Variation in Nutritional Components and Antioxidant Capacity of Different Cultivars and Organs of Basella alba.

Basella alba is a frequently consumed leafy vegetable. However, research on its nutritional components is limited. This study aimed to explore the variation in the nutritional components and antioxidant capacity of different cultivars and organs of Basella alba. Here, we primarily chose classical spectrophotometry and high-performance liquid chromatography (HPLC) to characterize the variation in nutritional components and antioxidant capacity among different organs (inflorescences, green fruits, black fruits, leaves, and stems) of eight typical cultivars of Basella alba. The determination indices (and methods) included the total soluble sugar (anthrone colorimetry), total soluble protein (the Bradford method), total chlorophyll (the ethanol-extracting method), total carotenoids (the ethanol-extracting method), total ascorbic acid (the HPLC method), total proanthocyanidins (the p-dimethylaminocinnamaldehyde method), total flavonoids (AlCl3 colorimetry), total phenolics (the Folin method), and antioxidant capacity (the FRAP and ABTS methods). The results indicated that M5 and M6 exhibited advantages in their nutrient contents and antioxidant capacities. Additionally, the inflorescences demonstrated the highest total ascorbic acid and total phenolic contents, while the green and black fruits exhibited relatively high levels of total proanthocyanidins and antioxidant capacity. In a comparison between the green and black fruits, the green fruits showed higher levels of total chlorophyll (0.77-1.85 mg g-1 DW), total proanthocyanidins (0.62-2.34 mg g-1 DW), total phenolics (15.28-27.35 mg g-1 DW), and ABTS (43.39-59.16%), while the black fruits exhibited higher levels of total soluble protein (65.45-89.48 mg g-1 DW) and total soluble sugar (56.40-207.62 mg g-1 DW) in most cultivars. Chlorophyll, carotenoids, and flavonoids were predominantly found in the leaves of most cultivars, whereas the total soluble sugar contents were highest in the stems of most cultivars. Overall, our findings underscore the significant influence of the cultivars on the nutritional composition of Basella alba. Moreover, we observed notable variations in the nutrient contents among the different organs of the eight cultivars, and proanthocyanidins may contribute significantly to the antioxidant activity of the fruits. On the whole, this study provides a theoretical basis for the genetic breeding of Basella alba and dietary nutrition and serves as a reference for the comprehensive utilization of this vegetable.

Identification and reactivity of pigments in prominent vegetable leaves of Basella alba L. var. 'Rubra' (Malabar spinach).

The unique profiles of betacyanins as well as their stability and antioxidant activity in purple leaf extracts of the fast-growing, soft-stemmed vine Basella alba L. var. 'Rubra', known as Malabar spinach, are partly characterized for the first time. The distribution of gomphrenin and its acylated derivatives in the leaves is completely different from the profiles of the pigments in the fruits. The most abundant acylated pigment in leaves (24%) turned out 6'-O-E-sinapoyl-gomphrenin (gandolin), however, the most significant difference in the pigment profiles is a presence of two novel pigments tentatively identified as highly abundant 6'-O-(3,4-dimethoxy-E-cinnamoyl)-gomphrenin and 6'-O-(3,4,5-trimethoxy-E-cinnamoyl)-gomphrenin as well as their isoforms. Significant degradation of the pigments in the fruit extracts under the impact of selected metal cations and UV-Vis irradiation as well as high protective activity of the leaf extract matrix were observed. Partial chromatographic purification of the leaf extract resulted in an increase of the pigment concentration which was correlated positively with the increased antioxidant activity of obtained fractions.

Basella alba L. (Malabar Spinach) as an Abundant Source of Betacyanins: Identification, Stability, and Bioactivity Studies on Natural and Processed Fruit Pigments.

The antioxidant and anti-inflammatory activities of acylated and decarboxylated gomphrenins, as well as Basella alba L. fruit extract, were investigated in relation to gomphrenin, known for its high biological potential. The most abundant natural acylated gomphrenins, namely, 6'-O-E-caffeoyl-gomphrenin (malabarin) and 6'-O-E-4-coumaroyl-gomphrenin (globosin), were isolated from B. alba extract for the studies. In addition, controlled thermal decarboxylation of gomphrenin in the purified B. alba extract at 65-75 °C resulted in the formation of the most prevalent decarboxylated products, including 17-decarboxy-gomphrenin and 2,17-bidecarboxy-gomphrenin, along with their isoforms. The structures of the decarboxylated pigments were confirmed by NMR analyses. Exploring the matrix effect on pigment reactivity revealed a tremendous increase in the stability of all betacyanins after the initial stage of extract purification using a cation exchanger under various conditions. This indicates the removal of a substantial portion of the unfavorable matrix from the extract, which presumably contains reactive species that could otherwise degrade the pigments. Furthermore, the high concentration of citrates played a significant role in favoring the formation of 2-decarboxy-gomphrenin to a considerable extent. In vitro screening experiments revealed that the tested compounds demonstrated strong anti-inflammatory properties in lipopolysaccharide (LPS)-activated human macrophages. This effect encompassed the selective inhibition of cytokine and chemokine release from activated macrophages, modulation of the chemotactic activity of immune cells, and the regulation of tissue remodeling mediators' release.

