Search results for: papaya leaf
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 714

Search results for: papaya leaf

474 Incidence and Molecular Mechanism of Human Pathogenic Bacterial Interaction with Phylloplane of Solanum lycopersicum

Authors: Indu Gaur, Neha Bhadauria, Shilpi Shilpi, Susmita Goswami, Prem D. Sharma, Prabir K. Paul

Abstract:

The concept of organic agriculture has been accepted as novelty in Indian society, but there is no data available on the human pathogens colonizing plant parts due to such practices. Also, the pattern and mechanism of their colonization need to be understood in order to devise possible strategies for their prevention. In the present study, human pathogenic bacteria were isolated from organically grown tomato plants and five of them were identified as Klebsiella pneumoniae, Enterobacter ludwigii, Serratia fonticola, Stenotrophomonas maltophilia and Chryseobacterium jejuense. Tomato plants were grown in controlled aseptic conditions with 25±1˚C, 70% humidity and 12 hour L/D photoperiod. Six weeks old plants were divided into 6 groups of 25 plants each and treated as follows: Group 1: K. pneumonia, Group 2: E. ludwigii, Group 3: S. fonticola, Group 4: S. maltophilia, Group 5: C. jejuense, Group 6: Sterile distilled water (control). The inoculums for all treatments were prepared by overnight growth with uniform concentration of 108 cells/ml. Leaf samples from above groups were collected at 0.5, 2, 4, 6 and 24 hours post inoculation for the colony forming unit counts (CFU/cm2 of leaf area) of individual pathogens using leaf impression method. These CFU counts were used for the in vivo colonization assay and adherence assay of individual pathogens. Also, resistance of these pathogens to at least 12 antibiotics was studied. Based on these findings S. fonticola was found to be most prominently colonizing the phylloplane of tomato and was further studied. Tomato plants grown in controlled aseptic conditions same as mentioned above were divided into 2 groups of 25 plants each and treated as follows: Group 1: S. fonticola, Group 2: Sterile distilled water (control). Leaf samples from above groups were collected at 0, 24, 48, 72 and 96 hours post inoculation and homogenized in suitable buffers for surface and cell wall protein isolation. Protein samples thus obtained were subjected to isocratic SDS-gel electrophoresis and analyzed. It was observed that presence of S. fonticola could induce the expression of at least 3 additional cell wall proteins at different time intervals. Surface proteins also showed variation in the expression pattern at different sampling intervals. Further identification of these proteins by MALDI-MS and bioinformatics tools revealed the gene(s) involved in the interaction of S. fonticola with tomato phylloplane.

Keywords: cell wall proteins, human pathogenic bacteria, phylloplane, solanum lycopersicum

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473 Synthesis of Silver Nanoparticles Adsorbent from Phytolacca Dodecandra ‘Endod’ Leaf to Water Treatment, at Almeda Textile Factory, Tigray Ethiopia

Authors: Letemariam Gebreslassie Gebrekidan

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Water pollution is one of the most feared problems in modern societies, especially in developing countries like Ethiopia. Nanoparticles with controlled size and composition are of fundamental and technological interest as they provide solutions to technological and environmental challenges in the areas of solar energy conversion, catalysis, medicine, and water treatment. The synthesis of metallic nanoparticles is an active area of academic and, more importantly, application research in nanotechnology. Adsorption is a process in which pollutants are absorbed on a solid surface. A molecule (pollutant) adhered to the solid surface is called an adsorbate, and the solid surface is an adsorbent. Adsorption is controlled by various parameters such as temperature, the nature of the adsorbate and adsorbent, and the presence of other pollutants along with the experimental conditions (pH, concentration of pollutants, contact time, particle size, and temperature). Depending on the main problem of water pollution, this research is available on the adsorption of wastewater using silver nanoparticles extracted from phytolacca Dodecandra leaf. AgNP was synthesized from a 1mM aqueous solution of silver nitrate (AgNO3) and Phytolacca Dodecandra leaf extract at room temperature. The synthesized nanoparticles were characterized using UV/visible Spectrometer, FTIR and XRD. In the UV-Vis spectrum, The Surface Plasmon resonance (SPR) peak was observed at 414 nm, which confirmed the synthesis of AgNPs. FTIR spectroscopy, recorded from 4000 cm-1 to 400 cm-1, indicated the presence of a capping agent with the nanoparticles. From the XRD results, the average crystalline size was estimated to be 20 nm Confirming the nanoparticle nature of the obtained sample. Thus, the present method leads to the formation of silver nanoparticles with well-defined dimensions. The effects of different parameters like solution pH, adsorbent dose, contact time and initial concentration of dye were studied. The concentration of MB is 0.01 mg/L and 0.002 mg/L before and after adsorption, respectively. The wastewater containing MB was well purified using AgNP adsorbent.

Keywords: wastewater, silver nanoparticle, Characterization, adsorption, parameter

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472 Investigating Prostaglandin E2 and Intracellular Oxidative Stress Levels in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages upon Treatment with Strobilanthes crispus

Authors: Anna Pick Kiong Ling, Jia May Chin, Rhun Yian Koh, Ying Pei Wong

Abstract:

Background: Uncontrolled inflammation may cause serious inflammatory diseases if left untreated. Non-steroidal anti-inflammatory drug (NSAIDs) is commonly used to inhibit pro-inflammatory enzymes, thus, reduce inflammation. However, long term administration of NSAIDs leads to various complications. Medicinal plants are getting more attention as it is believed to be more compatible with human body. One of them is a flavonoid-containing medicinal plants, Strobilanthes crispus which has been traditionally claimed to possess anti-inflammatory and antioxidant activities. Nevertheless, its anti-inflammatory activities are yet to be scientifically documented. Objectives: This study aimed to examine the anti-inflammatory activity of S. crispus by investigating its effects on intracellular oxidative stress and prostaglandin E2 (PGE2) levels. Materials and Methods: In this study, the Maximum Non-toxic Dose (MNTD) of methanol extract of both leaves and stems of S. crispus was first determined using 3-(4,5-dimethylthiazolyl-2)-2,5-diphenytetrazolium Bromide (MTT) assay. The effects of S. crispus extracts at MNTD and half MNTD (½MNTD) on intracellular ROS as well as PGE2 levels in 1.0 µg/mL LPS-stimulated RAW 264.7 macrophages were then be measured using DCFH-DA and a competitive enzyme immunoassay kit, respectively. Results: The MNTD of leaf extract was determined as 700µg/mL while for stem was as low as 1.4µg/mL. When LPS-stimulated RAW 264.7 macrophages were subjected to the MNTD of S. crispus leaf extract, both intracellular ROS and PGE2 levels were significantly reduced. In contrast, stem extract at both MNTD and ½MNTD did not significantly reduce the PGE2 level, but significantly increased the intracellular ROS level. Conclusion: The methanol leaf extract of S. crispus may possess anti-inflammatory properties as it is able to significantly reduce the intracellular ROS and PGE2 levels of LPS-stimulated cells. Nevertheless, further studies such as investigating the interleukin, nitric oxide and cytokine tumor necrosis factor-α (TNFα) levels has to be conducted to further confirm the anti-inflammatory properties of S. crispus.

