مجله زیست شناسی ایران
سال بیست و سوم شماره 5 (آذر و دی 1389)

There are a growing number of recombinant proteins that have been expressed in milk and one can consider putting any gene of interest under the control of the regulatory elements of a milk protein gene in a dairy farm animal. Nuclear transfer (NT) using donor cells are an efficient means for generation of transgenic animals. In this study in order to produce a transgenic goat with capability of secretion of recombinant human coagulation factor IX (rhcfIX) in the milk, a gene construct (pBC1-hfIX) was made by insertion of (rhcfIX) cDNA after goat beta-casein promoter into the expression cassette. Then, cell lines of goat fetal and adult fibroblasts were transfected by pBC1-hfIX. Fetal fibroblast cells isolated from day 35 to day 40 fetuses. Next, positive clones (the fibroblast cells that contain the gene construct) were selected and confirmed by PCR. The transfected fetal fibroblast cells were allowed to reach confluency and further cultured in 15% FBS in DMEM for an additional 72 h (full confluency) to synchronize G0/G1 cell cycle stage cells. Immature oocytes were obtained from slaughterhouse and matured before enucleation and NT. The reconstructed oocytes electrofused with donor somatic cells, and activated with ionomycin and 6DMAP. Reconstructed embryos were transferred into synchronized recipients. A total of 64 embryos derived from transgenic fetal fibroblast cells were transferred into 12 recipients. Two recipients (16.6%) were confirmed pregnant at Day 42 by ultrasound. Of these, two kids derived from the fetal fibroblast cell line. Presence of the coagulation factor IX gene was confirmed by polymerase chain reaction, sequencing, and fluorescent in situ hybridization analyses. These results demonstrate that goat fetal fibroblasts with human coagulation IX factor gene can direct full-term development following NT. This is the first live born of transgenic farm animal in Iran.

Dunaliella salina is unicellular green micro-algae living in lakes and marine habitats containing high salt concentration. This alga produces different cellular compounds such as beta-carotene under environmental stresses. In this research, the effect of both salt stress and salicylic acid on growth rate, photosynthesis pigments and beta-caroten content of the cells were investigated. Results showed in all experiments, the salicylic acid or combination of salicylic acid and salt stress decreased cell division rate. The presence of salicylic acid reduced the effect of salt stress on beta-carotene and chlorophyll content of the cells. The result indicated that salt stress can increase beta-carotene content. Both salt stress and salicylic acid due to decrease in cell division decreased chlorophyll and beta-carotene per volume unit.

Chromatin is a complex of DNA, RNA, histones and nonhistone chromosomal proteins. Identification of these proteins help us for better understanding of different mechanisms of gene expression. There are several methods for extraction of histones and nonhistone proteins from animal cells, but less information of a comparable state is available for plants. Also, biochemical evidence has shown that the DNA repeats length in micrococcal nuclease digests of the chromatin of higher eukaryotes varies widely. In this study, potato leaf cells, cell suspensions and calli were extracted with different methods using, acids and salts. Then the proteins were analyzed on SDS polyacrylamide gel electrophoresis and immunoblotted against histones and HMG antisera. The results show that histones H3 and H4 are relatively the same as their animal counterparts but H2A and H2B in potato are larger than their animal counterparts and exist as a several molecular weight variants. Also HMG proteins exhibit four main proteins in total tissue extract potato leaves and their electrophoretic pattern is different from mammalian HMG proteins. The potato leaf HMG proteins could not be detected by immunoblotting against thymus HMG antisera. Examination of extensive Micrococcal nuclease digest suggests that the DNA repeat length is approximately 158 base pairs which are shorter than the DNA repeat length of mammals and also other plants. It is concluded that there is little homology between the chromatin proteins and DNA repeat length of the animals and potato leaf cells.