Biosynthesis of CoCr2O4/ZnO nanocomposites using Basella alba L. leaves extracts with enhanced photocatalytic degradation of malachite green in aqueous media.

The use of chemical materials to tackle environmental concerns has undergone significant evolution, particularly in the pursuit of strategies for removing pollutants from wastewater as part of environmental remediation an increasingly crucial research topic. Employing green photocatalysts stands out as an efficient and cost-effective approach, playing a key role in promoting sustainable environmental remediation. This study introduces the modification of zinc oxide with cobalt chromite (CoCr2O4/ZnO) through a green synthesis method employing Basella alba L. leaves extract (BALE). Utilizing various characterization techniques, including FT-IR, UV-Vis DRS, XRD, SEM-EDS, and TEM, key features of ZnO, CoCr2O4, and CoCr2O4/ZnO nanocomposites were identified. The optical band gaps for ZnO, CoCr2O4, and CoCr2O4/ZnO nanocomposites were determined as 3.16, 1.71, and 2.80 eV, respectively, where it was shown that the band gap of the ZnO was reduced significantly. CoCr2O4/ZnO nanocomposites displayed a cubic shape of CoCr2O4 on the surface of ZnO, with a particle size of 23.84 ± 8.08 nm. The photocatalytic activity was assessed through the degradation of malachite green under visible light irradiation, where the CoCr2O4/ZnO nanocomposites exhibited superior photodegradation efficiency at 90.91%, surpassing ZnO alone (57.09%). This improvement in photocatalytic activity is attributed to a reduced band gap energy and a high rate constant value of 9.57 × 10-3 min-1, demonstrating pseudo-first-order reaction kinetics. In summary, this research presents the development of a ZnO-based photocatalyst with exceptional performance, especially in the visible light spectrum, making it a promising candidate for applications in wastewater removal.

Determination of biological activities of malabar spinach (Basellaalba) fruit extracts and molecular docking against COX-II enzyme.

To achieve the health benefit from the natural of Basella. Albafruit. This study intended to figure out the bioactive compounds in the two varieties of B. alba (native and hybrid) fruit extract and measurement its biological activities like antioxidant, anti-inflammatory, cytotoxic activities and a molecular docking were performed to observed the pharmaceutical impact on the anti-inflammatory Cyclooxygenase-2 (COX-2) enzyme. The cold extractions along with GC-MS were used for the extraction of and analysis of phytoconstituents from B. alba fruit. The hemolytic inhibitory and BSA (Bovine serum albumin)-denaturation assay, DPPH(2,2-diphenyl-1-picrylhydrazyl) and H2O2-free radical scavenging analysis, and brine shrimp lethalness test were performed to measure the biological activities of the extracted The biological activities assay results showed that the ethanol extract of native malabar spinach exhibited dose-dependent antioxidant activity. The IC50 value 21.55 ± 1.51 µg/mL was for DPPH scavenging assay and 23.36 ± 0.36 µg/mL was for H2O2 scavenging analysis. In anti-inflammatory activity assessment study, the IC50 values of the ethanol extracts were 20.52 ± 0.91 µg/mL for BSA inhibition and 20.43 ± 1.30 µg/mL for RBC hemolytic inhibitory study. In this study, cytotoxicity test results reveal that aqueous extract exhibited no cytotoxicity as compared to ethanol and ethyl acetate extract (LD50 = 875.27 µg/mL). Conversely, the current study insist the in silico analysis, to find out the anti-inflammatory activity of the investigated two fruit varieties due to pharmacokinetics analysis, toxicity properties analysis, ADMETand molecular docking. The result of this study signified that both (native and hybrid) malabar spinach fruit varieties contain phytoconstituents with potent antioxidant, anti-inflammatory, and cytotoxic action.Moreover, the in vitro and in silico results suggest that the native and hybrid fruit varieties of the extracts could be a superior striver for future appraisal as a prospective therapeutically active ingredient.

Antioxidant and antiproliferative activity of Basella alba against colorectal cancer.

Basella alba, a green leafy vegetable with remarkable nutraceutical potential is widely used since ancient times to maintain a healthy colon. This plant has been investigated for its medicinal potential due to the increase in young adult cases of colorectal cancer each year. This study was accomplished to investigate Basella alba methanolic extract (BaME) antioxidant and anticancer properties. BaME consisted of a substantial amount of both phenolic and flavonoid compounds which exhibited significant antioxidant reactivity. Both colon cancer cell lines experienced a cell cycle arrest at the G0/G1 phase after receiving treatment with BaME, which inhibited pRb and cyclin D1 and raised p21 expression levels. This was associated with the survival pathway molecule inhibition and downregulation of E2F-1. The results of the current investigation confirm that BaME inhibits CRC cell survival and expansion. To conclude, the bioactive principles in the extract act as potential antioxidants and antiproliferative agents against colorectal cancer.