Keywords: anti-inflammatory, natural products, prostaglandin E2, reactive oxygen species

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471 Activity of Some Plant Extracts on the Larvae and Eggs of Culex quinquefasciatus in the Laboratory

Authors: A. A. El Maghrbi

Abstract:

The control of vectors like mosquitoes based on the application of chemical insecticides but due to its adverse effect on the environment, and development of resistance by most of species of mosquitoes including vectors of important diseases. Ethanol and acetone extracts of nine species of plants (Allium tuberosum, Apium leptophylum, Carica papaya, Cymbopogon citratus, Euphorbia cotinofolia, Melia azedarach, Ocimum canum, Ricinus common, and Tagetes erecta) were tested in respect of their influence on the eggs and larvae of Culex quinquifasciatus in concentration 100, 10 and 1 mg/L. In relation to the survival of larvae, ethanol extract of O. canum and acetone extract of A.tuberosum in 100 mg/L have larvicide activity against L4 of Cx. quinquifasciatus. For hatching of eggs, ethanol and acetone extract of A.tuberosum (100 and 10 mg/L) and acetone extract of C.citratus (100 mg/L) produced reduction in the number of eggs hatched of Cx. quinquifasciatus. Our results indicate that each extract of the plant have potential to control mosquito population and suggest that further studies are needed in this field.

Keywords: Cx. quinquefasciatus, plant extract, ethanol, acetone, larvae, eggs

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470 Physicochemical Analysis of Soxhlet Extracted Oils from Selected Northern Nigerian Seeds

Authors: Abdulhamid Abubakar, Sani Ibrahim, Fakai I. Musa

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The aim of the present study is to investigate the potential use of the selected seed oils. The oil was extracted using Soxhlet apparatus and the physicochemical characteristics of the oil determined using standard methods. The following results were obtained for the physicochemical parameters analysed: for Egusi seed oil, Oil yield 53.20%, Saponification value 178.03±1.25 mgKOH/g, iodine value 49.10±0.32 g I2/100 g, acid value 4.30±0.86 mgKOH/g, and Peroxide value 5.80±0.27 meq/kg were obtained. For Pawpaw seed oil, Oil yield 40.10%, Saponification value 24.13±3.93 mgKOH/g, iodine value 24.87±0.19 g I2/100g, acid value 9.46±0.40 mgKOH/g, and Peroxide value 3.12±1.22 meq/kg were obtained. For Sweet orange seed oil, oil yield 43.10%, Saponification value 106.30±2.37 mgKOH/g, Iodine value 37.08±0.04 g I2/100g, acid value 7.59±0.77 mgKOH/g, and Peroxide value 2.21±0.46 meq/kg were obtained. From the obtained values of the determined parameters, the oils can be extracted from the three selected seeds in commercial quantities and that the egusi and sweet orange seed oils may be utilized in the industrial soap production.

Keywords: Carica papaya, Citrus sinensis, physicochemical, iodine value, peroxide value

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469 Spectroscopic Determination of Functionalized Active Principles from Coleus aromaticus Benth Leaf Extract Using Ionic Liquids

Authors: Zharama M. Llarena

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Green chemistry for plant extraction of active principles is the main interest of many researchers concerned with climate change. While classical organic solvents are detrimental to our environment, greener alternatives to ionic liquids are very promising for sustainable organic chemistry. This study focused on the determination of functional groups observed in the main constituents from the ionic liquid extracts of Coleus aromaticus Benth leaves using FT-IR Spectroscopy. Moreover, this research aimed to determine the best ionic liquid that can separate functionalized plant constituents from the leaves Coleus aromaticus Benth using Fourier Transform Infrared Spectroscopy. Coleus aromaticus Benth leaf extract in different ionic liquids, elucidated pharmacologically important functional groups present in major constituents of the plant, namely, rosmarinic acid, caffeic acid and chlorogenic acid. In connection to distinctive appearance of functional groups in the spectrum and highest % transmittance, potassium chloride-glycerol is the best ionic liquid for green extraction.

Keywords: chlorogenic acid, coleus aromaticus, ionic liquid, rosmarinic acid

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468 Sustainable Biostimulant and Bioprotective Compound for the Control of Fungal Diseases in Agricultural Crops

Authors: Geisa Lima Mesquita Zambrosi, Maisa Ciampi Guillardi, Flávia Rodrigues Patrício, Oliveiro Guerreiro Filho

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Certified agricultural products are important components of the food industry. However, certifiers have been expanding the list of restricted or prohibited pesticides, limiting the options of products for phytosanitary control of plant diseases, but without offering alternatives to the farmers. Soybean and coffee leaf rust, brown eye spots, and Phoma leaf spots are the main fungal diseases that pose a serious threat to soybean and coffee cultivation worldwide. In conventional farming systems, these diseases are controlled by using synthetic fungicides, which, in addition to intensifying the occurrence of fungal resistance, are highly toxic to the environment, farmers, and consumers. In organic, agroecological, or regenerative farming systems, product options for plant protection are limited, being available only copper-based compounds, and biodefensivesornon-standard homemade products. Therefore, there is a growing demand for effective bioprotectors with low environmental impact for adoption in more sustainable agricultural systems. Then, to contribute to covering such a gap, we have developed a compound based on plant extracts and metallic elements for foliar application. This product has both biostimulant and bioprotective action, which promotes sustainable disease control, increases productivity as well as reduces damage to the environment. The product's components have complementary mechanisms that promote protection against the disease by directly acting on the pathogens and activating the plant's natural defense system. The protective ability of the product against three coffee diseases (coffee leaf rust, brown eye spot, and Phoma leaf spot) and against soybean rust disease was evaluated, in addition to its ability to promote plant growth. Our goal is to offer an effective alternative to control the main coffee fungal diseases and soybean fungal diseases, with a biostimulant effect and low toxicity. The proposed product can also be part of the integrated management of coffee and soybean diseases in conventional farming associated with chemical and biological pesticides, offering the market a sustainable coffee and soybean with high added value and low residue content. Experiments were carried out under controlled conditions to evaluate the effectiveness of the product in controlling rust, phoma, and cercosporiosis in comparison to control-inoculated plants that did not receive the product. The in vitro and in vivo effects of the product on the pathogen were evaluated using light microscopy and scanning electron microscopy, respectively. The fungistatic action of the product was demonstrated by a reduction of 85% and 95% in spore germination and disease symptoms severity on the leaves of coffee plants, respectively. The formulation had both a protective effect, acting to prevent infection by coffee leaf rust, and a curative effect, reducing the rust symptoms after its establishment.

Keywords: plant disease, natural fungicide, plant health, sustainability, alternative disease management

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467 Properties and Antimicrobial Activity of Fish Protein Isolate/Fish Skin Gelatin Film Containing Basil Leaf Essential Oil and Zinc Oxide Nanoparticles

Authors: Yasir Ali Arfat

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Composite films based on fish protein isolate (FPI) and fish skin gelatin (FSG) blend incorporated with 50 and 100% (w/w, protein) basil leaf essential oil (BEO) in the absence and presence of 3% (w/w, protein) ZnO nanoparticles (ZnONP) were prepared and characterised. Tensile strength (TS) decreased, whilst elongation at break (EAB) increased as BEO level increased (p < 0.05). However, ZnONP addition resulted in higher TS but lower EAB (p < 0.05). The lowest water vapour permeability (WVP) was observed for the film incorporated with 100% BEO and 3% ZnONP (p < 0.05). BEO and ZnONP incorporation decreased transparency of FPI/FSG films (p < 0.05). FTIR spectra indicated that films added with BEO exhibited higher hydrophobicity. Both BEO and ZnONP had a marked impact on thermal stability of the films. Microstructural study revealed that presence of ZnONP prevented bilayer formation of film containing 100% BEO. FPI/FSG films incorporated with 100% BEO, especially in combination with ZnONP, exhibited strong antibacterial activity against food pathogenic and spoilage bacteria and thus could be used as an active food packaging material to ensure safety and to extend the shelf-life of packaged foods.

