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Soheila Afkar, Faranak Hadi, Ali Ashraf Jafari,
Volume 8, Issue 2 (3-2022)
Abstract
Festuca is one of the largest genera of the grass family, which has more than 600 species with different ploidy levels. The aim of this study was to estimate the genetic diversity within 22 populations of three species of Festuca (Festuca arundinacea, F.rubra and F.ovina) using a seed storage protein electrophoresis pattern. These species showed a significant variation in the number of protein bands from 5-13. The highest number of bands was found in G17 (F.rubra) and the lowest number of protein bands was in G5 (F.ovina). Band number 14 was only observed in G3. It is suggested that this band can be considered as a specific band for the identification of this genotype. According to the results of AMOVA analysis, there is a high level of genetic diversity within the species rather than between species that can be due to the out-crossing nature of this genus. According to observed differences for variation parameters among the three studied species, it is concluded that they have dissimilar genetic structures. The results of cluster analysis based on seed storage protein profiles in evaluated genotypes using Euclidean distance matrix and UPGMA method showed four groups. The lowest similarity coefficient was between G14 and G15 (F.arundinacea) with G6 (F.ovina). Hence, it is suggested that they evolved from a different evolutionary process and it is suggested to use them as the parents of new synthetic varieties. The observed diversity in the seed protein pattern in the three species of Festuca, can be explained by allogamy-induced-heterozygosity, species difference or population collection from various regions.
Shahla Hosseini, Mohammad Reza Rahgozar, Hedieh Badakhshan,
Volume 8, Issue 2 (3-2022)
Abstract
Genus Allium L. contains very taxonomically complex sections, especially the subgenus Melanocrommyum. The systematic position of the species in each section has been revised many times over time. In the present study, the relationship between 32 ecotypes belonging to 10 different species of Allium was investigated using ISSR markers. The nine primers used produced 166 polymorphic bands (average 18 bands per primer). Among the primers used, ISSR873 primer with 27 bands made the most, and ISSR4 primer with two bands had the lowest polymorphic bands. The PIC of the markers ranged from 0.04 to 0.43. Cluster analysis by UPGMA method based on molecular markers divided the studied ecotypes into four groups. The clustering and principal coordinate analysis results showed that most morphologically similar species were grouped in closed clusters. According to Dice similarity coefficient, the highest percentage of similarity was shown between Allium stipitatum and Allium saralicum ecotypes (72 percent) from the Melanocrommyum subgenus, and the lowest similarity was obtained between Allium tripedale and Allium iranicum ecotypes (12 percent). The ecotypes with the lowest similarity percentage belong to the subgenus Allium and Nectaroscordum, which are placed in separate clusters. Based on the results, the ecotypes of Pseudoprason, Melanocrommyum, and Procerallium sections showed the highest affinity. In general, it can be concluded that ISSR markers are useful for classifying Allium species and have sufficient potential for phylogenetic studies of species. In addition, due to significant genetic diversity among the studied ecotypes of wild Allium species, this diversity can be used in future breeding programs of crop.
Mahmood Aslanparviz, Varahram Rashidi, Mansour Omidi, Alireza Etminan, Alireza Ahmadzadeh,
Volume 8, Issue 2 (3-2022)
Abstract
Evaluation of genetic diversity is the key principal for plant breeding, providing an opportunity to discover novel characters and alleles for breeders. In the present study, 69 durum wheat genotypes were investigated for genetic diversity using several inter-simple sequence repeat (ISSR) markers. Sixteen ISSR primers amplified a total of 163 fragments, which out 160 fragments were polymorphic. The mean values of polymorphic information content (PIC), resolving power (Rp) and marker index (MI) indicated that the used ISSR primers could be exploited for further assessing relationships among investigated genotypes and population structure analysis. The results of the molecular analysis of variance showed that the genetic variation within populations is more than between them. Based on genetic variation parameters, the highest number of observed alleles (Na), Shannon’s information index (I) and the percentage of polymorphic loci (PPL) were found in Iranian landraces. Cluster analysis and population structure grouped all investigated genotypes into three main clusters and six subpopulations, respectively. In conclusion, our results revealed the high rate of genetic diversity within Iranian landraces, so this germplasm can be used as a valuable gene source for the selection of parent lines and use of them in durum wheat breeding programs.