First report of Nigrospora musae causing white leaf spot of Basella alba L in China.

Basella alba L, an edible annual twining herb of the genus Basella and the family Basella, has been widely introduced and cultivated in China. Basella alba L. as a leaf vegetable, is rich in vitamins A and C, iron, and calcium (FAO 1988). In May 2022, severe white leaf spots were observed in plantation located in Shuangfeng County (27°41'36" N, 111°56'60" E), Hunan Province, China. More than 50 Basella alba L plants were surveyed with over 80% disease incidence in an area of 300 square meters of greenhouse. The symptoms on leaves were initially small purplish-brown lesions from leaf margins or tips, with lesions expanded, the middle of the lesions was yellowish-white to yellowish-brown, slightly dented. The edge of lesions was purplish-brown, with obvious boundary between the diseased parts and the non-diseased ones. A total of 20 symptomatic samples were randomly collected. Lesion margins were surface sterilized in 2% sodium hypochlorite for 1 min, rinsed with sterile distilled water for three times, dried, placed on potato dextrose agar (PDA), and incubated at 25°C and 60% relative humidity in the dark for 3 days. Hyphal sections from colony edges were transferred to new PDA plates. Six isolates were obtained. Colonies were fast-growing, massive sparse aerial hyphae, initially white, turning gray and black after 7 days. Hyphae were branched, septa, and transparent. To induce sporulation, colonies were transferred to sodium carboxymethyl cellulose (CMC) plates (Z. M. Wen., & X. Y. Luo 1991). Conidia were single-celled, dark black, oblate, or nearly spherical, and measured 10.2 to 15.1 µm × 9.7 to 16.0 µm in diameter (n=50). For molecular identification, the rDNA internal transcribed spacer (ITS), the ß-tubulin gene (TUB), and the translation elongation factor 1-alpha gene (TEF1) were amplified from genomic DNA by primers ITS1/ITS4 (White et al. 1990), Bt2a/Bt2b (Glass & Donaldson. 1995), and EF1-728F/EF1-986R (Carbone & Kohn, 1999). The sequences of six isolates (L1, L2, L7, L10, L11, L12) were deposited in GenBank with accession numbers OP703335, OP703336, OP703337, OP703338, OP703339, OP703340 (ITS), OP784252, OP784157, OP784253, OP784254, OP784255, OP784256 (TEF-1α), and OP724156, OP724158, OP779771, OP779772, OP779773, OP779774 (TUB2). A blast search of sequences showed the ITS, TEF-1α, and TUB2 sequences had >98% identity with homologue sequences from Nigrospora musae isolates BRJ2 (OP451019.1), CBS 319.34 (KY019419.1) and LC6385 (KY019567.1), respectively. These morphological features and molecular identification indicated that the pathogen possessed identical characteristics as Nigrospora musae (Wang, 2017). Pathogenicity test was carried out in plants. Strains were cultured on CMC plates for 14 days, then the mycelium was scraped to make conidial suspension (1×106 conidia/mL). After 5-6 leaves of the Basella alba L were sprouted, conidial suspension was sprayed directly on the leaves, with leaves sprayed by sterile distilled water as the control. All plants were kept in the greenhouse with temperature at 25/30°C (night/day) and 75% relative humidity. After 7 days, symptoms were observed on inoculated leaves of plants, which were the same as previously described samples, while the control plants showed no symptoms. The test was repeated three times with similar results. The strains reisolated from the inoculated leaves were morphologically identical to Nigrospora musae, conforming to Koch's postulates. symptoms of Nigrospora musae is similar to that of the other leaf diseases of Basella alba L reported in China. (H. P. Jiang.2000; S. Tan.1996). To our knowledge, this is the first report of Nigrospora musae causing white leaf spot of Basella alba L in China. The pathogen may severely threat the production of Basella alba L. The information on identification of this fungus may be helpful to the control and prevention of the disease. References: 1. FAO. 1988. Page 103 in: Traditional Food Plants: A Resource Book for Promoting the Exploitation and Consumption of Food Plants in Arid, Semi-arid and Sub-humid Lands of Eastern Africa. FAO Food and Nutrition Paper 42. FAO, Rome, Italy. 2. Z. M. Wen., & X. Y. Luo. Fusarium graminearum spore production medium filtering [J]. Chinese journal of food hygiene, 1991 (04): 11-13. 3. White, T. J., et al. 1990. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA. 4. Carbone, I., et al. 1999. Mycologia. 91: 553-556. 5. Glass, N. L., and Donaldson, G. C. 1995. Appl. Environ. Microbiol. 61: 1323. 6. Wang, 2017. Phylogenetic reassessment of Nigrospora: Ubiquitous endophytes, plant, and human pathogens. 7. H. P. Jiang., et al. Occurrence and comprehensive control of white leaf spot of Basella alba L [J]. Plant Protection Technology and Extension, 2000(02):19. 8. S. Tan. The symptoms and control measures of white leaf spot of Basella alba L [J]. Anhui Agricultural, 1996(08):15. *Indicates the corresponding author. Kaifa Guo, E-mail: andygkf@126.com.

