Amaryllidaceae Research Articles

Chemical constituents from the arial of Crinum asiaticum L.

Crinum asiaticum L. (locally known Nang hoa trang), belonging to the Crinum genus of the Amaryllidaceae family, has been widely used in oriental and traditional medicine around the world for the treatment of various diseases and exerts diverse promising pharmacological effects. To find out new inhibitors for the treatment of inflammatory and Alzheimer’s diseases, a methanol extract of the aerial part of C. asiaticum was investigated to afford six compounds, chrysophanol (1), emodin (2), physcion (3), thymine (4), thymidine (5), and 5-(hydroxymethyl)-2-furfural (6) by using various chromatographic separations. Their structures were identified by the extensive analysis of 1H- and 13C-NMR data as well as a comparison with those of previously reported data. This is the first report of these compounds (1-6) from C. asiaticum. In addition, all compounds evaluated the inhibitory activities of NO production and acetylcholinesterase (AChE). Among them, emodin (2) significantly inhibited NO production with an IC50 of 5.13±0.42 μM. This result indicated that emodin (2) may be beneficial in the treatment of anti-inflammatory diseases. Besides, all compounds (1-6) also showed weak inhibitory effects on the AChE enzyme with a range of IC50 values from 82.98±9.19 to 237.63±13.80 µM.

Identification of Garlic Genotypes for Resistant Sources against Purple Blotch Disease

Garlic is an herbaceous annual bulbous plant in the family Amaryllidaceae grown for its pungent and edible bulb. Among the various biotic factors hindering production and productivity, purple blotch plays a predominant role in changing the crops morpho-physiological characters, thus deteriorating the metabolic activity in turn affecting the yield considerably. Fungicides have been recommended for the control of the disease, however use of fungicides is expensive and not eco-friendly. The best approach to control the disease is by using resistant genotypes. Hence, 37 genotypes were field screened at AINRPOG (All India Network Research Project on Onion and Garlic), MARS (Main Agricultural Research Station), Dharwad against purple blotch disease under artificial epiphytotic conditions during Kharif in year 2019 to identify the resistant sources. The results revealed that twenty genotypes were found to be resistant (1-10%) and five genotypes were immune (0%) however, ten genotypes were found moderately resistant (11-20%) in reaction. The remaining two genotypes viz., HRG Local and DOGR409 were found moderately susceptible (21-40%) with a maximum grade of 3 on (0-5) scale. None of the genotypes showed susceptible or highly susceptible reactions to purple blotch. Breeders can make use of these identified resistant lines in developing high yield purple blotch disease resistant varieties.

Screening of Anti-carcinogenic Properties of Phytocompounds from Allium ascalonicum for Treating Breast Cancer Through In Silico and In Vitro Approaches.

Plants, rich in phytocompounds, have been in usage since time immemorial for treating various diseases, namely, cancer. One such plant species, Allium ascalonicum(Shallot) belonging to Amaryllidaceae family is being studied here for its anti-carcinogenic properties against breast cancer. GC-MS characterization of A. ascalonicum exhibited 48 phytocompounds containing five peak phytocompounds and 13 phytocompounds with anti-carcinogenic properties. These 13 anti-carcinogenic phytocompounds were docked with three hormonal receptors involved in breast cancer malignancy, namely, ERα, PR, and human EGFR with tamoxifen as standard for in silico analysis. The results exhibited three phytocompounds that had better binding scores compared to that of the standard drug, tamoxifen. Lyophilized powder of aqueous A. ascalonicum extract, also referred as ASE, was used for in vitro approaches. Antioxidant study using DPPH assay revealed that the highest percentage of FRSA in ASE, nearly 51%, was observed at 50µg/ml concentration. Cytotoxicity study on MCF-7 cell line using MTT assay demonstrated IC50 value at 1400µg/ml and anti-proliferative study using Trypan blue assay for the determination of percentage viability of MCF-7 cells at IC50concentration was observed to be 49%. Anti-mitotic activity using Vigna radiata seed germination assay revealed clear morphological differences in a dose-dependent manner between the seeds grown at various concentrations of ASE with nearly 56.5% growth inhibition observed at 1500µg/ml concentration. Hence, this research work proves that Allium ascalonicumhas very good anti-carcinogenic properties and this can be confirmed further through in vivo animal model studies and it can also be formulated as a promising drug to treat breast cancer. GC-MS characterization of Allium ascalonicum demonstrated the presence of five peak compounds and thirteen anti-carcinogenic compounds. The thirteen anti-carcinogenic compounds were docked with three target proteins (in silico analysis) involved in breast cancer malignancy and identified the presence of three potential phytocompounds that can be used for treating breast cancer. In vitro approaches also confirmed the presence of anti-carcinogenic properties such as antioxidative potential, cytotoxic, anti-proliferative, and anti-mitotic effects. Hence, Allium ascalonicum can be taken further to in vivo studies so that it can be formulated to treat breast cancer.

