JOURNAL OF AGRICULTURAL BIOTECHNOLOGY
Year:2017 | Volume:9 | Issue:1|Papers Count:8

The Insulin-like growth factor I (IGF-I) has a similar structure with Insulin hormone.This hormone which is transcribed by (IGF-I) gene has an important functional in growth and development of different body tissues. The aim of this study was to investigate the polymorphism of this gene and its relationship with the estimated breeding values of the birth type, birth weight and fleece weight in Raieni Cashmere goats. Breeding values were estimated using records on 13020 Raieni cashmere goats originated from 336 sire and 3617 dams. Univariate animal model was applied for the genetic analysis of the investigated traits using ASReml. To study IGF-1 gene polymorphism, 94 Raieni cashmere goats were selected based on their estimated breeding values for these traits. PCR was used for amplification of 363 bp fragment of exon 4 of IGF-I gene. Thereafter the animal’s genotype was determined using PCR-RFLP. Three genotypes including AA, AB and BB were observed. The frequency of these genotypes was 0.22, 0.65 and 0.13, respectively. No significant associations were found between these genotypes and the investigated production and reproduction traits. The results of this study show that, there is important genetic diversity for the IGF-I in the investigated population but to study the association between the polymorphism of this gene and the studied traits, more data should be used as currently few animals were genotyped.

Endophytes are symbiont microorganisms within apoplast or symplast of plants that promote growth, control phytopathogens, increase plant resistance to stress and improve plant growth conditions. The aim of this study was identification of endophytes from six rice cultivars in Khouzestan province and assessment of their effects on growth of the host plant.For this purpose, a suspension was prepared from sterilized seeds, and their endophytes were isolated in LB broth. The ability of isolates in nitrogen fixation, auxin production and phosphate solubilizing was discovered. The seeds of Champa cultivar were then infected by endophytes and growth and physiologic outcomes of this treatment were studied. As a result of this study six endophytes belong to Bacillus, Staphylococcus and Erwinia genus were isolated from them only the Erwinia sp. was able to fix N2, dissolve phosphate and produced auxin more than others. Treatment with Erwinia sp. significantly increased the fresh and dry weight of aerial parts and root and aerial part length and density of hair roots. The auxin and carbohydrate content of treated rice with Erwinia sp. had significant difference than control.No significant difference was observed for chlorophyll a and b chlorophylls in different treatments and untreated plants produced more a and b chlorophylls. While there was significant difference for carotenoid between endophyte treated samples and untreated ones.Finally based on the obtained results it can be suggested that Erwinia sp. as an endophyte has significant effect on increasing of Champa cultivar performance and improvement main characteristics of this cultivar.

Transgenic plants are suitable bioreactors for the production of recombinant proteins at high levels and qualities. Cloning of a gene encoding the target protein in proper expression vectors is one of the most important steps for successful integration and expression of target protein in plants. The purpose of this study was to clone insulin-like growth factor -1 (IGF1) cDNA in plant expression vectors. To this end, we amplified amplify IGF1 cDNA using a pair of specific primer followed by its cloning in pMCS5 basic vector. After sequencing, because of several advantages of cereals for production of biopharmaceuticals, such as high cultivation area and biomass, possibility of grain storage and usage as food and feed, we constructed suitable vectors for IGF1 integration and expression in cereals. In addition, as transgenic chloroplasts contain higher potential to produce elevated levels of functional recombinant proteins, the IGF1 cDNA was recloned in two chloroplast-specific vectors. In one of these vectors, selectable marker gene is flanked by two direct repeats of LoxP sequences, providing the marker gene excision after production of transgenic plants via Cre/Lox system.