Survey of Viruses Infecting Basella alba in Hawaii.

Malabar spinach plants (Basella alba, Basellaceae) with leaves exhibiting symptoms of mosaic, rugosity, and malformation were found in a community garden on Oahu, HI in 2018. Preliminary studies using enzyme-linked immunosorbent assay and reverse-transcription (RT)-PCR identified Basella rugose mosaic virus (BaRMV) in symptomatic plants. However, nucleotide sequence analysis of RT-PCR amplicons indicated that additional potyviruses were also present in the symptomatic Malabar spinach. High-throughput sequencing (HTS) analysis was conducted on ribosomal RNA-depleted composite RNA samples of potyvirus-positive plants from three locations. Assembled contigs shared sequences similar to BaRMV, chilli veinal mottle virus (ChiVMV), Alternanthera mosaic virus (AltMV), Basella alba endornavirus (BaEV), broad bean wilt virus 2 (BBWV2), and Iresine viroid 1. Virus- and viroid-specific primers were designed based on HTS sequencing results and used in RT-PCR and Sanger sequencing to confirm the presence of these viruses and the viroid. We tested 63 additional samples from six community gardens for a survey of viruses in Malabar spinach and found that 21 of them were positive for BaRMV, 57 for ChiVMV, 21 for AltMV, 19 for BaEV, and 14 for BBWV2. This is the first characterization of the virome from B. alba.

Basella alba stem extract integrated poly (vinyl alcohol)/chitosan composite films: A promising bio-material for wound healing.

Natural extract-based bio-composite material for wound healing is gaining much attention due to risk of infection and high cost of commercial wound dressing film causes serious problem on the human well-being. Herein, the study outlines the preparation of Poly (vinyl alcohol)/Chitosan/Basella alba stem extract (BAE) based bio-composite film through solvent casting technique and well characterized for wound healing application. Incorporation of BAE into Poly (vinyl alcohol)/Chitosan matrix has shown existence of secondary interactions confirmed by FT-IR analysis. Good morphology, thermal stability and significant improvement in flexibility (∼63.38 %) of the films were confirmed by SEM, TGA and Mechanical test results, respectively. Hydrophilic property (∼9.04 %), water vapor transmission rate (∼70.07 %), swelling ability (∼14.7 %) and degradation rate (∼14.04 %) were enhanced with increase in BAE content. In-vitro studies have shown good antibacterial activity against foremost infectious bacterial strains S. aureus and E. coli. Additionally, BAE integrated Poly (vinyl alcohol)/Chitosan film has amplified anti-inflammatory (∼79.38 %) property, hemocompatibility and excellent biocompatibility (94.9 %) was displayed by cytotoxicity results. Moreover, in-vitro scratch assay and cell adhesion test results illustrated prominent wound healing (96.5 %) and adhesion. Overall results of the present work proclaim that developed bio-composite film could be utilized as a biomaterial in wound care applications.

First Report of Athelia rolfsii Causing Damping-Off and Leaf Blight on Basella alba in Malaysia.