Prolactin is a polypeptide hormone of the anterior pituitary gland that was originally named for its ability to promote lactation. Today we know that it has over 300 different function in vertebrates including osmoregulation, growth and development, metabolism, behavior, reproduction and immunoregulation. Recently, the role of Prolactin in immune system is investigated. Glucans are glucose polymers that are major constituents of the cell wall of fungi, plant and certain bacteria. In the present study, we used GH3/B6 cell line which is known to secrete Prolactin and growth hormone. We also studied the expression of β-glucan receptor, dectin-1, on this cells. Intracellular Prolactin levels were assayed by ELISA. Cells were treated with 5,150 and 250μgr/ml concentrations of β-glucan for incubation time of 48 hours. TRH (50 nM) was used as positive control and dextran (50nM), was our negative control. Therefore, Total RNA was extracted from spleen and GH3/B6 cells in the presence of β-glucan (250μgr/ml). Then, RT-PCR and PCR methods were done for verifying the expression of Dectin-1. Our result showed that 150 and 250 μgr/ml concentrations of β-glucan are capable to stimulation Prolactin synthesis significantly from GH3/B6 cells in comparison with control(P<0.05). The results showed that spleen, the positive control, expressed Dectin-1 as expected. Our results also showed that Dectin-1 was expressed in GH3/B6 cells at very low level, and this expression was increased under the effect of β-glucan. Therefore, β-glucan may induce GH3/B6 cells throughout Dectin-1 which lead increasing prolactin secretion from these cells.

The problem of the deterioration of monuments made of rock is particularly relevant incountries like Iran being rich in such cultural heritage. Stone surface of monuments can be considered as extreme environments that are exposed to high solar radiation, changing temperatures, low nutrient content and sporadic availability of sufficient moisture. These surfaces are colonized by diverse microbial communities which are frequently dominated by stress-tolerant Ascomycota. The ruins of Persepolis are covered by a well microbial growth. Lichens and fungi are known to actively decompose stone surfaces through both physical and chemical processes. Physical alteration of stones was studied by using light microscope, stereomicroscope, scanning electron microscope, maceration technique and periodic acid Schiff staining. In this work, penetration of hyphal bundle as well as individual fungi was observed. Biopitting, Etching, and Crumbling were the most biodeteriorating patterns which were detected in stones of Persepolis. Furthermore, rock inhabiting fungi from Persepolis was isolated. Morphologic, microscopic and molecular characterization of fungal isolates was carried out by using the classic method, DNA extraction and PCR. The results show that the black fungal isolates belong to the Chaetothyriomycetes and Lecanoromycetes which embrace most of black rock inhabiting fungi of Ascomycota. According to molecular analysis,Sarcinomyces sp., Bellemerea alpine, Rhinocladiella sp. and Cladophialophoraminourae was identified. These microorganisms can affect the substratum.

Pseudomonas is a gram negative bacterium which is able to biodegrade crude oil in contaminated soil and water. This bacterium can biodegrade the toxic organic materials to more simple and non-toxic materials. The respiratory chain of this bacterium contains different types of cytochromes which uses oxygen as final electron acceptor. The important cytochromes are a, b, c and o. The goal of this investigation is to study the changes of the different component of the Pseudomonas respiratory chain in the presence of different carbon sources. The methods include isolation of Pseudomonas aeruginosa from the soil and allow the bacterium to grow in the presence of different carbon sources and investigate respiratory chain changes. Our results showed that the respiratory chain of Pseudomonas aeruginosa in media with different carbon sources express different properties. In different media, cytochrome b and c synthesis increased more than other cytochromes, this is an indicative of cytochrome bo complex presence. Observing the bacterium spectrum in hexadecane and crude oil media showed that the bacterium also uses cytochrome aa3 as the final electron acceptor. These results suggest that the bacterium use oxygen as final electron acceptor in presence of crude oil and its derivatives.

A study on Cylindrospermopsis raciborskii was undertaken in the Anzali Lagoon, a shallow coastal wetland, during the growth seasons in spring and summer of 1992 and 2005. Water samples were collected on a monthly basis from four sampling stations for quantitative and qualitative analyses of phytoplankton species. A Ruthner sampler was used to collect water samples from a depth of 20 cm for the quantitative analyses of phytoplankton while water samples for the identification of plankton species were collected from the entire water column using a plankton net sampler with a mesh size of 25 microns. All samples were fixed in 2% formalin solution. C. raciborski, a freshwater species of the subtropical and pantropical regions was observed in the samples along with 19 other cyanophyta species. Masses of C. raciborskii consisted of filaments that were cylindrical, straight and narrowed towards the end. Cells were cylindrical or barrel shaped 2.5-12(16) × (1.7)2-4 μm. The cross walls were narrow, end cells conical or round. Heterocytes were drop-like with ovoid or pointed terminals. Akinetes were cylindrical or ovoid (7)8.5 - 18(22) μm. Formation of blooms is a distinctive feature of this species and populations of this species produce a kind of cyanotoxin. This species was found abundantly in the western region of the Anzali Lagoon where the pH was above 8. The species C. raciborskii comprising more than 70% of the species composition in the months of July and August was considered the dominant species in this region of the Anzali Lagoon. Owing to the lack of proper environmental management policies, a large amount of sewage (industrial, agricultural and domestic) enters the Anzali Lagoon everyday causing an increase in the organic load and eutrophication of this lagoon thus providing favorable conditions for the occurrence of heavy blooms of C. raciborskii. This will ultimately result in the mass mortality of aquatic organisms.