To investigate the effects of dietary zinc and betaine (Bet) substitution to methionine (Met) on hepatic betaine - homocysteine methyltransferase (BHMT) and lipogenic (malic enzyme and fatty acid synthase) genes expression in laying hens under heat stress by using real-time qPCR, an experiment was performed with 288 Hy-line W-36 leghorn (at 50 to 62 weeks of age) in complete randomized design (CRD) with a 3×2×2 factorial arrangement of treatments including twelve treatments, 3 replicates and 8 hens in each replicate. There were 6 dietary treatments: three doses of Bet (0, 13 and 26%) substitute to Met combined with three levels of zinc (90 and 100 mg/kg) supplemented to the basal diets that were combined with two environmental conditions (thermoneutral and heat stress conditions). Results showed that dietary supplemental zinc had no effect on BHMT, malic enzyme and fatty acid synthase (FAS) genes expression. The results confirmed that Bet substitution to methionine due to a 2.11-fold increase (P<0.01) compared to the control in BHMT gene expression whereas; it due to a 2.47 and 1.98- fold decrease (P<0.01) compared to the control in malic enzyme and FAS genes expression.Moreover, BHMT, malic enzyme and FAS genes expression was significantly reduced 1.58, 1.96 and 2.15 fold by heat stress when compared with control group, respectively (P<0.01).Therefore, betaine can reduce the harmful effects of heat stress by increasing the expression of BHMT gene and reducing the expression of malic enzyme and FAS genes in laying hens under heat stress.

The use of resistant cultivars is one of the safest and most profitable methods of controlling post including alfalfa weevil. After farm and greenhouse evaluating, Tafresh 42 genotype for molecular studies selected and evaluated to examine the pattern of protein expression in stress and non-stress conditions to alfalfa weevil. The Changes in protein expression were investigated using two-dimensional electrophoresis gels. Clustering analysis, principal component analysis and discriminant analysis were used in statistic studies. The results showed that 11 protein spots had expression changes among 218 protein spots. The most expression pattern of proteins with a similar increase in Tafresh 42, was related to a specific group of proteins which had 45.46% variations in the expression response to the stress. From the point of protein expression, cluster analysis caused a dividing of proteins into three main clusters indicating the presence of proteins with a similar performance in each cluster. Therefore, the presence of these proteins in a common biologic pathway is confirmed. The analysis of discriminant function confirmed 100% the results from clustering that indicated protein conformity with the conditions of the experiment. It is concluded that by using statistical analysis and software, it is possible to evaluate significant expression changes induced damage alfalfa weevil in proteome level and determine the index changes, and after that genetic engineering is utilized for enhancing the level of resistance in the plant under studying.

Pseudomonas putida is a bacterium that has ability to growth on limited substrates that mainly is alkanes. The ability to use wide range of hydrocarbons is advantage of this bacterium to other bacteria community. P. putida have alkane hydroxylase system (alk-B) for alkane biodegradation.In this study alk-B gene from P. putida was amplified. Blunt cloning first done in pBluescript plasmid then sticky end cloning was carried out in pET-26 vector. For expression of this recombinant gene E. coli BL-21 bacterium were used. The activity of recombinant enzyme was done by reduction of absorbance by oxidation of NADPH. The results of this study show that alk- B gene was inserted in pBluescript plasmid. Colony PCR confirmed this insertion. This gene successfully cloned in pET-26a vector. By increasing the time after induction by IPTG expression of alk-B protein were increased. Recombinant protein has activity. Transfer of this gene to fast growth bacterium and wide substrate confirm production of robust biocatalysts in the future.

Considerable progress has been made toward identifying and cloning genes involved in plant defense responses - one of the strategies to improve agricultural crops. The polygalacturonases (PGs) are among the first enzymes with different isomers produced during infections and colonization of phytopathogenic fungus in plant cell walls. Conversely, some of plant species have evolved polygalacturonase inhibitor proteins (PGIP) that specifically recognize and inhibit fungal PGs. This was exploited in order to construct a chimeric gene that contains pgip1 and pgip2 genes from Phaseolus vulgaris. For expression the fusion gene and pgip2 were transformed into tobacco by agroinfiltration method and the caplet plate assay activity of fusion protein has shown that the chimeric protein had high inhibitory activity against fungal PG enzyme. It can be used for the production of transgenic plants and planning a mutational strategy aimed to improve the recognition of fusion protein properties for inhibition of different PGs at the same time.