Variety Of Mutants Research Articles

Strictinin, a Major Ingredient in Yunnan Kucha Tea Possessing Inhibitory Activity on the Infection of Mouse Hepatitis Virus to Mouse L Cells.

Theacrine and strictinin of Yunnan Kucha tea prepared from a mutant variety of wild Pu'er tea plants were two major ingredients responsible for the anti-influenza activity. As the COVID-19 outbreak is still lurking, developing safe and cost-effective therapeutics is an urgent need. This study aimed to evaluate the effects of these tea compounds on the infection of mouse hepatitis virus (MHV), a β-coronavirus serving as a surrogate for SARS-CoV. Treatment with strictinin (100 μM), but not theacrine, completely eliminated MHV infection, as indicated by a pronounced reduction in plaque formation, nucleocapsid protein expression, and progeny production of MHV. Subsequently, a time-of-drug addition protocol, including pre-, co-, or post-treatment, was exploited to further evaluate the possible mechanism of antiviral activity mediated by strictinin, and remdesivir, a potential drug for the treatment of SARS-CoV-2, was used as a positive control against MHV infection. The results showed that all three treatments of remdesivir (20 μM) completely blocked MHV infection. In contrast, no significant effect on MHV infection was observed when cells were pre-treated with strictinin (100 μM) prior to infection, while significant inhibition of MHV infection was observed when strictinin was introduced upon viral adsorption (co-treatment) and after viral entry (post-treatment). Of note, as compared with the co-treatment group, the inhibitory effect of strictinin was more striking in the post-treatment group. These results indicate that strictinin suppresses MHV infection by multiple mechanisms; it possibly interferes with viral entry and also critical step(s) of viral infection. Evidently, strictinin significantly inhibited MHV infection and might be a suitable ingredient for protection against coronavirus infection.

A tyrosine-based YXXΦ motif regulates the degradation of aquaporin-4 via both lysosomal and proteasomal pathways and is functionally inhibited by a 10-amino-acid sequence within its C-terminus.

Aquaporin-4 (AQP4) is a dominant water channel in the brain and is expressed on astrocytic end-feet, mediating water homeostasis in the brain. AQP4 is a target of an inflammatory autoimmune disease, neuromyelitis optica spectrum disorders (NMOSD), that causes demyelination. An autoantibody recognizing the extracellular domains of AQP4, called NMO-IgG, is critically implicated in the pathogenesis of the disease. Complement-dependent cytotoxicity (CDC) and antibody-dependent cellular cytotoxicity (ADCC) in astrocytes are the primary causes of the disease, preceding demyelination and neuronal damage. Additionally, some cytotoxic effects of binding of NMO-IgG to AQP4, independent of CDC/ADCC, have been proposed. Antibody-induced endocytosis of AQP4 is thought to be involved in CDC/ADCC-independent cytotoxicity induced by the binding of NMO-IgG to AQP4. To clarify the mechanism responsible for antibody-induced endocytosis of AQP4, we investigated the subcellular localization and trafficking of AQP4, focusing on its C-terminal domain, by making a variety of deletion and substitution mutants of mouse AQP4. We found that a tyrosine-based YXXΦ motif in the C-terminal domain of AQP4 plays a critical role in the steady-state subcellular localization/turnover and antibody-induced endocytosis/lysosomal degradation of AQP4. Our results indicate that the YXXΦ motif has to escape the inhibitory effect of the C-terminal 10-amino-acid sequence and be located at an appropriate distance from the plasma membrane to act as a signal for lysosomal degradation of AQP4. In addition to lysosomal degradation, we demonstrated that the YXXΦ motif also functions as a signal to degrade AQP4 using proteasomes under specific conditions.

Suicidal Phenotype of Proofreading-Deficient Herpes Simplex Virus 1 Polymerase Mutants.