Due to comparison of different aspects with respect to plant species diversity, khanikan lowland forest was studied. In order to investigate of plant covers sixty plots (400 m2 for each) were taken by a systematic random sampled method. Plant species were identified using Braun - Blanquette method. Biodiversity indices of Simpson, Shannon Wiener, McIntosh, Margalef, Menhenic, Peet and Hill had been used for analysis. Results showed that the highest diversity and richness values were related to McIntosh and Margalef indices, respectively. The maximum and minimum of McIntosh indices devoted in West (70.713) and South (35.472) aspects. Also, the most and least Margalef indices is related to Southwest (1.827) and Southeast (1.346) aspects, respectively. The maximum of richness indices is related to Peet index in South, Northwest and Southeast aspects and Hill index in North, East, West, Northeast and Southwest aspects. Analysis of variance showed no significant difference among diversity indices with respect to different aspects.

Salvia leriifolia (Lamiaceae) is an endemic plant of Khorasan and Semnan province with antinociceptive, anti-inflammatory, antidiabetic and antioxidant properties. In this research, antibacterial activity of S. leriifolia root was investigated at different stages of growth and development and finally the best of harvesting time maximum antibacterial activity was determined. For this purpose, plant roots were harvested at vegetative (mid March), flowering (mid April) and ripening seed phases (late May). Different concentrations of metanolic extract (5000, 10000, 15000 and 20000 ppm) of roots were applied on five different bacteria (Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli and Proteus mirabilis) according to disk diffusion method. Inhibition zones were measured after 24 hours. Inhibitory effect of extracts at different stages were compared together and with compared certain antibiotics (Penicillin, Ampicilin and Tetracaiclin). Statistical analysis was performed according to the JMP software. The results showed that S.aureus and P.aeruginosa were the most sensitive and the most resistant bacteria, respectively. Maximum antibacterial activity of root was coincident with vegetative phase. In this phase, antibacterial activity of root was significantly higher than the flowering and ripening seed phases. Inhibitory effect of root extract at vegetative phase on S.aureus was higher than Penicillin and similar with tetracaiclin. Therefore, it seems that at vegetative phase (March) is the best time for obtaining the maximum antioxidant activity of root.

Carotenoids are unsaturated fatty acids composed of isopernoid units. Lycopene a carotenoid, with open-chain hydrocarbon exists in vegetables and fruits and recognized as a strong antioxidant in animal and plant systems. This compound has been received much attention for wide applications in pharmaceuticals, foods and health industries. The purpose of this study is to optimize extraction and quantification of Lycopene from Watermelon.Different ratio of solvents such as hexane, acetone, ethanol, tetrahydrofuran and as Di isopropyl ether were used to extract the freshly squeezed tissues of Watermelon. The extracted Lycopene was quantified using UV- visible spectrophotometry as well as HPLC methods. The best ratio of solvents for Lycopene extraction from Watermelon was found in hexane/ acetone/ethanol (16:7:7 V/V/V). Using this solvent, the amount of lycopene detected by HPLC 2.62 mg/l and by UV- visible spectrophotometry This was 2.32 mg/l. A proper ratios of the solvent, high quantify lycopene was extracted from Watermelon.

Fifty –eight species of the genus Salvia (Lamiaceae) are found in Iran of which 17 are endemic. The rate of endemism in the Salvia in Iran is 29 %. In this research, stem, leaf and petiole anatomical characteristics of eighteen Salvia species in West Azarbaijan were studied. Species were separated from each other by means of anatomical characters. These species had differences in the number of rays between the vascular bundles in the stem, the structure and the number of the vascular bundles in the petiole. Characters as: type of leaves, trichome features, size and type of stomata were applicable in delimiting species.