Basella alba is an evergreen perennial vine that grows as an edible leafy vegetable in Malaysia (Nordin et al. 2007). During January 2021, a cottony white hypha associated with aggregates of white to brown sclerotia and symptoms of damping-off were visualized on the stem base of B. alba at the soil surface in an isolated field (~0.03 ha) located in the district of Penampang, Sabah province, Malaysia (5°56'51.0"N 116°04'31.8"E). Yellowing and wilting of leaves, and defoliation were observed after four days of the development of damping-off. Survey was conducted on 100 plants which 30 were found infected. The disease severity (90%) on stems was estimated using IMAGEJ. Symptomatic stem tissues were surface sterilized with 75% of ethanol for 1 min, washed with 2% of sodium hypochlorite solution for 1 min, rinsed thrice with sterile distilled water, air dried and plated on potato dextrose agar (PDA). Plates were incubated for 7 days at 25°C in the dark. After 7 days, fungi were isolated; colony color was white and had a cottony appearance. On day 14, white to brown sclerotia 1.0 to 2.2 mm in diameter were produced. Hyaline septate hyphae with clamp connections and multiple nuclei were seen. Conidia and conidiophores were absent from the colony on PDA. Genomic DNA of fungi was extracted based on Khoo et al. (2022a and 2022b). PCR amplification (Khoo et al. 2022b) was performed using primer set ITS1/ITS4, EF983/EF2218 and LR0R/LR05 to amplify the internal transcribed spacer (ITS) region of rDNA, partial translation elongation factor 1 alpha (TEF-1α) gene and partial large subunit ribosomal RNA (LSU rRNA) gene, respectively (Vilgalys and Hester 1990; White et al. 1990; Carbone and Kohn, 1999; Rehner 2001). Phylogenetic analysis indicated that the isolates formed a supported clade to the related Athelia rolfsii sequences. The sequencing result (GenBank Accession Nos. OK465460, OP809607, OP857217) had a 99% identity over 625 bp, 941 bp, and 1,101 bp with the corresponding gene sequence of A. rolfsii (GenBank Accession Nos. MN622806, AY635773, MW322687) after analysis in BLASTn program. Pathogenicity test was performed based on Le (2011). Three 8-week-old B. alba plants cultivated on sterilized soil were inoculated with 5-mm mycelia plugs from 7-day-old culture. A plug was put on the upper soil surface layer 2 cm away from the base of the stem of B. alba plant before fully covered with a layer of sterilized soil. Plants that were inoculated with sterile PDA plugs served as the control treatment. Plastic bags were used to cover the plants after inoculation for 24 h before keeping them in a glasshouse under ambient conditions. Water-soaked and brown lesions were visualized on the stem base of all inoculated plants after four days of inoculation. Symptom of damping-off and leaf blight was observed after another 3 days. No symptoms developed on the mock controls. The pathogenicity test was repeated twice. Re-isolation was performed from the symptomatic tissues of inoculated plants and mock controls. The isolates reisolated from the symptomatic tissues were verified as A. rolfsii based on morphology and molecular characterization, thus fulfilling Koch's postulates. No pathogens were isolated from the mock controls. To our knowledge, this is the first report of A. rolfsii causing damping-off and leaf blight on B. alba in Malaysia, as well as worldwide. Our findings documented the wider geographical and host range of A. rolfsii and indicate its potential threat to B. alba production in Malaysia.

Structural Studies on Diverse Betacyanin Classes in Matured Pigment-Rich Fruits of Basella alba L. and Basella alba L. var. 'Rubra' (Malabar Spinach).

Identification of betacyanins in Basella alba L. and Basella alba L. var. 'Rubra' fruits was performed by low- and high-resolution mass spectrometry (LRMS and HRMS) as well as 1H, 13C and two-dimensional NMR which revealed hitherto completely not known betacyanin classes in the plant kingdom. Especially, the presence of unique nitrogenous acyl moieties in the structures of the pigments was ascertained by the HRMS Orbitrap detection. Except for detected polar betacyanin glycosylated derivatives, presence of a series of previously not reported pigments such as malonylated betanidin 6-O-ß-glusosides with their acyl migration isomers along with the evidence of the 3''-hydroxy-butyrylated betacyanins is reported. The first complete NMR data were obtained for novel and principal acylated gomphrenins with hydroxycinnamic acids: 6'-O-E-caffeoyl-gomphrenin (malabarin), 6'-O-E-sinapoyl-gomphrenin (gandolin), 6'-O-E-4-coumaroyl-gomphrenin (globosin) and 6'-O-E-feruloyl-gomphrenin (basellin).

Seed Priming with ZnO and Fe3O4 Nanoparticles Alleviate the Lead Toxicity in Basella alba L. through Reduced Lead Uptake and Regulation of ROS.

The increased lead (Pb) content in the environment has an impact on all living beings, including plant growth and quality. The present study aims to investigate the protective roles of zinc (Zn)- and iron (Fe)- nanoparticles (NPs) in alleviating stress symptoms caused by lead (Pb) exposure in Basella alba seedlings. For this purpose, 15 different treatment combinations of seed priming with two NPs at 0 and 200 mg L−1, and five Pb levels (0, 4, 8, 15, 20 mM) were chosen. Pb stress (20 mM) was found to reduce seed germination by 72.8% and seedling growth, particularly root length, by 92% when compared to the control. Under different Pb concentrations, seed priming with ZnNPs (200 mg L−1) and FeNPs (200 mg L−1) increased seed germination by 34.7% and 54.9%, respectively, and root length by 152.9% and 252.9%, respectively. In 20 mM Pb stress, NPs primed seedling showed decrease in Pb content by 33.7% with ZnNPs and 32.6% with FeNPs. Increased Pb stress resulted in increased reactive oxygen species (ROS) generation (H2O2) and lipid peroxidation (MDA) compared to non-Pb stressed seedlings. However, increased antioxidants in the NPs treatments such as SOD, CAT, POD and proline content, scavenged these ROS. Considering all the parameters under study, priming alleviated Pb stress in the following order: FeNPs > ZnNPs > hydropriming > control. To summarise, seed priming with Zn- and Fe-NPs has the potential to alleviate Pb toxicity via reduced Pb uptake, ROS generation and lipid peroxidation as well as increased proline content and activation of antioxidant enzymatic system.