Herpes simplex virus 1 (HSV-1) encodes a family B DNA polymerase (Pol) capable of exonucleolytic proofreading whose functions have been extensively studied in the past. Early studies on the in vitro activity of purified Pol protein found that the enzymatic functions of the holoenzyme are largely separate. Consequently, exonuclease activity can be reduced or abolished by certain point mutations within catalytically important regions, with no or only minor effects on polymerase activity. Despite unimpaired polymerase activity, the recovery of HSV-1 mutants with a catalytically inactive exonuclease has been so far unsuccessful. Hence, mutations such as D368A, which abolish exonuclease activity, are believed to be lethal. Here, we show that HSV-1 can be recovered in the absence of Pol intrinsic exonuclease activity and demonstrate that a lack of proofreading causes the rapid accumulation of likely detrimental mutations. Although mutations that abolish exonuclease activity do not appear to be lethal, the lack of proofreading yields viruses with a suicidal phenotype that cease to replicate within few passages following reconstitution. Hence, we conclude that high replication fidelity conferred by proofreading is essential to maintain HSV-1 genome integrity and that a lack of exonuclease activity produces an initially viable but rapidly suicidal phenotype. However, stably replicating viruses with reduced exonuclease activity and therefore elevated mutation rates can be generated by mutating a catalytically less important site located within a conserved exonuclease domain. IMPORTANCE Recovery of fully exonuclease-deficient herpes simplex virus 1 (HSV-1) DNA polymerase mutants has been so far unsuccessful. However, exonuclease activity is not known to be directly essential for virus replication, and the lethal phenotype of certain HSV-1 polymerase mutants is thus attributed to factors other than exonuclease activity. Here, we showed that the recovery of a variety of exonuclease-deficient HSV-1 polymerase mutants is possible and that these mutants are initially replication competent. We, however, observed a progressive loss of mutant viability upon cell culture passaging, which coincided with the rapid accumulation of mutations in exonuclease-deficient viruses. We thus concluded that a lack of DNA proofreading in exonuclease-deficient viruses causes an initially viable but rapidly suicidal hypermutator phenotype and, consequently, the extinction of mutant viruses within few generations following recovery. This would make the absence of exonuclease activity the primary reason for the long-reported difficulties in culturing exonuclease-deficient HSV-1 mutants.

Climate Resilient Varietal Development of Diverse Crops through Mutation Breeding in Bangladesh

The capability to quickly generate a considerable range of genetic variation in plants through mutation enables the mutagenesis approach extremely successful in developing climate-resistant cultivars across the entire world. In Bangladesh, plant breeders are confronted with a significant challenge in producing resilient and high-yielding cultivars rapidly capable of adapting to the changing environment in this densely populated region. Based on Mutant varieties data of Bangladesh which are available in the domain of FAO/IAEA Mutant Varieties Database (MVD), this review attempted to highlight the relevance of mutation breeding in crop variety development to adapt to changes occurring in diverse ecosystems of Bangladesh during this era of ongoing climate change. So far, seventy-eight mutant varieties of important crops have been released in Bangladesh, including rice, wheat, mustard, sesame, jute, rapeseed, tomato, black gram, chickpea, grass pea, lentil, mung bean, groundnut, and onion. Significant improvements in agronomic and botanical qualities have been made to maximize crop production. Salinity-tolerant varieties for southern coastal regions, drought-tolerant varieties for drought-prone locations, early mature varieties for flash flood escape in haor regions, submerged tolerant varieties for flooding regions and various major disease and insect resistant varieties have been generated through mutation breeding. These mutant varieties with climate-resilient traits are sustainable under different climatic circumstances, playing a key role in assuring food security in Bangladesh. Therefore, it is anticipated that mutant breeding will contribute significantly to meeting the challenges that food production will confront in the near future due to a changing climate.

