Iranian Journal of Genetics and Plant Breeding
Volume:11 Issue: 1, Apr 2022

The basic fibroblast growth factor (bFGF) has the potential applications as a therapeutic agent for tissue repair, skin wound healing, and recovery of the neurodegenerative diseases. In this study, after the optimization of all steps in transient transformation by Agrobacterium, the bFGF gene expression was examined in fenugreek leaves infiltrated with different Agrobacterium strains under optimal conditions. These include the AS concentration of 200 μM, bacterial density (OD) of 1.0 OD600, and co-cultivation time for 10 days. The transient expression of the insert was verified in the infiltrated leaves by qRT-PCR analysis. The production of the recombinant bFGF protein was also rectified by Western blot analysis. In addition, the production titers of the recombinant bFGF protein were measured by enzyme-linked immunosorbent assay (ELISA) in all infiltrated leaf samples. Although, there were no significant differences in relative levels of bFGF transcript among the different treatments, but the accumulation levels of recombinant bFGF were significantly affected by Agrobacterium strains. It is possibly due to the post-transcriptional gene silencing. The highest accumulation level of recombinant protein was obtained in leaf samples infiltrated by the EHA105 strain, estimated to be about 3.85 ng bFGF g−1 FW.

After the identification of resistant, semi-susceptible, and susceptible genotypes among 42 alfalfa (Medicago sativa L.) genotypes concerning farm traits, nine genotypes were selected. Saponins obtained as a result of homogenous suspension of alfalfa tissue under maximum alfalfa weevil (Hypera postica Gyll.) attack were extracted using methanol and were purified using water-saturated butanol. The resulting solution was immobilized using the active enzyme extracted from the midgut of the pest and the thermally deactivated enzyme. In addition to the isolation of secondary glycosidic metabolites extracted from alfalfa, the enzymatic hydrolysis activity of the pest in the glycosides of genotypes were monitored using thin layer chromatography. The results of the enzymatic digestion test using TLC indicated that other than glycosides1 epigenin, other compounds were digested in the solution. The flavonoidal band of epigenin glycoside1 in all five resistant genotypes, including Tak Buteh and Ranger, was high in density and nearly equal. While maintaining the stability of the molecular structure, this compound was isolated and revealed at Rf=0.45 on a TLC plate. Therefore, midgut enzymes of alfalfa weevil were not able to digest the above compound in alfalfa. In two susceptible genotypes of Poly Cross Shiraz and Mahali Neyshaburi, the saponin band of soysaponin1 at Rf=37 had the highest density. In natural field conditions and at the time of biological stress, the lowest concentrations of saponin, soysaponin 1, and the highest concentration of flavonoids, epigenin glycoside 1, had a significant biological role in resistance to pests, and reduced damage to the cultivars under study.

Grain quality in rice plays a critical role in consumer acceptance. This research aimed to investigate the grain physicochemical and cooking characteristics of 18 Iranian rice genotypes under both normal conditions and 35 days of drought stress. Significant differences were observed in the studied traits specially percentage of total conversion, head rice, and broken rice indicating differences in the extent of grain retention and damage during processing among the genotypes under both normal and drought stress conditions. The drought stress markedly influenced the expression of nine cooking and nutritional properties and resulted in decreased total conversion percentage, head rice percentage, and cooked head rice length, while the percentage of broken rice increased considerably across all studied genotypes. Conversely, the impact of drought stress on the rough rice length/width ratio, head rice length/width ratio, and elongation ratio were negligible. Most drought-tolerant promising mutant lines exhibited superior grain physicochemical and cooking properties under both drought stress and normal conditions compared to four well-known Iranian rice landraces and cultivars. Evaluation of the grain physicochemical and cooking properties suggested that two drought-tolerant promising mutant lines, namely TM-B-7-1 and HM-250-E-1-1, could be suitable for final cultivar registration experiments.

Drought is one of the main abiotic stresses limiting wheat growth and productivity worldwide. The main objective of this work was to determine population structure and marker-trait association (MTA) of 13 morpho-physiological traits of bread wheat for drought-tolerance breeding. To this end, twenty-five diverse wheat cultivars and promising lines were genotyped using AFLP. The phenotype evaluation steps of studding wheat genotypes were performed under normal and drought-stress conditions during 3 years. Low heritability estimates were obtained for spike length, heading date (DTH), and shoot biomass (24.87-28.8%) while, a high heritability was observed for the number of kernels per spike (KPS) (89.21-90.55%). The results exhibited high polymorphic level ranged from 84.62 to 100%, proving that AFLP method can be an effective tool in assessing genetic variability in any wheat breeding programs. Population structure analysis showed five subpopulations with at least 65% membership ancestry to their allocated sub-clusters, which was highly consistent with the results of cluster analysis. Mixed linear method association analysis identified 66 significant MTAs with p-values 10−06 to 10−04, justifying 7.8 to 38.7% of the phenotypic variation, observed under both environmental conditions. There were two pleiotropic markers for grain yield (GY) and KPS under normal and one pleiotropic marker for GY, thousand kernel weight (TKW) and KPS under stress conditions. The common MTAs were detected for DTH, plant height, peduncle length, and TKW under both environmental conditions. The identified linked markers with GY and its components in this study could be desirable candidate genes for future studies and marker assisted selection to develop drought-tolerant genotypes in wheat breeding programs.