First report of Epicoccum sorghinum causing leaf spot on Basella alba in Malaysia.

Basella alba (family Basellaceae) is a perennial vine that serves as an edible leaf vegetable in Malaysia. In May 2021, red spots were observed on leaf samples of B. alba in Lido, Sabah Province (5°56'39.1"N, 116°04'47.6"E). The disease severity was about 20% with 10% incidence. The spots enlarged and coalesced into larger necrotic spots. Small pieces (5 x 5 mm) of infected leaves were excised from three plants, and then surface disinfected based on Khoo et al. (2022). One fungal isolate (Lido01) was isolated and cultured on potato dextrose agar (PDA) at 25°C. A single isolate with cottony aerial mycelia and pink concentric rings was observed on the upper surface of the culture. Unicellular and multicellular chlamydospores were observed, and measured 7.1 to 14.3. × 17.8 to 74.5 µm. Conidia were unicellular, hyaline, oval, and measured 3.8 to 5.2 x 1.7 to 2.7 µm (n= 20). Pycnidia were spheroid, and measured 66.2 to 114.3 x 44.1 to 86.1 µm (n= 20). Genomic DNA was extracted from fresh mycelia according to the extraction method of Khoo et al. (2022a and 2022b). ITS1/ITS4, LR0R/LR7, ACT512F/ACT783R, and T10/Bt2b primers were used to amplify the internal transcribed spacer (ITS), large subunit (LSU), actin (ACT), and tubulin (TUB) genes, respectively (O'Donnell and Cigelnik, 1997; Chen et al. 2021). PCR products were Sanger sequenced by Apical Scientific Sdn. Bhd. (Serdang, Malaysia). Sequences of isolate Lido01 were deposited in GenBank as OM501130 (ITS), OM501128 (LSU), OM513916 (ACT) and OM513917 (TUB). Respective gene sequences of this isolate showed 100% homology to ITS sequence of isolate BPL01 (OM453926) (507/507 bp), LSU sequence of isolate BPL01 (OM453925) (1328/1328 bp), ACT sequence of isolate CZ01 (MN956831) (275/275 bp) and TUB sequence of isolate BJ-F1 (MF987525) (556/556 bp). The sequences of Lido01 established a supported clade (99% bootstrap value) to the related Epicoccum sorghinum type sequences, according to phylogenetic analysis using maximum likelihood based on the concatenated ITS, ACT, and TUB sequences. Morphological characters also matched the description of E. sorghinum (Li et al. 2020). Koch's postulates were tested as described by Chai et al. (2017) with modification by spray inoculation (106 spores/ml) on the leaves of three healthy one-month-old B. alba, while sterilized distilled water served as the control treatment. Monitoring and incubation were performed in a greenhouse based on Iftikhar et al. (2022). All inoculated leaves developed symptoms as described above by 8 days post-inoculation, whereas no symptoms occurred on controls, thus fulfilling Koch's postulates. The experiment was repeated twice. The reisolated pathogen was morphologically and genetically identical to E. sorghinum. E. sorghinum was reported causing leaf spot on Brassica parachinensis (Yu et al. 2019), Camellia sinensis (Bao et al. 2019), Myrica rubra (Li et al. 2020), Oryza sativa (Liu et al. 2020) and Zea mays (Chen et al. 2021). To our knowledge, this is the first report of E. sorghinum causing leaf spot on B. alba in Malaysia. Our findings have expanded the geographic range and host range of E. sorghinum in Malaysia, though the host range of this isolate is not known.

Basella alba, Grewia asiatica, Solanum nigrum and Ficus carica fruit extracts ameliorate the testicular histopathology induced by CCl4 exposure in albino mice: An experimental study.

Background: Carbon tetrachloride (CCl 4 ) is extensively used in various industries and induces oxidative stress in humans. Unfortunately, it is one of the neglected potent male reproductive toxicants. Objective: The present research reports the testicular histopathology of CCl 4 and ameliorations by four medicinal fruit pulp extracts (FPEs) in mice. Materials and Methods: Sixty male albino mice were divided into six groups (10/group) as per the following: 1. Vehicle control (Vc); 2. CCl 4 (C); 3-6. CCl 4 +Basella alba (CBa), CCl 4 +Solanum nigrum (CSn), CCl 4 +Ficus carica (CFc) and CCl 4 +Grewia asiatica (CGa). Except for the control group, CCl 4 (0.1 mL of 0.2 mL kg - 1 ) was given to the animals in corn oil. The four plant extracts (0.1 mL each) were respectively given to the relevant FPE group animals for the next five consecutive days, while the animals in the Vc and CCl 4 groups received water instead of FPE. Results: The CCl 4 exposure led to various histometric and histological alterations (loss of interstitial tissue and various dislodged tailless spermatids with enlarged heads) that were recovered in all except Solanum nigrum FPE mice post-treatment. The micrometric data of testicular sections also indicated significant decline in the number of spermatogonia, while the cross-sectional area of the sperm heads remained significantly higher in the CSn and C groups. Conclusion: Except for Solanum nigrum,the three FPEs, especially Ficus carica,showed rehabilitative properties against CCl 4 exposure-related modifications in testicular histopathologies.

