Functional characterization of a novel synthetic herbicide resistance gene in a model plant

As crop losses can occur owing to the abrupt growth of uncontrollable weeds in the field, research is currently being conducted in Pakistan to eradicate herbs. To carry out our proposed research domain, we have designed a novel synthetically modified EPSPS gene that has a potent role in herbicide-resistance development. The novel codon-optimized synthesized mEPSPS sequence was inserted into the pXCSG-mYFP plant expression vector. Results of colony PCR (1400 bp) confirmed the integration of genes into bacteria. For functional validation of pXCSG-mYFP-EPSPS, transient expression in tobacco (Nicotiana benthamiana) in comparison with PBSF-16 was done. Benth infiltration results showed that transient expression was successfully confirmed through ELISA and western blot analysis via anti-YFP antibody in tobacco leaves. For further validation of transient expression, the stable transformation results of the pXCSG-mYFP-EPSPS vector showed that successful transformation was done via two days of co-cultivation followed by selection and regeneration of transformed tobacco plants. The regenerated tobacco plants were then confirmed through gene-specific-based PCR. After PCR-based confirmation, western blot further validates the 26 kDa anti-YFP antibodies expression in transformed tobacco plants. Another important finding of the study was the bar and PAT gene real-time expression elucidating that the bar gene was 4.9-fold more expressive under the 35S promoter than the PAT gene under the nos promoter

Genome-wide identification and expression analysis of the Glutamine synthetase family genes in Zea mays under drought stress

Glutamine synthetase (GS) enzyme is involved in nitrogen metabolism by the assimilation of NH4+ with glutamate and glutamine, and hence play their role in plant development, biotic and abiotic stress responses, secondary metabolite synthesis and crop yield. However, in maize, the expression levels of ZmGS genes under drought stress are not explored. Globally drought stress significantly affected maize production. Keeping in view the importance of GS genes in maize, we have checked their response under drought stress. The targeted 05 ZmGS genes in maize evolutionary analysis was performed for 81 GS protein sequences in different species, clustered into three clades based on their phylogenetic analysis. GS genes that were identified in maize are distributed on Chromosome 1, chrom4, chrom5, and chrom9. Conserved domain analysis, gene structure, motifs analysis, and alignment of ZmGSs demonstrated that all genes showed a conserved pattern. Analysis of cis-regulatory elements in promoter region revealed that almost all the ZmGS genes had stress responsive and plant hormonal related cis elements and their number are different in all genes. We also identified 55 putative miRNAs targeting 5 ZmGS genes. Furthermore, GS genes of maize were identified as drought-responsive genes by using qRT-PCR expression analysis. The high expression level of GS genes was detected in the roots and leaves of Haq Nawaz compared to CIMMYT PAK maize cultivars. The analysis of all identified ZmGS genes using qRT-PCR data showed that they expressed in roots and shoots under normal and drought stress. The expression analysis of drought-responsive GS genes was found more in H. Nawaz than in C. Pak maize cultivar under drought stress. Therefore, the H. Nawaz cultivar could have the potential to tolerate mild to moderate drought stress. Our study offers valuable data on the functioning of GS genes in maize under drought stress and provides a foundation for further analysis of the function of GS gene family under abiotic stresses

Inter Simple Sequence Repeat-Based Genetic Divergence and Varietal Identification of Banana in Pakistan

Banana is one of the major cash and fruit crops of Pakistan. The lack of information concerning genetic diversity and purity within locally cultivated banana varieties is a major bottleneck in improving its genetics. Due to the existence of a narrow genetic background, it’s quite important to find genomic variations in banana varieties. DNA marker-based techniques have been used to effectively characterize banana varieties. In the current study, Inter Simple Sequence Repeat (ISSR) markers were used to characterize banana cultivars and to assess the genetic diversity of 14 local banana varieties grown in Pakistan. Out of the 45 primers used, 40 primers revealed reproducible results and produced 121 polymorphic bands, which contributed a ratio of 47.87 polymorphism. The ISSR UBC-835 and UBC-834 possessed the highest PIC ranged between (86–88%) in banana varieties, while the lowest PIC (46%) was detected in the case of UBC−857 marker with (100–1500 bp) PCR product size. Pairwise Jaccard’s similarity coefficient values were also calculated, and these were ranged from 0.56–0.88. Multivariate analysis divided 14 banana varieties into two distinct groups—A and B respectively—and furthermore into subgroups, clusters, and sub−clusters. Our results indicated that at the molecular level, the banana varieties in group—A were found to be 66% similar whereas in group B were 88% similar. Nei’s genetic diversity, PCA analysis, and a minimum spanning tree depicted Fenjiao, Dajiao, and NIGAB-2 as the most diverse members as compared to all other varieties of the three populations. Out of 14 varieties used, 11 varieties were uniquely identified by 54 polymorphic ISSR bands of different sizes. Some varieties like NIGAB-2 and NIGAB-3 were uniquely identified only with one band while others were tagged by multiple unique bands. In future, this study will be utilized to establish a molecular-based protocol for the identification of banana varieties

The effect of flaxseed oil consumption on blood pressure among patients with metabolic syndrome and related disorders: A systematic review and meta-analysis of randomized clinical trials

We systematically reviewed randomized clinical trials (RCTs) to elucidate the overall effects of flaxseed oil consumption on blood pressure (BP) in patients with metabolic syndrome and related disorders. PubMed, Scopus, Cochrane Library, and ISI Web of Science databases were systematically searched until March 31, 2020, to find RCTs that examined the effect of flaxseed oil consumption on BP. Weighed mean difference (WMD) was pooled using a random-effects model. Standard methods were used for the assessment of heterogeneity, sensitivity analysis, and publication bias. Meta-analysis of five trials (6 arms) showed significant reductions in systolic (WMD: −3.86 mmHg, 95% CI: −7.59 to −0.13, p = .04) BP (SBP) after flaxseed oil consumption. However, the overall effect illustrated no significant change in diastolic (WMD: −1.71 mmHg, 95% CI: −3.67 to 0.26, p = .09) BP (DBP) in the intervention group compared with the control group. Our findings revealed that flaxseed oil consumption has favorable effects on SBP in patients with metabolic syndrome and related disorders. However, further investigations are needed to provide more reliable evidence