مجله سلول و بافت
سال هشتم شماره 2 (تابستان 1396)

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 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.
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).
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.

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 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.
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).
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.

Aim: In this study, the effects of simulated microgravity on the apoptosis and expression of p75NTR gene were investigated in the adipose derived stem cells before and after the neural differentiation.
Material and Human adipose derived stem cells were isolated, cultured and differentiated. A single-axis clinostat apparatus was used to simulate microgravity for 6, 24 and 72 hours. Real time PCR technique was used for gene expression analysis after extraction of RNA of samples. Cell viability was assessed by MTT assay and apoptosis rate was calculated by Annexin V staining.
Our results showed that microgravity led to a significant decrease in p75NTR gene expression in adipose derived stem cells. However, microgravity had no significant effect on viability of cells before and after differentiation, but apoptosis in undifferentiated cells was decreased in contrast to controls.
Due to reduction of apoptosis and increment of differentiation potential of cells, microgravity can be introduced as a powerful tool and also new condition for cell culture and neural differentiation for achievement to effective cell therapy.

Aim: in the present study, morphology and also histology of six Nepeta species were investigated.
Material and six species of the genus were collected from different parts of Iran. From each species, one population and from each populations three flowering stems were randomly elected. The mature intact leaf of each sample was fixed in FAA solution, and then transverse hand sections of them were double stained and examined under light microscopy. For scanning electron microscopy, a small part of leaves were coated with gold then, samples transferred to SEM for taking micrograph. The used software was SPSS.
Thirteen glandular and non-glandular trichomes were observed in the studied species. The morphology and density of trichomes varied between species and ANOVA test showed significant differences in some trichome types between the studied species. The peltate and capitate were the most important glandular trichomes. In addition, non- glandular hairs were existed in two forms: branched and non-branches. The branched ones were found in only one species.
on the basis of variations in glandular trichomes, it is expected that the amount of essential oil is variable between the studied species. In addition, the abilities of species are different in maintenance of essential oil. Moreover, the trichomes can use a good trait for improvement of Nepeta taxonomy.

Aim: The present research attempts to study the effect of different concentrations of zinc oxide (ZnO) nanoparticles on the germination, the amount of photosynthetic pigments and sugar content and analyze the ultrastructure of Ricinus communis plant leaves.
Material and Experiment were performed under controlled greenhouse conditions, and designed completely randomly with three incidents. The plants were exposed to various concentrations (0, 10, 100, 500 and1000) mg/l of zinc oxide nanoparticles. The ultrastructural Characteristics of plant leaves were made with the use of TEM electron microscope in the experimental plants of 1000 mg/l.
Treatment of the plant with (ZnO) Nanoparticels at concentration of 10 mg/l caused increased and higher concentrations significantly reduced the rate and percentage of germination, as well as the radicle and plumule length and the photosynthetic pigments. The amount of soluble sugars in leaves increased significantly with the increase of nanoparticle concentration. The images TEM electron microscope revealed the concentration of zinc oxide nanoparticles and cell membrane rupture, as well as a deformation and decrease of the number of chloroplasts in the1000 mg/l treated plants, compared with the control plants.
The Zn absorption by the plant increases by increasing the concentration of (ZnO) nanoparticles. At high concentrations due to toxicity, germination parameters in the plant decrease, which leads to oxidative stress in the plant. The plant in response to this stress increases its sugar content. In these conditions, the amount of photosynthetic pigments decreased and caused ultrastructural ruptures in plant cells.