Keywords: bionanocomposite, fish protein isolate, fish skin gelatin, basil essential oil, ZnO nanoparticles, antimicrobial packaging

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466 Safety Assessment and Prophylactic Efficacy of Moringa stenopetala Leaf Extract Through Mitigation of Oxidative Stress in BV-2 Microglial Cell

Authors: Stephen Adeniyi Adefegha, Vitor Mostardeiro, Vera Maria Morsch, Ademir F. Morel, Ivana Beatrice Manica Da Cruz, Sabrina Somacal Maria Rosa Chitolina Schetinger

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Moringa stenopetala is often consumed as food and used in folkloric medicine for the management of several diseases. Purpose: This study was set up in order to assess the effect of aqueous extract of Moringa stenopetala on cell viability and oxidative stress biomarkers in BV-2 microglial cells. Aqueous extracts of M. stenopetala were prepared, lyophilized and reconstituted in 0.5% dimethylsulphoxide (DMSO). Cells were treated with M. stenopetala extracts (0.1 - 100 µg/ml) for cell viability and nitric oxide (NO) production tests. However, M. stenopetala extract (50 µg/ml) was used in the treatment of cells for the determination of protein carbonyl content and reactive oxygen species (ROS) level. Incubation of BV-2 microglia cell with M. stenopetala extract maintained cell viability, diminished NO and ROS levels, and reduced protein carbonyl contents Chlorogenic acid, rutin, kaempferol and quercetin derivatives were the main phenolic compounds identified in M. stenopetala leaf extract. These phenolic compounds present in M. stenopetala may be responsible for the mitigation of oxidative stress in BV-2 microglial cells.

Keywords: oxidative stress, BV-2 microglial cell, Moringa stenopetala, cell viability, antioxidant

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465 Nephroprotective Effect of Asparagus falcatus Leaf Extract on Adriamycin Induced Nephrotoxicity in Wistar Rats: A Dose Response Study

Authors: A. M. S. S. Amarasiri, A. P. Attanayake, K. A. P. W. Jayatilaka, L. K. B. Mudduwa

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Adriamycin (ADR) is an effective anthracyclin antitumor drug, but its clinical use is limited due to renal toxicity. The leaves of Asparagus falcatus (Family: Liliaceae) have been used in the management of renal diseases since antiquity. In the present investigation, the aqueous leaf extract of A. falcatus was evaluated for acute nephroprotective activity in ADR induced nephrotoxic rats. Nephrotoxicity was induced in healthy male Wistar rats by intraperitoneal administration of ADR 20 mg/kg. The lyophilized powder of the aqueous refluxed (4h) leaf extract of A. falcatus was administered orally at three selected doses; 200, 400 and 600 mg/kg for three consecutive days. Fosinopril sodium (0.09 mg/kg) was used as the standard drug. Administration of the plant extract and the standard drug was commenced 24 hours after the induction of nephrotoxicity to rats. The nephroprotective effect was determined by selected biochemical parameters and by the assessment of histopathology on H and E stained kidney sections. The results were compared to a group of control rats with ADR induced nephrotoxicity. A group of rats administered with the equivalent volume of normal saline served as the healthy control. Administration of ADR 20 mg/kg produced a significant increase in the concentrations of serum creatinine (61%) and urine protein (73%) followed by a significant decrease in serum total protein (21%) and albumin (44%) of the plant extract treated animals compared to the healthy control group (p < 0.05). The aqueous extract of Asparagus falcatus at the three doses; 200, 400 and 600 mg/kg and the standard drug were found to decrease the elevation of concentrations of serum creatinine (33%, 51%, 54% and 42%) and urine protein (8%, 63%, 80% and 86%) respectively. The serum concentrations of total protein (12%, 17%, 29% and 12%) and albumin (3%, 17%, 17% and 16%) were significantly increased compared to the nephrotoxic control group respectively. Assessment of histopathology on H and E stained kidney sections demonstrated that ADR induced renal injury, as evidenced by loss of brush border, cytoplasmic vacuolization, pyknosis in renal tubular epithelial cells, haemorrhages, glomerular congestion and presence of hyaline casts. Treatment with the plant extract and the standard drug resulted in attenuation of the morphological destruction in rats. The results of the present study revealed that the aqueous leaf extract of A. falcatus possesses significant nephroprotective activity against adriamycin induced acute nephrotoxicity. The improved kidney functions were supported with the results of selected biochemical parameters and histological changes observed on H and E stained sections of the kidney tissues in Wistar rats.

Keywords: adriamycin induced nephrotoxicity, asparagus falcatus, biochemical assessment, histopathological assessment, nephroprotective activity

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464 Impact of the Achyranthes aspera (Amaranthaceae) Extracts on the Survival and Histological Architecture of the Midgut Epithelial Tissue of Early Fourth Instars of Aedes aegypti (Diptera: Culicidae)

Authors: Aarti Sharma, Sarita Kumar, Pushplata Tripathi

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Aedes aegypti L. is one of the most important insect vectors in the world transmitting several diseases of concern; dengue fever, dengue haemorrhagic fever and yellow fever. Though since ages the control of dengue vector is primarily relied upon the use of synthetic chemical insecticides, the continued and indiscriminate use of insecticides for their control has received wide public apprehension because of multifarious problems including insecticide resistance, resurgence of pest species, environmental pollution, toxic hazards to humans and non-target organisms. These problems have necessitated the need to explore and develop alternative strategies using eco-friendly and bio-degradable plant products. Bio-insecticides, despite being the focus of research nowadays, have not been investigated much regarding their physiological effects on the mosquitoes. Thus, the present studies were carried out to investigate the anti-mosquito potential of the leaf and stem hexane extracts of Achyranthes aspera against early fourth instars of Aedes aegypti L and their effects on the histological architecture of their midgut. The larvicidal bioassays conducted with the A. aspera leaf hexane extracts revealed the respective LC30, LC50 and LC90 values of 66.545 ppm, 82.555 ppm, 139.817 ppm while the assays with stem hexane extracts resulted in respective values of 54.982 ppm, 68.133 ppm, 115.075 ppm. The studies clearly indicate the efficacy of extracts as larvicidal agents against Ae. aegypti, the stem extracts being found more effective than the leaf extracts. When the larvae assayed with extracts were investigated for the modifications in the histo-architecture of the midgut, the studies showed significant damage, shrinkage, distortion and vacuolization of gut tissues and peritrophic membrane causing disintegration of epithelial cells and cytoplasmic organelles; extent of toxicity and damage varied depending upon the concentration and exposure time period. These changes revealed appreciable stomach poison potential of A. aspera extracts against Ae. aegypti larvae, which may have also caused adverse impact on the growth and development of larvae. These effects were also found to be more pronounced with the stem extract than the leaf extract. Our findings may prove significant suggesting the use of A. aspera extract as a bio-insecticide against early fourth instar larvae of Ae. aegypti. Further studies are needed to identify the bioactive component in the extracts and to ascertain the use of component in the fields as anti-mosquito control agent.