Abdul Karim Tahmasebi, Reza Darvishzadeh, Amir Fayaz Moghaddam, Esmaeil Gholinezhad, Hossein Abdi,
Volume 8, Issue 2 (3-2022)
Abstract
The selection of genotypes based on multiple traits is a fundamental issue and an important part of the process of plant breeding. In the present study, the efficiency of selection indices based on phenological, morphological and physiological traits was studied to improve sesame grain yield. The evaluation of 25 sesame populations was realized in a completely randomized design with 10 replications under Urmia conditions in 2017.The results showed that phenotypic and genotypic correlations between grain yield and No. of capsules per plant, No. of grains per capsule, No. of branches, leaf temperature, leaf index and biological weight were positive and significant. By regression and path analysis, the No. of capsules and No. of branches were identified as the variables of the first-order cause and biological weight, harvest index, leaf index, plant height and chlorophyll as the second-order cause variables, among which only plant height had a direct negative effect. In order to obtain selection indices, two optimal and basic methods and ten different vectors of economic values of traits were used. The vectors were based on the analysis of correlation, regression, path and broad sense heritability. The third and fourth indices, in which the first-order cause entered the model, showed high relative efficiency and in terms of these two indices, and the sesame populations with code number of 12, 17, 18 and 19 populations were identified as the most desirable populations. Finally, it is suggested that the efficiency of these selection indices be evaluated in the field
Reza Mir Derikvand, Seyede Sajad Sohrabi, Seyyed Mohsen Sohrabi, Kamran Samiei,
Volume 8, Issue 2 (3-2022)
Abstract
Rizan Elyasi, Mohammad Majdi, Abdolbaset Azizi,
Volume 8, Issue 2 (3-2022)
Abstract
Black cumin (Nigella sativa) is a medicinal plant of the Ranunculacea family which raised attention due to its pharmaceutical properties. Medical significance of N. sativa mainly attributed to its oxygenated monoterpenes which are biosynthesized via the methyl erythritol phosphate (MEP) pathway located in plastids. In this study, the essential oil components of leaves, flowers, and developmental stages of seed including half black seeds, soft black seeds, and hard black seeds were analyzed in N. sativa. Whereas no terpene was detected in flowers and leaves, seeds were found to be the major site of biosynthesis and accumulation of terpenes, and the amount of terpene compounds changed during seed maturation. The essential oil consists of monoterpenes (more than 99%) and sesquiterpenes (less than 1%). In order to improve our understanding of monoterpene metabolism, the partial sequence of a hypothetical monoterpene synthase (NsTPS2) was isolated from N. sativa plant using RACE-PCR technique. This monoterpene synthase was identified from RNA sequencing data from soft black seeds. Except of the highly conserved DDXXD motif in NsTPS2 which is necessary to validate monoterpene synthases, no other conserved regions of other identified monoterpene synthases were observed. Dendrogram analysis revealed that NsTPS2 had the highest homology with a terpene synthase (72.89%) from Aconitum carmichaelii and these two sequences were grouped in the same group. Nigella sativa and Aconitum carmichaelii both belong to the Ranunculacea family. This indicates that the genetic information of plants of the Ranunculacea family can be used to isolate different monoterpene synthase. The results of this research can be useful in genetic manipulation and metabolic engineering of Nigella sativa.