Screening of Endophytic Bacteria of Leucojum aestivum 'Gravety Giant' as a Potential Source of Alkaloids and as Antagonist to Some Plant Fungal Pathogens.

Leucojum aestivum is a medicinal plant belonging to the Amaryllidaceae family well known as a producer of alkaloids such as galanthamine and lycorine. However, the endophytic microbes that colonize different plant tissues without causing any damage have not been reported in this plant. Here, we explored the different endophytic bacterial communities isolated from different surface disinfected tissues of L. aestivum ‘Gravety giant’ and screened bacterial isolates producing alkaloids and their potential use as biocontrol agent against wheat pathogens. For that purpose, endophytic bacteria were isolated from bulbs, roots and shoots of L. aestivum. After taxonomical characterization, these microorganisms were screened for their ability to produce alkaloids using high-performance thin-layer chromatography (HPTLC) and untargeted liquid chromatography-Mass Spectrometry/Mass Spectrometry (LC-MS/MS) strategies. We isolated 138 bacteria belonging to four phyla and 42 genera, mainly from roots and shoots. The most abundant genera were Rahnella in shoot, Patulibacter in bulb and Bacillus in roots. Among the different bacterial isolates, the methanolic extracts of Luteibacter rhizovicinus (LaBFB3301) and Commamonas denitrificans (LaBFS2103) slightly delayed the growth of F. graminearum colonies in in vitro dual tests against F. graminearum and M. nivale strains with 15.5% and 19.9% inhibition rates, respectively. These isolates are able to produce an indolic alkaloid tryptophol (C10H11NO, [M + H]+ 162.0913). These endophytic bacteria might be investigated to characterize the plant protection effect and the plant growth promotion effect.

Antiviral alkaloid principles of the plant family Amaryllidaceae

BackgroundViral-borne diseases are amongst the oldest diseases known to mankind. They are responsible for some of the most ravaging effects wrought on human health and well-being. The use of plants against these ailments is entrenched in both traditional and secular medicine around the globe. Their natural abundance and chemical diversity have also boosted their appeal in drug discovery. AimThe plant family Amaryllidaceae is distinguished for its alkaloid principles, some of which are of considerable interest in the clinical arena. This account is the outcome of a literature review undertaken to establish the applicability of these substances as antiviral agents. MethodsThe survey utilized the search engines Google Scholar, PubMed, SciFinder, Scopus and Web of Science engaging the word ‘antiviral’ in conjunction with ‘Amaryllidaceae’ and ‘Amaryllidaceae alkaloid’. The search returned over five hundred hits, of which around eighty were of relevance to the theme of the text. ResultsOver eighty isoquinoline alkaloids have been screened against nearly fifty pathogens from fourteen viral families, the majority of which were RNA viruses. Potent activities were reported in some instances, such as that of trans-dihydronarciclasine against Yellow fever virus (IC50 0.003 μg/ml), with minimal effects being manifested on host cells. There were also promising results obtained from in vivo studies, in most cases without lethal effects on test subjects. Structure-activity relationship studies afforded useful insight to the antiviral pharmacophore, with the phenanthridone alkaloid nucleus shown to be the most enabling. Although the mechanistic basis to these activities pertained mostly to inhibition of DNA, RNA and protein synthesis, evidence was also forthcoming about the inhibitory action of some of the alkaloids against viral neuraminidase, protease and reverse transcriptase. In silico methods of analysis have offered further perspectives of how some of the alkaloids interact at the active sites of their targets. ConclusionThe Amaryllidaceae offers a viable platform for plant-based antiviral drug discovery. Its cause is strengthened not only by its wide proliferation and exploitation of its members in alternative forms of medicine, but also by its rich chemical diversity which has already spawned useful antiviral drug leads.