Transposon Mutagenesis for enhancement of nitrogen fixing ability of Rhizobium japonicum infecting Vigna radiata

This study developed mutant library of 800 mutants with transposition frequency of 3.7 x 10- 6. Transposon mutagenesis is an easy and highly effective method for generating bacteria with improved characteristics and gene knockouts. The slow growing Rhizobium japonicum isolated from root nodules of locally grown mung bean plant. The isolated strain was subjected for transposon mutagenesis to produce 800 mutants. These 800 mutants were screened for nodulation. After screening of these 800 mutants, the100 mutants showed pink coloured nodulation to mung bean plant. Out of 100 mutants 10 were promising for nodulation and nitrogen fixation. Bacterial surface polysaccharides including Exopolysaccharides (EPS) and Lipopolysaccharides (LPS) are constituents of the bacterial cell wall. They have been reported in numerous studies as symbiotically important; defects in EPS production tend to result in arrest at the infection thread stage which is the early stage of nodulation. EPS and LPS were isolated from wild variety and mutant variety. The polysaccharides production in mutant and wild variety were compared. This study was aimed to investigate the role of EPS and LPS on nodulation and enhancement of nitrogen fixation of Rhizobium japonicum infecting Vigna radiata.

Determinants of adoption and preferences for Aman rice mutant variety Binadhan-7 cultivation in Bangladesh

This study was conducted in fourteen agricultural regions of Bangladesh namely-Mymensingh, Jashore, Cumilla, Bogura, Rajshahi, Sylhet, Dinajpur, Rangpur, Dhaka, Khulna, Chattagram, Rangamati, Barishal and Faridpur. The specific objectives of the present study were: i) to examine the determinants to adoption of rice mutant variety Binadhan-7; and ii) to identify the major preferences and constraints of rice mutant variety Binadhan-7 cultivation; A multistage stratified random sampling technique was used to collect the data. Marginal coefficients indicate that if male farmers increased by 100%, the probability of adopting Binadhan-7 variety would increase at 38 times more likely to adopt the variety. If the farm size of Binadhan-7 increased by 100%, the probability of adopting the variety would be increased by 0.07%. A farmer who has access to agricultural extension service is about 39 times more likely to adopt the variety. Again, if the yield increased by 100%, adopting the varieties would increase by 0.08%. The marginal coefficients of duration and soil fertility are negatively significant, indicating that if these two variables increased by 100%, the probability of adopting the varieties would decrease by 0.18% and 28%, respectively. Among the preferences, the highest preference was 88.93% for short duration followed by poverty reduction (88.57%), and lastly high yielding (81.43%). Among the constraints, the highest constraint was 80.71% for high labour price followed by lack of godown (69.64%), marketing problem (67.50%), destroy by bird (66.79%) and lack of quality seed (48.93%).

Changes on mutant pot chrysanthemums (Dendranthema × grandiflora Tzelev.)

Chrysanthemum (Dendranthema x grandiflora Tzvelev) is one of the most important ornamental plants in the world that has the richest mutant varieties with numerous colors. The objective of this study is to determine the effective mutagen dose (EMD50) for creating variations by gamma irradiation. It is aimed to get a mutagenesis protocol that could develop new mutants in pot chrysanthemums. To determine the EMD50, rooted cuttings of brownish-red color ‘Brandevil’ variety were irradiated by gamma radiation at 0, 10, 20, 30, 40, 50 Gray (Gy) doses. According to the shoot lengths, EMD50 was calculated as 27 Gy. The mutation frequency was calculated as 4.8%. Some changes were observed for flower numbers per plant, plant heights and widths, shapes and colors of both flowers and leaves. The color changes varied from brownish-red to yellow and orange. Two different colors appeared in the same pot at some genotypes as well as form changes of flowers. The similarity of the mutants was determined by the hierarchical cluster dendrogram involving five groups. Various colors were obtained for leaves and flowers. Remarkable mutations of the selected mutants were multiplied by tissue culture.

