Liver-boosting potential: chicory compound-mediated silver nanoparticles for hepatoprotection-biochemical and histopathological insights.

Background: Liver disease is a serious health concern in today's world, posing a challenge to both healthcare providers and pharmaceutical companies. Most synthetic drugs and chemicals cause liver damage accounting for approximately 10% of acute hepatitis and 50% of acute liver failure. Purpose: The present study aimed to evaluate the hepato-protective activity of an extract of chicory formulation assisted by silver nanoparticles against carbon tetra chloride (CCl4)-induced hepatic damage in rat's liver. Methods: Rats of the Wistar strain (Rattus norvegicus) were used to test the in vivo hepato-protective efficacy at various doses. Rats were randomly divided into nine groups, each containing six rats. The groups were as follows: first group (control), second group (CCl4), third group, silymarin (20 mg/kg of body weight), fourth group (CCl4+chicory) (1.75 mg/kg of b. wt), fifth group (CCl4 + chicory at the dose of 2.35 mg/kg), sixth group (CCl4 + chicory of 3.25 mg/kg), seventh group (CCl4 +AgNPs 1.75 mg/kg of b. wt.), eighth group (CCl4 + AgNPs 2.35 mg/kg of body weight), and ninth group (CCl4 + AgNPs 3.25 mg/kg of b. wt.). Blood samples were taken 24 h after the last administration (i.e., 30th day). The blood samples were analyzed for different serum enzymes such as ALP (alkaline phosphatase), ALT (alanine transaminase), bilirubin (Blr), triglyceride, and cholesterol. Histology liver sections were performed. Results: Treatment with AgNPs and chicory extract showed significant hepato-protective activity in a dose-dependent manner. In three doses, the chicory extract at a rate of 3.25 mg/kg of body weight significantly reduced elevated levels of biochemical markers in comparison to CCl4-intoxicated rats. Histology of the liver sections from CCl4-treated rats revealed inflammation of hepatocytes, necrosis, cytoplasmic degeneration, vacuolization, and a deformed central vein. The chicory formulation extract exhibited a remarkable recovery percentage in the liver architecture that was higher than the drug (i.e., silymarin). While treatment with AgNPs also repaired the degenerative changes and restored the normal form of the liver, chicory formulation extract possessed more hepato-protective potential as compared to AgNPs by regulating biochemical and histo-pathological parameters. Conclusion: This study can be used as confirmation of the hepato-protective potential of chicory compounds for possible use in the development programs of drugs to treat liver diseases.

Ethno-Dentistry of Medicinal Plants Used in North Waziristan, Pakistan.

OBJECTIVE: This study aimed at recording therapeutic plant species used by inhabitants to treat dental disorders in the district of North Waziristan, Pakistan. The indigenous people of the studied area are dependent on medicinal plants for their basic health care needs including dental care. METHODS: Ethnomedicinal data were collected using a semi-structured questionnaires, and in addition 130 local informants were interviewed. The collected data were evaluated using various quantitative indices, including use value (UV), relative frequency of citation (RFC), fidelity level (FL%), and Jaccard Index (JI). RESULTS: A total of 69 plants belonging to 48 plant families used in dental disorders were identified. The Lamiaceae was the leading family that shared 7 species, followed by Solanaceae (4 spp).The dominant life form used was herbs (47.83%), folowed by leaves (43.90%) in preparing remedies for different dental disorders. Decoction was the most common mode of preparation (34.21%), followed by pastes (21.05%). The highest RFC (0.36) was reported for Bergenia ciliata, followed by Salvadora oleoides (0.35). The majority of the plants (36 spp) were utilised as herbal medicine to treat toothache, followed by 13 species for periodontal (gum) infections, 11 species used for teeth cleaning, and 9 species for halitosis (bad breath). CONCLUSIONS: This study is the first-ever record of ethnomedicinal applications for the treatment of dental diseases from Pakistan. Some of the forgeoing hebal medications should be further evalauted for the development of pahrmaceutical bio-products for the treatment of dental disorders.

Advanced methods of algal pigments extraction: A review.

