Wasba Sarfraz¹, Maria Khalid², Awais Rasheed¹, Tariq Mahmood^1*

¹Department of Plant Sciences, Quaid-i-Azam University, Islamabad 45320, Pakistan

²Atta-ur-Rehman School of Biological Sciences (ASAB), National University of Science & Technology (NUST), Islamabad, Pakistan

*Corresponding author’s email: tmahmood@qau.edu.pk

Received: 27 December 2023 / Accepted: 10 July 2024 / Published Online: 13 January 2025

Abstract

Rapid change in global environmental conditions will increase the severity of salinity and drought stresses. Wheat (Triticum aestivum L.) is a major staple crop worldwide and its production is affected by these two major abiotic stresses. Use of breeding and transgenic techniques facilitated the development of tolerant varieties able to withstand in drought and saline environments. This work investigated the role of Solanum lycopersicum proteinase inhibitor II (PI-II), SlPI-II, gene for mounting tolerance under salinity and drought stress in transgenic wheat. Significant increase of 6.41-fold (TL1) was observed in transgenic plants at 1 day post 100m NaCl treatment. A gradual increase in the expression was observed at 200 mM NaCl treatment. At 300mM treatment, 3.78-fold (TL5) increase in the expression level was recorded. The maximum fold change of 5.01 and 3.99 was observed in 6 days post treatment at 100 mM and 200 mM in TL1, respectively. The extended saline treatment (14 days post treatment) in transgenic lines resulted the higher SlPI-II induction than control wheat plants which showed the efficacy of this gene in salinity tolerance. SlPI-II expression in 15% PEG treated plants showed transcript up to 7.31 fold increase during 1 day post treatment, while at 25% PEG transgene expression ranged from 3.45 to 2.27 fold increase. Conclusively, the role of SlPI-II gene in drought tolerance was due to sustained transcript increment during PEG treatment. We demonstrated that modern day transgenic approaches utilizing plant-based proteinase inhibitors will provide the new opportunities for the development of tolerant varieties in future.

Keywords: Wheat, NaCl, Proteinase inhibitor, Transgenic wheat, PEG6000

Thi Thuy Tien Nguyen¹*, Hien Trang Nguyen¹, Thanh Long Le ¹, Thy Dan Huyen Nguyen¹

¹Department of Engineering and Food Technology, University of Agriculture and Forestry, Hue University, Hue, Thua Thien Hue, 530000, Vietnam

Abstract

Forty-eight Streptomyces strains were screened for antifungal activities against P. digitatum P2 and P. italicum I6 isolated from oranges infected with green mold and blue mold diseases, respectively, using the dual culture method. Among 48 screened strains of Streptomyces, S. murinus NARZ showed the strongest activity by completely inhibiting the growth of both pathogens. The antifungal activities of S. murinus NARZ were evaluated based on the percentage inhibition of radial growth (PIRG, %) using culture filtrate (CF) and ethyl acetate crude extract. The EC₅₀ values of the 7-day S. murinus CF were 13.25% and 33.72% against P. digitatum P2 and P. italicum I6, respectively. The EC₅₀ and MIC₉₀ values of the crude extract were 45.03 µg/mL and 193.15 µg/mL for P. digitatum P2, and 68.25 µg/mL and 295.60 µg/mL for P. italicum I6, respectively. Notably, out of the six International Streptomyces Project Media (ISP2, ISP3, ISP4, ISP4G, ISP4Y, and ISP8), ISP3 medium was optimal for S. murinus NARZ cultivation to produce antifungal metabolites, with 50% CF yielding 100% inhibition against both Penicillium pathogens. Furthermore, the CF had notable thermal resistance; the PIRGs decreased from 100% to 74.22% for P. digitatum P2 and to 64.59% for P. italicum P6 after treatment at 121°C for 15 min. The tests showed that oranges sprayed with 100% CF of S. murinus NARZ remained symptom-free of blue and green mold diseases after a seven-day incubation following artificial inoculation with Penicillium species. This study highlights the potential of Streptomyces strains, particularly S. murinus NARZ, as effective biocontrol agents against postharvest fungal diseases in oranges.