Indução de mutagênese em crisântemo

Abstract Crop genetic diversity has a significant role in improving new plants through breeding. The chrysanthemum contains the most mutant varieties, making mutation breeding one of the most widely utilized breeding procedures for ornamental plants. The goal of this research is to use gamma irradiation to induce genetic variation and mutation breeding to improve chrysanthemum features. In vitro bud explants of the white ‘Bacardi’ type were treated with gamma rays at 20 Gy on this scope. The explants were subcultured until M1V4 growing period occurred, and observations were made during blooming on this time. Variable flower head widths, distinction on plant heights and widths, numerous flower numbers, color and size variations of ray florets were among the mutagenic changes observed in plants and flowers. Ray florets varied in length, width, number of rows, and color. The mutation frequency of the population was estimated 1.1% and yellow-colored florets were developed whereas the control group remained white. The dendrogram was grouped into five groups with 1, 28, 31, and 41 mutants in each based on the plant height and width, plant stem height and width, number of shoots and flowers per plant, flower head width, ray florets’ number- height- color, number of leaves, leaf length and width, and weight of flowering stems. The yellow-colored mutants were located in the first, second, and fourth groups. The advantageous mutations could result in improving new varieties. Gamma radiation is an effective mutagen for creating new chrysanthemum types when applied to in vitro bud explants.

A Sensitive GC-MS Method for Quantitation of Lipid A Backbone Components and Terminal Phosphate Modifications.

Lipid A, the hydrophobic anchor of lipopolysaccharide (LPS) present in the outer membrane of Gram-negative bacteria, serves as a target for cationic antimicrobial peptides, such as polymyxins. Membrane stress from polymyxins results in activation of two-component regulatory systems that produce lipid A modifying enzymes. These enzymes add neutral moieties, such as aminoarabinose (AraN) and ethanolamine (EtN) to lipid A terminal phosphates that mask the phosphate's negative charge and inhibit electrostatic interaction with the cationic polymyxins. Currently, these modifications may be detected by MALDI-TOF MS; however, this analysis is only semiquantitative. Herein we describe a GC-MS method to quantitate lipid A backbone components, glucosamine (GlcN) and inorganic phosphate (Pi), along with terminal phosphate modifications AraN and EtN. In this assay, lipid A is isolated from Gram-negative bacterial samples, hydrolyzed into its individual moieties, and derivatized via methoximation followed by silylation prior to analysis via GC-MS. Changes in AraN and EtN quantity were characterized using a variety of regulatory mutants of Salmonella, revealing differences that were not detected using MALDI-TOF MS analysis. Additionally, an increase in the abundance of AraN and EtN modifications were observed when resistant Enterobacter and Escherichia coli strains were grown in the presence of colistin (polymyxin E). Lastly, increased levels of Pi were found in bisphosphorylated lipid A compared to monophosphorylated lipid A samples. Because lipid A modifications serve as indicators of polymyxin resistance in Gram-negative bacteria, this method provides the capacity to monitor polymyxin resistance by quantification of lipid A modification using GC-MS.

Novel resistance to the Bymovirus BaMMV established by targeted mutagenesis of the PDIL5-1 susceptibility gene in barley.

The Potyviridae are the largest family of plant-pathogenic viruses. Members of this family are the soil-borne bymoviruses barley yellow mosaic virus (BaYMV) and barley mild mosaic virus (BaMMV), which, upon infection of young winter barley seedlings in autumn, can cause yield losses as high as 50%. Resistance breeding plays a major role in coping with these pathogens. However, some viral strains have overcome the most widely used resistance. Thus, there is a need for novel sources of resistance. In ancient landraces and wild relatives of cultivated barley, alleles of the susceptibility factor PROTEIN DISULFIDE ISOMERASE LIKE 5-1 (PDIL5-1) were identified to confer resistance to all known strains of BaYMV and BaMMV. Although the gene is highly conserved throughout all eukaryotes, barley is thus far the only species for which PDIL5-1-based virus resistance has been reported. Whereas introgression by crossing to the European winter barley breeding pool is tedious, time-consuming and additionally associated with unwanted linkage drag, the present study exemplifies an approach to targeted mutagenesis of two barley cultivars employing CRISPR-associated endonuclease technology to induce site-directed mutations similar to those described for PDIL5-1 alleles that render certain landraces resistant. Homozygous primary mutants were produced in winter barley, and transgene-free homozygous M2 mutants were produced in spring barley. A variety of mutants carrying novel PDIL5-1 alleles were mechanically inoculated with BaMMV, by which all frameshift mutations and certain in-frame mutations were demonstrated to confer resistance to this virus. Under greenhouse conditions, virus-resistant mutants showed no adverse effects in terms of growth and yield.