Structural characterization of the carbohydrate and protein part of arabinogalactan protein from Basella alba stem and antiadhesive activity of polysaccharides from B. alba against Helicobacter pylori.

BACKGROUND: Increasing drug resistance of Helicobacter pylori has highlighted the search for natural compounds with antiadhesive properties, interrupting the adhesion of H. pylori to stomach epithelia. Basella alba, a plant widely used in Asian traditional medicine, was investigated for its antiadhesive activity against H. pylori. METHODS: B. alba extract FE was prepared by aqueous extraction. Polysaccharides were isolated from FE by ethanol precipitation and arabinogalactan-protein (AGP) was isolated with Yariv reagent. Carbohydrate analyses was performed by standard methods and sequence analysis of the protein part of AGP by LC-MS. In vitro adhesion assay of fluorescent-labelled H. pylori J99 to human AGS cells was performed by flow cytometric analysis. RESULTS: Raw polysaccharides (BA1) were isolated and 9% of BA1 were identified as AGP (53.1% neutral carbohydrates L-arabinose, D-galactose, rhamnose, 5.4% galacturonic acid, 41.5% protein). After deglycosylation of AGP, the protein part (two bands at 15 and 25 kDa in tricine SDS-PAGE) was shown to contain peptides like ribulose-bisphosphate-carboxylase-large-chain. Histological localization within the stem tissue of B. alba revealed that AGP was mainly located at the procambium ring. Functional assays indicated that neither FE nor BA1 had significant influence on viability of AGS cells or on H. pylori. FE inhibited the bacterial adhesion of H. pylori to AGS cells in a dose dependent manner. Best anti-adhesive effect of ~67% was observed with BA1 at 2 mg/mL. CONCLUSION: The data obtained from this study characterize in part the mucilage and isolated polysaccharides of B. alba. As the polysaccharides interact with the bacterial adhesion, a potential uses a supplemental antiadhesive entity against the recurrence of H. pylori after eradication therapy may be discussed.

Suppression and management of Meloidogyne incognita in soil using Trichoderma harzianum NFCF160 and Trichoderma virens Isf-77

Aims@#Meloidogyne incognita adversely affects numerous crop plants worldwide. Therefore, the modern world has been moving towards biocontrol methods to prevent nematode attacks. This study was aimed to (i) investigate the potential use of Trichoderma harzianum NFCF160 and T. virens Isf-77 in managing M. incognita in soil and (ii) identify trapping mechanisms employed by both Trichoderma strains to suppress M. incognita.@*Methodology and results@#Three weeks old, Basella alba L. plants were subjected to five different treatments. The above and below ground growth parameters and the galling indices of these plants were measured every four weeks for three sampling times. Trapping mechanisms employed by Trichoderma strains were examined following plate assays. Plants treated with T. harzianum NFCF160 and T. virens Isf-77 had significantly higher values for the total number of leaves (34 ± 2.84) and (27 ± 2.61), fresh weight of the shoot (81 ± 9.51 g) and (91 ± 9.70 g), dry weight of the shoot (71 ± 5.24 g) and (62 ± 5.81 g), respectively eight weeks after inoculation of M. incognita. Significantly low galling indices (2 and 2) were recorded in B. alba treated with Trichoderma strains. Both Trichoderma strains exhibited various nematode-trapping mechanisms, such as non-constricting rings and adhesive spores.@*Conclusion, significance and impact of study@#This investigation highlighted the potential of both Trichoderma strains as biocontrol agents to control M. incognita effect in sustainable agriculture.

Shelf Life Extension of Chilled Pork by Optimal Ultrasonicated Ceylon Spinach (Basella alba) Extracts: Physicochemical and Microbial Properties.

The effect of ultrasonication on the antioxidant and antibacterial properties of Ceylon spinach (Basella alba) extracts (CE) and the shelf life of chilled pork with CE were studied. The CE were ultrasonicated at different power levels (60-100%) for 10-40 min in an ultrasonic bath with the rise of antioxidant activities (p ≤ 0.05) proportional to the ultrasonication time. The additional investigation of antibacterial activities showed that the ultrasonicated extracts (100 mg/mL) could inhibit and inactivate Staphylococcus aureus and Escherichia coli with the optimal condition of 80% power for 40 min. For shelf-life testing, fresh pork treated with the ultrasonicated extracts at 100 and 120 mg/mL had lower values of thiobarbituric acid reactive substances (TBARS) than the control (without dipping). For food safety as measured by the total microbial count, the fresh pork dipped with 100-120 mg/mL CE extract could be kept at 0 °C for 7 days, 2 to 3 days longer than control meat at 0 and 4 °C, respectively. A sensory evaluation using a nine-point hedonic scale showed that fresh pork dipped with 100-mg/mL CE extracts was accepted by consumers. It is suggested that CE extracts can be applied in the food industry to enhance the quality and extend the shelf life of meat products.