Fungal biotransformations of anticholinesterase norbelladine derivatives to obtain new products and mimic mammalian metabolism

Norbelladine is the natural precursor of all alkaloids of the Amaryllidaceae family. These compounds have been reported to exert interesting biological activities. Particularly, galantamine is currently used in the palliative treatment of Alzheimer’s disease for its cholinergic effect. The biotransformation of two norbelladine analogues exhibiting anticholinesterase activity was studied using 11 fungi belonging to Aspergillus , Rhizopus , Cunninghamella and Fusarium genera. The substrates were refractory to biotransformation by all fungi screened, except by A. clavatus . Based on GC-MS analyses, we demonstrated that its enzymes were able to catalyse the oxidative cleavage of the C-N bond of the secondary amine of 4’- O -methylnorbelladine. On the other hand, the hindered phenolic hydroxy group at the 3’ position of the brominated derivative was regioselectively methylated. The latter compound was chemically synthesised for better characterisation and the biological assays showed that this metabolite exerted a lower inhibition of BChE and lost the anti-AChE activity. The metabolic pathways involved here were useful to mimic phase I and II xenobiotic metabolism in mammals and thus predict the products that could be formed. A new source of biocatalytic tools to obtain new protoalkaloid derivatives was also discovered. • Norbelladine derivatives have shown promising anticholinesterase activities. • Fungal model is useful to predict phase I and II metabolism in mammals. • Norbelladine analogues remained untransformed with most of the fungi tested. • Two norbelladine analogues were metabolised by Aspergillus clavatus . • Biotransformation of the brominated derivative led to loss of anti-AChE activity.

Micro-morpho-anatomical transitions at various stages of in vitro development of Crinum malabaricum Lekhak and Yadav: A critically endangered medicinal plant.

Crinum malabaricum Lekhak & Yadav is a recently discovered and critically endangered aquatic bulbous plant of the family Amaryllidaceae. It gained attention as a wild source of the acetylcholinesterase inhibiting alkaloid 'galanthamine' used to treat Alzheimer and Parkinson diseases. The bulbs of this plant contain the highest amount of galanthamine among Crinum species. In vitro regeneration systems were developed to produce quality uniform plantlets of C. malabaricum. Bright field light microscopy was used to analyse micro-morpho-anatomical developments taking place in the leaves and roots during in vitro, ex vitro and in vivo transitions of plantlets. Leaves and roots of plants raised in vitro possessed a higher degree of microscopic structural anomalies, such as underdeveloped epicuticular wax deposition, immature and non-functional stomata, more aquiferous parenchyma with a reduced lumen. Roots developed in vitro were characterized by extremely large, uneven cortical cells and reduced intercellular spaces. The vascular tissues were under-developed and only primary vascular tissues were observed. As a result of ex vitro acclimation, there was a significant acceleration in the improvement of tissue systems in leaves and roots. Such plantlets can tolerate elevated temperatures and light under in vivo conditions. Thus, the microscopic evaluation of the structural trajectory in different stages of plantlet development provides an understanding of the acclimation process and structural adaptations, which could help enhance survival of in vitro raised plantlets under ex vitro and in vivo conditions.