Keywords: Achyranthes aspera, Aedes aegypti, histological architecture, larvicidal, midgut, stomach poison

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463 In vitro Susceptibility of Madurella mycetomatis to the Extracts of Anogeissus leiocarpus Leaves

Authors: Ikram Mohamed Eltayeb Elsiddig, Abdel Khalig Muddather, Hiba Abdel Rahman Ali, Saad Mohamed Hussein Ayoub

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Anogeissusleiocarpus (Combretaceae) is well known for its medicinal uses in African traditional medicine, for treating many human diseases mainly skin diseases and infections.Mycetoma disease is a fungal and/ or bacterial skin infection, mainly cause by Madurella mycetomatis fungus.This study was carried out in vitro to investigate the antifungal activity of Anogeissusleiocarpus leaf extracts against the isolated pathogenicMadurellamycetomatis, by using the NCCLS modified method compared to Ketoconazole standard drug and MTT assay. The bioactive fraction was subjected to chemical analysis implementing different chromatographic analytical methods (TLC, HPLC, and LC-MS/MS). The results showed significance antifungal activity of A. leiocarpus leaf extractsagainst the isolated pathogenicM. mycetomatis, compared to negative and positive controls. The chloroform fraction showed the highest antifungal activity.The chromatographic analysis of the chloroform fraction with the highest activity showed the presence of important bioactive compounds such as ellagic and flavellagic acids derivatives, flavonoids and stilbenoid, which are well known for their antifungal activity.

Keywords: Anogeissus leiocarpus, crude extracts and fractions of Anogeissus leiocarpus, in vitrosusceptibility of Madurella mycetomatis, Madurella mycetomatis

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462 Preparation, Characterisation, and Antibacterial Activity of Green-Biosynthesised Silver Nanoparticles Using Clinacanthus Nutans Extract

Authors: Salahaedin Waiezi, Nik Ahmad Nizam Nik Malek, Hassan Abdelmagid Elzamzami, Shahrulnizahana Mohammad Din

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A green and safe approach to the synthesis of silver nanoparticles (AgNP) can be performed using plant leaf extract as the reducing agent. Hence, this paper reports the biosynthesis of AgNP using Clinacanthus nutans plant extract. C. nutans is known as belalai gajah in Malaysia and is widely used as a medicinal herb locally. The biosynthesized AgNP, using C. nutans aqueous extract at pH 10, with the reaction temperature of 70°C and 48 h reaction time, was characterized by UV-Vis spectroscopy, X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), energy dispersive X-ray (EDX), and transmission electron microscope (TEM). A peak appeared in the UV-Vis spectra at around 400 nm, while XRD confirmed the crystal structure of AgNP, with the average size between 20 to 30 nm, as shown in FESEM and TEM. The antibacterial activity of the biosynthesized AgNP, which was performed using the disc diffusion technique (DDT) indicated effective inhibition against Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. In contrast, minimal antibacterial activity was detected against Enterococcus faecalis and methicillin-resistant Staphylococcus aureus (MRSA). In general, AgNP produced using C. nutans leaf extract possesses potential antibacterial activity.

Keywords: silver nanoparticles, Clinacanthus nutans, antibacterial agent, biosynthesis

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461 The Expression Patterns of Thai Moderately Salt Tolerant Rice and High Salt Tolerant Rice in Response to Salt Stress

Authors: Kongngern K., Homwonk C., Theerakulpisut P., Roytrakul R.

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Rice cultivation is crucial globally, especially in Asia. Soil salinity poses a significant challenge for agricultural lands. Understanding the expression patterns of different rice varieties under salt stress can provide insights for developing more salt-tolerant cultivars. This study aims to compare the expression patterns of two rice varieties, Thai moderately salt-tolerant rice (Leaung Anan) and high salt-tolerant rice (Pokkali), in response to salt stress. By analyzing protein expression, the research seeks to identify key proteins associated with salt tolerance in rice. The expression patterns of the two rice varieties under salt stress were analyzed using 1D-SDS-PAGE, NanoLC-MS/MS, and MEV software. These methods enabled the researchers to assess the differential expression of proteins in the leaf sheaths of the rice plants. These results indicate that the study identified 18 proteins, exhibited significantly different expression patterns between the two rice cultivars under salt stress. Notably, certain proteins, such as Os05g0364500 and pr1-like protein, showed contrasting expression profiles in the two varieties. The up-regulated proteins, predominantly observed in the salt-tolerant rice, may contribute to the survival of rice plants under salt stress and may provide valuable insights for breeding programs aiming to enhance salt tolerancein rice cultivars.

Keywords: mass spectrometry, proteomics, rice leaf sheaths, salt stress

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460 Evaluation of Chromium Fortified-Parboiled Rice Coated with Herbal Extracts: Resistant Starch, and Glycemic Index

Authors: Wisnu Adi Yulianto, Chatarina Lilis Suryani, Mamilisti Susiati, Hendy Indra Permana

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Parboiled rice was developed to produce rice that has low glycemic index, especially for diabetics. Yet, parboiled rice is not enough because diabetics also lack of chromium. The sign of chromium (Cr) deficiency in diabetics is impaired glucose tolerance. Cr fortification was done for increasing Cr content in rice. Naturally-occurring compounds that have been proven to improve insulin sensitivity include Cr and polyphenol found in cinnamon, pandan and bay leaf. This research aimed to evaluate content of resistant starch and glycemic index of Cr - fortified - parboiled rice (Cr-PR) coated with herbal extracts. Variety of unhulled rice and forticant used in the experiment were Ciherang and CrCl3, respectively. Three herbal extracts used were cinnamon, pandan and bay leaf. Each concentration of herbal extracts in the amount of 3%, 6%, and 9% were added in the coating substance to coat Cr-PR. Resistant starch (RS) content was determined by enzymatic process through glucooxydase method. Testing of the GI was conducted on 18 non-diabetic volunteers. RS content of Cr-PR coated with herbal extracts ranged between 8.27 – 8.84 % (dry weight). Cr-PR coated with all herbal extracts of 3% concentration had higher RS content than the ones with herbal extracts of 6% and 9% concentration (P <0.05). Value of the rice GI ranged 29 - 40. The lowest GI (29-30) was attained by the rice coated with enrichment of 6-9% cinnamon extract.

Keywords: coating, Cr-fortified-parboiled rice, glycemic index, herbal extracts, resistant starch

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459 Identification of Tissue-Specific Transcription Factors in C. roseus with Emphasis to the TIA Biosynthetic Pathway

Authors: F. M. El-Domyati, A. Atef, S. Edris, N. O. Gadalla, M. A. Al-Kordy, A. M. Ramadan, Y. M. Saad, H. S. Al-Zahrani, A. Bahieldin

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Transcriptome retrieved from SRA database of different tissues and treatments of C. roseus was assembled in order to detect tissue-specific transcription factors (TFs) and TFs possibly related to terpenoid indole alkaloids (TIA) pathway. A number of 290 TF-like transcripts along with 12 transcripts related to TIA biosynthetic pathway were divided in terms of co-expression in the different tissues, treatments and genotypes. Three transcripts encoding peroxidases 1 and 12 were downregulated in hairy root, while upregulated in mature leaf. Eight different transcripts of the TIA pathway co-expressed with TFs either functioning downstream tryptophan biosynthesis, e.g., tdc, str1 and sgd, or upstream vindoline biosynthesis, e.g., t16h, omt, nmt, d4h and dat. The results showed no differential expression of TF transcripts in hairy roots knocked down for tdc gene (TDCi) as compared to their wild type controls. There were several evidences of tissue-specific expression of TF transcripts in flower, mature leaf, root/hairy root, stem, seedling, hairy root and immature/mature leaves. Regulation included transcription factor families, e.g., bHLH, MYB and WRKY mostly induced by ABA and/or JA (or MeJA) and regulated during abiotic or biotic stress. The information of tissue-specific regulation and co-expression of TFs and genes in the TIA pathway can be utilized in manipulating alkaloid biosynthesis in C. roseus.