Algae are exclusively aquatic photosynthetic organisms that are microscopic or macroscopic, unicellular or multicellular and distributed across the globe. They are a potential source of food, feed, medicine and natural pigments. A variety of natural pigments are available from algae including chlorophyll a, b, c d, phycobiliproteins, carotenes and xanthophylls. The xanthophylls include acyloxyfucoxanthin, alloxanthin, astaxanthin, crocoxanthin, diadinoxanthin, diatoxanthin, fucoxanthin, loroxanthin, monadoxanthin, neoxanthin, nostoxanthin, perdinin, Prasinoxanthin, siphonaxanthin, vaucheriaxanthin, violaxanthin, lutein, zeaxanthin, ß-cryptoxanthin, while carotenes include echinenone, α-carotene, ß-carotene, γ-carotene, lycopene, phytoene, phytofluene. These pigments have applications as pharmaceuticals and nutraceuticals and in the food industry for beverages and animal feed production. The conventional methods for the extraction of pigments are solid-liquid extraction, liquid-liquid extraction and soxhlet extraction. All these methods are less efficient, time-consuming and have higher solvent consumption. For a standardized extraction of natural pigments from algal biomass advanced procedures are in practice which includes Supercritical fluid extraction, Pressurized liquid extraction, Microwave-assisted extraction, Pulsed electric field, Moderate electric field, Ultrahigh pressure extraction, Ultrasound-assisted extraction, Subcritical dimethyl ether extraction, Enzyme assisted extraction and Natural deep eutectic solvents. In the present review, these methods for pigment extraction from algae are discussed in detail.

Ethnic practices in treating skin diseases: The traditional dermatologist's role.

This study documents the role of traditional medicinal and therapeutic plants in treating various skin ailments by indigenous communities of the tribal district of North Waziristan, Pakistan. A total of 130 informants and traditional dermatologists were interviewed. They employed 77 plant species belonging to 49 families to treat various skin ailments. The leading life form reported was herbs (41 species), whereas the dominant family was Lamiaceae (5 species). Leaves were the most commonly used plant part (37 species). The most preferred mode of preparation was paste (30.38%), and the dominant mode of administration was topical (69.23%). Important medicinal plants reported in this study are recommended for further phytochemical screening for bioactive constituents, which may lead to novel drug discoveries.

Ethnogynaecological Knowledge of Traditional Medicinal Plants Used by the Indigenous Communities of North Waziristan, Pakistan.

Background: Since the beginning of civilization, medicinal plants have been used in human healthcare systems. Studies have been conducted worldwide to evaluate their efficacy, and some of the results have triggered the development of plant-based medications. Rural women in Pakistan frequently experience gynaecological disorders due to malnutrition and heavy physical work during pregnancy. Due to the low economic status, the remoteness of the area, and the lack of modern health services, herbal therapy for gynaecological disorders is common among the indigenous tribes of the study area. Methods: Field surveys were carried out from April 2018 to October 2020 to collect data regarding medicinal plants used for different gynaecological disorders. A semistructured questionnaire was used to collect ethnogynaecological data. Results: In total, 67 medicinal plant species belonging to 38 families are being used to treat 26 different gynaecological problems. The herbaceous growth form and the Lamiaceae family were recorded with the maximum number of plant species (42 species and 7 species, respectively). Leaves are the most highly utilized plant part, with 16 species. In the case preparation method, decoction was the dominant method (25 species, 36.76%). The informants reported the maximum number of species for the treatment of irregular menstrual flow as 11 species (15.28%). The highest relative frequency of citation (RFC) value was obtained for Acacia modesta (0.37), and the use value (UV) for Tecomella undulata (0.85). The highest informants' consensus factor (ICF) value (1.0) was obtained for emmenagogue and tonic each after delivery. The highest consensus index (CI%) value was calculated for Acacia modesta (36.92%). The Lamiaceae had the highest family importance value (FIV) (98.46%). Conclusion: This is the first ever quantitative study focusing mainly on ethnogynaecological study conducted in the tribal areas of North Waziristan which highlights the importance of traditional herbal remedies for their basic medical requirements. The results of this study would serve as a baseline for advanced phytochemical and pharmacological screening, as well as conservationists for further studies.