Beneficiation of organic black onion and lemon peel by-products through extraction and investigation of bioactive compounds

Much research on the shallot phytochemical is currently employed in various disciplines, such as medical reasons. Shallot (Allium ascalonicum) is a plant species belonging to the family Amaryllidaceae. It is a native plant grown in the North and Northeast region of Thailand. The present study aimed to determine the extracts' yield and antioxidant properties of black shallots made from fresh shallots. The black shallots were extracted with digestion by water, 47.5% and 95% ethanol at 50 ℃ for 1, 3 and 5 hours. The result discovered that black shallot extraction with digestion by water for 5 hours showed the highest weight of extracts (3.620 g). The determination of antioxidant activity by DPPH was observed, and extraction by water for 5 hours gave the highest percentage of inhibition. It was equal to 84.61%, which is not different from the digestion of 95% ethanol for 1 hour, with 83.48%. Similarly, the most outstanding value of IC50 was obtained from extraction with 95% ethanol for 1 and 5 hours with 21.30 mg/ml and 20.52 mg/ml. In terms of monitoring, the digestion method evaluated the productivity of pectin extracts from citrus peel. Hydrochloric acid (0.100, 0.050 and 0.025 M) was used as a solvent at 95 ℃ for 30, 60 and 90 minutes. The pectin extract by 0.1 M hydrochloric acid for 90 minutes gave the highest extract (5.237 g). The preliminary testing of cosmetic production was done by mixing black shallot extracts and pectin from citrus peels, and the two substances can be combined. Still, without adding preservatives, they could cause contamination in cosmetic samples.

Lycorine inhibits angiogenesis by docking to PDGFRα

Lycorine (Lyc) is a natural alkaloid derived from medicinal plants of the Amaryllidaceae family. Lyc has been reported to inhibit the recurrence and metastasis of different kinds of tumors. However, Lyc’s effect on angiogenesis and its specific mechanism are still not clear. This study was designed to test the antiangiogenesis effect of Lyc and to explore the possible mechanisms. We performed cell experiments to confirm Lyc’s inhibitory effect on angiogenesis and employed sunitinib as a positive control. Moreover, the synergistic effect of Lyc and sunitinib was also explored. Next, we conducted bioinformatics analyses to predict the potential targets of Lyc and verified them by western blotting and immunofluorescence. Molecular docking, kinase activity assays, Biacore assays and cellular thermal shift assays (CETSAs) were applied to elucidate the mechanism by which Lyc inhibited target activity. Lyc inhibited angiogenesis in human umbilical vein endothelial cells (HUVECs). Employing bioinformatics, we found that Lyc’s target was PDGFRα and that Lyc attenuated PDGFRα phosphorylation. We also found that Lyc inhibited PDGFRα activation by docking to it to restrain its activity. Additionally, Lyc significantly inhibited PDGF-AA-induced angiogenesis. This study provides new insights into the molecular functions of Lyc and indicates its potential as a therapeutic agent for tumor angiogenesis.

Complete Genomic RNA Sequence of Tuberose Mild Mosaic Virus and Tuberose Mild Mottle Virus Acquired by High-Throughput Sequencing.

Tuberose (Polianthes tuberosa) is an ornamental flowering crop of the Amaryllidaceae family. Tuberose mild mosaic virus (TuMMV) and tuberose mild mottle virus (TuMMoV), members of the genus Potyvirus, are ubiquitously distributed in most tuberose growing countries worldwide with low biological incidence. Here, we report the first coding-complete genomic RNA of TuMMV and TuMMoV obtained through high-throughput sequencing (HTS) and further, the presence of both the viruses were confirmed using virus-specific primers in RT-PCR assays. Excluding the poly (A) tail, the coding-complete genomic RNA of TuMMV and TuMMoV was 9485 and 9462 nucleotides (nts) in length, respectively, and contained a single large open reading frame (ORF). Polyprotein encoded by both the viral genomes contained nine putative cleavage sites. BLASTn analysis of TuMMV and TuMMoV genomes showed 72.40–76.80% and 67.95–77% nucleotide sequence similarities, respectively, with the existing potyviral sequences. Phylogenetic analysis based on genome sequences showed that TuMMV and TuMMoV clustered in a distinct clade to other potyviruses. Further studies are required to understand the mechanism of symptom development, distribution, genetic variability, and their possible threat to tuberose production in India.

Alkaloid Composition and Biological Activities of the Amaryllidaceae Species Ismene amancaes (Ker Gawl.) Herb.