Transcriptome analysis of branches reveals candidate genes involved in anthocyanin biosynthesis of ‘Red Bartlett’ pear (Pyrus communis L.)

The color of the peel is the most direct reflection of fruit quality, and is also an important factor in attracting consumers. The ‘Red Bartlett’ pear (Pyrus communis L.) is a red bud sport mutant variety of ‘Bartlett’ pear. It was found that the anthocyanin content in branches and pericarps of ‘Red Bartlett’ pears were significantly higher than that of ‘Bartlett’ pears. The present study aimed to clarify the underlying mechanisms of anthocyanin accumulation in ‘Red Bartlett’ using a transcriptome analysis. A weighted gene coexpression network analysis (WGCNA) identified 495 differentially expressed genes positively associated with anthocyanin. Additionally, 12 anthocyanin-related structural genes and 13 transcription factors, including WRKY, ERF, bHLH, ASIL2, and HSF, were identified through screening of 495 DEGs. The expression levels of one UFGT (Pbr014258.1) and one GST (Pbr012649.1) were also upregulated in the pericarps of ‘Red Bartlett’ and ‘Starkrimson’, more so than in ‘Bartlett’ and ‘Clapps Favourite’. Overall, this study clarified that the red color mutation of ‘Red Bartlett’ was closely linked to increased anthocyanin biosynthesis and transport. This study provided an abundance of valuable information on anthocyanin biosynthesis for future breeding of red-skinned pears.

Determination of the boundary lipids of sticholysins using tryptophan quenching

Sticholysins are α-pore-forming toxins produced by the sea-anemone Stichodactyla helianthus. These toxins exert their activity by forming pores on sphingomyelin-containing membranes. Recognition of sphingomyelin by sticholysins is required to start the process of pore formation. Sphingomyelin recognition is coupled with membrane binding and followed by membrane penetration and oligomerization. Many features of these processes are known. However, the extent of contact with each of the different kinds of lipids present in the membrane has received little attention. To delve into this question, we have used a phosphatidylcholine analogue labeled at one of its acyl chains with a doxyl moiety, a known quencher of tryptophan emission. Here we present evidence for the contact of sticholysins with phosphatidylcholine lipids in the sticholysin oligomer, and for how each sticholysin isotoxin is affected differently by the inclusion of cholesterol in the membrane. Furthermore, using phosphatidylcholine analogs that were labeled at different positions of their structure (acyl chains and headgroup) in combination with a variety of sticholysin mutants, we also investigated the depth of the tryptophan residues of sticholysins in the bilayer. Our results indicate that the position of the tryptophan residues relative to the membrane normal is deeper when cholesterol is absent from the membrane.

Localizing Molecules in Plant Cell Walls Using Fluorescence Microscopy.

Autofluorescence of plant tissues can be used as a label-free method to detect a range of phenolic-based cell wall components including lignin, suberin, and ferulate using widefield or confocal fluorescence microscopy. Likewise, fluorescently labeled antibodies can be used to localize specific carbohydrate molecules including arabinoxylan, β-1,4 galactan, glucomannan, glucuronoxylan, pectins, and xyloglucan. When combined, these two methods allow detailed study of topochemistry in different plant tissues for phenotyping of mutant varieties and plant biology studies. This article describes the protocols for fluorescent detection and imaging of molecules in plant cell walls using autofluorescence and immunofluorescence.

Analysis of the Adoption Rate of BATAN Mutant Rice Varieties in the Musi Rawas Area

The adoption of innovations of superior mutant rice varieties expected to help communities overcome hunger and food insecurity by providing easier access to food for poor people in rural areas. The purpose of this study is to determine the level of adoption of farmers for irradiated rice and analyze the factors that influence farmers in adopting irradiated rice. This research was conducted from October to November 2018 in Musi Rawas Regency using a survey method. Data obtained from interviews with 75 farmers who have applied the mutant variety. Survey data obtained were analyzed using SEM. The results showed that the technology adoption rate of mutant rice varieties reached the experimental stage; and There are three latent constructs that are significantly (ρ <0.01) positively correlated, namely: farmer preference (X2) to adoption rate (Y1), support of extension agent (X5) to farmer characteristics (X1) and farmer preference (X2).