Comparative Study of Antimicrobial and Antioxidant Potential of Olea ferruginea Fruit Extract and Its Mediated Selenium Nanoparticles.

Nanotechnology, the science of the recent era, has diverse applications in agriculture. Selenium (Se) is a non-metal and an essential micronutrient for animals and humans. In this study, selenium nanoparticles (SeNPs) were biosynthesized by using Olea ferruginea fruit extracts. The size, shape, chemical nature, and identification of functional groups involved in the synthesis of SeNPs were studied by UV-visible spectroscopy, Scanning Electron Microscope (SEM), and Fourier Transform Infra-Red (FTIR) spectrometry. SeNP synthesis was confirmed by an absorption peak at 258 nm by UV-visible spectroscopy. SEM showed that SeNPs were spherical, smooth, and between 60 and 80 nm in size. FTIR spectrometry confirmed the presence of terpenes, alcohols, ketones, aldehydes, and esters as well as phyto-constituents, such as alkaloids and flavonoids, that possibly act as reducing or capping agents of SeNPs in an aqueous solution of Olea ferruginea. Antimicrobial activity was examined against bacterial pathogens, such as Klebsiella pneumonia, Escherichia coli, Staphylococcus aureus, and Staphylococcus epidermitis, as well as fungal pathogens, such as Aspergillus niger and Fusarium oxysporum, by using the well-diffusion method. Antioxidant activity was observed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, ABTs assay, and reducing power assay. At a higher concentration of 400 ppm, biosynthesized SeNPs showed an inhibition zone of 20.5 mm, 20 mm, 21 mm, and 18.5 mm against Klebsiella pneumonia, Escherichia coli, Staphylococcus aureus, and Staphylococcus epidermitis, respectively. Similarly, SeNPs also demonstrated a zone of inhibition against Aspergillus niger and Fusarium oxysporum of 17.5 and 21 mm, respectively. In contrast to Olea ferruginea fruit extracts, Olea ferruginea-mediated SeNPs demonstrated strong antimicrobial activity. By performing the DPPH, ABTs, and reducing power assay, SeNPs showed 85.2 ± 0.009, 81.12 ± 0.007, and 80.37 ± 0.0035% radical scavenging potential, respectively. The present study could contribute to the drug development and nutraceutical industries.

The potential of Bacillus subtilis and phosphorus in improving the growth of wheat under chromium stress.

AIM: Hexavalent chromium (Cr+6 ) is one of the most toxic heavy metals that have deteriorating effects on the growth and quality of the end product of wheat. Consequently, this research was designed to evaluate the role of Bacillus subtilis and phosphorus fertilizer on wheat facing Cr+6 stress. METHODS AND RESULTS: The soil was incubated with Bacillus subtilis and phosphorus fertilizer before sowing. The statistical analysis of the data showed that the co-application of B. subtilis and phosphorus yielded considerably more significant (p < 0.05) results compared with an individual application of the respective treatments. The co-treatment improved the morphological, physiological and biochemical parameters of plants compared with untreated controls. The increase in shoot length, root length, shoot fresh weight and root fresh weight was 38.17%, 29.31%, 47.89% and 45.85%, respectively, compared with untreated stress-facing plants. The application of B. subtilis and phosphorus enhanced osmolytes content (proline 39.98% and sugar 41.30%), relative water content and stability maintenance of proteins (86.65%) and cell membranes (66.66%). Furthermore, augmented production of antioxidants by 67.71% (superoxide dismutase), 95.39% (ascorbate peroxidase) and 60.88% (catalase), respectively, were observed in the Cr+6 - stressed plants after co-application of B. subtilis and phosphorus. CONCLUSION: It was observed that the accumulation of Cr+6 was reduced by 54.24%, 59.19% and 90.26% in the shoot, root and wheat grains, respectively. Thus, the combined application of B. subtilis and phosphorus has the potential to reduce the heavy metal toxicity in crops. SIGNIFICANCE AND IMPACT OF THE STUDY: This study explored the usefulness of Bacillus subtilis and phosphorus application on wheat in heavy metal stress. It is a step toward the combinatorial use of plant growth-promoting rhizobacteria with nutrients to improve the ecosystems' health.