Natural products have always played a significant role in the search for new drugs. One of the most relevant alkaloid-containing plant groups is the Amaryllidaceae family, a source of exclusive structures with a wide variety of pharmacological activities. The aim of this work was to determine the alkaloid composition and biological potential of an extract from the bulbs of an endemic Peruvian Amaryllidaceae species Ismene amancaes (Ker Gawl.) Herb. The alkaloid profiling was carried out by GC-MS, which revealed the presence of 13 compounds, 2 of them unidentified. The plant extract was found to contain high amounts of lycoramine, a galanthamine-type alkaloid. The extract also presented low inhibitory potential against the enzymes AChE and BuChE, with IC50 values of 14.6 ± 0.6 and 37.6 ± 1.4 μg·mL−1, respectively, and good to moderate inhibitory activity against the protozoan Plasmodium falciparum strain FCR-3 (chloroquine-resistant), with IC50 values of 3.78 ± 0.3 μg·mL−1. This is the first report of the alkaloid profile of a plant of the Ismene genus, which could be an interesting source of bioactive compounds.

SUGAR AND PROTEIN CONTENT, HYDROGEN PEROXIDE RADICAL SCAVENGING AND TOTAL ANTIOXIDANT CAPACITY OF Polianthes tuberosa L FLOWERS GROWN IN ALGERIA

Tuberose flowers (Polianthes tuberosa L.) belong to the Amaryllidaceae family and they are widely enhanced through their absolute and essential oils, especially in the perfume industry. However, the sugar and proteins of tuberose are still unknown. For this reason, this work aimed to quantify them, particularly in flowers, besides to the evaluation of their hydrogen peroxide radical scavenging and total antioxidant capacity. The obtained results revealed 79.51±0.08 mg glucose equivalent/g extract of sugar and 0.097±0.001 mg bovine serum albumin equivalent/g extract of proteins. These metabolites were quantified using phenol-sulfuric acid and Coomassie Blue methods, respectively. Furthermore, the antioxidant activity of flower extract was investigated using total antioxidant capacity (TAC) and hydrogen peroxide radical scavenging. The TAC assay indicated the inhibitory concentration of 50% of free radicals (IC50) was 0,09±0,01 mg/mL of flower extract, however, the scavenging ability against H2O2 showed the IC50 of 7,09±0,15 mg/mL of tuberose flower extract. This work contributes to the knowledge of tuberose flowers content in proteins and sugar, in addition to the total antioxidant and the H2O2 scavenging capacities.

Perilaku Mencari Polen dan Identifikasi Polen Tetragonula laeviceps pada dua Kebun Tanaman Obat

Unit Konservasi Budidaya Biofarmaka (UKBB) and Kampoeng Djamoe Organik Martha Tilaar (KaDO) are two medicinal plantation that possibly as resources for stingless bees nectar and pollen. This study was aimed to describe the foraging behavior of Tetragonula laeviceps in medicinal plantations, determine the plant species which become the pollen resources, and the distance between the colony and medicinal plants. Observation on foraging behavior were divided into three activities, i.e. flying out from the nest, returning the nest with pollen, and returning the nest without pollen. We found a similar pattern of numbers of bees returning to the nest with pollen in both UKBB and KaDO that occurred in the morning with the highest between 9 and 10 am and gradually decreased in the afternoon. The bees in UKBB showed highly number flying out from the nest in around 3 pm which might be the activites of resin collection. We identified 11 and 16 pollen types at the UKBB and the KaDO, respectively. The pollen in both locations are from the families Amaryllidaceae, Anacardiaceae, Clusiaceae, Cucurbitaceae, Ebenaceae, and Sapindaceae. Our results revealed that the distance range of medicinal plants from the bees hives was 13.4-113,3 m. Thus, this data indicate that the hives should be located in the distance of the bee forage. The medicinal pollen identified in the T. laeviceps pollen pot confirmed the role of the bees as the ecosystem service in the medicinal plantations.
 