Mitra Khademi, Marzih Varasteh-Shams, Farhad Nazarian-Firouzabadi,
Volume 9, Issue 1 (9-2022)
Abstract
Hairy and adventitious roots are efficient systems for expressing recombinant proteins. In the present study, the amount of DrsB1-CBDAvr4 recombinant protein in hairy and adventitious root systems was compared. To this end, the effect of different factors on the optimization of culture conditions to obtain adventitious and hairy roots was evaluated in three separate experiments by assessment of biomass production in T1 transgenic plants expressing DrsB1-CBDAvr4 recombinant protein. The efficacy of Agrobacterium rhizogenesis in producing hairy roots was ensured using rolC gene specific primers. The insertion of DrsB1-CBDAvr4 recombinant peptide transgene in the genome of hairy and adventitious roots was confirmed by PCR analysis. Also, the level of DrsB1-CBDAvr4 protein was measured in hairy and adventitious roots by ELISA analysis. Analysis of the variance of data showed that the highest number of roots and the longest roots were obtained in MS media supplemented with 1 mg/L NAA and 0.5 mg/L IBA. The results of adventitious root biomass showed that liquid MS medium containing 1 mg/L of NAA hormone had a significant effect (P <0.01) on biomass production. More biomass was obtained in MS medium supplemented with 1mg/L NAA, whereas a lower fresh and dry weight was obtained in a 1/4 MS medium with no NAA. The results also showed that ATCC15834 strain with MS media supplemented with 3% sucrose, 10 minutes inoculation time was high efficiency to induce hairy roots in tobacco plants. The results of ELISA analysis showed that clones obtained from both roots showed a significant difference in terms of total protein content. The amount of recombinant protein from hairy roots was much higher than that of adventitious roots.
Razieh Ghorbani, Raheleh Ghasemzadeh, Hadi Alipour,
Volume 9, Issue 1 (9-2022)
Abstract
In order to identify loci controlling seedling morpho-physiologic characteristics in 88 bread wheat cultivars, a greenhouse experiment based on simple alpha lattice was conducted under both normal and 120 mM (12 ds/m) salt stress condition of the Faculty of Agriculture, Urmia University in 2020-2021 cropping season. Chlorophyll a, b and carotenoid content, proline, plant fresh and dry weight, plant height and leaf relative water content (RWC), Na+, K+ and K+/Na+ concentrations were measured. After genotyping by sequencing with Ion Torrent technology and removal of SNPs with more than 20% of missing data and minor allele frequency less than 5%, a total of 5869 SNP markers were identified. Based on association mapping with the mixed linear model (MLM) method, a total of 25 marker-trait associations were detected under normal conditions. The A and D genomes had the highest and lowest number of significant marker-trait associations (MTAs). Among the studied traits under normal conditions, chlorophyll a had the highest number of MTAs on 1A, 3B, 3D, 5B, 7A chromosomes with eight MTAs. A total of 21 MTAs were identified under salt stress conditions which the genome B and D had the highest and lowest number of MTAs, respectively. Five MTAs were identified for plant fresh weight, which were located on chromosomes 4A and 6B. The results of this study provide valuable information about the loci associated with the studied traits, which can be used in marker assisted selection in wheat breeding programs after confirmation in biparental populations and additional experiments.
Halime Arbabi, Mojtaba Keykhasaber, Leila Fahmideh, Valiollah Ghasemi Omran,
Volume 9, Issue 2 (3-2023)
Abstract
Luffa (Luffa cylindrica) is a plant from the Cucurbitaceae family that grows mostly in tropical and subtropical regions, as well as in most regions of Iran. In this research, the genetic diversity of nine native and non-native genotypes of L. cylindrica was investigated through the evaluation of the chloroplast trnH-psbA intergenic region (IGS). After sampling the young leaves, DNA extraction was performed by using the Dellaporta method, and PCR was conducted by using IGS intergenic region primers. After sequencing of the amplified products, their quality was determined using Chromas software and then aligned using ClustalW method by BioEdit and MEGA7 softwares. Next, the dendrogram of phylogenic relationships was drawn and the matrix of the difference and similarity of the sequences were determined. In the present research, by analyzing the relationships between studied samples, based on the trnH-psbA (IGS) marker, a strong intraspecies variation was observed in native and non-native L. cylindrical genotypes. The genetic distance matrix between the samples examined in this research ranged from 0 to 6.865 with an overall average distance of 2.53. The average value of synonymous and non-synonymous substitutions (dN/dS) for the IGS sequence was ds/dn = 0.68, which indicates positive and pure line selections in the process of natural selection of studied genotypes. The results of this research showed that trnH-psbA is a suitable marker for evaluation of the intraspecific diversity of luffa species.
