Annum Jabar^1, Muhammad Athar², Kashf Mehmood⁴, Taha Ishfaq², Saqib Bashir ^2,3*, Zafar Iqbal⁵, Mureed Hussain², Anaam Zahra¹, Javaria Sherani⁶, Shahbaz Khan⁷*, Mohamed A. El- Sheikh⁸, Temoor Ahmed^9,10

¹Department of Botany, Ghazi University, Dera Ghazi Khan, Pakistan

²Department of Soil and Environmental Sciences, Ghazi University, Dera Ghazi Khan, Pakistan

³CAS Key Laboratory of Soil Environment and Pollution Remediation, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008, China

⁴Department of Biological Science, Superior University, Lahore, Pakistan

⁵Department of Botany, Sargodha University, Sargodha, Pakistan

⁶Department of Horticulture, Ghazi University, Dera Ghazi Khan, Pakistan

⁷Colorado Water Center, Colorado State University, Fort Collins, Colorado, USA

⁸Botany and Microbiology Department, College of Science, King Saud University, Riyadh, Saudi Arabia

⁹Department of Life Sciences, Western Caspian University, Baku, Azerbaijan

¹⁰MEU Research Unit, Middle East University, Amman, Jordan

Abstract

Recently, conversion of organic waste into useful products (organic fertilizers) is an emergent option for soil health restoration and sustainable ecosystem. In developing countries, excessive use of untreated wastewater and agrochemicals caused serious impact on food security and soil health. In this regard, the incorporation of recycled organic byproducts like compost (CP) and vermicompost (VC) have significant contribution in soil restoration by providing carbon and nutrients in polluted soil. A pot study was carried out to investigate the influence of CP and VC on maize growth and yield under copper stress. Maize was sown as a test plant in pots with 7 different treatments along with three repeats, such as T1: Control (CK); T2: 1% compost (CP 1%); T3: 2% Compost (CP 2%); T4: 5% Compost (CP 5%); T5: 1% vermicompost (VC 1%); T6: 2% Vermicompost (VC 2%) and T7: 5% Vermicompost (VC 5%). The current findings exhibited that incorporation of CP and VC prominently enhanced maize growth, biomass, plant height, chlorophyll contents, NPK status in soil and plant tissues. In addition, the results revealed that soil pH was prominently reduced by 0.54 and 0.59 units when CP and VC were mixed in Cu polluted soil. The addition of CP and VC at 5% rate presented the profound reduction in soil Cu by 24.41% and 43.02% respectively over control. Whereas Cu uptake by maize tissues was also reduced by 45% and 47% when CP and VC were incorporated at 5% rate. Overall, among all the treatments and application rates VC at 5% exhibited prominent results over control as well as other soil additives.

Keywords: Copper, Compost, Vermicompost, Immobilization, Maize

Mahmoud Ahmed Amer^1*, Zaheer Khalid¹, Khadim Hussain¹, Muhammad Amir¹, Muhammad Zaman¹, Ibrahim Mohammed Al-Shahwan¹, Mohammed Ali Al-Saleh^2*

¹Plant Protection Department, College of Food and Agriculture Sciences, King Saud University, Riyadh, Saudi Arabia

²Chair of Date Palm Research, Plant Protection Department, College of Food and Agriculture Sciences, King Saud University, Riyadh, Saudi Arabia

Abstract

Tomato leaf curl disease is the most harmful disease of tomato caused by different begomovirus species mainly Tomato yellow leaf curl virus (TYLCV) and Tomato leaf curl Sudan virus (TLCSDV). Both are single-stranded DNA begomoviruses that cause leaf curling, stunting, vein banding, yellowing and results in significant crop losses in tomato in Saudi Arabia. In the first objective of this study, infectious clones of TLCSDV and TYLCV were constructed by cloning and subcloning of partial dimer genome containing two origins of replication on PstI-KpnI/KpnI sites in binary plasmid vector pGreenII 0000. To assess the functionality of agro-infectious clones, those were transformed into Agrobacterium tumefaciens GV3101 strain and infiltrated into the leaves of model host plant Nicotiana benthamiana. Three weeks post inoculation begomovirus symptoms were observed and infectivity of our infectious clones were confirmed by PCR using begomovirus diagnostic primers (AC1048 and AV494). In the second phase, 13 available commercial cultivars of tomato crop grown in open fields as well as greenhouse were inoculated with agro-infectious clones of TLCSDV and TYLCV to assess their tolerance against these important pathogens of tomato. The obtained results observed after 30 days of inoculation showed all cultivars were susceptible to both begomoviruses showing variable symptoms severity. These results were confirmed by PCR using AC1048 and AV494 primer. The significance of response of different cultivars against infection of TLCSDV and TYLCV were discussed.