During an ecological study in biomasses of Zayandehrood River, five stations were selected. During autumn 2004 to summer 2005 sampling were repeated three times with Grab Walkman method so the amount of organic matter was observed. In this survey, samples were separated and counted. The result showed that the dominant aquatic organism in the river is oligochaetes which were observed in all stations. After oligochaetes, Chironomidae were the most abundant. Tubificidae was seen in all stations and its maximum density was 3635 organisms in each square meter. Maximum biomass of tubificidae was observed in summer with the average figure of 4.06 g/m2 and minimum biomass was observed in winter with average biomass of 0.89g/m2.Maximum and minimum biomass of Chironomidae were recorded in spring and autumn with average biomass of 2.24 g/m2and 0.33 g/m2 respectively. Maximum (3.15 g/m2) and minimum (0.29 g/m2) biomass of benthic organisms were observed in stations 5 and 1.

Savadkooh is one of the cityies of Mazandaran Province, and Zirab is one of its towns. The weather of Zirab is relatively cold, and the relative humidity is not high. The average annual temperature is 17ºC and the average rainfall is around 600mm. This study has been carried out in Shahid Beheshti University Science Park of Zirab. Sampling has been done all over the area, during sunny days, especially one or two days after rainfall, and in the spring, summer 2005, and In this investigation on Rhoplocera suborder from Lepidoptera order, 24 species have bee identified. All of them be included in four families of the Papilionoidae superfamily as follows: One species in Papili, 7 species, and subspecies in Pieridae family, 3 Species, in Lycenidae family, 13 species, and subspecies in Nymphalidae family. The specimens heve been kept in zoology collection of Researeh Institute of Environmenta Sciences of Shahid Behoshti University.

The effect of water deprivation on camel blood parameters was conducted on four lactating camel which were used within latin squre template with four treatment and four repeat (4×4). Treatments consisted of the camels received water daily (treatment 1”control”), after 2 days (treatment 2), after 4 days (treatment 3), after 6 days (treatment 4) respectively, for 2 hr. The experiment was rotatorly accomplished during 4 phases that each phase included 15 days as a adaptation period and 12 days as a test days. During experiment, camel food ration was invariant. blood samples were taken once end of per 12 days period of experimental. camels’ weight, water intake, dry mater intake, blood parameters (Red blood cells, White blood cells, Hemoglobin, Hematocrit and Glucose) were measured. camels lost about 10 percent of their body weight during 6 days water deprivation (P<0.01). Dry matter intake significantly decreased in fourth group to other groups (P<.05).Water intake were different between groups, significantly (P<.01). But blood parameters did not indicate any difference between groups, significantly (P>.05). it can be concluded that 6 days water deprivation dose not have any ill-effect on camel blood parameters.

This investigation was carried out to compare performance of seven new silkworm lines including Japanese origin lines of Xinhang1, Xinhang2, Xinhang3, 101433 and Chinese origin ones including Koming1, Koming2, Y at three rearing periods using generalized linear model procedure. Survived larvae number, survived pupae number, pupation rate, and produced cocoon number were studied as economic characters. Also, good cocoon percentage, middle quality cocoon percentage, low quality cocoon percentage, and double cocoon percentage were studied as economic characters. Furthermore, total best cocoon weight, total cocoon weight by 10000 larvae, larval duration, total produced cocoon weight, single cocoon weight, single cocoon shell weight and cocoon shell percentage were studied as economic characters. Results from this study showed that Chinese lines have a higher genetic resistance than Japanese ones, thus Chinese varieties can be used as donor of resistance genes in crossbreeding programs. Total best cocoon weight for Chinese group was higher than Japanese ones in spring 1381 (P<0.05).Total cocoon weight by 10000 larvae was significantly different between Japanese group (12851 gr) and Chinese one (15355 gr) only in autumn rearing (P<0.05). Thus, for Chinese varieties have higher potent for unfavorite conditions. Generation had significant effect on total produced cocoon number, best cocoon percentage, and middle quality cocoon percentage. Also, generation had significant effect on low quality cocoon percentage, total best cocoon weight, and total cocoon weight by 10000 larvae (P<0.0001) and larval duration (P<0.001), while it had not significant effect on double cocoon percentage. At total rearing periods, Xinhang3 (34.16%) and Y (19.42%) had higher and lowest middle quality cocoon percentage. At autumn rearing, Xinhang2 (15.23%) and at spring rearing, 101433 (7.18%) had more low quality cocoon percentage.