 Keywords: foraging range, medicinal plants, pollination, pollen, Tetragonula

Behavioral disruption of two orchard hemipteran pests by garlic essential oil

AbstractEssential oils (EOs) are promising sources of effective bioactive compounds for insect pest control, especially in heavily treated agrosystems like orchards. Plants belonging to the Amaryllidaceae family – onion, garlic (Allium sativum L.), and others – present an interesting potential in apple (Malus domestica Borkh.) and pear (Pyrus communis L., both Rosaceae) tree integrated pest management due to the repellent and deterrent properties of their chemical compounds. This study explores the potential disruption by garlic EOs of the behavior of two hemipteran pests towards their host trees: rosy apple aphid, Dysaphis plantaginea (Passerini) (Aphididae), on cultivated apple, and pear psyllid, Cacopsylla pyri (L.) (Psyllidae), on pear. Effects on orientation and feeding behavior were evaluated using choice tests and the electropenetrography (EPG) technique. The results revealed that garlic EOs delivered through rubber dispensers were intrinsically repellent, but when combined with the attractive host plant, the repellents and attractants canceled each other out for the two studied hemipterans. Garlic EOs caused disruptions in the feeding behavior of the two insects, particularly the psyllids, which showed a decrease in phloem sap intake. This study highlights the mechanisms disrupting the process of host‐plant colonization, in particular the possible masking of host‐locating cues or the involvement of confusing chemical stimuli due to EO volatiles.

Carbon dioxide expanded liquid: an effective solvent for the extraction of quercetin from South African medicinal plants

BackgroundQuercetin is one of the most important bioflavonoids having positive effects on the biological processes and human health. Typically, it is extracted from plant matrices using conventional methods such as maceration, sonication, infusion, and Soxhlet extraction with high solvent consumption. Our study aimed to optimize the environmentally friendly carbon dioxide-based method for the extraction of quercetin from quince fruit with an emphasis on extraction yield, repeatability, and short extraction time.ResultsA two-step design of experiments was used for the optimization of the key parameters affecting physicochemical properties, including CO2/co-solvent ratio, co-solvent type, temperature, and pressure. Finally, gas expanded liquid combining CO2/ethanol/H2O in a ratio of 10/81/9 (v/v/v) provided the best extraction yield. Extraction temperature 66 °C and pressure 22.3 MPa were the most suitable conditions after careful optimization, although both parameters did not significantly affect the process. It was confirmed by experiments in various pressure and temperature conditions and statistical comparison of obtained data. The optimized extraction procedure at a flow rate of 3 mL/min took 30 min. The repeatability of the extraction method exhibited an RSD of 20.8%.ConclusionsThe optimized procedure enabled very fast extraction in 30 min using environmentally friendly solvents and it was successfully applied to 16 different plant samples, including 14 bulbs and 2 fruits from South Africa. The quercetin content in extracts was quantified using ultra-high performance liquid chromatography (UHPLC) with tandem mass spectrometry. UHPLC hyphenated with high-resolution mass spectrometry was used to confirm chemical identity of quercetin in the analyzed samples. We quantified quercetin in 11 samples of all 16 tested plants. The quercetin was found in Agapanthus praecox from the Amaryllidaceae family and its presence in this specie was reported for the first time.Graphical

Investigations on the Biological Activity of Allium sativum Agglutinin (ASA) Isolated from Garlic.

Garlic (Allium sativum) from the family Amaryllidaceae is widely used in culinary and is reported to have potential anticancer, anti-diabetic, antimicrobial, and cardioprotective activities. Allium sativum agglutinin (ASA) is a bulb-type lectin (BTL) domaincontaining lectin isolated from garlic and has been studied for its various biological functions. Previous studies have reported the anti-cancer effects of ASA on histiocytic lymphoma (U937), promyelocytic leukemia (HL60), and oral cancer (KB). In this study, we have purified and characterized ASA and evaluated it for its anticancer effects on other cancer cell lines. MTT assay and FACS analysis was done to corroborate the anticancer findings against cervical (HeLa) and lung cancer (A549) cell lines. IC50 value of 37 μg/ml in HeLa and a weak activity (26.4 ± 1.9% cellular inhibition at 100μg/ml treatment) in A549 were found in the MTT assay. FACS analysis further corroborated these findings and showed the apoptotic effects of ASA in these cell lines. Anticancer activity for members of bulb-type lectin (BTL) domain-containing lectins has been widely reported, and we hope that our study forms a basis for the development of ASA as a therapeutic agent.