Keywords: SRA database, bHLH, MYB, WRKY, co-expression

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458 A Neural Network Classifier for Estimation of the Degree of Infestation by Late Blight on Tomato Leaves

Authors: Gizelle K. Vianna, Gabriel V. Cunha, Gustavo S. Oliveira

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Foliage diseases in plants can cause a reduction in both quality and quantity of agricultural production. Intelligent detection of plant diseases is an essential research topic as it may help monitoring large fields of crops by automatically detecting the symptoms of foliage diseases. This work investigates ways to recognize the late blight disease from the analysis of tomato digital images, collected directly from the field. A pair of multilayer perceptron neural network analyzes the digital images, using data from both RGB and HSL color models, and classifies each image pixel. One neural network is responsible for the identification of healthy regions of the tomato leaf, while the other identifies the injured regions. The outputs of both networks are combined to generate the final classification of each pixel from the image and the pixel classes are used to repaint the original tomato images by using a color representation that highlights the injuries on the plant. The new images will have only green, red or black pixels, if they came from healthy or injured portions of the leaf, or from the background of the image, respectively. The system presented an accuracy of 97% in detection and estimation of the level of damage on the tomato leaves caused by late blight.

Keywords: artificial neural networks, digital image processing, pattern recognition, phytosanitary

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457 Nonlinear Porous Diffusion Modeling of Ionic Agrochemicals in Astomatous Plant Cuticle Aqueous Pores: A Mechanistic Approach

Authors: Eloise C. Tredenick, Troy W. Farrell, W. Alison Forster, Steven T. P. Psaltis

Abstract:

The agriculture industry requires improved efficacy of sprays being applied to crops. More efficacious sprays provide many environmental and financial benefits. The plant leaf cuticle is known to be the main barrier to diffusion of agrochemicals within the leaf. The importance of a mathematical model to simulate uptake of agrochemicals in plant cuticles has been noted, as the results of each uptake experiments are specific to each formulation of active ingredient and plant species. In this work we develop a mathematical model and numerical simulation for the uptake of ionic agrochemicals through aqueous pores in plant cuticles. We propose a nonlinear porous diffusion model of ionic agrochemicals in isolated cuticles, which provides additions to a simple diffusion model through the incorporation of parameters capable of simulating plant species' variations, evaporation of surface droplet solutions and swelling of the aqueous pores with water. The model could feasibly be adapted to other ionic active ingredients diffusing through other plant species' cuticles. We validate our theoretical results against appropriate experimental data, discuss the key sensitivities in the model and relate theoretical predictions to appropriate physical mechanisms.

Keywords: aqueous pores, ionic active ingredient, mathematical model, plant cuticle, porous diffusion

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456 Extraction, Isolation and Comparative Phtochemical Study of Aegle Marmelos, Calendula Officinalis and Fenugreek

Authors: Nitin Rajan, Kashif Shakeel, Shashank Tiwari, Shachan Sagar

Abstract:

Background: - Aegle Marmelos (Bael) leaf extract is taken twice daily to treat ophthalmia, ulcers, and intestinal worms, among other ailments. Poultice made from bael leaf is used in the treatment of eye conditions. The leaf juice has a variety of therapeutic applications, with the most notable being the treatment of diabetes. Fenugreek is used to cure red spots around the eyes, as well as to soften the throat and chest and to give relief from coughing. The use of this plant in the form of infusion, powder, pomade, and decoction has been extremely popular in Iranian traditional medicine. The plant may be used to wash one's vaginal linings. This plant is used as an emollient in the lack of appetite, treatment of pellagra, and gastrointestinal problems, as well as a general tonic. Calendula officinalis leaves are used to treat varicose veins on the outside of the body by infusing them. In Europe, the leaves are diaphoretic and resolvent in nature, while the blooms are employed as an emmenagogue and antispasmodic stimulant in Canada and the United States. The flowers were decocted and served as a posset drink when smallpox and measles were common in England, and the fresh juice was used to treat jaundice. Objective: - This study is done to compare the physicochemical parameter of the alcoholic extract of the leaves of Aegle Marmelos, Calendula Officinalis, and Fenugreek. Materials and Methods: Extraction and Isolation of Aegle Marmelos, Calendula Officinalis, Fenugreek, were done. Preliminary phytochemical study for alkaloids, cardiac glycosides, flavonoids, glycosides, phenols, resins, saponins, steroids, tannins, terpenoids of the extract was done individual by using the standard procedure. Result: - The phytochemical screening of Aegle Marmelos, Calendula Officinalis, and Fenugreek shows the presence of alkaloids, carbohydrates, total phenolics, total flavonoids, tannins, saponins gum. Conclusion: - In this study, we have found that crude aqueous and organic solvent extracts of Aegle Marmelos, Calendula Officinalis, and Fenugreek leaves contain some important bioactive compounds and it justifies their use in the traditional medicines for the treatment of different diseases.

Keywords: Aegle Marmelos, Calendula Officinalis, Fenugreek, physiochemical parameter

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455 Ant and Spider Diversity in a Rural Landscape of the Vhembe Biosphere, South Africa

Authors: Evans V. Mauda, Stefan H. Foord, Thinandavha C. Munyai

Abstract:

The greatest threat to biodiversity is a loss of habitat through landscape fragmentation and attrition. Land use changes are therefore among the most immediate drivers of species diversity. Urbanization and agriculture are the main drivers of habitat loss and transformation in the Savanna biomes of South Africa. Agricultural expansion and the intensification in particular, take place at the expense of biodiversity and will probably be the primary driver of biodiversity loss in this century. Arthropods show measurable behavioural responses to changing land mosaics at the smallest scale and heterogeneous environments are therefore predicted to support more complex and diverse biological assemblages. Ants are premier soil turners, channelers of energy and dominate insect fauna, while spiders are a mega-diverse group that can regulate other invertebrate populations. This study aims to quantify the response of these two taxa in a rural-urban mosaic of a rapidly developing communal area. The study took place in and around two villages in the north-eastern corner of South Africa. Two replicates for each of the dominant land use categories, viz. urban settlements, dryland cultivation and cattle rangelands, were set out in each of the villages and sampled during the dry and wet seasons for a total of 2 villages × 3 land use categories × 2 seasons = 24 assemblages. Local scale variables measured included vertical and horizontal habitat structure as well as structural and chemical composition of the soil. Ant richness was not affected by land use but local scale variables such as vertical vegetation structure (+) and leaf litter cover (+), although vegetation complexity at lower levels was negatively associated with ant richness. However, ant richness was largely shaped by regional and temporal processes invoking the importance of dispersal and historical processes. Spider species richness was mostly affected by land use and local conditions highlighting their landscape elements. Spider richness did not vary much between villages and across seasons and seems to be less dependent on context or history. There was a considerable amount of variation in spider richness that was not explained and this could be related to factors which were not measured in this study such as temperature and competition. For both ant and spider assemblages the constrained ordination explained 18 % of variation in these taxa. Three environmental variables (leaf litter cover, active carbon and rock cover) were important in explaining ant assemblage structure, while two (sand and leaf litter cover) were important for spider assemblage structure. This study highlights the importance of disturbance (land use activities) and leaf litter with the associated effects on ant and spider assemblages across the study area.

