Real-time expression and in silico characterization of pea genes involved in salt and water-deficit stress.

BACKGROUND: To tolerate salt and water-deficit stress, the plant adapts to the adverse environment by regulating its metabolism and expressing certain stress-induced metabolic pathways. This research analyzed the relative expression of four pea genes (P5CR, PAL1, SOD, and POX) in three pea varieties (Climax, Green grass, and Meteor) under different levels of salt and water-deficit stress. METHODS AND RESULTS: The experiments on salt stress and water-deficit stress were carried out within greenhouse settings under controlled environment. The saturation percentage was employed to create artificial salinity conditions: Control without NaCl treatment, Treatment 1: 50 mM NaCl treatment, Treatment 2: 75 mM NaCl treatment, and Treatment 3: 100 mM NaCl treatment. Field capacity (FC) was used for the development of artificial water-deficit treatments in the pots, i.e., Treatment 1 (Control; water application 100% of FC), Treatment 2 (water application 75% of FC), and Treatment 3 (water application 50% of FC). Pea genes involved in biosynthetic pathways of proline, flavonoids, and enzymatic antioxidant enzymes including P5CR, PAL1, SOD, and POX were selected based on literature. Quantitative real-time PCR using cDNA as a template was used to analyze the gene expression. Pea genes were analyzed for phylogenetic analysis in closely related crops having similarity percent identity 80% and above. In silico characterization of selected proteins including the family classification was done by the NCBI CDD and INTERPRO online servers. Results from RT-qPCR analysis showed increased expression of P5CR, PAL1, and POX genes, while SOD gene expression decreased under both stresses. Climax exhibited superior stress tolerance with elevated expression of P5CR and PAL1, while Meteor showed better tolerance through increased POX expression. Phylogenetic analysis revealed common ancestry with other species like chickpea, red clover, mung bean, and barrel clover, suggesting the cross relationship among these plant species. Conserved domain analysis of respective proteins revealed that these proteins contain PLNO 2688, PLN02457, Cu-Zn Superoxide dismutase, and secretory peroxidase conserved domains. Furthermore, protein family classification indicated that the oxidation-reduction process is the most common chemical process involved in these stresses given to pea plant which validates the relationship of these proteins. CONCLUSIONS: Salt and water-deficit stresses trigger distinct metabolic pathways, leading to the up-regulation of specific genes and the synthesis of corresponding proteins. These findings further emphasize the conservation of stress-tolerance-related genes and proteins across various plant species. This knowledge enhances our understanding of plant adaptation to stress and offers opportunities for developing strategies to improve stress resilience in crops, thereby addressing global food security challenges.

Proteomic Analysis of Medicinal Plant Calotropis Gigantea by In Silico Peptide Mass Fingerprinting.

Medicinal plants are the basic source of medicinal compounds traditionally used for the treatment of human diseases. Calotropis gigantea is a medicinal plant belonging to the family of Apocynaceae in the plant kingdom and subfamily Asclepiadaceae usually bearing multiple medicinal properties to cure a variety of diseases. BACKGROUND: The Peptide Mass Fingerprinting (PMF) identifies the proteins from a reference protein database by comparing the amino acid sequence that is previously stored in the database and identified. OBJECTIVE: The purpose of the study is to identify the peptides having anti-cancerous properties by in silico peptide mass fingerprinting. METHODS: The calculation of in silico peptide masses is done through the ExPASy PeptideMass and these masses are used to identify the peptides from the MASCOT online server. Anticancer probability is calculated by iACP server, docking of active peptides is done by CABS-dock the server. RESULTS: The anti-cancer peptides are identified with the MASCOT peptide mass fingerprinting server, the identified peptides are screened and only the anti-cancer are selected. De-novo peptide structure prediction is used for 3D structure prediction by PEP-FOLD 3 server. The docking results confirm strong bonding with the interacting amino acids of the receptor protein of breast cancer BRCA1 which shows the best peptide binding to the active chain, the human leukemia protein docking with peptides shows the accurate binding. CONCLUSION: These peptides are stable and functional and are the best way for the treatment of cancer and many other deadly diseases.

Use Of Steroids In Rhinoplasty With Lateral Osteotomies For Reducing Post Operative Oedema.

BACKGROUND: TPostoperative periorbital oedema is a commonly encountered side effect of rhinoplasties in which lateral osteotomies have been incorporated. It dissatisfies the surgeon as well as the patient. Osteotomies are done at the end of all soft tissue manipulation to reduce the development of oedema. The aim of this study was to determine the efficacy of intravenous dexamethasone in reducing oedema in patients who undergo rhinoplasty with lateral osteotomies. METHODS: A Prospective randomized controlled trial was done at department of plastic and reconstructive surgery, Shifa International Hospital Islamabad. Sixty patients age between 16-55 requiring open rhinoplasty were taken for this study and divided in two groups. One group received dexamethasone 8mg intravenously preoperatively and second dose 4 hours postoperatively. The second group did not receive anything. Both groups were assessed on first post-operative day and 7th day for periorbital oedema. RESULTS: The overall decrease in oedema in patients who received steroid was by 50% while in control group was 33.3%. By the 7th day control group 13.3% patients had grade III oedema as compared to 3.33% in steroid group. Chi test was applied and p-value of 0.0289 was obtained which was found to be statistically very significant. CONCLUSIONS: Dexamethasone used in minimal dosage showed significant advantage in reducing periorbital oedema after rhinoplasty with no evidence of any side effects secondary to steroid administration.