Analysis of the Differentially Expressed Proteins and Metabolic Pathways of Honeybush (Cyclopia subternata) in Response to Water Deficit Stress.

Honeybush (Cyclopia spp.) is a rich source of antioxidant properties and phenolic compounds. Water availability plays a crucial role in plant metabolic processes, and it contributes to overall quality. Thus, this study aimed to investigate changes in molecular functions, cellular components, and biological processes of Cyclopia subternata exposed to different water stress conditions, which include well-watered (as Control, T1), semi-water stressed (T2), and water-deprived (T3) potted plants. Samples were also collected from a well-watered commercial farm first cultivated in 2013 (T13) and then cultivated in 2017 (T17) and 2019 (T19). Differentially expressed proteins extracted from C. subternata leaves were identified using LC-MS/MS spectrometry. A total of 11 differentially expressed proteins (DEPs) were identified using Fisher's exact test (p < 0.00100). Only α-glucan phosphorylase was found to be statistically common between T17 and T19 (p < 0.00100). Notably, α-glucan phosphorylase was upregulated in the older vegetation (T17) and downregulated in T19 by 1.41-fold. This result suggests that α-glucan phosphorylase was needed in T17 to support the metabolic pathway. In T19, five DEPs were upregulated, while the other six were downregulated. Based on gene ontology, the DEPs in the stressed plant were associated with cellular and metabolic processes, response to stimulus, binding, catalytic activity, and cellular anatomical entity. Differentially expressed proteins were clustered based on the Kyoto Encyclopedia of Genes and Genomes (KEGG), and sequences were linked to metabolic pathways via enzyme code and KEGG ortholog. Most proteins were involved in photosynthesis, phenylpropanoid biosynthesis, thiamine, and purine metabolism. This study revealed the presence of trans-cinnamate 4-monooxygenase, an intermediate for the biosynthesis of a large number of substances, such as phenylpropanoids and flavonoids.

Entomopathogenic nematodes associated with organic honeybush (Cyclopia spp.) cultivation in South Africa.

Entomopathogenic nematodes (EPNs) of the genera Steinernema and Heterorhabditis have demonstrated great potential as useful bio-control agents in the management of certain important soil-inhabiting insect pests of agricultural crops. In a survey of nematodes associated with organic honeybush cultivation, soil samples were obtained from nine organic honeybush plots, which are located in the Bredasdorp area of the Western Cape province of South Africa. The EPNs were isolated from soil by baiting with larvae of Galleria mellonella (greater wax moth) and identified by the amplification of the internal transcribed spacer region using the primer set TW81 and AB28. EPNs were abundant in the honeybush orchards, accounting for about 50% of the sampled fields and five EPN species were identified including Heterorhabditis bacteriophora, Heterorhabditis safricana, Steinernema khoisanae,. Steinernema nguyeni, Oscheius sp. and an unknown EPN, therefore suggesting a substantial diversity of EPNs in the sampled fields. A 100% mortality of infected G. mellonella larvae was recorded within 48 h of exposure to the nematodes. However, although these EPNs have been previously reported in South Africa, it is the first time they are found in such diversity on a conservative tillage management system in organic honeybush cultivation.

Diversity and population distribution of nematodes associated with honeybush (Cyclopia spp.) and rooibos (Aspalathus linearis) in the Western Cape province of South Africa.

Nematodes are important soil organisms that constitute a key component of the soil ecosystem. A plant-parasitic survey was conducted to identify the diversity of nematodes associated with two endemic tea plants, honeybush (Cyclopia spp.) and rooibos (Aspalathus linearis) in the Western Cape province of South Africa. A total of 20 farmlands were surveyed and soil samples were collected from the rhizosphere of plants, for nematode isolation and identification based on morphological characters. Confirmation of the species of plant-parasitic nematodes was done using molecular-based tools. Nematodes were classified into various feeding groups based on their colonizer-persister (c-p) values. Plant-feeding nematodes identified from the honeybush tea plants include; Criconema mutabile, Meloidogyne hapla, M. javanica, and Xiphinema oxycaudatum, while Hoplolaimus sp., Neodolichorhynchus estherae and Pratylechus bolivianus were pathogenic on the rooibos monocultures. Bacterial and fungal feeders (Cephalobidae and Rhabditidae) were also abundant and frequently encountered in all samples. The study provides information on the diversity of nematodes associated with the indigenous herbal tea plants of South Africa.