Ethnomedicinal appraisal of plants used for the treatment of gastrointestinal complaints by tribal communities living in Diamir district, Western Himalayas, Pakistan.

Majority of the mountain dwelling communities living in the Himalayas rely on traditional herbal medicines for primary healthcare needs. Present study was conducted in fairy meadows and allied valleys in District Diamir, Gilgit Baltistan autonomous territory in northern Pakistan. Documentation of traditional medicinal knowledge (TMK) of local communities for the treatment of gastrointestinal disorders was carried out as a component of a wider medico-botanical expedition conducted in the entire base camp of the great Nanga Parbat peak during 2016-19. Various ethnobotanical parameters i.e. use value (UV), informant consensus factor (ICF), Fidelity level (FL), direct matrix ranking test (DMRT) and preference ranking (PR) were applied to evaluate the data collected during field surveys. The plants were also subjected to a comparative review for novelty assessment. A total of 61 medicinal plant species belonging to 55 genera and 35 families are reported here for the treatment of GIDs. Compositae was the leading family with 8 (13%) species. Fourteen gastrointestinal disorders were cured with 32% taxon were reported for stomachic followed by diarrhea (15%) and constipation (14%). Highest use reports (5) and use citations (207) were reported for Mentha longifolia L. while highest UV (1.79) was obtained for Artemisia maritima L. Hylotelephium telephioides (Ledeb.), A. maritima, M. longifolia, M. piperita L., Allium cepa L., and A. annua L. exhibited 100% FL. Highest ICF was calculated against dysentery and flatulence. DMRT ranked Prunus persica L. first for its multipurpose uses. Taking constipation as a reference gastrointestinal disease, PR for ten plant species was calculated where H. telephioides was ranked first followed by A. maritima. Present study concluded that 19 out of 61 plant species were documented for the first time with novel medicinal uses to cure GIDs. These plant species could act as potential reservoirs of novel lead compounds for the treatments of gastrointestinal disorders.

Light and scanning electron microscopic characterization of aflatoxins producing Aspergillus flavus in the maize crop.

Maize (Zea mays L.) is considered as one of the main cereals, used as a source of food, forage, and processed products. The loss of maize productivity is reported due to effect on roots, stalks, ears, and kernels mainly caused by many fungi. Among these fungal pathogens of maize, Aspergillus flavus (A. flavus) are the most prevalent that produces highly toxigenic aflatoxins that are highly carcinogenic to the consumers. The present study is confined to isolate and characterize the A. flavus from maize seeds for accurate identification that can be helpful for determination and management of aflatoxins in maize crop. Eighty stored seed samples of maize were collected from warehouses where seeds are stored for food and feeding purposes. For the isolation of A. flavus, Potato Dextrose Agar was used. Isolated fungi were identified macro and microscopically using light microscope and scanning electron microscope. A total of 212 Aspergillus isolates were identified based on macro-morphological and micro-morphological characteristics. The results showed that A. flavus colonies were granular, flat with yellow-green to deep yellow-green colony color having a white border and compact, spherical spore heads. Rapid rate of growth was observed maturing in about 3-5 days. In microscopic features, A. flavus have apically swollen conidiophores with various conidia bearing cells in long and dry chains. Spherical conidial heads were split into several columns ranging 300-400 µm in diameter. This will be helpful for farmers, researchers and traders in future for correct identification of sources of aflatoxins. RESEARCH HIGHLIGHTS: Maize seed samples were collected from Pothohar region of Pakistan. The fungi were isolated on PDA. Aspergillus flavus was identified macro-morphologically by observing growth rate, colony color and texture. Furthermore, these fungi were identified micro-morphologically by using light and scanning electron microscope. The 212 Aspergillus flavus strains were isolated and identified.

Neuroprotective Effect of Otostegia limbata Against PTZ-Induced Mice Model of Epilepsy by Attenuated Expression of p-NFκB and TNF-α.