 Keywords: Tomato, TLCSDV, TYLCV, Agroinfiltration, Infectious clone, Cultivars, Virus resistance

Sana Riaz¹, Ghulam Mustafa^1*, Muhammad Sarwar Khan¹, Muhammad Amjad Abbas²

¹Centre of Agricultural Biochemistry and Biotechnology (CABB), University of Agriculture, Faisalabad, Pakistan

²Department of Plant Pathology, University of Agriculture, Faisalabad, Pakistan

Abstract

Colletotrichum falcatum is the most infectious pathogen of sugarcane which not only affects crop productivity but also lowers sugar recovery. Vegetative mode of propagation, long breeding cycle and anatomy of the plant further vulnarize it to pathogen infection. Owing to limitations in the control of pathogen with chemicals and conventional methods, mycoparasitic agents are more effective and efficient. Indigenous Trichoderma strains were used in these studies to see their impact on the control of this infectious pathogen under in vitro and in planta conditions. Both, the pathogen and Trichoderma isolates were purified and confirmed through molecular tools. Phylogenetic analysis was carried out to assess their genetic relatedness with other Colletotrichum species. In vitro dual plate culture assay was performed to assess mycoparasitic potential of all of the isolates. Trichoderma harzianum showed 85.5% inhibition in the growth of Colletotrichum falcatum whereas Trichoderma viride showed 81.1% growth inhibition. Further, in planta infection assay was conducted in red rot susceptible sugarcane genotype SPF-234 to seek for its potential to suppress pathogen infestation. Six months old plants were inoculated with C. falcatum alone and in combination with T. viride by plug method of inoculation. The infected canes were dissected out and observed for red discoloration. Both of the biocontrol agents inhibited growth of the pathogen yet T. harzianum appeared to be more effective for the control of aforementioned pathogen. Hence, Trichoderma harzianum isolates can effectively be used for the control of red rot infection in sugarcane leading to enhanced crop production and sugar recovery.

Keywords: Red rot, Biocontrol agent, Sugarcane, Dual plate culture assay, In planta infection assay

Mahmood Rasool^1*, Khalid I. Alhassan², Sajjad Karim¹, Absarul Haque³, Mohammed H.Z. Mutwakil², Mohammed Alharthi⁴, Adeel G. Chaudhary¹, Peter Natesan Pushparaj¹

¹Center of Excellence in Genomic Medicine Research, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia

²Department of Biological Science, King Abdulaziz University, Jeddah, Saudi Arabia

³King Fahd Medical Research Center, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia

⁴Faculty of Medicine, King Abdulaziz University Hospital, King Abdulaziz University, Jeddah, Saudi Arabia

Abstract

The aim of our research was to identify molecular targets that can be targeted by drugs and patient-specific models for personalized medicine for colorectal cancer (CRC). Here, we obtained high-throughput RNA sequencing data from Gene Expression Omnibus (GEO) with accession number GSE205787 and analyzed it using next-generation knowledge discovery tools such as BioJupies and Ingenuity Pathway Analysis (IPA) software. Differentially expressed genes (DEGs) were identified by comparing the raw counts from 47 CRC patient-derived spheroids (CRC-CSCs) with those from normal spheroids from the epithelium of the colon and rectum of healthy individuals, using BioJupies tools. IPA was used to identify differentially regulated canonical pathways, upstream regulators of CRC, non-directional networks, diseases, and biofunctions, as well as to conduct subsequent perturbation analysis using the Molecular Prediction Analysis (MAP) tool. Our study demonstrates that several KEGG pathways, including the AMPK, Phospholipase D, MAPK, and PI3-AKT signaling pathways, were significantly downregulated in the CRC-CSC group. Additionally, Wnt signaling and FGFR pathways were significantly upregulated. Moreover, according to Wikipathways, the EGF/EGFR signaling pathway, MAPK signaling pathway, G-protein signaling pathway, and Focal Adhesion-PI3-AKT pathway were downregulated in the CRC-CSC group. Furthermore, based on the Reactome, the Metabolism, Vesicle-mediated transport, RAF signaling, and G-alpha (12/13) signaling pathways were also downregulated in the CRC-CSC group. Utilizing innovative drug combination approaches and innovative drug delivery techniques, CRC treatments can be enhanced by modulating the FGFR, EGFR, and AMPK signaling pathways, which may ultimately lead to improved patient outcomes.