An ethnobotanical survey of medicinal plants used by traditional healers for the treatment of cancer in Hammanskraal and Winterveld, Tshwane Metropolitan Municipality, South Africa.

Background: Knowledge of medicinal plants used by the traditional healers are mostly confined among the locals and the adherents, hence, proper enquiry and documentation can help the ever dynamic scientific world to find permanent cure to the menace of such deadly diseases such as cancer. This study aimed at (1) specifically recording medicinal plants traditionally used for the treatment of cancer in Hammanskraal and Winterveld, South Africa, and (2) documenting the different methods of preparation and administration of those medicinal plants as recorded during the study. Method: An open-ended semi-structured questionnaire was administered to 90 willing traditional healers in Hammanskraal and Winterveld area, Tshwane Municipality, South Africa to document plants used for cancer treatments. The study was conducted over a period of six months (July - December, 2018). Descriptive statistics was used to present the obtained data. Results: The study recorded twenty-eight plant species belonging to 18 families for the treatment of different types of cancer. Plant species in the Fabaceae family particularly Lessertia frutescens (L.) Goldblatt and J.C. Manning, Senna italica Mill and Trifolium pratense L. were the most prominently mentioned (highest citation frequency) by the traditional healers for lung, and skin cancer treatment. Based on the citation frequency, the most treated cancer by the traditional healers is in the order: skin cancer > lung cancer > breast cancer > prostate cancer > cervical cancer. The method of preparation included decoction (32.3%), infusion (29%), paste (16.1%) and maceration (22.6%). Conclusion: In addition to the documentation of indigenous knowledge related to the use of medicinal plants in the traditional management of cancer in South Africa, this study opens a vista for investigations into the phytochemical and pharmacological properties of the documented plants.

Vigna subterranea ammonium transporter gene (VsAMT1): Some bioinformatics insights.

Ammonium transporters (AMTs) play a role in the uptake of ammonium, the form in which nitrogen is preferentially absorbed by plants. Vigna subterranea (VsAMT1) and Solanum tuberosum (StAMT1) AMT1s were characterized using molecular biology and bioinformatics methods. AMT1-specific primers were designed and used to amplify the AMT1 internal regions. Nucleotide sequencing, alignment and phylogenetic analysis assigned VsAMT1 and StAMT1 to the AMT1 family. The deduced amino acid sequences showed that VsAMT1 is 92% and 89% similar to Phaseolus vulgaris PvAMT1.1 and Glycine max AMT1 respectively, while StAMT1 is 92% similar to Solanum lycopersicum LeAMT1.1, and correspond to the 5th-10th trans-membrane domains. Residues VsAMT1 D23 and StAMT1 D15 are predicted to be essential for ammonium transport, while mutations of VsAMT1 W1A-L and S87A and StAMT1 S76A may further enhance ammonium transport. In addition to nitrogen uptake from the roots, VsAMT1 may also contribute to interactions with rhizobia.

Completely green synthesis of dextrose reduced silver nanoparticles, its antimicrobial and sensing properties.

We herein report the green synthesis of highly monodispersed, water soluble, stable and smaller sized dextrose reduced gelatin capped-silver nanoparticles (Ag-NPs) via an eco-friendly, completely green method. The synthesis involves the use of silver nitrate, gelatin, dextrose and water as the silver precursor, stabilizing agent, reducing agent and solvent respectively. By varying the reaction time, the temporal evolution of the growth, optical, antimicrobial and sensing properties of the as-synthesised Ag-NPs were investigated. The nanoparticles were characterized using UV-vis absorption spectroscopy, Fourier transform infra-red spectroscopy (FT-IR), X-ray diffraction (XRD), transmission electron microscopy (TEM) and high resolution transmission electron microscopy (HR-TEM). The absorption maxima of the as-synthesized materials at different reaction time showed characteristic silver surface plasmon resonance (SPR) peak. The as-synthesised Ag-NPs show better antibacterial efficacy than the antibiotics; ciproflaxin and imipenem against Pseudomonas aeruginosa with minimum inhibition concentration (MIC) of 6 µg/mL, and better efficacy than imipenem against Escherichia coli with MIC of 10 µg/mL. The minimum bactericidal concentration (MBC) of the as-synthesised Ag-NPs is 12.5 µg/mL. The sensitivity of the dextrose reduced gelatin-capped Ag-NPs towards hydrogen peroxide indicated that the sensor has a very good sensitivity and a linear response over wide concentration range of 10(-1)-10(-6)M H2O2.