Abstract 946: Utility of intestinal organoids to model serrated colon cancer in vitro

Abstract Serrated colorectal cancer (CRC) is an aggressive and treatment-resistant form of colorectal carcinogenesis that accounts for one-fourth of the patients with CRC1. At the molecular level, serrated CRC is characterized by epigenetic modifications termed the CpG Island Methylator Phenotype (CIMP) and the presence of somatic mutations that activate the mitogen-activated protein kinase (MAPK) pathway1,2,3. The BRAFV600E is an early genetic change in serrated polyps, but it is not sufficient to drive tumor formation. Combinations of mutations and epigenetic modifications in P16INK4A, MLH1, ZNFR3, RNF43, and TGFBR2 are often seen in most patients4. Therefore, a combinatorial approach to develop BrafV600E-driven serrated CRC organoids harboring a variety of the above mutants will enable in vitro studies of the tumorigenic potential of each combination as well as allow engraftment into animals. In turn, the latter will allow for applying preclinical in-vivo pharmacology modalities in vitro, thereby bypassing the time required to create a specific knock-in/out strain for each genetic modification desired. Here, we describe a pipeline to directly genetically manipulate specific organoid systems in vitro. We first established organoid cultures from the small and large intestine of unique Tet-inducible RNAi mice where efficient gene knock-down can contribute to microsatellite instability. Using CRISPR/Cas9, we then edited these to generate CRC organoid models containing the BrafV600E mutation in combination with knock outs or knock-ins of cell cycle regulators and negative regulators of the Wnt pathway. These deficiencies mimic the progressive mutational changes upon acquisition BrafV600E. Our data lay the groundwork for future use of RNAi technology in combination with sequential gene editing for in vitro pharmacology and modeling diseases with complex genetic architecture both in vitro and in vivo. Citation Format: Prem K. Premsrirut, Huaien Wang, Eduardo J. Garcia Reino, Isabella Breen, Ana A. Vasileva. Utility of intestinal organoids to model serrated colon cancer in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 946.

Comparative Analysis of the Metabolome and Transcriptome between the Green and Yellow-Green Regions of Variegated Leaves in a Mutant Variety of the Tree Species Pteroceltis tatarinowii.

In nature, many different factors cause plants to develop variegated leaves. To explore the mechanism of variegated leaf formation in Pteroceltis tatarinowii, a mutant variety (‘Jinyuyuan’), which was induced by ethylmethylsulfone, was selected, and its morphological structure, physiology, biochemistry, transcription and metabolism were analysed. According to differences in colour values, the colours were divided into two regions: a green region and a yellow-green region. The chlorophyll content of the two regions was significantly different. Moreover, the yellow-green regions of the leaves were significantly thinner than the green regions. The chloroplast ultrastructure in the yellow-green region revealed small chloroplasts, large vacuoles, small starch grains, obviously increased numbers of osmophilic grains, loose lamellae of the inner capsule and thin lamellae. Moreover, the yellow-green region was accompanied by oxidative stress, and the activity of the oxidative phosphorylation pathway related to oxidative activity in the transcriptome showed an upward trend. Vitamin B6 and proline contents also increased, indicating that the antioxidant activity of cells in the yellow-green region increased. Transcriptomic and metabolomic analysis showed that the differentially expressed genes (DEGs) related to chlorophyll synthesis and metabolism led to a decrease in the photosynthesis and then a decrease in the assimilation ability and contents of sucrose, starch and other assimilates. Amino acid synthesis and metabolism, lipid synthesis and the activity of metabolic pathways were obviously downregulated, and the contents of differentially accumulated metabolites associated with amino acids and lipids were also reduced. At the same time, 31 out of 32 DEGs involved in the flavonoid synthesis pathway were downregulated, which affected leaf colour. We hypothesized that the variegated leaves of P. tatarinowii ‘Jinyuyuan’ are caused by transcriptional and post-transcriptional regulation. Mutations in pigment and flavonoid synthesis pathway genes and transcription factor genes directly affect both pigment and flavonoid synthesis and degradation rate, which in turn affect carbon assimilation, carbon fixation, related protein synthesis and enzyme activity, lipid synthesis and degradation and the activity of other metabolic pathways, eventually leading to the formation of different colour regions.