Saeid Navabpour, Horeyeh Najafi,
Volume 9, Issue 2 (3-2023)
Abstract
Environmental stress is one of the main factors that reduce the growth and performance of crops and threatening human food security. This study was conducted in order to investigate the effect of drought stress on the changes in biochemical traits and the level of expression of a MYB transcription factor gene in two wheat cultivars (Tajan and Zagros), under drought stress. The experiment was conducted as a factorial based on a completely randomized design with 3 replications. Drought treatments were applied at three levels of 40, 70 and 100% of field capacity 4 weeks after germination. Twenty days after the application of stress, leaves and roots were sampled in order to investigate the expression of MYB genes and measuring some biochemical traits. The results of examining the chlorophyll content under stress showed that the content of chlorophyll a and b decreased with increasing of stress intensity in different genotypes. The rate of reduction of chlorophyll a and b in Tajan genotype under severe stress was higher than Zagros genotype. Also, TBARM content under severe drought stress was significantly higher than moderate stress condition and this increase was seen in Tajen genotype more than Zagros genotype. qRT-PCR analysis showed that the MYB genes showed an increase in expression under drought stress. Furthermore, Zagros genotype, which is considered as a tolerant cultivar to drought stress, had a higher MYB expression level than Tajan cultivar for both genes, suggesting this cultivar for future breeding programs, also considering the importance MYB family genes during drought stress, the results can be used in molecular breeding and pyramiding breeding projects.
Khadijeh Abbaszadeh, Reza Shirzadian-Khorramabad, Mohammad Mahdi Sohani, Zahra Hajiahmadi,
Volume 9, Issue 2 (3-2023)
Abstract
Salinity stress affects morpho-physiological and biochemical traits of plants. The transgenic Bt plants play a significant role in pest control, but their response and ability to cope with environmental stresses still need to be evaluated. Therefore, effect of salinity stress at 0, 50, 100, 150, and 200 mM on morphological, physiological, and molecular traits of T3 transgenic tomato plants containing cry1Ab gene (CH-Falat-Bt) was investigated and compared with that of the non-transgenic control (CH-Falat). Evaluation of the morphological traits (leaf area, root length, fresh and dry weight of roots) at different salinity levels revealed that CH-Falat-Bt transgenic plants are more tolerant to salinity stress compared to CH-Falat non-transgenic plants. The chlorophyll content at 150 and 200 mM salinity levels was 12 and 9% plants, respectively. Moreover, the amount of RWC, carotenoids, proline and soluble sugars increased significantly in transgenic plants as salinity levels increased. The relative expression of SOS1 and SOS2 genes showed a significant increase in all salinity levels in CH-Falat-Bt transgenic plants compared to CH-Falat non-transgenic plants. The amount of electrolyte leakage in the transgenic plants was significantly reduced compared to the non-transgenic plants. The results of morphological, physiological, and molecular investigations of CH-Falat-Bt transgenic plants confirmed that the undesirable effects of salinity stress on transgenic plants is much less than non-transgenic ones. in general CH-Falat-Bt transgenic plants are more tolerant to different applied salinity levels than the wild variety.
Seyyed Mohsen Sohrabi, Seyed Karim Mousavi,
Volume 9, Issue 2 (3-2023)
Abstract
Chickpea (Cicer arietinum L.) is one of the most important crops in the world. After bean and pea, chickpea is the most important cold season legume. Weeds are one of the most important threats to chickpea production worldwide. Due to the sensitivity of chickpea to herbicides, the majority of herbicides are used pre-emergence and the use of post-emergence herbicides is limited, and therefore weeds cause a significant decrease in chickpea yield. Therefore, herbicide-tolerant chickpea cultivars that have a higher flexibility for post-emergence herbicide application are needed to improve the chickpea yield. In this study, using seed bioassay and PCR method, resistance mechanism of Iranian chickpea cultivars to Pursuit herbicide was investigated. The results showed a significant genotypic and phenotypic variation among Iranian chickpea cultivars for tolerance to the Pursuit herbicide. The results did not show a difference between the target genes of Pursuit herbicide, ALS1 and ALS2, in all investigated cultivars with that of the reference sequences in the GenBank. This proves that the resistance observed in different chickpea cultivars to the herbicide Pursuit is not associated with the target site resistance mechanism and probably follows a non-target resistance mechanism. The superior genotypes of this study (Bivanij, Aksou, Mansour, TDS-Maragheh90-400 and TDS-Maragheh90-358) can be recommended to farmers and also suggested as parents to produce natural herbicide resistant chickpea plants in breeding programs.