Epilepsy is a chronic neurological disorder characterized by recurrent unprovoked seizures. Currently available antiepileptic drugs have severe side effects and do not offer complete cure. Herbal remedies have been used for centuries to treat many neurodegenerative disorders. Otostegia limbata L. belongs to the largest and medicinally important family Lamiaceae and is distributed in hilly areas of Pakistan. This study was designed to assess the antioxidant, anti-inflammatory, and anticonvulsant potential of O. limbata. The methanolic extract showed significant antioxidant activity assessed by (1,1-diphenyl 2-picrylhydrazyl) free-radical scavenging assay, nitric oxide scavenging, and iron chelation antioxidant assays. The methanolic extract was evaluated for its anticonvulsant effect, employing the pentylenetetrazole (PTZ)-induced mice model of epilepsy. Three different doses of O. limbata (100, 200, and 300 mg/kg) were administered orally 30 min before PTZ [50 mg/kg, intraperitoneal (i.p.)] injection, while diazepam was used as a positive control. The extract at 300 mg/kg significantly decreased the duration and increased the latency of the PTZ-induced seizures. The expression of inflammatory cytokines tumor necrosis factor α (p-TNF-α) and phosphorylated transcription factor nuclear factor kappa B (p-NF-κB), in the cortex and hippocampus of the brains of treated mice were analyzed through enzyme-linked immunosorbent assay and western blot analysis. The morphological changes and number of surviving neurons were recorded through hematoxylin and eosin staining. The seizure score and survival rate of the treated group showed considerable differences as compared to the PTZ group. TNF-α and p-NF-K b expression were downregulated as compared to the PTZ group. The anticonvulsant effect may be the outcome of the antioxidant potential and high levels of phenols and flavonoids detected in the methanolic plant extract through Fourier transform infrared spectrophotometer and gas chromatography-mass spectrometry analysis.

Ethnobotanical study of Mandi Ahmad Abad, District Okara, Pakistan.

This study hypothesized that native people have unique traditional knowledge of plant resources in the rural areas and basic objective was the documentation of this valuable inheritance. Ethnobotanical data was collected from a remote rural area of Mandi Ahmad Abad, Union council number NA-144 Tehsil Depalpur District Okara, Pakistan. A total of 94 informants were randomly interviewed to collect data about local names of plant species, mode of administration, recipes and ailments, and ethnobotanical uses through semi-structured questionnaire, interviews and group discussion methods. The collected data was statistically analyzed by calculating use value (UV), frequency of citation (FC), relative frequency of citation (RFC), factor of informant consensus (FIC), family importance value, and relative importance (RI). This study is also compared with ethnobotanical literature by using Jaccard's index (JI) for similarity analysis. A total of 126 species belonging to 52 families were documented. The Poaceae (13spp.), Leguminosae (12spp.), Solanaceae (10spp.) and Cucurbitaceae (10spp.) were dominant families. Highest used value (UV = 0.22) was obtained for Azadirachta indica. The minimum used value (UV) was showed by Alhagi maurorum, Eclipta prostrata, Hibiscus rosa-sinensis, Solanum virginianum and Trianthema potulacastrum (UV = 0.01). Hepatitis, stomach ulcer, bowel disorders, urinary problems, psoriasis, cancer, and leucoderma were the most treated ailments with ICF value of 1, followed by leucorrhea (ICF: 0.89), and vomiting (ICF: 0.86). The highest Jaccard's similarity index value (JI = 0.329) showed that plant species reported in our study was more similar with Arid regions of Northern Punjab, Pakistan. This novel ethnobotanical report concluded that traditional knowledge about use of medicinal plants is decreasing due to allopathic medicines. Immediate steps should be taken for conservation and documentation of traditional knowledge of plants especially those having medicinal properties.

Ethnoveterinary Practices of Medicinal Plants Among Tribes of Tribal District of North Waziristan, Khyber Pakhtunkhwa, Pakistan.