Keywords: ants, assemblages, biosphere, diversity, land use, spiders, urbanization

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454 The Isolation of Enterobacter Ludwigii Strain T976 from Nicotiana Tabacum L. Yunyan 97 and Its Application Study

Authors: Gao Qin, Hu Liwei, Dong Xiangzhou, Zhu Qifa, Cheng Tingming, Zhao Limei, Yang Mengmeng, Zhai Zhen, Dai Huaxin, Liang Taibo, Zhang Shixiang, Xue Chaoqun

Abstract:

The functional strain T976 for starch degradation was isolated from Nicotiana tabacum L. Yunyan 97 tobacco leaves, the ratio of starch hydrolysis transparent circle diameter to colony diameter of the strain was 4.14, 16S rDNA sequencing identified these strains as Enterobacter ludwigii. Then Enterobacter ludwigii T976 was fermented and spaying Yunyan 97 plant in vigorous growing stage. The results of once spraying fermentation broth of Enterobacter ludwigii T976 showed that starch content of upper leaves decreased slightly, from 3.77% to 3.1%, the reducing sugar content increased from 4.39% to 5.53%, and the total sugar content increased from 5.82% to 7.39%. The chemical content was also checked after three time spraying. The starch content of middle leaves decreased from 5.63% to 3.74%, while the content of total sugar and reducing sugar decreased slightly. And the starch content of upper leaves decreased from 7.62% to 4.78%, the total sugar and reducing sugar decreased slightly, and starch content of middle leaf decreased from 6.27% to 3.62%, the total sugar and reducing sugar did not change much, and other chemical components were in a suitable range.

Keywords: nicotiana tabacum, yunyan 97, leaf, starch, degradation, enterobacter ludwigii

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453 Spinetoram10% WG+Sulfoxaflor 30% WG: A Promising Green Chemistry to Manage Pest Complex in Bt Cotton

Authors: Siddharudha B. Patil

Abstract:

Cotton is a premier commercial fibre crop of India subjected to ravages of insect pests. Sucking pests viz thrips, Thrips tabaci,(lind) leaf hopper Amrsca devastance,(dist) miridbug, Poppiocapsidea beseratense (Dist) and bollworms continue to inflict damage Bt Cotton right from seeding stage. Their infestation impact cotton yield to an extent of 30-40 percent. Chemical control is still adoptable as one of the techniques for combating these pests. Presently, growers have many challenges in selecting effective chemicals which fit in with an integrated pest management. Spinetoram has broad spectrum with excellent insecticidal activity against both sucking pests and bollworms. Hence, it is expected to make a great contribution to stable production and quality improvement of agricultural products. Spinetoram is a derivative of biologically active substances (Spinosyns) produced by soil actinomycetes, Saccharopolypara spinosa which is semi synthetic active ingredient representing Spinosyn chemical class of insecticide and has demonstrated higher level of efficacy with reduced risk on beneficial arthropods. The efforts were made in the present study to test the efficacy of Spinetoram against sucking pests and bollworms in comparison with other insecticides in Bt Cotton under field condition. Field experiment was laid out during 2013-14 and 2014-15 at Agricultural Research station Dharwad (Karnataka-India) in a randomized block design comprising eight treatments and three replications. Bt cotton genotype, Bunny BG-II was sown in a plot size of 5.4 m x5.4 m. Recommend agronomical practices were followed. The Spinetoram 12% SC alone and incombination with sulfaxaflore with varied dosages against pest complex was tested. Performance was compared with Spinosad 45% SC and thiamethoxam 25% WG. The results of consecutive seasons revealed that nonsignificant difference in thrips and leafhopper population and varied significantly after 3 days of imposition. Among the treatments, combiproduct, Spinetoram 10%WG + Sulfoxaflor 30% WG@ 140 gai/ha registered lowest population of thrips (3.91/3 leaves) and leaf hoppers (1.08/3 leaves) followed by its lower dosages viz 120 gai/ha (4.86/3 leaves and 1.14/3 leaves of thrips and leaf hoppers, respectively) and 100 gai/ha (6.02 and 1.23./3 leaves of thrips and leaf hoppers respectively) being at par, significantly superior to rest of the treatments. On the contrary, the population of thrips, leaf hopper and miridbugs in untreated control was on higher side. Similarly the higher dosage of Spinetoram 10% WG+ Sulfoxaflor 30% WG (140 gai/ha) proved its bioefficacy by registering lowest miridbug incidence of 1.70/25 squares, followed by its lower dosage (1.78 and 1.83/25 squares respectively) Further observation made on bollworms incidence revealed that the higher dosage of Spinetoram 10% WG+Sulfoxaflor 30% WG (140 gai/ha) registered lowest percentage of boll damage (7.22%), more number of good opened bolls (36.89/plant) and higher seed cotton yield (19.45q/ha) followed by rest of its lower dosages, Spinetoram 12% SC alone and Spinosad 45% SC being at par significantly superior to rest of the treatments. However, significantly higher boll damage (15.13%) and lower seed cotton yield (14.45 q/ha) was registered in untreated control. Thus Spinetoram10% WG+Sulfoxaflor 30% WG can be a promising option for pest management in Bt Cotton.

Keywords: Spinetoram10% WG+Sulfoxaflor 30% WG, sucking pests, bollworms, Bt cotton, management

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452 Biosynthesis of Silver Nanoparticles from Leaf Extract of Tithonia diversifolia and Its Antimicrobial Properties

Authors: Babatunde Oluwole Ogunsile, Omosola Monisola Fasoranti

Abstract:

High costs and toxicological hazards associated with the physicochemical methods of producing nanoparticles have limited their widespread use in clinical and biomedical applications. An ethically sound alternative is the utilization of plant bioresources as a low cost and eco–friendly biological approach. Silver nanoparticles (AgNPs) were synthesized from aqueous leaf extract of Tithonia diversifolia plant. The UV-Vis Spectrophotometer was used to monitor the formation of the AgNPs at different time intervals and different ratios of plant extract to the AgNO₃ solution. The biosynthesized AgNPs were characterized by FTIR, X-ray Diffraction (XRD) and Scanning Electron Microscope (SEM). Antimicrobial activities of the AgNPs were investigated against ten human pathogens using agar well diffusion method. The AgNPs yields were modeled using a second-order factorial design. The result showed that the rate of formation of the AgNPs increased with respect to time while the optimum ratio of plant extract to the AgNO₃ solution was 1:1. The hydroxyl group was strongly involved in the bioreduction of the silver salt as indicated by the FTIR spectra. The synthesized AgNPs were crystalline in nature, with a uniformly distributed network of the web-like structure. The factorial model predicted the nanoparticles yields with minimal errors. The nanoparticles were active against all the tested pathogens and thus have great potentials as antimicrobial agents.

