MODERN GENETICS JOURNAL (MGJ)
Year:2012 | Volume:7 | Issue:3 (30)|Papers Count:19

Medicinal plant had uses as a major source of medicine since thousand years ago. Using approach such as cell, organ and tissue culture, as well as genetic engineering and molecular markers, biotechnology is able to extend application and effacing of medicinal plant as renewable sources in production of medicine. Cell, tissue and organ culture, has facilitated the massive and rapid propagation of a large number of medicine plant. Propagated plants through in vitro methods are free-disease and uniform in terms of quality and genetics. Cryopreservation (Long-term Conversation in liquid nitrogen) is a useful approach in preserving endangered plant spices. Over recent years, cell and organ suspension culture has gone notices for providing secondary metabolites and studying metabolite biosynthesis pathway. So far, cell culture for a wide range medicinal plants have been investigated, and fundamental compounds such as flavonoids, tannin, alkaloids have been produce. Some of the most significant factors in increasing production of secondary metabolites in cell culture are as follows. Genetic engineering has played in important role in identification and genetic modification of enzymes, involved in metabolic biosynthesis of secondary metabolites with limited efficiency yield. Due to their independence on age, physiological and environmental conditions, molecular markers are plant special and studying genetic diversity, classification of inheriting pool and determination of genetic map. This article is a review of different biotechnological approaches which are effective in improving efficiency and production of high quality.

Human P-glycoprotein (P-gp) is encoded by the MDR1 gene, which is located on chromosomal region 7q21 and consists of 28 exons. To date, over 50 single nucleotide polymorphisms (SNPs) have been reported for the MDR1 gene this gene is very polymorphic and is known that these polymorphisms not only may affect the expression level of this gene but also may alter the pharmacokinetics of substrate drugs. We aim to analyze the MDR1 gene C3435T and C1236T polymorphisms in Iranian population. The genotyping was performed by the ARMS and RFLP methods in 120 Iranian individuals and we use Real time RT-PCR to measure the expression level of MDR1 gene. The frequencies of the wild-type C3435 and mutant T3435 alleles were 43 and 57%, respectively and the distributions of CC, CT, and TT genotypes were35, 44 and 21%, respectively. For C1236T polymorphism C and T alleles frequencies were 44 and 56% respectively and CC, CT and TT genotypes frequencies were 13, 63 and 24%. There was no considerable association between MDR1 expression level and C3435T or C1236T polymorphism in our results. Our study provides a framework for future studies concerning the role of polymorphic variants of MDR1 gene in the drug responses in various diseases.

In this study 71 specimens were collected from the Haraz, Shirud and Gazafrud rivers in Mazandaran and Guilan Provinces. The mtDNA was extracted from fish fin using Phenol-Chloroform method. Genetic variation and probable population differentiation of Alburnus chalcoides were studied based on the mitochondrial cytochrome b gene. The specific primers were designed for A. chalcoides and the PCR experiments were performed. Nine restriction endonuclease enzymes were applied for RFLP analysis (BamHI, EcoRV, HaeIII, HincII, HinfI, PpuMI, RsaI, Tsp45I, and TaqI). PCR products (831 bp) and DNA digests were subjected to Agarose gel electrophoresis to separate fragments according to their molecular weight. A part from TagI the rest patterns were identified similar to all specimens from Gazafrud River. Regarding to this patterns, it can be concluded that polymorphism phenomena cannot be observed by above mentioned enzymes and cytochrome b gene, and there is no separate population of A. chalcoides in this study.

Common beans (Phaseolus vulgaris L.) are susceptible to drought stress especially in reproductive stage. A series of genes are identified recently in which their transcription either increase or decrease in response to drought stress. Out of these genes, identified in common bean, could be referred to LEA gene. The present study was conducted at Shahrekord University aimed to analyze LEA gene expression in response to drought stress at vegetative and reproductive stages in 11 beans genotypes. Water deficit stress was induced at vegetative stage with the appearance of the third trifoliate and at reproductive stage when flower buds were passing through meiosis. The leaves and flower buds were sampled from control and water-stressed plants at vegetative and reproductive stages, respectively. Results indicated that drought stress at either of vegetative and reproductive stages increased the expression of the gene LEA, but the level of expression was not similar in all genotypes. The increment in the expression of LEA gene was 75 and 95 fold in leaf tissues of Goynok98 under stress at vegetative stage in comparison with the gene expression of its control and control of the susceptible line G-14088, respectively. However in flower buds of line KS-21191, the increment in the expression of LEA gene due to stress was 366 and 250 fold of that in its control and control of the susceptible line, respectively. The expression trend of LEA gene was similar in cultivars Kara and Daneshkadeh. The lines KS-21189 and KS- 21191also showed similar expression trend. Considering the trend and the level of expression for LEA gene, it could probably be concluded that the line KS-21191 is more tolerant to stress than other studied genotypes.