Keywords: Antifungal activities, Biocontrol, Postharvest diseases, Penicillium, Streptomyces murinus

Piangpassorn Kowwilaisang, Montinee Teerarak*, Chamroon Laosinwattana

School of Agricultural Technology, King Mongkut’s Institute of Technology Ladkrabang, Bangkok, 10520, Thailand

Abstract

Plant essential oils have limited water solubility, for which problem oil-in-water emulsions provide a good solution. The aim of this work was to prepare a ginger essential oil (Zingiber officinale Rosc.; GEO) coarse emulsion, microemulsion, and nanoemulsion all with the same formula, determine their characteristics, and compare their antioxidant activities and utility as holding solutions for Davallia fronds. A coarse emulsion was firstly formed by a magnetic stirrer of GEO with a 1:2 by weight mixture of Tween 20 and Span 20 at weight ratio of 1:8, respectively. The coarse emulsion droplets (810.0 nm) were then broken by sonication and high-pressure homogenization to create the microemulsion (426.1 nm) and nanoemulsion (76.4 nm), respectively. The three emulsions exhibited V-potential values more negative than -30 mV, indicating them to be stable. All three emulsions demonstrated DPPH^• and ABTS^•+ free radical scavenging capacities significantly higher than those of GEO in ethanol. Meanwhile, the nanoemulsion significantly improved the Fe²⁺ chelating effect Finally, in a bioefficacy experiment with Davallia fronds, both the microemulsion at concentrations of 5 and 10 mg/mL and the coarse emulsion at 10 mg/mL were found to extend frond vase life. The nanoemulsion demonstrated superior frond longevity at low concentration (5 mg/mL), but at higher concentration (10 mg/mL), vase life and leaf chlorophyll were not improved and malondialdehyde formation increased. Based on the current investigation, emulsification significantly enhances the antioxidant activities of GEO. Vase solutions containing high concentrations of coarse emulsion (10 mg/mL), microemulsion (5 and 10 mg/mL), and low concentration of nanoemulsion (5 mg/mL) increased Davallia frond longevity from 6.3 days to up to 11.9 days. Our findings suggest that there is an optimal concentration range for using essential oil emulsions in different delivery systems as a preservation solution in cut fronds.

Keywords: Cut leaf, Delivery vehicle, Fern, Oil-in-water, Holding solution

Saima Malik¹, Imran Mukhtar¹, Humaira Muzaffar¹, Laaraib Nawaz¹, Haseeb Anwar^1*

¹Health Biology Research Lab (HBRL), Department of Physiology, Government College University, Faisalabad, Pakistan

Abstract

The gut microbiome potentially modulates pharmacokinetics of orally administered drugs. Homologous transporting proteins in epical membrane of the enterocytes and cell membrane of the residing microbial cells of the host may compete for absorption of the orally administered drugs. Microbial cells residing the small intestine of the host may uptake/bio-accumulate some of the quantity of the dose of orally administered drug. This project is aimed to observe absorption/bio-accumulation behavior of enalapril by the gut microbiome when enalapril was administered orally in pure form and in the presence of excipients (Tablet; commercial preparation). Currently, no data confirms specific transport system for enalapril uptake by gut microbiome in absence and presence of excipients as well. Two in-vivo trials, enalapril pure drug treated trial and enalapril commercial tablet treated trial were conducted in parallel. Each trial was conducted in adult Wistar albino rats (n=42) divided into seven groups having same number of rats in each group (n=6); one control group and six drug treated groups administered orally with single dose of enalapril 10mg/kgbwt. Rats (n=6) were subsequently sacrificed at different intestinal transit times of 1, 2, 3, 4, 5 and 6 hours post drug administration to harvest microbial mass pellet from digesta. Pellet was lysed to expose microbial lysate and pursued through HPLC. The microbiome absorbed enalapril at 4hour transit time (103±7.31µg) significantly (p≤0.05) higher as compared to 5hour transit time (73.2±5.17µg). Percent dose recovery from microbiome was significantly (p≤0.05) higher at 4hour transit time (4.15±0.05%) as compared to 5hour transit time (3.14±0.18%) post drug administration. Independent of presence of excipients, from both formulations enalapril was absorbed in equal amount competitively by the intestinal microbiome through the homologous transport mechanism present in the enterocytes of the host. Conclusively, enalapril serves as a substrate of gut microbiome independent of dosage form when administered orally.