Lycorine Alkaloid and Crinum americanum L. (Amaryllidaceae) Extracts Display Antifungal Activity on Clinically Relevant Candida Species

Candida species are the main fungal agents causing infectious conditions in hospital patients. The development of new drugs with antifungal potential, increased efficacy, and reduced toxicity is essential to face the challenge of fungal resistance to standard treatments. The aim of this study is to evaluate the in vitro antifungal effects of two crude extracts of Crinum americanum L., a rich alkaloid fraction and lycorine alkaloid, on the Candida species. As such, we used a disk diffusion susceptibility test, determined the minimum inhibitory concentration (MIC), and characterized the components of the extracts using Electrospray Ionization Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (ESI FT-ICR MS). The extracts were found to have antifungal activity against various Candida species. The chemical characterization of the extracts indicated the presence of alkaloids such as lycorine and crinine. The Amaryllidaceae family has a promising antifungal potential. Furthermore, it was found that the alkaloid lycorine directly contributes to the effects that were observed for the extracts and fraction of C. americanum.

Effect of Different Abiotic Stresses on Allium Cepa Plants: Results from a Field Study of Two Different Seasons

Abiotic factors have the capacity to influence the nature and quantity of secondary metabolites. Alteration of environmental factors during plant growth can affect the production of bioactive phytoconstituents. A six-month field study was carried out in two different seasons to assess the effect of different abiotic stresses on bioactive compounds, i.e. flavonoids and phenols and plant yield of selected Allium species, i.e. Allium cepa L. (AC). (Family Amaryllidaceae). AC plants were grown for six months from November to April and for another six months from May to October on three different soil types (i.e. black, red, clay-loamy) and various stresses (salinity, water deprivation, flooding, fertilizer, metal, shade) were applied on the plants. At the end of the season, plants were collected, dried, and plant yield was determined. Hydroalcoholic extracts of all samples were prepared. Plant yield, TFC, TPC were determined in all the extracts. The results show varied response of AC plants to different soil types, stresses, and seasons. AC plants cultivated on red soil with fertilization give higher biomass yield in both seasons. AC plants grown in black soil under metal stress have highest TFC, TPC. Hence these conditions may be recommended for incorporation in cultivation practices of these valuable medicinal plants to ensure commercial supply of plants with higher phenol and flavonoid content and better plant yield.

Complete genome organization and characterization of Hippeastrum latent virus.

The complete genome sequences of two carlaviruses were determined by high-throughput sequencing of RNA extracted from ringspot and mosaic, disease symptoms on leaves of spider lily plants (Crinum asiaticum, family Amaryllidaceae) growing as landscape plants in Hawaii. One, named Nerine latent virus (NeLV)-Hawaii with a genome of 8281 nucleotide exhibited the highest nucleotide identity and amino acid similarity of 95.5% and 96.0%, respectively, to the genome sequence of an isolate of NeLV from Narcissus sp. in Australia (JQ395044). The second, named Hippeastrum latent virus (HiLV)-Hawaii with a genome of 8497 nucleotides exhibited the highest nucleotide identity and amino acid similarity, 84.3% and 88.7%, respectively, to the sequence of a previously uncharacterized HiLV isolate from a potted flowering plant, Amaryllis (Hippeastrum hybridum Hort) in Taiwan (DQ098905). The amino acid sequence similarities of replicase (Rep) and coat protein (CP) between HiLV-Hawaii and NeLV-Hawaii were 44.8% and 38.4%, respectively. Results of viral protein Rep and CP amino acid sequence comparisons from various carlaviruses provide evidence that HiLV and NeLV, previously classified as synonymous viruses are in fact unique viruses. This is thefirst report for the complete sequence, organization, and phylogenetic characterization of HiLV and the first detection of HiLV both in C. asiaticum and in the USA.

First report of Colletotrichum siamense Causing Anthracnose on Crinum asiaticum in Malaysia.

