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Fariba Ranjbar, Babak Abdollahi Mandoulakani, Raheleh Ghasemzadeh,
Volume 10, Issue 1 (9-2023)
Abstract
To evaluate the expression pattern of genes encoding antioxidant enzymes catalase, ascorbate peroxidase and polyphenol oxidase under iron deficiency conditions in Fe- efficient (Pishtaz) and -inefficient (Falat) bread wheat cultivars, a CRD (completely randomized design) based factorial experiment was conducted with three replications. The cultivars were grown under iron deficiency (Less than 1.5 mg Fe/kg soil) and compared with normal conditions (10 mg Fe/kg soil). The relative expression levels of the above-mentioned genes were measured using Real-time PCR technique in the leaves and roots of the cultivars at two growth stages: vegetative (one month after germination) and reproductive (30% of heading). The results revealed a remarkable enhancement in calatalse expression in the roots of both cultivars in the vegetatative stage but it was higher in Fe-efficient cultivar than -inefficient one. The expression of this gene was decreased in leaves at the same stage as well as in the roots of both cultivars in the vegetative stage. The expression level of ascorbate peroxidase gene in the reproductive stage in the roots of Fe-inefficient cultivar was higher than that of -efficient one. In the vegetative stage, the expression of this gene increased in the leaves and roots of Fe-efficient cultivar, but it was decresed in Fe-inefficient cultivar. The relative expression level of polyphenol oxidase gene in the vegetative stage under iron deficiency conditions in the leaf increased almost three times, compared to the roots, while the expression of this gene decreased in the reproductive stage in both leaves and roots. By increasing the expression of both catalase and ascorbate peroxidase genes in the roots of both cultivars in the reproductive stage under iron deficiency conditions, it seems that bread wheat cultivars might reduce the deletrious effects of stress and maintain yield through transferring much iron to the seeds in the seed filling stage. The findings of the present study may increase our understanding of the important role of genes encoding antioxidant enzymes in Fe deficiency stress conditions.
Fatemeh Derikvand, Eidi Bazgir, Mostafa Darvishnia, Hossein Mirzaei Najafgholi,
Volume 10, Issue 1 (9-2023)
Abstract
Apple is one of the most important economic products of Iran and the world. Apple brown rot disease (Monilinia laxa) is one of the important diseases that causes yield loss in pre-harvest and post-harvest stages. In this research, the amount of changes in some defense compounds of apple fruit, including peroxidase and catalase enzymes, following inoculation with M. laxa was assessed. Extraction and measurement of peroxidase and catalase enzymes were done at 0, 3, 6, 9 and 12 days post inoculation with M. laxa. Also, in this study, the changes in the expression of PR1 and PR8 genes in response to brown rot disease in apple fruit was recorded at 12, 24, 48 and 96 hours along with controls. The results of the analysis of variance of resistance genes expression at different time points were significant. After 48 hours, the expression of PR1 and PR8 genes was observed to be the highest compared to the control. The expression of PR1 and PR8 genes was observed to be 3 and 8 times that of the control, respectively. In this research, the expression of PR1 and R8 genes was assessed for the first time following M. laxa inoculation in apples. Results of the present study showed that reseistance genes as well as the antioxidant enzymes can help to improve resistance against apple brown rot disease as an important storage pathogens for long-term storage.
Ramezanali Pourali, Mohammadhadi Pahlevani, Khalil Zeinalnejad,
Volume 10, Issue 2 (2-2024)
Abstract
Increasing grain yield and improving the quality of bread are among the most important goals of wheat breeding programs in Iran. Understanding the genetic control of traits and finding molecular or morphological markers associated with them are also prerequisites for any genetic engineering program. In this study, 100 progenies of a 10 × 10 diallel cross were used to analyse the genetics of grain yield and bakery values using STS markers associated with HMWG subunits. This research was carried out during 2018 and 2019 cropping season at Gorgan University of Agricultural Sciences and Natural Resources experimental fields. In the first year, 10 wheat cultivars, including Gonbad, Morvarid, Kalate, Ehsan, Sirvan, Baharan, Chamran2, Shush, Mehrgan and Brat collected from different geographical regions of Iran were planted and crossed in the field. In the second year, the parents and crosses were planted in the form of a randomized complete block design with three replications. The grain yield, number of spikes per plant, number of seeds per spike, seed weight, days to emergence and plant height were recorded. The results of this study indicated significant genetic differences between the parents. Narrow-sense heritability analysis revealed that the crossing of cultivars is the best breeding method to enhance seed yield, number of spikes per plant and days to emergence. Also, to improve the number of seeds per spike, seed weight and plant height, classical breeding methods may offer higher efficiency. Marvarid and Gonbad were ranked 1st and 2nd, respectively with respect to general combining abilities for grain yield, attributed to their positive and significant general combining ability effects. The highest specific combining ability was observed for Ehsan×Gonbad, Marvarid×Chamran 2 and Shush×Sirvan crosses. The results of molecular markers analysis showed that the STS markers were able to identify the difference in the baking value of cultivars. The quality score of the cultivars ranged 6 and 10 and to this end, Kalate and Brat were the top cultivars. Therefore, due to superiority in terms of both quantity and quality for yield, these cultivars can be used as parents with desirable genes for future breeding programs. Overall, the STS markers employed in this study proved to be valuable markers for enhancing the genetic background of bread wheat, particularly when employing marker-assisted selection for bakery value.