In order to increase global wheat production, it is necessary to examine different ways of increasing yield. One of the solutions is to identify the genes that control different stress tolerance indices. This research aims to identify the genes controlling quantitative traits under normal conditions and salt stress in the germination stage. The experiments were carried out in a factorial form using a completely randomized design with 3 replications, on 107 lines resulted from the crossing of Gonbad and Zagros cultivars at Gonbad Kavous University, 2021. A linkage map was obtained using 519 SSR, 8 CAAT, 33 IJS, 47 iPBS, 3 IRAP, 17 RAPD, 8 SCoT and 12 ISSR markers on 21 wheat chromosomes. The length of the linkage map was 4918.94 cM and the distance between two adjacent markers about 5.55 cM. A total of 84 QTLs were detected in normal and salinity stress conditions (control, 6 dS/m, 12 dS/m), of which seven QTLs were related to control condition, 42 QTLs were related to 6 dS/m salinity stress and 35 QTLs were related to 12 dS/m stress condition. qLR-B3, qMGT-A5, qR/SDW-B2, qR/SDW-A3, qR/SDW-B7, qLS-A5 and qILVS-A5 were detected under control condition. qSLI-D6 was identified as a major QTL for SLI under 6 dS/m salinity stress by explaining more than 44% of the phenotypic variation of the trait. In the 12 dS/m salinity stress, several gene loci of large effect QTLs were detected, among which qIWVS-B3 explained more than 55% of the phenotypic diversity of the trait. After validation, the results of this research can introduce suitable candidates for marker-assisted selection programs in the population of Iranian wheat RILs.

Brassica napus is a versatile crop with oil and protein-rich seeds, used in food, industry, medicine, and animal feed. However, heat stress limits its productivity, making it essential to identify genes and pathways involved in stress response. We analyzed differentially expressed genes (DEGs) in B. napus under heat stress using bioinformatics tools to identify key genes and pathways. Firstly, DEGs were analyzed for gene interactions using the STRING database and visualized using Cytoscape. To identify key genes involved in heat stress response in B. napus L., we employed the CytoHubba tool for hub gene identification. Additionally, we conducted Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses to gain insights into the functional roles and potential biological pathways associated with these genes. We also used MEME Suite to analyze the promoter regions of hub genes. Our results showed decreased activity of the b6-f complex, a key component of the electron transport chain, under heat stress. We also identified significantly enriched calcium transporter ATPase and heat shock protein family (HSP20). KEGG and cluster analyses highlighted the importance of membrane lipids, galactose metabolism, and protein processing in the endoplasmic reticulum in stress and signal transduction. Our study provided key genes, including transcription factors and chaperones for developing heat-resistant plants via genetic modification. However, these promising results were obtained through rigorous bioinformatics analysis and require further validation using experimental approaches, such as gene editing, phenotypic characterization, and field trials.

In order to produce haploid plants through ovary culture in cucumber (Cucumis sativus L.), a factorial experiment based on randomized design was conducted with three replicates under in vitro conditions. The ovaries of Sina, a hybrid variety of cucumber were harvested one week after flowering. They were cultured in the MS medium containing different concentrations of (0, 0.8 and 1.2 mg/L) TDZ and BAP, respectively. The best calli and the highest percentage of calli were obtained in the culture medium containing 0.8 mg/L TDZ and BAP. Then, 3 week-old explants were transferred to the differentiation medium; MS medium, supplemented with NAA, IBA, IAA, 2,4-D, Pic at 0.05 mg/L and 3 cytokinins including TDZ, BAP, Kin with 0.5, 1 and 1.5 mg/L, respectively, with a total of 15 treatments and 3 replicates. They were subcultured once every two weeks. The highest percentage of callus formation was obtained in the presence of 0.05 mg/L NAA and 1.5 mg/L TDZ. The highest percentage of embryogenesis (93.33%) was observed in the presence of IAA at 0.05 mg/L and 0.5 mg/L Kin. Application of IBA at 0.05 mg/L with 0.5 mg/L Kin increased the embryogenic calli formation. Also, the highest regeneration percentage (83.33%) was found in all 4 treatments including 0.05 mg/L IBA, with 0.5 mg/L TDZ, 0.05 mg/L IBA with 0.5. 0 mg/L BAP, 0.05 mg/L 2,4-D with 1.5 mg/L BAP and 0.05 mg/L Pic, with 1.5 mg/L BAP. After transferring the explants to the second step culture media, the embryo-like structures (ELS) developed into plantlets. Finally, complete plantlets were obtained after rooting the plantlets on the MS medium with no supplement. In addition, the results of chromosome counting showed that among 10 regenerated plantlets, one plantlet was haploid, and 9 plantlets were diploid.