Keywords: Enalapril, Microbiome, Microbial lysate, Percent drug recovery

Amir Manzoor¹, Muhammad Anjum Aqueel^*1, Shams Ul Islam¹, Eldessoky S. Dessoky², Muhammad Haseeb Ahsan¹, Bashir Ahmad^1,3, Muhammad Usman Yousuf¹, Muhammad Saqib⁴, Muhammad Fahim Raza⁵, Rashid Iqbal^*4,6

¹Department of Entomology, Faculty of Agriculture and Environment, The Islamia University of Bahawalpur, Pakistan

²Department of Biology, College of Science, Taif University, Taif, Saudi Arabia

³PARC-Arid Zone Research Institute, Bahawalpur, Pakistan

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

⁵Department of Veterinary Pathology, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, S7N 5B4, Canada

⁶Department of Life Sciences, Western Caspian University, Baku, Azerbaijan

Abstract

Artificial diets are necessary for managed honeybees (Apis mellifera L.) in arid regions when naturally occurring flora becomes scarce. The current study was planned to evaluate different artificial diets in terms of consumability and digestibility of A. mellifera and their effects on weight of queen bee, worker bee longevity and number of broods for colony survival and management in dearth period. Different artificial diets were administered separately to 33 colonies in three replications. The best combination was 25 g, each of soya flour, dry apricot powder, date paste and powder sugar, mixed in 8 ml of vegetable oil. Honeybees consumed most of diet-T₈ in all weeks (average 49.53 g), followed by diet-T₇ (average 38.81 g) and diet-T₆ (average 34.55 g); while the minimum consumption was of diet-T₁ (average 6.67 g). In diet digestibility experiment, diet-T₂ showed the highest digestibility (73.9%) followed by diet-T₁ (71.3%) and diet-T₄ (66.7%) with the lowest digestibility in diet-T₉ (59%). The protein present in hemolymph was highest in diet-T₈ (26.9 ug/ul) followed by diet-T₇ (24.1 ug/ul) and diet-T₆ (22.9 ug/ul) with the lowest in diet-T₁ (17.7 ug/ul). As a response to the feeding on the prepared diets, the queen’s bee weight was highest in diet-T₈ (198.65 ± 0.36 mg) followed by diet-T₇ and diet-T₆ while the lowest in diet-T₀ (control group, 182.28 ± 0.55 mg). Similarly, the worker bee longevity was highest in diet-T₈ (53 ± 2.14 days) followed by diet-T₇ and diet-T₆ and lowest in diet-T₀ (control group, 34 ± 1.73 days). In the brood experiment, the number of eggs, larvae and pupae was highest in diet-T₈ (85 ± 2.03 eggs, 66 ± 1.15 larvae and 78 ± 1.15 pupae) while lowest in diet-T₀ (51 ± 1.5 eggs, 26 ± 1.5 larvae, 23 ± 1.5 pupae) after 28 days of diet feeding. Therefore, diet-T₈ can be used as substitute diet for the honeybees during dearth period for the survival and colony management of A. mellifera

Keywords: Apis mellifera diet, Substitute diet, Dearth period management, Diet digestibility, Weight of queen, Worker bee longevity, Number of broods

Adeel Yunus¹, Nighat Sultana^1*, Alia Gul², Abdullah^3,4

¹Department of Biochemistry, Hazara University, Mansehra, Pakistan

²Department of Botany, Hazara University, Mansehra, Pakistan

³State Key Laboratory of Chinese Medicine Modernization, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China

⁴Haihe Laboratory of Modern Chinese Medicine, Tianjin, 301617, China

Abstract

The plant family Athyriaceae Alston consists of 3 genera and 650 species dispersed in different regions of the world, including Athyrium Roth, Diplazium Sw., and Deparia Hook. & Grev. The family has diverse morphological characteristics, ranging from creeping rhizomes to ascending or erect scales at apices. In the present study, the chloroplast (cp) genome of Diplazium polypodioides Blume was de novo assembled and compared with twelve other reported genomes of the species of the family Athyriaceae. The cp genome of D. polypodioides was 152,009 bp and showed a quadripartite structure in which a large single copy (82,389 bp) and a small single copy (22,303 bp) were separated by a pair of long inverted repeats (IRa and IRb: 23,659 bp each). We identified 116 genes, including 4 rRNAs, 29 tRNAs, and 84 protein-coding genes, with 15 genes duplicated in inverted repeats. The cp genome sizes of the thirteen analyzed species ranged from 150,797 bp (Diplazium striatum Desv.) to 152,009 bp (D. polypodioides). Despite high variability in SSRs and oligonucleotide repeats, the species showed similarities in GC content, contraction and expansion of inverted repeats, codon usage, amino acid frequency, and substitutions. Transition substitutions were more common than transversion substitutions across all species. Phylogenetic analysis of 84 protein-coding genes revealed monophyletic relationships among the limited species of three genera, namely, Diplazium, Athyrium, and Deparia. In addition, Diplazium was more closely related to Athyrium than to Deparia. Our study provides preliminary insights into the evolutionary dynamics of the cp genome in Athyriaceae and clarifies its phylogenetic relationships.