Keywords: Colorectal cancer, Cancer stem cell spheroids, AMP‐activated protein kinase signaling, Fibroblast Growth Factor Receptor, Epidermal Growth Factor Receptor, BioJupies, Ingenuity Pathway Analysis

Uğur Yegül^*

Department of Agricultural Machinery and Technologies Engineering, Faculty of Agriculture, Ankara University, 06135 Ankara, Türkiye

Abstract

The present study aimed to develop a kit to collect data on important parameters for cultivation in a hydroponic farming environment and send and store these data online. This Study was carried out between February and August 2022. The area where the experiment was conducted is fully controlled (coordinates 39.962013 and 32.867491) and established within Ankara University, Ankara, Türkiye. The kit developed for indoor use in agriculture consisted of a microcontroller, different sensors, and hardware components. For all the hardware to be combined and work properly, a closed box was designed using SolidWorks solid modeling software and fabricated with a 3D printer. The code developed for the kit to fulfill the desired function was written in C++ and transferred to the microcontroller via Arduino software using a personal computer. This kit can measure T (temperature), H (humidity: %), carbon dioxide (CO₂), total volatile organic compounds (TVOC), LUX (luminous intensity: lux), Ultra-violet (UV) W m^-2, P (air pressure: pascal), and AQ (air quality: ppm). The developed kit can transmit and store the data simultaneously on the Internet. IoT technologies need further agricultural studies, and more data to be obtained can contribute to resolving more problems. The kit obtained in the study can be used in domestic agriculture and various agricultural activities.

Keywords: Internet of things, Indoor farming, Arduino, Sensor, Environmental parameters

Hammad Anwar¹, Moazzam Jamil¹, Azhar Hussain^1*, Abubakar Dar¹, Maqshoof Ahmad¹, Saleh H. Salmen², Mohammad Javed Ansari³, Rashid Iqbal^4,5*

¹Department of Soil Science, the Islamia University, Bahawalpur-63100, Pakistan

²Department of Botany and Microbiology, College of Science, King Saud University, Riyadh, Saudi Arabia

³Department of Botany, Hindu College Moradabad, Mahatma Jyotiba Phule Rohilkhand University, Bareilly, India

⁴Department of Agronomy, Faculty of Agriculture and Environment, The Islamia University of Bahawalpur, Bahawalpur 63100, Pakistan

⁵Department of Life Sciences, Western Caspian University, Baku, Azerbaijan

Abstract

Zinc (Zn) is the most limiting micronutrient responsible for malnutrition. World Health Organization (WHO) reported deficiency of Zn is the 5^th most significant cause of death and disease in underdeveloped world. However, 70% Pakistani soils are Zn deficient and responsible for Zn deficiency in crops. The present study aimed to mitigate Zn deficiency and improve them a nd Zn use efficiencies through synergizing dry region Zn solubilizing plant growth promoting rhizobacteria (PGPR) by coating on Zn coated urea. Pre-isolated dry region Zn solubilizing isolates were evaluated for zinc solubilization, urease activity, siderophores production, organic acid production and ACC-deaminase activity. Four effective strains Bacillus amyloliquefaciens (IUB-34), Klebsiella variicola (IUB-96), Klebsiella variicola (IUB-80) and Klebsiella pneumoniae subsp. pneumoniae (IUB-93) and their consortium coated on Zn coated urea. This improved product was tested for N and Zn release pattern, growth promotion and Zn biofortification in pot trial on wheat. Results showed that SPAD chlorophyll value, root, shoot length and their dry weight was significantly improved (p≤ 0.05) by 19.4, 20.3, 45.9, 27.3 and 39.5%, respectively, over control. Similarly, N, P, K, Zn, Fe in grains and 100-grain weight was significantly increased (p≤ 0.05) by 97.5, 23.5, 61.1, 63, 32 and 50.5%, respectively, over control. The results confirmed that dry region Zn solubilizing bacterial consortium coated on Zn coated urea is an efficient method for the biofortification of Zn in wheat grains and can effectively overcome Zn deficiency in humans.

