Imunosuppresive Activity of Momordica charantia L. fruit extract on the NF-κB pathway in Drosophila melanogaster

The activation of the NF-kB pathway leading to the production of proinflammatory cytokines is a critical feature in innate antiviral immunity. However, in SARS-CoV-2 infection, a high number of cases with a prolonged late-stage stimulation of cytokine production that leads to a cytokine storm phenotype, an undesirable, dangerous immune-related state that can cause multiple organ failures, have been reported. To treat this, immunosuppressants with selective action on the innate NF-kB pathway are urgently required. Bitter gourd (Momordica charantia L.) has been reported to yield anti-inflammatory activity, thus might be a potential candidate for such effort. In this study, we carried out experimental procedures on the PGRP-LB mutant line of Drosophila melanogaster to examine the immunosuppressive effect of Momordica charantia L. fruit extract (MCFE) on the NF-kB pathway. Initial phytochemical screening revealed that Momordica charantia L. fruit extract contains alkaloids, flavonoids, tannins, and saponins. Furthermore, the phenotypical analysis demonstrated that MCFE could improve the survival and locomotor of the PGRP-LB mutant line of Drosophila melanogaster in a concentration-dependent manner. Additional gene expression analysis revealed that the expression of dpt and dro, two important downstream genes in the Imd (NF-kB) pathway of D. melanogaster, was significantly reduced, in a different expression profile, in response to MCFE treatment. However, it is important to note that while the expression of dpt was dramatically repressed in all extract-treated groups, the expression of dro occurred in a concentration-dependent manner. These results strongly support the notion that Momordica charantia L. can reduce the expression of proinflammatory cytokines downstream of the NF-kB pathway, hence potential to be used as a source candidate to harvest prospective immunosuppressive compounds to alleviate the cytokine storm condition.

Chromatin spatial organization of wild type and mutant peanuts reveals high-resolution genomic architecture and interaction alterations

Background Three-dimensional (3D) chromatin organization provides a critical foundation to investigate gene expression regulation and cellular homeostasis. Results Here, we present the first 3D genome architecture maps in wild type and mutant allotetraploid peanut lines, which illustrate A/B compartments, topologically associated domains (TADs), and widespread chromatin interactions. Most peanut chromosomal arms (52.3%) have active regions (A compartments) with relatively high gene density and high transcriptional levels. About 2.0% of chromosomal regions switch from inactive to active (B-to-A) in the mutant line, harboring 58 differentially expressed genes enriched in flavonoid biosynthesis and circadian rhythm functions. The mutant peanut line shows a higher number of genome-wide cis-interactions than its wild-type. The present study reveals a new TAD in the mutant line that generates different chromatin loops and harbors a specific upstream AP2EREBP-binding motif which might upregulate the expression of the GA2ox gene and decrease active gibberellin (GA) content, presumably making the mutant plant dwarf. Conclusions Our findings will shed new light on the relationship between 3D chromatin architecture and transcriptional regulation in plants.

Distinct Morphological and Behavioural Alterations in ENU-Induced Heterozygous Trpc7K810Stop Mutant Mice

Trpc7 (transient receptor potential cation channel, subfamily C, member 7; 862 amino acids) knockout mice are described showing no clear phenotypic alterations, therefore, the functional relevance of the gene remains unclear. A complementary approach for the functional analysis of a given gene is the examination of individuals harbouring a mutant allele of the gene. In the phenotype-driven Munich ENU mouse mutagenesis project, a high number of phenotypic parameters was used for establishing novel mouse models on the genetic background of C3H inbred mice. The phenotypically dominant mutant line SMA002 was established and further examined. Analysis of the causative mutation as well as the phenotypic characterization of the mutant line were carried out. The causative mutation was detected in the gene Trpc7 which leads to the production of a truncated protein due to the novel stop codon at amino acid position 810 thereby affecting the highly conserved cytoplasmic C terminus of the protein. Trpc7 heterozygous mutant mice of both sexes were viable and fertile, but showed distinct morphological and behavioural alterations which is in contrast to the published phenotype of Trpc7 knockout mice. Thus, the Trpc7K810Stop mutation leads to a dominant negative effect of the mutant protein.