 Keywords: Athyrium, Athyriaceae, Deparia, Diplazium polypodioides, Pteridophytes, Phylogenetics

Khaoula Habbadi^1*, Meriem Guartoumi El Idrissi¹, Salma El Iraqui El Houssaini¹, Ilyass Maafa², Faical Aoujil¹, Abdellatif Benbouazza¹, El Hassan Achbani¹, Moha Ferrahi³

¹Phytobacteriolgy and Biological Control Laboratory, Regional Center of Agricultural Research of Meknes, National Institute of Agricultural Research, Avenue Ennasr, BP 415 Rabat Principal, 10090 Rabat, Morocco

²International Center for Agricultural Research in the Dry Areas (ICARDA), Rabat 10080, Morocco

³Department of Breeding and Conservation of Genetic Resources, National Institute for Agricultural Research (INRA), Rabat 10020, Morocco

Abstract

Considering the imperative challenges posed by climate change, particularly the escalating instances of drought stress, optimizing wheat production stands as a critical goal for agricultural development. This experimental study delves into the pivotal role of Plant Growth-Promoting Rhizobacteria (PGPR) and Water Retention Agents (WRA) in bolstering the resilience of bread wheat crops to drought conditions. The study encompasses germination tests, field observations, and a comprehensive analysis of agronomic parameters, emphasizing the significance of PGPR and WRA in mitigating the adverse effects of water scarcity on wheat crops. The germination tests in Petri dishes reveal a substantial enhancement with the application of PGPR strains, showing higher rates (95% for Serratia nematodiphila strain GAB111 and 92% for Pseudomonas koreensis strain GAJ222) compared to the control. PGPR significantly increased root and shoot lengths, with significant increases observed. Field observations show the climatic impact on wheat development, with an accelerated growth cycle due to high temperatures and arid conditions. Further analyses reveal that WRA and PGPR significantly affect agronomic parameters such as the number of leaves, tillers, ears, and chlorophyll content. The study also assesses the impact on leaf temperature, wheat yield, plant-root development, and soil parameters such as organic matter and nutrient content. The combined use of WRA and PGPR shows promising results, highlighting their potential synergistic effects on wheat growth and development, especially in challenging environmental conditions. These findings offer practical solutions for enhancing wheat resilience and have broader implications for the sustainable development of agricultural systems confronting increasing climate-induced stresses.

Keywords: Climate change, Wheat resilience, Aagronomic parameters, PGPR, WRA

Naphat Somala¹, Nutcha Manichart¹, Muanfan Thongbang¹, Pattharin Wichittrakarn², Chamroon Laosinwattana^1*, Montinee Teerarak¹

¹School of Agricultural Technology, King Mongkut’s Institute of Technology Ladkrabang, Bangkok, 10520, Thailand

²King Mongkut Chaokhunthahan Hospital, King Mongkut’s Institute of Technology Ladkrabang, Bangkok, 10520, Thailand

Abstract

Fungal extract-based natural herbicides are a promising avenue for the development of sustainable weed management. The study investigates the herbicidal activity of fungal extracts against the prominent aquatic invasive weed Eichhornia crassipes (water hyacinth). The ethanol crude fraction derived from Diaporthe sp. strain EC010 demonstrated the highest phytotoxicity. Chemical characterization using gas chromatography-mass spectrometry revealed 2(3H)-furanone, dihydro-4-hydroxy- (22.81%), linoelaidic acid (6.87%), and hordenine (6.62%) as major constituents. Phytotoxicity was evaluated by wrapping bioassay under greenhouse conditions. Observable foliar damage, such as necrosis, chlorosis, and depigmentation, occurred within 1 day after treatment (DAT). The rapid onset of visible damage within 1 DAT and the detailed analysis of tissue damage are particularly noteworthy. Lesion progression reflected the phytotoxicity of the extract, increasing to 77.33% visible phytotoxicity at 14 DAT for the highest concentration (8.0% w/v). Microscopic analysis revealed disintegration and complete deformation of epidermal and parenchymal tissues, and treated plants featured extensive aerenchyma spaces. Furthermore, the Diaporthe sp. extract decreased chlorophyll a, b, and carotenoid concentrations while increasing electrolyte leakage and malondialdehyde, indicative of weed deterioration. The current work offers valuable insights for sustainable and eco-friendly strategies in the management of water hyacinth populations in a lentic ecosystem. The findings suggest that Diaporthe sp. extract could serve as a natural herbicide, offering an environmentally friendly alternative to synthetic chemicals in managing water hyacinth.