Pomegranate (Punica granatum L.) is one of the oldest known fruits and the most important horticultural plants in Iran with a considerable distribution and diversity. Different molecular markers have been used to determine the genetic diversity among some pomegranate cultivars such as RAPD, AFLP and SSR, indicated a low level of polymorphism among its genotypes in spite of morphological diversity. In current study petA-psaJ region (petA, psbJ, psbL, psbF, psbE, petL, petG, tRNApro, trnAtrp, psaJ) was used to sequence and phylogenetic analysis of pomegranate and myrtle. Genomic DNA was extracted from the wild, cultivated and ornamental pomegranate genotypes and a wild type of myrtle. Sequencing of PCR products showed that this region is consisted by 5400 bp and psbE-petL region with 4.98% diversity can be used as an appropriate marker for detection intra-specific variation of pomegranate. Comparing pomegranate with the other plants based on petA-psaJ region revealed a considerable genetic relation between pomegranate from Onagraceae and Mythraceae families.

The number of Bactrian camels has been decreasing dramatically during last decades in Iran, whereas very little is known about its genetic diversity. All of these animals are found in northwest of Iran (Dasht-e-Moghan). In the present research, we used 7 microsatellite DNA markers (LCA33, LCA56, LCA63, LCA77, VOLP03, VOLP67, YWLL08) to characterise 110 camels. Blood samples were collected from existing camels in Dasht-e-Moghan and stored at -20oC for further analysis. DNA was extracted using modified salting out method. Out of 7 loci, polymorphism were foun at 6 loci in this population. Locus LCA77 was monomorphic. All loci deviated from Hardy-Weinberger equilibrium (0.001). Genetic diversity expressed as mean number of effective allele numbe (NE), observed (HO) and expected heterozygosity (HE) and PIC were estimated 2.622±0.534, 0.714±0.184, 0.489±0.114 and 0.557±0.237, respectively. In this population, LCA33 and VOLP67 loci had the lowest (0.165) and the highest (0.771) heterozygosity and lowest (0.151) and highest (0.742) PIC value, respectively. These results suggest that however Iranian Bactrian camels has the property genetic diversity but for more clarification of genetic structure of this population, it is necessary to investigate additional microsatellite DNA markers.

Drought is one of the most important constraints on the growth and productivity of barley (Hordeum vulgare L.). A proteome analysis based on 2-D gel electrophoresis was performed in order to identify drought stress responsive proteins. Pot experiments were arranged in randomized block factorial design with two watering treatments, control and drought conditions, and three barley genotypes. Plants were well-watered until the four leaf stage and then exposed to severe drought stress for 10 days. The overall effect of drought was highly significant (P<0.01), as determined by dry matter, fresh matter, relative water content and proline free content. To study protein expression changes in response to drought stress, proteome analysis was performed based on DIGE technology with the inclusion of an internal standard. About 768 spots were detected on CyDyes gels, of these 37 spots showed significantly changes as genotypes or genotype ×irrigation levels effects. Finally mass spectrometry analyzing using MALDI-TOF-TOF led to the identification of 19 proteins. The functional of these proteins were identified as defense, energy, metabolism, cell structure, signal transduction and cell differentiation.

One of the hypotheses of cell damage after cryopreservation is the deleterious effects of oxygen free radicals generated during freezing and thawing. This study evaluated the protective effect of Vitamin E supplementation as antioxidant on post-thaw DNA integrity of beluga (Huso huso) sperm. Collected sperm was divided into three parts and using DMSO alone as control group or in combination with vitamin E supplementation in 600 and 1200 mM has been diluted and was maintained for 30 days in liquid nitrogen storage tank. We used the Comet assay, a microelectrophoretic method to detect DNA damage in response to cryopreservation process in beluga spermatozoa. This technique was used to assess DNA integrity in the cells by measuring damages reflected as strand break. According to the results DNA damage rate in control freeze-thawed sperm was significantly higher than levels measured in fresh sperm. Adding the vitamin E had a dose dependent effect on the amount of DNA damage, as in 600 mM concentration, non-significant reduction in the level of damage than the control group was observed, but in 1200 mM, probably due to the pro-oxidant properties of the high-dose antioxidants significant increase in damage was assessed. In addition, the results indicated that the extent of damage to sperm motility caused by freeze-thawing was correlated with DNA breakage.