Seyede Maryam Seyed Seyed Hassan Pour, Leila Nejadsadeghi, Zahra Sadat Shobbar, Danial Kahrizi,
Volume 10, Issue 2 (2-2024)
Abstract
Camelina )Camelina sativa (is an annual, self-pollinating, allohexaploid plant with diploid inheritance belonging to the Brassicaceae family. Camelina exhibits a remarkable degree of similarity to the model plant Arabidopsis thaliana. WRKY transcription factors are among important gene families in plants that play crucial roles in regulating growth and development and in response to diverse stresses. In this research, using bioinformatics analysis and databases, members of the WRKY gene family were identified and their various characteristics were investigated. Overall, the genome of the Camelina plant was found to harbor 214 members of the WRKY gene family. All 214 WRKY genes were found to possess the conserved WRKY functional domain, along with a variety of motifs within their structural composition. Phylogenetic analysis divided the identified members of Camelina WRKY genes into four main groups. Examination of the chromosomal positions revealed that the 214 identified WRKY genes exhibited an uneven distribution across the chromosomes. In order to validate the identified genes, the expression of two genes (Csa11g065620 and Csa07g035970) orthologs of two genes involved in drought stress in Arabidopsis (WRKY8 and WRKY57), were investigated in a drought tolerant (DH 91) and a drought sensitive (DH 101) lines. The results of the gene expression analysis showed that both genes had high expression in drought stress conditions in tolerant line in comparison to normal conditions, whereas no significant expression was found in drought sensitive line. The findings of the present study offer valuable insights for evolutionary investigations and enhance our understanding of the functional roles of the WRKY gene family in Camelina, thereby laying a foundation for future research endeavors in this field.
Seyedeh Sanaz Ramezanpour, Hassan Soltanloo, Saied Navabpour,
Volume 10, Issue 2 (2-2024)
Abstract
To evaluate the effect of fungus Blumeria graminis (powdery mildew disease) on expression of genes associated with resistance reactions in barley, a susceptible cultivar (Afzal), a semi-susceptible genotype (Line 67) and a resistant genotype (Line 104) were selected. Following inoculation with Blumeria graminis at seedling stage, sampling was performed at different time points (0-10 days). Changes in gene expression levels were measured by qRT-PCR analysis. Analysis of molecular data showed that the genes encoding chitinase and glucanase as the key enzymes in fungal cell wall degradation, had higher expression levels in the resistant genotype (Line 104). The transcript level of chitinase in semi-susceptible genotype (Line 67) was lower than that of the resistant genotype (Line 104) and higher than that of the susceptible cultivar. Most transcripts of chitinase gene were seen at 12 hours post inoculation in the resistant genotype (Line 104), whereas the lowest expression level was recorded at the same time in the susceptible cultivar. The expression levels of the other two genes (glucanase and peroxidase) were higher in the resistant genotype (Line 104) than those in the susceptible cultivar. Increasing in MAPK transcripts in resistant genotype (Line 104) and its depletion in susceptible cultivar confirmed MAPK role in Hypersensitive response (HR) and defense responses of barley infected with powdery mildew disease. Based on the findings of this study, it appears that the HR in the resistant genotype initiated as early as six hours post inoculation, effectively hindering the penetration and dissemination of the pathogen within the plant. Such reaction was not observed in the semi-susceptible and susceptible barley plants, possibly due to delayed in responses, allowing the pathogen ample time to penetrate and propagate within the host plant. The results of this research can be used to evaluate the resistance level of cultivars and also to evaluate the resistance in the seedling stage of promising lines.
Maryam Ehsasatvatan, Bahram Baghban Kohnehrouz,
Volume 10, Issue 2 (2-2024)
Abstract
The global prevalence of type 2 diabetes mellitus is continuously increasing, and there is currently no definitive cure for type 2 diabetes. The potent glucagon-like peptide 1 (GLP-1), a natural small incretin hormone, enhances insulin secretion in a glucose-dependent manner. However, the exceedingly short half-life of GLP-1 limits its therapeutic applications. Albumin-binding DARPin can be used to increase the serum half-lives of therapeutic proteins, peptides, and small compounds. In this study, a long-acting GLP-1 agonist with oral delivery potential containing a protease-resistant GLP-1, an albumin-binding DARPin, and Penetratin as a fusion protein was expressed in a bioencapsulated form within tobacco chloroplasts to confer digestive system protection in plant cells. The successful transformation of tobacco chloroplasts with trivalent fusion protein-coding genes was conducted using a pPRV111A chloroplastic expression vector and a gene gun. Homoplasmic transplastomic plants were obtained after three rounds of selection in selection medium containing 500 mg/L spectinomycin and streptomycin. Transgene integration and homoplasmic status in the transplastomic plants were confirmed by PCR and Southern blot analyses. Western blot analysis confirmed the accumulation of the mGLP1-DARPin-Pen fusion protein in the chloroplasts of the transplastomic plants. The fusion protein content estimated by ELISA was 21.8% of the total soluble protein content in the transplastomic plants. The successful expression of the designed fusion protein indicated that the production of functional GLP-1 in plants may facilitate the development of a low-cost, orally deliverable form of this protein for the treatment of type 2 diabetes.
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