Studies on the spectrometric analysis of metallic silver nanoparticles (Ag NPs) using Basella alba leaf for the antibacterial activities.

In this present investigation, an aqueous Basella alba leaves extract was used to synthesize AgNPs. The green synthesis approach is carried out in our work due to non-toxic, less cost, and ecofriendly methods. FTIR spectra are used to confirm the biomolecules present in B.alba leaves extract along with AgNPs and these compounds are responsible for Ag particle from micro to nanostructure. The FCC structure and crystalline nature of the AgNPs are analyzed with the help of XRD and TEM techniques respectively. DLS and Zeta potential techniques are carried out to find the size and stability of AgNPs respectively and UV is used to verify the presence of AgNPs in synthesized samples employing SPR peaks around 435 nm. The antioxidant studies expose eminent scavenging activity which ranges from 13.71% to maximum 67.88%. Green synthesized AgNPs possess well organized biological activities concerning antioxidant and antibacterial, which can be used in some biologically applications.

Deciphering the molecular pathways of apoptosis using purified fractions from leaf extract of Basella alba through studying the regulation of apoptosis related genes.

Apoptosis plays a pivotal role in the exclusion of abnormal cells without any ruin of surrounding healthy cells. Generally, it occurs through an orderly and autonomously process which is controlled by proper function of various genes. Therefore, the current experiments detect the expression level/pattern of those genes to confirm the involvement of extrinsic and intrinsic pathway using Basella alba leaf (BAL). Several fractions after gel filtration chromatography of BAL extract have been pooled to evaluates its apoptosis induction potentiality on Ehrlich's Ascites Carcinoma (EAC) cells through conducting a number of bio-assays such as cell growth inhibition assay, fluorescence and optical microscopy, DNA fragmentation assay and gene expression analysis etc. The pooled fractions of BAL showed 12-56% inhibitory effect on EAC cell line at the concentration range of 25-400 µg/ml that was determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. They also exhibited excellent cell growth inhibition at in vivo and in vitro condition when treated with 10, 20 and 40 mg/kg day. After administration of six consequent days, significant morphological features of apoptosis were observed in EAC cells under both fluorescence and optical microscope which was further supported by DNA fragmentation assay. The polymerase chain reaction amplification of bax, bcl-2 (B-cell lymphoma 2), p53, tumor necrosis factor-α, Fas, NF-kß (Nuclear factor-Kappa-B), PARP-1 (Poly (ADP-ribose) polymerase), Cyt-c cas-8, cas-9 and cas-3 revealed that the experimental sample able to induce apoptosis in both extrinsic and intrinsic pathways through altering the gene expression. The current findings suggest that sample from BAL occupy wonderful competence to induce cell apoptosis and become an ideal resource for cancer treatment.

Technical feasibility of conventional and nonconventional vegetables' intercropping under organic fertilization / Viabilidade técnica do cultivo consorciado de hortaliças convencionais e não convencionais sob fertilização orgânica

This work aimed to evaluate the technical feasibility of lettuce, Indian spinach and taro intercropping under organic fertilizer. The experiment was performed at Água Limpa Farm, belonging to the University of Brasília, located in Brasília - DF, from October 2014 to June 2015. A completely randomized block design with seven treatments and four replications was used. Each experimental plot had 18 m2 (4.5 m x 4.0 m), totaling 28 plots. The following treatments were assessed: lettuce monoculture, Indian spinach monoculture, taro monoculture, lettuce/Indian spinach intercropping, lettuce/taro intercropping, Indian spinach/taro intercropping, and lettuce/Indian spinach/taro intercropping. In all treatments, the following spacings were used: 0.25 x 0.25 m (lettuce), 1.0 x 0.6 m (Indian spinach), and 1.0 x 0.3 m (taro). Crop production was evaluated from sample plants taken from the central part of each plot. Two harvests of lettuce, two of Indian spinach and one of taro were carried out during intercropping. In the first cycle, lettuce highest mean yield values were observed in the lettuce monoculture (3.5 kg m-2) and lettuce/Indian spinach intercropping (3.4 kg m-2). The crop reached the commercial size in both production cycles, except when intercropped with Indian spinach in the second cycle. The highest Indian spinach mean fresh mass was recorded in the lettuce/Indian spinach intercropping (974.7 g plant-1; first harvest) and monoculture (327.5 g plant-1; second harvest). The yield of taro rhizomes was significantly higher when intercropped with lettuce, showing a mean value of 16.8 kg m-2. The intercropping arrangements with lettuce increased yield of nonconventional vegetables. The conventional and nonconventional vegetables' intercropping technique increased species diversity in the area without interfering in the agronomic performance and quality of the crops.