Domestic animals play a vital role in the development of human civilization. Plants are utilized as remedies for a variety of domestic animals, in addition to humans. The tribes of North Waziristan are extremely familiar with the therapeutic potential of medicinal plants as ethnoveterinary medicines. The present study was carried out during 2018-2019 to record ethnoveterinary knowledge of the local plants that are being used by the tribal communities of North Waziristan, Khyber Pakhtunkhwa, Pakistan. In all, 56 medicinal plant species belonging to 42 families were identified, which were reported to treat 45 different animal diseases. These included 32 herbs, 12 shrubs, and 12 trees. Among the plant families, Asteraceae contributed the most species (5 spp.), followed by Amaranthaceae (4 spp.), Solanaceae (4 species), and Alliaceae, Araceae, and Lamiaceae (2 spp. each). The most common ethnoveterinary applications were documented for the treatment of blood in urine, bone injury, colic, indigestion, postpartum retention, skin diseases, constipation, increased milk production, mastitis, foot, and mouth diseases.

Palyno-morphological attributes of some selected plant species of family Asteraceae from district Dera Ismail Khan, KPK, Pakistan.

The current study aimed to investigate the palyno-morphological features of Asteraceous species from District Dera Ismail Khan, Khyber Pakhtunkhwa, Pakistan. A total of 13 species were collected, pressed, identified, and examined by using light and scanning electron microscopy. Both qualitative and quantitative pollen characters were observed, that is, equatorial and polar diameter, number of pores and colpi, exine thickness, exine sculpturing, pollen shape, and P/E ratio. Pollen shapes observed in studied taxa were spheroidal, suboblate, oblate-spheroidal, and lophate-spheroidal. The aperturation patterns of pollens vary from tricolporate to tetracolporate. The maximum polar and equatorial diameter of 75-100 (87.5 ± 13.6) µm and 87.5-117.5 (102.5 ± 16.4) µm, respectively, was recorded in Echinops echinatus, while minimum polar and equatorial diameter of 20-22.5 (21 ± 1.36) µm and 22.5-25 (23.5 ± 1.36) µm, respectively, was recorded in Aster subulatus. Six types of exine sculpturing patterns were observed; echinate, micro-echinate, echinate-microreticulate, microreticulate, echinate-perforate, and scabrate. Maximum exine thickness of 8.75 µm was recorded in E. echinatus and minimum of 1.25 µm in Launaea mucronata. The pollen morphology has valuable significance in understanding the taxonomy of different plant groups and plays a central role in the correct identification and classification of Asteraceous flora at species, generic, and tribe levels.

Formulation and evaluation of antioxidant and antityrosinase activity of Polygonum amplexicaule herbal gel.

Medicinal plants are long been used for pharmaceutical and cosmetic industry. Among medicinal plants, Polygonum amplexicaule of family polygonaceae has traditional use in medicines and skin care. P. amplexicaule belongs to genus Polygonum that contains several important phytochemicals and considered as a rich source of antioxidants. The present study was designed to formulate herbal gel containing P. amplexicaule extract and evaluate its different physical properties as well as antioxidants and antityrosinase activities. Chitosan gel base was used as gelling agent and different gel formulations were prepared by different concentrations of extracts and polymers. Physical properties like pH, colour, odour, appearance and homogeneity, spreadability, extrudability and stability were optimized and analysed. A stable gel formulation containing 1% chitosan gel base and 5% plant extract was prepared that showed good appearance and homogeneity, easily spread ability and excellent extrudability. This gel formulation was tested for antioxidant and skin whitening properties by DPPH free radical scavenging assay and tyrosinase inhibition assay respectively and ascorbic acid was used as reference standard. DPPH scavenging activity with an IC50 value of 0.446 mg/mL and tyrosinase inhibition activity with an IC50 value of 0.805 mg/mL was observed and results indicated that this herbal gel formulation has a good potential for cosmetic use.

Effects of Pre-Anthesis Drought, Heat and Their Combination on the Growth, Yield and Physiology of diverse Wheat (Triticum aestivum L.) Genotypes Varying in Sensitivity to Heat and drought stress.