Hossein Abdi, Hadi Alipour, Iraj Bernousi, Jafar Jafarzadeh,
Volume 10, Issue 1 (9-2023)
Abstract
Evaluating the population structure is essential for understanding diversity patterns, choosing proper parents for crossing, accurate identification of genomic regions controlling traits, and evolutionary and kinship relationship studies. In this research, the genetic structure of a wheat population was studied in a panel consisting of 383 Iranian wheat genotypes of hexaploid (cultivars and landraces) and tetraploid species based on distance-based methods (principal component analysis and discriminant analysis of principal component). For this purpose, 16270 single nucleotide polymorphism (SNP) markers obtained by the GBS technique were used. According to the results, almost a quarter of the total variance was belonged to the diversity between populations, and the Fst coefficient between cultivars and landraces was equal to 0.15. In contrast, the above coefficient between tetraploid samples and hexaploid landraces was high and equal to 0.44. Genome D had the lowest value of Fst index and chromosome 4B showed the highest Fst coefficient, and other genetic diversity indices. Although the PCA biplot distinguished hexaploid wheat cultivars from landraces, it was unable to distinctly separate tetraploid genotypes from other genotypes. Accurate evaluation of the population structure with the DAPC method was able to identify and separate the predetermined successfully groups, suggesting that the DAPC approach maximizes the differentiation between groups and minimizes the changes within the group. Partial admixture between cultivars and landraces of hexaploid wheat can be related to gene exchange between these two groups or perhaps their wrong labeling at the time of collection. In general, the results of this study provided valuable information about the genetic differentiation of Iranian tetraploid and hexaploid wheat, which can be used in future wheat breeding programs. Further, protecting these genotypes in gene banks is necessary for different strategies.
Maryam Ghorbani, Kianoosh Cheghamirza, Saeed Abbasi, Zahra Aziziaram,
Volume 10, Issue 1 (9-2023)
Abstract
The current research was carried out to evaluate genetic diversity of 18 common bean cultivars and promising lines and to determine SSR and SCoT informative markers associated with 14 seed characteristics including the number of seeds per pod, 100 seed weight, seed length, seed width, the contents of crude protein, total soluble sugar, starch content, crude fat, iron, calcium, magnesium, zinc, uronic acid, and mineral ash. The polymorphism information content (PIC) values varied from 0.2 to 0.5 with an average of 0.39 for the SSR markers and from 0.19 to 0.42 with an average of 0.34 for the SCoT markers. The total average resolving power of SSR and SCoT markers were 1.54 and 5.34, respectively, indicating higher efficiency of SCoT markers than SSR markers for the diversity analysis. The common bean studied genotypes were clustered into three distinct groups for both markers based on the Complete Linkage method. Principal coordinate analysis (PCoA) for the SSR markers revealed that the first two principal components justified 59.05% of whole variation. For the SCoT marker, on the other hand, the fraction of variances explained by the first two principal components was equal to 25.43 indicating a better distribution of SCoT markers than SSR markers in the common bean genome. Analysis of molecular variance based on the grouping obtained from cluster analysis showed that the diversity within the group based on SSR and SCoT markers was equal to 89 and 78%, respectively. The results of regression analysis for the studied markers and the bean seed characteristics showed the existence of a significant relationships between a single marker with the several studied traits. This could indicate the association or linkage of the marker locations. Twelve out of 14 of the investigated common bean seed characteristics showed a significant relationship with at least one molecular marker.
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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