Aim: This study attempts to optimize callus induction and analyze of yeast extract and nano-silver elicitors on phenol/flavonoid content in black cumin under tissue culture conditions.
Material and The experiment was conducted a factorial design based on CRD with three replications. Factors included: explants (root, hypocotyledon, leaf and Cotyledon), 2, 4-D (1, 2, 4 and 8 Mg/L) and BAP (0.25, 0.5, and 1 Mg/L) in MS base medium. Elictor's including yeast extract (100, 250 and 500 Mg/L) and nano-silver (30, 60 and 90 Mg/L) in two time (3 and 7 days) periods.
The results showed that hypocotyledon explant and interaction effect of BAP (0.25 Mg/L) and 2, 4-D (4 Mg/L) were the most effective on the callus induction percent. Direct regeneration was caused by the root explant and the interaction effect of BAP (0.5 Mg/L) and 2, 4-D (1 Mg/L). The most effective treatment on the total phenol content was yeast extract (250 ppm) in a 7-day period. HPLC for quercetin (a flavonoid component) indicated that the most effective treatment was the interaction effect of nanoAg particles (30 Mg) and yeast extract (250 Mg) in a 3-day period.
The highest amount of callus induction is obtained from the hypocotyledon explants. The best direct regeneration is recorded in the root explants. To increase total phenol, it is necessary to use yeast extracts in a 7-day period and to increase flavonoids using the interaction effect of nano-silver (30 Mg) and yeast extract (250 Mg) over a 3-day period.

Aim: The aim of this study is to evaluate the spermatogonial stem cells differentiation into male gametes.
Materia and Spermatogonial stem cells were cultured in T25 flasks after the enzymatically isolation of the testicular biopsies through azoospermic patients. In the third passage, cells divided into 4 different groups and were treated for 1 to 4weeks under the effect of a medium containing extracts of sheep testes as inducer and then expression of sperm maturation genes: Acrosin and Protamine1 were investigated by using of western blotting technique.
After spermatogonial stem cells treatment by the extracts of sheep testes, variations were seen in cell shape and they convert into sperm- like. Moreover, Acrosin and Protamine1 expression were confirmed.
examination of induced cells showed that Acrosin and Protamin1 were expressed. Since Acrosin and Protamine1 are the major proteins of the spermiogenesis, it could be concluded that these cells had been completed spermatogenesis stage and started the spermiogenesis stage.

Aim: This study was conducted to determine the neuroprotective effects of mint extract on alpha motor neuron degeneration at the anterior horn of the spinal cord after sciatic nerve compression in rats.
Material and In this study, 30 male Wistar rats weighing 200-250 g were randomly divided into 5 groups including control, compression and treatment groups of 50,75 and 100 mg. In order to induce the compression, sciatic nerve was undergone to compress by using locking- scissors for 60 seconds. Hydroalcoholic extract of mint was injected intraperitoneally during the first and second weeks after the compression. After 28 days, rats were undergone by the perfusion method and after sampling the lumbar spinal cord, neuronal density was calculated by using dissector and stereological methods and the findings were compared together.
a significant decrease was observed in compression group compared to the control for the neuronal density and also a significant increase was seen in treatment groups rather than the compression group (p The results indicate that the mint extract has a neuroprotective effect that may be due to the antioxidant and anti-inflammatory properties of the extract of this plant.

Aim: In this research, a simple and rapid method (green synthesis) was applied for synthesis of silver nanoparticles (AgNPs) using Scrophularia striata fruit extract, so that the metabolites present in S.striata fruit extract caused to reduce silver ions to AgNPs in green synthesis process.
Material and UV–visible spectroscopy and scanning electron microscopy (SEM) were used to characterize the synthesized nanoparticles from S. striata extract. The antibacterial activity of the synthesized silver nanoparticles from S. striata extract was investigated against Gram-negative bacteria (Escherichia coli clinical, Escherichia coli ATCC, Salmonella typhi ATCC and Klebsiella pneumoniae) and Gram-positive bacteria (Staphylococcus aureus and Bacillus cereus). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined using microdilution technique. The antibacterial activity was determined by agar well diffusion method.
The results showed that with increased concentration of silver nanoparticles, antibacterial activity increased and in the concentration of 5 mM silver nanoparticles, antibacterial activity was observed against all bacteria, however the highest antibacterial activity of silver nanoparticles observed against Staphylococcus aureus (inhibition zone diameter with 32 mm). Also, in the low concentrations of 0.312 and 0.625 mM of silver nanoparticles, no inhibitory effects were observed on the Klebsiella pneumoniae and Salmonella typhi ATCC.
From the results, it is suggested that silver nanoparticles synthesized using S. striata fruit extract could be used as a suitable antibacterial agent against clinical pathogens.