A Non-Rogue Mutant Line Induced by ENU Mutagenesis in Paramutated Rogue Peas (Pisum sativum L.) Is Still Sensitive to the Rogue Paramutation

The spontaneously emerging rogue phenotype in peas (Pisum sativum L.), characterized by narrow and pointed leaf stipula and leaflets, was the first identified case of the epigenetic phenomenon paramutation. The crosses of homozygous or heterozygous (e.g., F1) rogue plants with non-rogue (wild type) plants, produce exclusively rogue plants in the first and all subsequent generations. The fact that the wild phenotype disappears forever, is in clear contradiction with the Mendelian rules of inheritance, a situation that impedes the positional cloning of genes involved in this epigenetic phenomenon. One way of overcoming this obstacle is the identification of plant genotypes harboring naturally occurring or artificially induced neutral alleles, non-sensitive to paramutation. So far, such alleles have never been described for the pea rogue paramutation. Here, we report the induction via 1-ethyl-1-nitrosourea (ENU) mutagenesis of a non-rogue revertant mutant in the rogue cv. Progreta, and the completely unusual fixation of the induced non-rogue phenotype through several generations. The reversion of the methylation status of two previously identified differentially methylated genomic sequences in the induced non-rogue mutant, confirms that the rogue paramutation is accompanied by alterations in DNA methylation. Nevertheless, unexpectedly, the induced non-rogue mutant showed to be still sensitive to paramutation.

Molecular profile of the D. melanogaster mutant genotype w1118 in the presence of variable amount of deuterium (D)

The Drosophila melanogaster w1118 mutant line was used to identify the effect of deuterium (D) on DNA synthesis. D concentrations ranged from 30ppm to 96.89% (low and very high amount respec-tively). Five generations of flies were bred on culture media prepared with 6 concentrations of D. For each generation the DNA was analyzed, and its variability was established. The results showed a small involvement of D in the successive synthesis of nuclear DNA.

Performance Evaluation of Rice Mutant Line against Two Widely Grown Rice Cultivar in Mymensingh Region of Bangladesh

Growth performance, yield components and total grain yield of a rice mutant lines were evaluated againest two popular rice varieties on farm experiments within five different upazila of Mymensingh region to find out the better cultivar on the besis of yield and duration. The experiment was laid out in a randomized complete block design (RCBD) with three replications. The study was conducted for two consecutive season that ensure the stability of the mutant line. The results revealed that the cultivar RM(2)-40-©-4-2-8 was recorded highest effective tiller and maximum filled grain in most of the location. Among the all cultivar RM(2)-40-©-4-2-8 was recorded maximum grain yield in most of the location for two consecutive year which was statistically significant from other cultivar.

Proteomic and Biochemical Analyses of the Mechanism of Tolerance in Mutant Soybean Responding to Flooding Stress

To investigate the mechanism of flooding tolerance of soybean, flooding-tolerant mutants derived from gamma-ray irradiated soybean were crossed with parent cultivar Enrei for removal of other factors besides the genes related to flooding tolerance in primary generated mutant soybean. Although the growth of the wild type was significantly suppressed by flooding compared with the non-flooding condition, that of the mutant lines was better than that of the wild type even if it was treated with flooding. A two-day-old mutant line was subjected to flooding for 2 days and proteins were analyzed using a gel-free/label-free proteomic technique. Oppositely changed proteins in abundance between the wild type and mutant line under flooding stress were associated in endoplasmic reticulum according to gene-ontology categorization. Immunoblot analysis confirmed that calnexin accumulation increased in both the wild type and mutant line; however, calreticulin accumulated in only the mutant line under flooding stress. Furthermore, although glycoproteins in the wild type decreased by flooding compared with the non-flooding condition, those in the mutant line increased even if it was under flooding stress. Alcohol dehydrogenase accumulated in the wild type and mutant line; however, this enzyme activity significantly increased and mildly increased in the wild type and mutant line, respectively, under flooding stress compared with the non-flooding condition. Cell death increased and decreased in the wild type and mutant line, respectively, by flooding stress. These results suggest that the regulation of cell death through the fermentation system and glycoprotein folding might be an important factor for the acquisition of flooding tolerance in mutant soybean.