Root rot disease is one of the most important sugar beet disease in Iran that infected by Pythium aphanidermatum Fitzp. For studying genetic diversity of this pathogen, 10-20 samples were collected from sugar beet fields in eight provinces of Iran. After purification, isolates were identified based on morphological characteristics of sexual and asexual organs and 19 isolates were selected in order to molecular diversity studies. ISSR analysis of all isolates was performed using eight primers. Results of phylogenetic tree of primers based on UPGMA method and Jacards similarity coefficient, were categorized isolates into five groups (64% similarity). The results of analysis of growth rate in two temperatures at 26±2Co and 29±2Co, by MSTATC and MVSP, was categorized isolates in to two groups with low growth and high growth rate. Analysis of morphological and molecular variation were indicated that isolates of this eight geographical regions have genetic relationship but there isn’t necessary relationship between growth rate of isolate and their geographical distribution.

Sturgeon fish are valuable species, especially for Caviar and also meat production purpose. Because of the long maturation period, sex determination at the early stages of their life could significantly increase the profit of the culturing system. Moreover understanding the sex determining system of the sturgeon is still questionable and it is necessary to distinguish the sex related fragments. In this study the genome of Iranian sturgeon Acipenser persicus was assayed with AFLP marker and sixteen primer combinations. Results revealed no sex specific markers. In addition, the primer combination named (P-ACA; M-CAG) had the highest number of fragments and polymorphism indices and could be used in other studies on the genome of this animal. Our results suggest that either the sex chromosomes do not exist or if exist, they are weakly differentiated in the sturgeon genome.

In this study, genomic DNA contents of part Iranian wild diploid wheat (Triticum monococcum) collection were measured through flowcytometry. This collection included 21 morphotypes of T. boeoticum subsp. boeoticum, 53 morphotypes of T. boeoticum subsp. thaoudar and 19 morphotypes of T. urartu. The pick indices of different samples were highly divergent to elucidate a variation in their genomic DNA content; however none of them was similar to polyploidy checks. Some accessions with low and high indices of flowcytometry pick were selected for karyotypic studies. Mitotic karyotype of each accession was studied by estimation of means of total length of chromosomes, arm ratios and centromeric index. Mitotic karyotyping analysis indicated that all the studied accessions encompass the expected chromosome number of diploid wheat, 2n=2x=14. Aneuploid karyotype was not found in any sample. Therefore, it seems that the variation in total genomic DNA content could be due to some changes in the structure of A genome chromosomes of wild Iranian Triticum monococcum. Variation in karyotipic characters, e.g. chromosome size, centromeric index and arm ratio, deduce evolutionary changes in the structure of chromosomes as plant species evolve and diverge. Further elucidation of the observed variation necessities complementary studies using chromosome banding techniques.

Tobacco is one of the agricultural and industrial plants which plays important role in economy and revenus of producer contries. Potato Virus Y (PVY) is one of the most Dangerous Viral diseases on tobacco plants and reduces the quality and quantity of tobacco crops. The best and most industrial method for PVY disease is production of resistant varieties. Improvement for resistance to this disease from classic methods is difficult and need many expancess. DNA markers Linked to resistance gene and have high polymorphism (Like SSR Markers) can be useful in acceleration of resistant varieties Breeding programs. This research has been done for studying the probability accesses to Marker-assisted Selection (MAS) for resistance trait to PVY disease in tobacco with SSR markers. For this aim, in Farvardin 1389, cultivation of parental seeds (VAM Variety) which resistant to PVY and (K326 Variety) as a susceptible parent to PVY were done. DNA of resistant and susceptible parents Leaves, were extracted, Separately., and after of performance polyacrylamid electrophoresis and Gel staining by silver nitrate solution ,among of them,28 pair primers have the most polymorphisms on parents, were selected and screened .Then, this 28 SSR markers were tested on resistant and susceptible Bulks to PVY disease .The final results showed that 3 SSR markers (PT20357), (PT30002), (PT20165) have the good polymorphism on resistant and susceptible Bulks to PVY. These markers can be useful to MAS for resistance or Tolerance to PVY in tobacco.