Keywords: antimicrobial activities, green synthesis, silver nanoparticles, Tithonia diversifolia

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451 Inversion of PROSPECT+SAIL Model for Estimating Vegetation Parameters from Hyperspectral Measurements with Application to Drought-Induced Impacts Detection

Authors: Bagher Bayat, Wouter Verhoef, Behnaz Arabi, Christiaan Van der Tol

Abstract:

The aim of this study was to follow the canopy reflectance patterns in response to soil water deficit and to detect trends of changes in biophysical and biochemical parameters of grass (Poa pratensis species). We used visual interpretation, imaging spectroscopy and radiative transfer model inversion to monitor the gradual manifestation of water stress effects in a laboratory setting. Plots of 21 cm x 14.5 cm surface area with Poa pratensis plants that formed a closed canopy were subjected to water stress for 50 days. In a regular weekly schedule, canopy reflectance was measured. In addition, Leaf Area Index (LAI), Chlorophyll (a+b) content (Cab) and Leaf Water Content (Cw) were measured at regular time intervals. The 1-D bidirectional canopy reflectance model SAIL, coupled with the leaf optical properties model PROSPECT, was inverted using hyperspectral measurements by means of an iterative optimization method to retrieve vegetation biophysical and biochemical parameters. The relationships between retrieved LAI, Cab, Cw, and Cs (Senescent material) with soil moisture content were established in two separated groups; stress and non-stressed. To differentiate the water stress condition from the non-stressed condition, a threshold was defined that was based on the laboratory produced Soil Water Characteristic (SWC) curve. All parameters retrieved by model inversion using canopy spectral data showed good correlation with soil water content in the water stress condition. These parameters co-varied with soil moisture content under the stress condition (Chl: R2= 0.91, Cw: R2= 0.97, Cs: R2= 0.88 and LAI: R2=0.48) at the canopy level. To validate the results, the relationship between vegetation parameters that were measured in the laboratory and soil moisture content was established. The results were totally in agreement with the modeling outputs and confirmed the results produced by radiative transfer model inversion and spectroscopy. Since water stress changes all parts of the spectrum, we concluded that analysis of the reflectance spectrum in the VIS-NIR-MIR region is a promising tool for monitoring water stress impacts on vegetation.

Keywords: hyperspectral remote sensing, model inversion, vegetation responses, water stress

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450 Systematics of Water Lilies (Genus Nymphaea L.) Using 18S rDNA Sequences

Authors: M. Nakkuntod, S. Srinarang, K.W. Hilu

Abstract:

Water lily (Nymphaea L.) is the largest genus of Nymphaeaceae. This family is composed of six genera (Nuphar, Ondinea, Euryale, Victoria, Barclaya, Nymphaea). Its members are nearly worldwide in tropical and temperate regions. The classification of some species in Nymphaea is ambiguous due to high variation in leaf and flower parts such as leaf margin, stamen appendage. Therefore, the phylogenetic relationships based on 18S rDNA were constructed to delimit this genus. DNAs of 52 specimens belonging to water lily family were extracted using modified conventional method containing cetyltrimethyl ammonium bromide (CTAB). The results showed that the amplified fragment is about 1600 base pairs in size. After analysis, the aligned sequences presented 9.36% for variable characters comprising 2.66% of parsimonious informative sites and 6.70% of singleton sites. Moreover, there are 6 regions of 1-2 base(s) for insertion/deletion. The phylogenetic trees based on maximum parsimony and maximum likelihood with high bootstrap support indicated that genus Nymphaea was a paraphyletic group because of Ondinea, Victoria and Euryale disruption. Within genus Nymphaea, subgenus Nymphaea is a basal lineage group which cooperated with Euryale and Victoria. The other four subgenera, namely Lotos, Hydrocallis, Brachyceras and Anecphya were included the same large clade which Ondinea was placed within Anecphya clade due to geographical sharing.

Keywords: nrDNA, phylogeny, taxonomy, waterlily

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449 Mass Rearing and Effects of Gamma Irradiation on the Pupal Mortality and Reproduction of Citrus Leaf Miner Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae)

Authors: Shiva Osouli, Maryam Atapour, Mehrdad Ahmadi, Shima Shokri

Abstract:

Citrus leaf miner (Phyllocnistis citrella Stainton) is native to Asia and one of the most serious pests of Iran’s citrus nursery stocks. In the present study, the possibility of insect mass rearing on four various citrus hosts and the effects of gamma irradiation on the pupal mortality and reproduction of this pest were studied. Trifoliate orange and grapefruit showed less infection, while the number of pupae in Valencia oranges and sweet lemons cages was so high. There was not any significant difference between weight of male and female pupae among different citrus hosts, but generally the weight of male pupae was less than females. Use of Valencia orange or sweet lemons seedlings in especial dark emergence and oviposition cages could be recommended for mass rearing of this pest. In this study, the effects of gamma radiation at doses 100 to 450 Gy on biological and reproductive parameters of the pest has been determined. The results show that mean percent of pupal mortality increased with increasing doses and reached to 28.67% at 450 Gy for male pupae and 38.367% for female pupae. Also, the mean values of this parameter were higher for irradiated female, which indicated the higher sensitivity of this sex. The gamma ray irradiation from 200 and 300 Gy caused decrease in male and female adult moth longevity, respectively. The eggs were laid by emerged females, and their hatchability was decreased by increasing gamma doses. The fecundity of females in both combinations of crosses (irradiated male × normal female and irradiated female × normal male) did not differ, but fertility of laid eggs by irradiated female × normal male affected seriously and the mean values of this parameter reached to zero at 300 Gy. The hatchability percentage of produced eggs by normal female × irradiated male at 300 Gy was 23.29% and reached to less than 2 % at 450 Gy as the highest tested dose. The results of this test show that females have more radio-sensitivity in comparison to males.

Keywords: citrus leaf miner, Phyllocnistis citrella, citrus hosts, mass rearing, Sterile Insect Technique (SIT)

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448 Determination of Genotypic Relationship among 12 Sugarcane (Saccharum officinarum) Varieties

Authors: Faith Eweluegim Enahoro-Ofagbe, Alika Eke Joseph

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Information on genetic variation within a population is crucial for utilizing heterozygosity for breeding programs that aim to improve crop species. The study was conducted to ascertain the genotypic similarities among twelve sugarcane (Saccharum officinarum) varieties to group them for purposes of hybridizations for cane yield improvement. The experiment was conducted at the University of Benin, Faculty of Agriculture Teaching and Research Farm, Benin City. Twelve sugarcane varieties obtained from National Cereals Research Institute, Badeggi, Niger State, Nigeria, were planted in three replications in a randomized complete block design. Each variety was planted on a five-row plot of 5.0 m in length. Data were collected on 12 agronomic traits, including; the number of millable cane, cane girth, internode length, number of male and female flowers (fuss), days to flag leaf, days to flowering, brix%, cane yield, and others. There were significant differences, according to the findings among the twelve genotypes for the number of days to flag leaf, number of male and female flowers (fuss), and cane yield. The relationship between the twelve sugarcane varieties was expressed using hierarchical cluster analysis. The twelve genotypes were grouped into three major clusters based on hierarchical classification. Cluster I had five genotypes, cluster II had four, and cluster III had three. Cluster III was dominated by varieties characterized by higher cane yield, number of leaves, internode length, brix%, number of millable stalks, stalk/stool, cane girth, and cane length. Cluster II contained genotypes with early maturity characteristics, such as early flowering, early flag leaf development, growth rate, and the number of female and male flowers (fuss). The maximum inter-cluster distance between clusters III and I indicated higher genetic diversity between the two groups. Hybridization between the two groups could result in transgressive recombinants for agronomically important traits.