Keywords: Zinc, Dry region, Wheat, Consortium, Zn coated urea, Biofortification

Yasmeen Rasheed¹, Nazia Ehsan¹, Muhammad Faisal Hayat¹, Asma Ashraf², Hammad Ahmad Khan¹, Aisha Khatoon³, Muhammad Umar Ijaz^1*, Yasir S. Raouf⁴, Abdelouahid Samadi^4*, Samir Chtita⁵

¹Department of Zoology, Wildlife and Fisheries, University of Agriculture, Faisalabad 38040, Pakistan

²Department of Zoology, Government College University, Faisalabad 38000, Pakistan

³Department of Pathology, University of Agriculture, Faisalabad 38040, Pakistan

⁴Department of Chemistry, College of Science, United Arab Emirates University, Ai-Ain P.O. Box 15551, United Arab Emirates

⁵Laboratory of Analytical and Molecular Chemistry, Faculty of Sciences Ben M’Sik, Hassan II University of Casablanca, P.O. Box No. 7955, Casablanca, Morocco

Abstract

Polyethylene microplastics (PEMPs) are one of the most toxic pollutants in our surroundings that induce damage to various organs including heart. Poncirin (PON) is a natural flavonoid that shows diverse pharmacological activities. This study was aimed to assess the alleviative potential of PON against PEMPs provoked cardiac damage in rats. Twenty-four rats were segregated into 4 groups including control, PEMPs (1.5 mg/kg) treated group, PEMPs (1.5 mg/kg) + PON (5mg/kg) exposed group and PON (5mg/kg) alone treated group. It was revealed that PEMPs exposure notably decreased the expression of Nrf2 and its associated antioxidant genes while upregulating the expression of Keap-1. Besides, PEMPs intoxication reduced the activities of superoxide dismutase (SOD), catalase (CAT), heme-oxygenase-1 (HO-1), peroxidase (GPx), glutathione reductase (GSR), and glutathione (GSH) content while increasing reactive oxygen species (ROS) and malondialdehyde (MDA) levels. Additionally, exposure to PEMPs resulted in upregulation of lactate dehydrogenase (LDH), creatine kinase MB (CK-MB), troponin I and phosphokinase (CPK).  Besides, PEMPs administration escalated the levels of TNF- α IL-6, NF-κB, TNF- α, IL-1β, and COX-2 activity. Moreover, the administration of PEMPs escalated the levels of Caspase-3 and Bax, while downregulating the levels of Bcl-2. Additionally, PEMPs exposure disrupted the architecture of cardiac tissues. Nonetheless, PON supplementation remarkably protected the cardiac tissues by regulating the aforementioned damages.

Keywords: Polyethylene microplastics, Poncirin, Cardiac damage, Oxidative stress, Inflammation

Muhmmad Kashif Shahzad Sarwar^1,2, Abdul Ghaffar³, Saghir Ahmad⁴, Shoaib Ur Rehman¹, Ummara Waheed^1*

¹Institute of Plant Breeding and Biotechnology, MNS University of Agriculture, Multan, Pakistan

²Cotton Research Station, Ayub Agricultural Research Institute, Faisalabad, Pakistan

³Departemnt of Agronomy, MNS University of Agriculture, Multan, Pakistan

⁴Cotton Research Institute, Multan, Pakistan

Abstract

Cotton diversity has long been studied using physiological and biochemical traits. This diversity has led to the development of various superior cotton cultivars over the year. At present and in face of climate change, development of high yielding and drought tolerant cotton varieties are necessary to fulfill the demand of ever-growing population of the world. In this study, Gossypium hirsutum L. germplasm (200) was evaluated under two irrigation regimes i.e., well-watered (W1) and limited water (W2) conditions. Various morphological and physiological traits were recorded under both irrigation regimes. A considerable reduction was recorded in W2 conditions in all the recorded traits except for glycine betaine, soluble sugars, and proline contents, highlighting the impact of drought on cotton germplasm. Cotton genotypes that maintained higher yield had positive correlation with biochemical traits. Out of 63 best performing genotype (superior parents based on the recorded data), FH-414, FH-415, FH-416, FH-326, FH-492, FH-Anmol, Gomal-105, Marvi, NIAB-878 and VH-327 were selected for hybridization to make crosses following Line x Tester fashion. F₁ hybrids (25 crosses) and 10 parents were again planted under W1 and W2 conditions. Out of 25 crosses, FH-326 × Marvi (CS5) and NIAB-878 × FH-414 (CS16) performed better under water deficit conditions. Quantitative real-time PCR was also performed using GhHH3 and GhIDD. CS5 and CS16 had higher expression of drought tolerance causing GhHH3 and GhIDD genes. The newly developed cotton crosses will pave the way for the development of high yielding drought tolerant cotton varieties in face of climate change.

