of shoot-derived calli induced from shoots reduced only in 100 mM NaCl and the dry weight of them was not significantly different. The necrosis percentage increased by increasing the salt concentration. Callus regeneration just occurred at the concentration of 50 mM of NaCl and all regenerated shoots well rooted on half strength MS medium, with 0.8% (w/v) sucrose and without growth regulators.

93-102
The Effect of Monte Carlo, Molecular Dynamic and Langevin Dynamic Simulation and Computational Calculations on Insulin-like Growth Factor- 1(IGF-1)

Authors: Reihaneh Sabbaghzadeh, Majid Monajjemi, Fatemeh Mollaamin and Shahrbano Oryan

Abstract

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Number of views: 487

Insulin-like growth factor (IGF-1) is an anti-apoptosis factor in multiple cell types associated with various cancers. Computational methods allow investigating the systems between 50–100 atoms in the frame of quantum mechanics and up to 50,000 atoms with molecular dynamics. Since there are specific interactions between the residues, the solvent could play an important role in the stability of the native structure. Therefore it is useful to carry out such simulations at atomistic detail. MC, MD and LD simulations of the IGF-1 were performed with the HyperChem7.0 program. The geometries, and the interaction energies, bonds, angles, stretch-bends, electrostatic and the VDW Interactions were carried out in solution and gas phase. We have computed the transition temperature for the IGF-1 molecule. Studying the changes occurred in the potential energy of the three force fields showed that Amber force field is better than MM+ and OPLS force field and also MD simulation, at least in this model, is more effective than MC and LD methods. After equilibration, the MD simulation was very stable, and the difference between the relation coefficients R2=0.8173 in gas and R2=0.7558 in water was compared. The Pearson correlation suggests that there is an inverse relationship (R=- 0.25) between in vitro temperature and stability of the structure.