Transcriptome mining of genes in Zanthoxylum armatum revealed ZaMYB86 as a negative regulator of prickly development

Zanthoxylum armatum DC. is an important economic tree species. Prickle is a type of trichome with special morphology, and there are a lot of prickles on the leaves of Z. armatum, which seriously restricts the development of Z. armatum industry. In this study, the leaves of Z. armatum cv. Zhuye (ZY) and its budding variety ‘Rongchangwuci’ (WC) (A less prickly mutant variety) at different developmental stages were used as materials, and the transcriptome sequencing data were analyzed. A total of 96,931 differentially expressed genes (DEGs) were identified among the samples, among which 1560 were candidate DEGs that might be involved in hormone metabolism. The contents of JA, auxin and CK phytohormones in ZY leaves were significantly higher than those in WC leaves. Combined with weighted gene co-expression network analysis, eight genes (MYC, IAA, ARF, CRE/AHK, PP2C, ARR-A, AOS and LOX) were identified, including 25 transcripts, which might affect the metabolism of the three hormones and indirectly participate in the formation of prickles. Combining with the proteins successfully reported in other plants to regulate trichome formation, ZaMYB86, a transcription factor of R2R3 MYB family, was identified through local Blast and phylogenetic tree analysis, which might regulate prickle formation of Z. armatum. Overexpression of ZaMYB86 in mutant A. thaliana resulted in the reduction of trichomes in A. thaliana leaves, which further verified that ZaMYB86 was involved in the formation of pickles. Yeast two-hybrid results showed that ZaMYB86 interacted with ZaMYB5. Furthermore, ZaMYB5 was highly homologous to AtMYB5, a transcription factor that regulated trichomes development, in MYB DNA binding domain. Taken together, these results indicated that ZaMYB86 and ZaMYB5 act together to regulate the formation of prickles in Z. armatum. Our findings provided a new perspective for revealing the molecular mechanism of prickly formation.

Comparative retrotransposon analysis of mutant and non-mutant rice varieties grown at different salt concentrations

This study investigated the effects of retrotransposon movements on the regeneration of 15- and 30-day-old leaves and roots of four different rice varieties grown in tissue culture medium at different salt concentrations (0, 50, 100, and 200 mmol/L) by Inter Retrotransposon Amplified Polymorphism (IRAP) and Retrotransposon Microsatellite Amplified Polymorphism (REMAP) polymerase chain reaction (PCR)-based marker methods. The study used Hopi, Houba, Osr30, Tos17 primers for IRAP analysis. The (GA)9C ISSR primer was used with four IRAP primers in REMAP analyses. There were different band profiles and polymorphism results between different salts concentrations of the same variety and various rice varieties. We observed that the polymorphism ratios for Houba ranged from 0 to 98% in IRAP and REMAP analyzes in the calculation made with the Jaccard similarity index in mutant samples. We report that mutant varieties give more valuable IRAP and REMAP results than non-mutant varieties. The results reported in this study contribute to understanding the effect of salinity stress on rice in terms of retrotransposon-based molecular markers. These results suggest that all tested retrotransposons are still active, causing genomic polymorphism among rice plantlets.

Clinical Profile and Survival Outcome of Endometrial Cancer with p53 Mutation.