Keywords: Biochemical attributes, Yield and yield components, GhHH3, GhIDD, Drought

Usama bin Naeem¹, Muhammad Adil Rasheed^1*, Muhammad Ashraf¹, Muhammad Yasir Zahoor²

¹Department of Pharmacology and Toxicology, Faculty of Biosciences, University of Veterinary and Animal Sciences Lahore, Pakistan

²Insititue of Biochemistry and Biotechnology, Faculty of Biosciences, University of Veterinary and Animal Sciences Lahore, Pakistan

Abstract

One of the most dominant diseases in the world, particularly among women, is breast cancer. Breast cancer has tumor suppressor genes called CHEK2 and TP53. When there is a mutation in CHEK2 and TP53 genes there are more chances of breast cancer. This study aimed to investigate the already prepared and characterized nanoparticles loaded with Chitosan for Cell death, Mitochondrial Membrane and cell cycle arrest estimated through Flow Cytometry and gene expression analysis of CHEK2 and TP53 genes by real-time PCR. The Livak method was used to evaluate the results. The mean (± S.D) comparison between the control and target genes were used to calculate gene expression. Results showed that Ivermectin and Tamoxifen NPs (B+C) represented 34.8% cell death that is better than other combinations with propidium iodide stain while with Acridine orange stain Tamoxifen+Ivermectin (A+B) combination showed the remarkable and maximum of the all cell cycle arrest with value of 69.7% cell arrest at G0/G1 phase, 7.11% of cell arrest at S Phase and 7.05% of G2/M Phase arrest. It was demonstrated that the expression levels of CHEK2 and TP53 genes were significantly increased (P<0.001) in Ivermectin+Tamoxifen NPs (B+C) compared with control groups. It is concluded that Tamoxifen nanoparticles with Ivermectin showed strong anti-proliferative activity against breast cancer cells. The expression levels of nanoparticles containing Tamoxifen were significantly increased compared to the other treatments and control groups (P<0.001). Gene expression change with change in dose concentrations.

Keywords: Breast cancer, Apoptosis, Cell cycle arrest, Pharmacogenomic, Gene expression

Muhammad Khalid Mansoor*¹, Kashif Iqbal¹, Ali Hassan², Muhammad Saqib², Ali Zohaib¹, Sabiqaa Masood³

¹Department of Microbiology, Faculty of Veterinary & Animal Science, The Islamia University of Bahawalpur, Pakistan

²Department of Clinical Medicine & Surgery, University of Agriculture, Faisalabad, Pakistan

³Department of Parasitology, Faculty of Veterinary & Animal Science, The Islamia University of Bahawalpur, Pakistan

*Corresponding author’s email: khalid.mansoor@iub.edu.pk

Received: 13 September 2023 / Accepted: 5 April 2024 / Published Online: 13 December 2024

Abstract

Contagious Ecthyma (CE), also known as scabby mouth disease, is caused by an epitheliotropic parapoxvirus that primarily affects the goat and sheep populations worldwide. This study focused on investigating 12 outbreaks of CE in sheep and goat herds across various regions of Punjab, Pakistan. A total of 35 samples were collected between March 2021 and May 2022, with 34 out of 35 samples testing positive for parapoxvirus through PCR. Subsequently, 24 complete sequences of the major envelope protein B2L gene were successfully obtained. The nucleotide and amino acid sequences of the ORF virus B2L gene were analyzed. The 1206bp amplicons, after Sanger sequencing revealed an open reading frame of 1137bp encoding 378 amino acids. The minimum and maximum nucleotide differences of 0 and 34, respectively, were observed, while the percentage similarity at the nucleotide level and amino acid level ranged from 97.98% to 100% and 97.62% to 100%, respectively, among the ORF strains in this research study. The results of the phylogenetic analysis revealed that all 24 ORF virus isolates from Pakistan belonged to Group-I ORF viruses. The comparative homology of Pakistani ORF virus strains with Indian, Chinese, and Turkish isolates was 99.03%, 98.59%, and 98.15%, respectively. This study contributes to understanding the circulation of Group I ORF viruses in Pakistan and their relationship with strains from neighbouring countries. Furthermore, these findings may offer insights into the genotype of the causative agent responsible for the contagious pustular dermatitis (CPD) outbreak in Punjab, Pakistan.

Keywords: B2L gene, Contagious, Contagious pustular dermatitis, ORF