Independent and combined drought and heat stress negatively affect wheat yield and physiology. The present study was aimed to quantify effects of Drought [D], heat [H] and combined heat and drought [HD] during reproductive stage on wheat yield and to identify physio-biochemical traits which were strongly associated with improved yield and tolerance of wheat under stressful conditions. One hundred and eight elite diverse wheat genotypes were exposed to [H], [D] and [HD] treatments from heading till maturity. Grain yield was reduced by 56.47%, 53.05% and 44.66% under [HD], [H] and [D] treatment, respectively. The [HD] treatment affects the grain yield by reducing metabolism and mobilization of reserves to developing grains and leaves. Disintegration of membrane structure, chlorophyll and protein molecules was higher under [H] stress than [D] stress while water status of genotypes and sink strength was more affected by [D] than [H] stress. Multivariate analysis showed a strong correlation of chlorophyll content before and after anthesis, water-soluble carbohydrates (WSC), proline content (PC) and all other studies agronomic and physiological traits with grain yield while days to anthesis (DTA) and days to maturity (DTM) were negatively associated with grain yield under stress showing advantage of early maturity during stress. Traits having a major contribution in the first two principal components under different stress treatments may lead to improved varieties with heat and drought stress tolerance. To best of over knowledge, the present study is the first detailed study which used physiological and biochemical traits to explain the variation in grain yield and related traits in diverse wheat germplasm.

Genome-wide association analyses for yield and yield-related traits in bread wheat (Triticum aestivum L.) under pre-anthesis combined heat and drought stress in field conditions.

Understanding the genetic basis of heat and drought stress tolerance in wheat is prerequisite for wheat breeding program. In the present study, a wheat panel comprising of 192 elite bread wheat genotypes was phenotyped in eight environments for yield and related traits in field conditions. Four stress environments were created by implying four different treatments differing in sowing date and water availability, panel was evaluated for two years in field conditions. The panel was genotyped with 15K Illumina chip and 9236 polymorphic markers concentrated on B genome were employed in GWAS analysis. Consistent, fast LD decay was observed on D genome and structure analysis germplasm divided panel into three major populations. GWAS was performed using BLUEs values of combined environment data in R package GAPIT using log10(P) = 3.96 as significance threshold. The significance of association was further checked using FDR<0.05 threshold. The GWAS identified 487 loci associated with the traits and were significant at log10(p) threshold out of these 350 loci were significant at FDR threshold. For two stress indices 108 associations were significant at FDR threshold. Nine genomic regions were shared among all treatment, while multiple pleiotropic regions were present on chromosome 7D followed by unmapped chromosome. The present study validated many marker trait associations for yield and other traits, MTAs significant under combined drought and heat stress were novel. These regions are important and can be used for fine mapping and marker assisted selection to discover new genes responsible for heat and drought tolerance in wheat.

In vitro anticancer activities of Withania coagulans against HeLa, MCF-7, RD, RG2, and INS-1 cancer cells and phytochemical analysis.

BACKGROUND: The Pakistani Salt Range has a rich floral diversity including Withania coagulans from the Solanaceae family. METHODS: The crude methanolic extracts of the root, leaf, leaf stalk, and fruit of this plant were screened for their cytotoxic activity against human (HeLa, MCF-7, RD) and rat (RG2 and INS-1) cancer cell lines at 20 µg/mL and compared to methotrexate. The IC50 values indicated that leaf stalk and fruit extracts exert an 80% or higher cytotoxic activity against all cell lines at 24 hours. RESULTS: The leaf stalk extract showed the highest cytotoxic efficacy against all tested cell lines, with IC50 values ranging from 0.96 ± 0.01 µg/mL to 4.73 ± 0.05 µg/mL followed by the fruit extract with IC50 values of 0.69 ± 0.01-6.69 ± 0.06 µg/mL after 48-72 hours incubation. The leaf stalk and seed extracts were analyzed for polyphenols and flavonoids using RP-HPLC. The total flavonoid content (TFC) was calculated for all tested samples, and the highest TFC was recorded for the root extract (394.34 ± 1.26 µg/g). The total phenolic content (TPC) was found in the seed extract (307.86 ± 9.42 µg/g) of W. coagulans. The highest contents of myricetin (358.46 ± 2.91 µg/g) were noted in the leaf extract, and highest quercetin was recorded in the seed extract (21.43 ± 0.13 µg/g). The highest gallic acid concentration (83.62 ± 0.71 µg/g) was recorded in leaf stalk extract and p-hydroxybenzoic acid in the seed extract (157.46 ± 1.43 µg/g). CONCLUSION: The present study gives a scientific insight and comparative analysis of various plant parts in this medicinally important plant species from the Salt Range of Pakistan against both human and rat cancer cells.