Clinicopathologic classification of endometrial cancer imperfectly reflects the tumor biology. Pathologic categorization - especially in high-grade tumors - results in an imprecise estimation of the risk of disease, recurrence, and death. Molecular subtyping is emerging as the standard of care in diagnosis and treatment of endometrial cancers. Molecular markers are important prognostic factors in tumor dissemination and early recurrence of endometrial cancers. TP53 mutation is an important prognostic factor for both serous and endometrioid cancers. The study aims to compare the clinical profile and overall survival of endometrial cancers with and without p53 mutation. Sixty-three patients who underwent surgical staging for carcinoma endometrium were included in the study.TP53 mutation status was determined based on p53 expression by immunohistochemistry (IHC) as a p53 wild or p53 mutant type. Data were analyzed for the clinical profile, p53 mutation status on IHC, histological pattern, tumor grade, stage of the disease, lymph node spread, recurrence pattern, treatment received, 2-year disease-free survival, and overall survival. Recurrence was noted in 12.7% patients after 2-year follow-up, of which 75% patients had p53 mutation. Significant association was seen between p53 expression and high-grade tumors, stage, cervical involvement, and adnexal involvement. The 2-year overall survival of the p53 wild type was 97.2% and the p53 mutant type was 91.7%. The 2-year disease-free survival for the p53 wild type was 94.3% and the disease-free survival of the p53 mutant variety was 83.5%. The 2-year disease-free survival for endometrioid carcinoma with p53 wild type was 100% and p53 mutant variety was 86.2% (p value 0.033). About 15.9% (10) patients were reassigned to the high-risk group needing chemotherapy and radiation according to the ESGO ESTRO 2021 consensus classification, due to their p53 mutation status. IHC to assess somatic p53 mutation may be done in endometrial biopsies irrespective of their histology. This may help to identify that the aggressive tumors thereby help in tailoring surgery, planning adjuvant treatment, and follow-up.

Electrical impulse characterization along actin filaments in pathological conditions

We present an interactive Mathematica notebook that characterizes the electrical impulses along actin filaments in both muscle and non-muscle cells for a wide range of physiological and pathological conditions. The simplicity of the theoretical formulation, and high performance of the Mathematica software, enable the analysis of multiple conditions without computational restrictions. The program is based on a multi-scale (atomic → monomer → filament) approach capable of accounting for the atomistic details of a protein molecular structure, its biological environment, and their impact on the travel distance, velocity, and attenuation of monovalent ionic wave packets propagating along microfilaments. The interactive component allows investigators to choose the experimental conditions (intracellular Vs in vitro), nucleotide state (ATP Vs ADP), actin isoform (alpha, gamma, beta, and muscle or non-muscle cell), as well as a conformation model that covers a variety of mutants and wild-type (the control) actin filaments. We used the computational tool to analyze environmental changes such as temperature effects and pH changes of the surrounding solutions, as well as structural changes to an actin monomer due to radius changes. Additionally, we investigated for the first time the electrostatic consequences of actin mutations from different disease conditions. These studies may provide an unprecedented molecular understanding of why and how age, inheritance, and disease conditions induce dysfunctions in the biophysical mechanisms underlying the propagation of electrical signals along actin filaments. Program summaryProgram Title: EIAF (Electrical Impulse along Actin Filaments)CPC Library link to program files:https://doi.org/10.17632/k8vw4j6y75.1Developer's repository link:https://github.com/MarceloMarucho/SignalPropagationPathologicalConditionLicensing provisions: GPLv2Programming language: MathematicaNature of problem: Electrical properties of actin filaments in pathological conditions have yet to be studied, despite previous work showing influential consequences for physiological conditions on wild type actins.Solution method: The interactive Mathematica notebook is based on the Multi-scale Approach for electrical impulses along cytoskeletal filaments and uses non-linear considerations for the Boltzmann statistics of ionic distributions in the electrical double layer (EDL). Using the numerical methods incorporated in Mathematica’s default libraries, and an interactive GUI, the program allows users to conduct their own research by choosing the experimental conditions (intracellular Vs in vitro), nucleotide state (ATP Vs ADP), actin isoform (including muscle and non-muscles cells), as well as a conformation model that covers a variety of mutants and wild-type (the control) actin filament configurations.Additional comments including restrictions and unusual features: The program allows users to adjust pH values, input voltages, as well as study two default electrolyte conditions without rerunning the program. The program is fully editable, such that motivated users can change the default electrolytes to fit specific conditions in their research project in necessary.