Keywords: sugarcane, Saccharum officinarum, genotype, cluster analysis, principal components analysis

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447 Heterothic Effect of Some Quantitative Traits in F1 Diallel Hybrids of Various Tobacco Types

Authors: Jane Aleksoski

Abstract:

The mode of inheritance and heterotic effect were studied in ten F1 crosses obtained by one-way diallel crossing between five parental genotypes: MV-1, P 76/86, Adiyaman, Basma-Djebel, and P 66 9 7. The following quantitative traits were studied: the number of leaves per stalk, length of leaves from the middle belt of the stalk, and yield of green leaf mass per stalk and per hectare. The trial was set up in the experimental field of Scientific Tobacco Institute - Prilep, using a randomized block design with four replications in the period 2018-2019. Traditional cultural practices were applied during the growing season of tobacco in the field. The aim of this work was to study the mode of inheritance of the quantitative traits, to detect heterosis in the F1 generation, and to assess its economic viability. Analysis of variance determined statistically significant differences in traits between parents and their hybrids in the two-year investigation. The most common way of trait inheritance is partial-dominant, then intermediate. The negative heterotic effect on the number of leaves per stalk has P 76/86 x P 66 9 7. The hybrids MV-1 x Adiyaman, P 76/86 x Basma-Djebel, P 76/86 x P 66 9 7, and Basma-Djebel x P 66 9 7 have a positive heterotic effect on the length of the leaves. Oriental hybrids, where one of the parents is variety P 66 9 7, have positive heterosis in the yield of green leaf mass per stalk. The investigation provides very useful guidance for future successive selection activities.

Keywords: dominance, heterosis, inheritance, tobacco.

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446 Nutritional Value and Leaf Disease Resistance of Different Varieties of Wheat

Authors: Danutė Jablonskytė-Raščė, Vidas Damanauskas

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The wheat (Triticum) genus is divided into many species, of which only two are widely distributed in the world - common wheat (Triticum aestivum L.) and durum wheat (Triticum durum Desf.). Common (soft) wheat is the most common type of wheat in the world and the most suitable for the harsh climate of Lithuania, but the grains have lower protein content and poorer nutritional properties. Durum wheat is characterized by a high protein content of the grain, but it is a crop of warmer climates grown in southern countries, Italy, Spain, the United States, Egypt, etc. Today's important issue is food, its resources and quality. The research focuses on healthier food grown in our conditions, the quality of which recently depends a lot not only on the cultivation technology but also on the warming climate conditions. Climatic conditions change the distribution of fungi and their hosts. Plants that have grown in our climate for many years have adapted to the use of fungicides, so the aim is to study cereal varieties grown in warmer climates and compare them with our country's varieties, studying their nutritional value and the spread of fungal diseases. The field experiments of different varieties of wheat were conducted at Joniškėlis Experimental Station of the Lithuanian Research Centre for Agriculture and Forestry in 2023. The soil of the experimental site was Endocalcari-Endohypogleyic Cambisol (CMg-n-w-can). The research was designed to identify the resistance to leaf diseases and the nutritional value of various wheat varieties. This research aims to focus on healthier food grown in our conditions, the quality of which recently depends a lot not only on the cultivation technology but also on the conditions of the warming climate. The study found that hot and humid summer weather led to the spread of foliar diseases in wheat. Tan spot (Pyrenophora tritici-repentis) is mostly spread in wheat crops. This disease had an average prevalence of 86.90%. The wheat crop was sparse, so this year was unfavorable for the spread of powdery mildew (Blumeria graminis). Dry weather prevailed during the period of flowering of cereals, which prevented the spread of ear diseases. Examining the qualitative indicators of grain, it was found that durum wheat had the best parameters.

Keywords: varieties, wheat, leaf disease, grain quality

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445 Plant Regeneration via Somatic Embryogenesis and Agrobacterium-Mediated Transformation in Alfalfa (Medicago sativa L.)

Authors: Sarwan Dhir, Suma Basak, Dipika Parajulee

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Alfalfa is renowned for its nutritional and biopharmaceutical value as a perennial forage legume. However, establishing a rapid plant regeneration protocol using somatic embryogenesis and efficient transformation frequency are the crucial prerequisites for gene editing in alfalfa. This study was undertaken to establish and improve the protocol for somatic embryogenesis and subsequent plant regeneration. The experiments were conducted in response to natural sensitivity using various antibiotics such as cefotaxime, carbenicillin, gentamycin, hygromycin, and kanamycin. Using 3-week-old leaf tissue, somatic embryogenesis was initiated on Gamborg’s B5 basal (B5H) medium supplemented with 3% maltose, 0.9µM Kinetin, and 4.5µM 2,4-D. Embryogenic callus (EC) obtained from the B5H medium exhibited a high rate of somatic embryo formation (97.9%) after 3 weeks when the cultures were placed in the dark. Different developmental stages of somatic embryos and cotyledonary stages were then transferred to Murashige and Skoog’s (MS) basal medium under light, resulting in a 94% regeneration rate of plantlets. Our results indicate that leaf segments can grow (tolerate) up to 450 mg/L of cefotaxime and 400 mg/L of carbenicillin in the culture medium. However, the survival threshold for hygromycin at 12.5 mg/L, kanamycin at 250 mg/L, gentamycin at 50 mg/L, and timentin (300 mg/L). The experiment to improve the protocol for achieving efficient transient gene expression in alfalfa through genetic transformation with the Agrobacterium tumefaciens pCAMBIA1304 vector was also conducted. The vector contains two reporter genes such as β-glucuronidase (GUS) and green fluorescent protein (GFP), along with a selectable hygromycin B phosphotransferase gene (HPT), all driven under the CaMV 35s promoter. Various transformation parameters were optimized using 3-week-old in vitro-grown plantlets. The different parameters such as types of explant, leaf ages, preculture days, segment sizes, wounding types, bacterial concentrations, infection periods, co-cultivation periods, different concentrations of acetosyringone, silver nitrate, and calcium chloride were optimized for transient gene expression. The transient gene expression was confirmed via histochemical GUS and GFP visualization under fluorescent microscopy. The data were analyzed based on the semi-quantitative observation of the percentage and number of blue GUS spots on different days of agro-infection. The highest percentage of GUS positivity (76.2%) was observed in 3-week-old leaf segments wounded using a scalpel blade of 11 size- after 3 days of post-incubation at a bacterial concentration of 0.6, with 2 days of preculture, 30 min of bacterial-leaf segment co-cultivation, with the addition of 150 µM acetosyringone, 4 mM calcium chloride, and 75 µM silver nitrate. Our results suggest that various factors influence T-DNA delivery in the Agrobacterium-mediated transformation of alfalfa. The stable gene expression in the putative transgenic tissue was confirmed using PCR amplification of both marker genes, indicating that gene expression in explants was not solely due to Agrobacterium, but also from transformed cells. The improved protocol could be used for generating transgenic alfalfa plants using genome editing techniques such as CRISPR/Cas9.

Keywords: Medicago sativa l. (Alfalfa), agrobacterium tumefaciens, β-glucuronidase, green fluorescent protein, transient gene

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