Genome-wide association mapping in bread wheat subjected to independent and combined high temperature and drought stress.

Most investigations to date aiming to identify the genetic basis of the stress response of wheat (Triticum aestivum L.) have focused on the response to single stress agents such as high temperature or drought, even though in the natural situation, these stresses tend often to occur together. Here, a panel of 108 spring type bread wheat cultivars was phenotyped for 15 yield and yield related traits for two years (2014/15 and 2015/16) under non-stressed conditions, under high temperature stress, under drought and under a combined high temperature and drought regime. The mean loss in grain yield caused by all stress regimes was 51.33%. Analysis of variance (ANOVA) of yield trait showed significant differences among genotypes environments and their interactions (G×E). All the studied traits had higher heritability values which ranged from 0.35-0.94 under [C], 0.58-0.95 under [D], 0.62-0.93 under [H] and 0.60-0.95 under [HD]. GWAS was performed by using 9,646 informative SNP markers and based on these polymorphic SNPs population structure analysis divided whole germplasm into five major sub-populations. Mixed model association analysis detected 503 marker-trait associations (MTAs) at P ≤0.001 while 329 MTAs crossed FDR ≤ 0.05 for all traits with phenotypic variances (R2) ranged from 24.83% to 12.51%. Seven new pleiotropic SNPs on chromosome 7D and IAAV8258 (86.91cM) and wsnp_Ex_c7168_12311649 (57.93cM) on chromosome 5A were most stable association in present study. Furthermore, candidate genes Psy and Sr25 (TG0040) were also significant in present study, these genes were previously mapped on 7A and 7D. The region on 7D was assiociated with 7DL.7Ag translocation from Lophopyrum carring rust resistance Yr16 and many other genes. Similarly region on chromosome 7A which was associated with Psy gene was linked with grain yellow pigment content QTLs. Favourable alleles controlling grain yield were of vital importance and incorporation of these alleles after validation through marker assisted selection and fine mapping could be helpful in wheat yield improvement stress and non-stress conditions.

Pre-harvest aflatoxins and Aspergillus flavus contamination in variable germplasms of red chillies from Kunri, Pakistan.

Various cultivars of red chilli were collected from a small town named Kunri, located in the province Sindh, Pakistan. This town is a hub of red chilli production in Asia. A total of 69 samples belonging to 6 cultivars were obtained and analysed for the occurrence of aflatoxins and Aspergillus flavus, to explore the potential of resistant and susceptible germplasm. Aflatoxins were detected by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC), while A. flavus was isolated and identified using agar plate, blotter paper, deep freezing and dilution techniques. Molecular characterization using internal transcribed spacer (ITS) 1/4 and A. flavus specific FL1-F/R primers confirmed the identity of A. flavus. The data revealed that 67 and 75% samples contaminated with aflatoxin B1 (AFB1) and with A. flavus, respectively. A highly susceptible chilli cultivar was 'Nagina', showing 78.8% frequency of total aflatoxins (1.2-600 µg/kg) and a mean of 87.7 µg/kg for AFB1 and 121.9 µg/kg for total aflatoxins. A. flavus was detected with 93% frequency and 2.14 × 104 colony forming units. In contrast, cultivars 'Kunri' and 'Drooping Type' were found to be resistant, with low levels of aflatoxins and fungal counts. The study was conducted for the first time to explore two potential cultivars that were less susceptible towards A. flavus and aflatoxin contamination. These cultivars could be preferably cultivated and thereby boost Pakistan's chilli production.