Crinum asiaticum (family Amaryllidaceae), locally known as 'Pokok Bakung', is an ornamental medicinal plant grown in Malaysia. It contains chemical compounds used for antimicrobial, antioxidant, antitumor, antiemetic and wound healing (Patel, 2017). In July 2021, 'Pokok Bakung' leaves with anthracnose symptoms were collected from a park of Universiti Malaysia Sabah in the Sabah province. The disease severity was about 100% with 20% incidence. Red spots were primarily found on the leaf surfaces. Anthracnose developed as the disease progressed, and acervuli were observed in the spots. Small pieces of infected leaves (5 x 5 mm) were excised from spot margins, surface sterilized based on Khoo et al. (2022a), placed on potato dextrose agar (PDA) in Petri dishes, which were incubated for 5 days at 25°C in the dark. The colonies formed on the PDA plates were abundant with gray-white fluffy mycelia after 5 days, and the reverse view revealed brown. UMS01, a representative isolate, was used to morphologically and molecularly characterize the fungus. Conidia were one-celled, cylindrical, hyaline, smooth, and blunt at the ends, measuring 13.8 to 16.5 x 3.6 to 6.7 µm (n = 20). Appressoria ranged in size from 7.6 to 9.3 x 5.5 to 6.9 µm (n= 20) and were ovoid to clavate, spherical to irregular in shape and dark brown in color. Genomic DNA was extracted from fresh mycelia of isolate UMS01 based on Khoo et al. (2021) with the addition of mechanical disruption using a micro pestle before heating at 95°C. PCR amplification was performed based on Khoo et al. (2022a) using ITS1/ITS4, CL1C/CL2C, ACT-512F/ACT-783R, CHS-79F/CHS-354R, and GDF1/GDR1 primer pairs to amplify the internal transcribed spacer (ITS) region, calmodulin (CAL), actin (ACT), chitin synthase (CHS-1), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Weir et al. 2012). PCR products with positive amplicons were sent to Apical Scientific Sdn. Bhd. for sequencing. The sequences were deposited in GenBank under accession numbers OK458683 (ITS), OL953033 (CAL), OL953030 (ACT), OL953036 (CHS-1), and OL953039 (GAPDH). Before BLAST, the search set were adjusted to exclude model sequences (XM/XP) and the uncultured/environmental sample sequences, and limit to sequences from type material. They were 99-100% similar to the Colletotrichum siamense ITS (JX010171), CAL (JX009714), ACT (FJ907423) and CHS-1 (JX009865), and Colletotrichum changpingense GAPDH (MZ664048) type sequences. The GAPDH marker did not reliably resolve the relationships within the C. gloeosporioides complex (Vieira et al. 2020). Phylogenetic analysis using maximum likelihood based on the combined ITS, CAL, ACT, CHS-1 and GAPDH indicated that the isolate formed a supported clade (100% bootstrap value) to the most related C. siamense. Morphological and molecular characterization matched the description of C. siamense (Huang et al. 2021). Pathogenicity tests were performed to fulfil Koch's postulates by spraying a spore suspension (106 spores/ml) on the leaves of three healthy four-month-old 'Pokok Bakung' plants, while three additional plants were sprayed with water as a control. The inoculated plants were covered with plastics for 48 h at 25°C in the dark. Incubation was performed based on Iftikhar et al. (2022). Symptoms similar to those of the field collection occurred after 6 days post inoculation. No symptoms occurred on the control plants. The experiment was repeated two more times. The reisolated fungal isolates were identical to C. siamense morphologically and molecularly. Previously, C. siamense has been reported to cause anthracnose on Allamanda cathartica (Huang et al. 2021) and avocado (Li et al. 2022) in China, and 'Purple Dream' eggplant in Malaysia (Khoo et al. 2022b). Colletotrichum fructicola has been reported to cause anthracnose on C. asiaticum in China (Qing et al. 2020). To our knowledge, this is the first report of C. siamense causing anthracnose on C. asiaticum in Malaysia. Our findings expand the geographic range of C. siamense and indicate that it could be a potential threat limiting the growth and production of C. asiaticum in Malaysia.