Cell and Tissue Journal

Abstract Aim: Folate, also known as Vitamin B9, is a water-soluble nutrient present in natural food sources. It plays a crucial role in DNA synthesis and repair, as well as in methylation reactions. Folic acid (FA) is the synthetic form of folate found in enriched foods and supplements. The effects of folate deficiency after birth have not been extensively studied. MicroRNAs (miRNAs) are a type of non-coding RNAs that play essential roles in regulating gene expression. In recent years, miRNAs have been associated with various aspects of brain development, including neurogenesis, neuronal migration, axon and dendrite formation, and synaptogenesis. Additionally, altered expression and dysregulation of miRNAs have been linked to neurodevelopmental disorders. Methionine synthase (Mtr) is involved in amino acid metabolism and is responsible for converting the amino acid homocysteine to methionine. Mtr is crucial for brain development and is associated with myelin content. This project aimed to investigate the impact of maternal FA supplementation during pregnancy on the expression of mmu-miR-103-1-5p and Mtr in the cerebral cortex of mouse pups.
Materials and Methods: A total of 45 pregnant mice were divided into three groups, with 15 mice in each group. The first and second groups received FA at doses of 2 and 40 mg/kg of body weight daily by gavage throughout their entire pregnancy, respectively. The third group did not receive FA and served as the control. Subsequently, the cerebral cortex of the offspring was collected immediately after birth for the analysis of mmu-miR-103-1-5p and Mtr expression using real-time PCR. Statistical analysis was conducted using GraphPad Prism software.
Results: In the FA-treated group, the expression of mmu-miR-103-1-5p at a dose of 2 mg/kg body weight was 0.46±0.01, and at a dose of 40 mg/kg body weight, it was 0.27±0.02, whereas in the control group, it was 1.0007±0.03. Statistical analysis revealed a significant decrease in mmu-miR-103-1-5p expression at both the 2 mg/kg and 40 mg/kg doses compared to the controls (p

Abstract Aim: In this study, the effect of silence on the expression of the key gene expression of DBOX (which encodes the final enzyme for the synthesis of two alkaloids, Sanguinarin and papaverin) was used by VIGS technique in a species of poppy (Papaver somniferum L.).
Material and methods: A fragment of 350 pairs of alkali from the DBOX gene sequence (within the range of 1112-1462bp) was selected based on the highest number of siRNA production with 21 nucleotides length. After cloning this segment into the pTZ57R/T vector and transferring the vector to pTRV2 viral vector, Agrobacterium inoculation liquid containing silencer was injected into the poppy plants leaves. Primary transgenic plants were selected by PCR reaction using a protein-binding protein coding gene primer (CP) and secondary screening was performed by semi-quantitative PCR technique. In the next step, the samples with the maximum silence (lowest expression) of the gene were examined by real-time RT-PCR technique.
Results: Cloning accuracy in pTZ57R/T and pTRV2 plasmids were confirmed using PCR and enzymatic digestion. Based on the results of semi-quantitative PCR, 5 transgenic plants were selected with the lowest expression for DBOX gene. Based on semi-quantitative PCR results, 5 transgenic plants with the lowest expression were selected for DBOX gene. The results of real-time RT-PCR showed averagely decrease of 81% in the expression of DBOX gene transcriptions in transgenic plants compared to control plants (inoculated with the pTRV2 empty plasmid).
Conclusion: The results generally showed that the VIGS technique could successfully reduce the DBOX gene expression in poppy plants. In addition, the results obtained for this gene can be used to understand the biosynthetic pathway of poppy alkaloids and transgenic plants for metabolic engineering purposes.
 

Abstract Aim: In this study, the effect of silence on the expression of the key gene expression of DBOX (which encodes the final enzyme for the synthesis of two alkaloids, Sanguinarin and papaverin) was used by VIGS technique in a species of poppy (Papaver somniferum L.).
Material and methods: A fragment of 350 pairs of alkali from the DBOX gene sequence (within the range of 1112-1462bp) was selected based on the highest number of siRNA production with 21 nucleotides length. After cloning this segment into the pTZ57R/T vector and transferring the vector to pTRV2 viral vector, Agrobacterium inoculation liquid containing silencer was injected into the poppy plants leaves. Primary transgenic plants were selected by PCR reaction using a protein-binding protein coding gene primer (CP) and secondary screening was performed by semi-quantitative PCR technique. In the next step, the samples with the maximum silence (lowest expression) of the gene were examined by real-time RT-PCR technique.
Results: Cloning accuracy in pTZ57R/T and pTRV2 plasmids were confirmed using PCR and enzymatic digestion. Based on the results of semi-quantitative PCR, 5 transgenic plants were selected with the lowest expression for DBOX gene. Based on semi-quantitative PCR results, 5 transgenic plants with the lowest expression were selected for DBOX gene. The results of real-time RT-PCR showed averagely decrease of 81% in the expression of DBOX gene transcriptions in transgenic plants compared to control plants (inoculated with the pTRV2 empty plasmid).
Conclusion: The results generally showed that the VIGS technique could successfully reduce the DBOX gene expression in poppy plants. In addition, the results obtained for this gene can be used to understand the biosynthetic pathway of poppy alkaloids and transgenic plants for metabolic engineering purposes.
 

Abstract Aim: In this study, the effect of salinity stress on quantitative gene expression of sucrose synthase1, relative water content and the uptake ratio of sodium to potassium in the leaves of wheat were evaluated.
Material and Methods: Wheat seedlings treated with 100 and 200 mM sodium chloride and at times zero, 6, 12, 24 and 36 hours after treatment relative water content, the uptake ratio of sodium to potassium in the leaves and quantitative gene expression of Sus1 by qRT-PCR method were studied. 
Results: Results showed a significant decrease in relative water content, increase in the uptake ratio of sodium to potassium by leaves and increase in the relative expression of sucrose synthase1 gene at treated plants compared to control group.
Conclusion: Generally, increase in the expression of sucrose synthase 1 gene could be one of the possible mechanisms in the process of tolerance to salinity stress.