Keywords: Allelopathy, fungi extract, natural herbicide, water hyacinth

Rungravee Chaiyod¹, Benyapa Prakit¹, Kittiya Khongkool¹, Wankuson Chanasit², Monthon Lertworapreecha^2*

¹Biotechnology program, Faculty of Science and Digital Innovation, Thaksin University, Phatthalung Province, Thailand

²Microbial Technology for Agriculture, Food, and Environment Research Center, Faculty of Science and Digital Innovation, Thaksin University, Phatthalung Province, Thailand

Abstract

This study investigates the production and characterization of bacterial cellulose (BC) by Novacetimonas pomaceti KMPG_12, focusing on enhancing its prebiotic properties through size reduction via high-pressure homogenization (HPH). Optimal conditions for BC production were established using Response Surface Methodology (RSM), yielding a maximum dry weight of 9.78 g/L under conditions of 15% glucose, 0.4% peptone, and 4.5% alcohol over a 14-day cultivation period. The produced BC was subjected to HPH at 20,000 psi for eight cycles, significantly reducing fiber size, as confirmed by SEM analysis. The transformation from a compact, net-like structure to a more separated fibrous network at the nanometer scale was observed. FTIR analysis revealed modifications in the BC’s molecular framework post-HPH, indicating a disruption of hydrogen bonds and a transition towards a less crystalline structure. The prebiotic potential of HPH-treated BC was assessed through in vitro experiments with Bacillus amyloliquefaciens, showing a significant enhancement in bacterial proliferation (P=0.0437) compared to untreated BC. This study highlights the potential of BC, particularly when processed to the nanoscale, to serve as a functional ingredient that supports gut health and microbial growth, marking a significant advancement in the development of sustainable and effective prebiotic products.

Keywords: Bacterial cellulose, High pressure homogenization, Prebiotic, Acetic acid, Bacteria, Novacetimonas pomaceti

Xue Jing Wong¹, Shiamala Devi Ramaiya², Wan Hee Cheng¹, Zheng-Feng Wang³, Muhammad Syahmi Hishamuddin^4*, Shiou Yih Lee^1*

¹Faculty of Health and Life Sciences, INTI International University, 71800 Nilai, Negeri Sembilan, Malaysia

²Department of Crop Science, Faculty of Agricultural and Forestry Science, Universiti Putra Malaysia Bintulu Sarawak Campus, 97008 Bintulu, Sarawak, Malaysia

³Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, Guangdong, China

⁴Department of Forestry Science and Biodiversity, Faculty of Forestry and Environment, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia

Abstract

Coelostegia griffithii is a member of Malvaceae, which is native to the west Malaysia region. It has been exploited as a timber species to produce clogs, furniture, and construction materials. However, genetic studies of this species are limited. Thus, this study describes the first complete plastid genome (plastome) sequence of C. griffithii. The quadripartite-structured plastome was 163,159 bp long, consisting of large (95,536 bp) and small (20,435 bp) single-copy regions, which are separated by a pair of inverted repeats (IR) regions (each 23,594 bp). A total of 133 genes were annotated, including 88 protein-coding (CDS), 37 tRNA, and eight rRNA genes. Repeat analyses recorded 250 simple sequence repeats and 50 large repeats. The preferred amino acid often ended with codon A/T based on relative synonymous codon usage analysis. When compared to the plastid CDS of Reevesia thyrsoidea, a total of 12 genes displayed positive selection. There was no evidence of gene block rearrangement or inversion in comparison to four other closely related species of Helicteroideae. To ascertain its molecular placement, the phylogenetic analysis was carried out with the concatenated dataset of 79 shared unique CDS of 32 taxa of Malvaceae, using maximum likelihood (ML) and approximate Bayesian test (aBayes) methods. Both the ML and aBayes trees revealed a nearly resolved and well-resolved relationship within Malvaceae, respectively; C. griffithii is placed in the Helicteroideae clade and has a close relationship to the three Durio species.

 Keywords: Chloroplast genome, Durian, Durioneae, Genetic resources, Phylogenomics