Alhagi Maurorum Research Articles

Exploring the Genetic Basis of Drought Tolerance in Alhagi camelorum: A Comprehensive Transcriptome Study of Osmotic Stress Adaptations

Alhagi camelorum, a desert shrub known for its impressive drought tolerance, exhibits notable resilience under arid conditions. However, the underlying mechanisms driving its drought resistance remain largely unexplored. This study aims to investigate these mechanisms by exposing A. camelorum to osmotic stress using varying polyethylene glycol (PEG) concentrations (1%, 5%, 10%) in a controlled laboratory setting. Growth analysis revealed significant inhibition and phenotypic changes with increasing PEG levels. Transcriptomic analysis, including differentially expressed gene identification, GO enrichment analysis, and hierarchical cluster analysis of genes in roots and shoots, identified key pathways associated with drought adaptation, such as ABA-activated signaling, cell wall biogenesis, photosynthesis, and secondary metabolite biosynthesis. Notably, some genes involved in these pathways exhibited tissue-specific expression patterns and showed PEG concentration-dependent regulation. Key findings include the dose-dependent (R2 > 0.8) upregulation of a proline-rich protein (Asp01G030840) and a BURP domain-containing protein (Asp02G039780), as well as critical genes involved in cell wall biogenesis (encoding Pectinesterase inhibitor domain-containing and Fasciclin-like arabinogalactan protein), and secondary metabolite biosynthesis (encoding enzymes for terpenoid and flavonoid biosynthesis). The regulation of these genes is likely influenced by phytohormones such as ABA and other stress-related hormones, along with significant transcription factors like ABI4, TALE, MYB61, GRAS, and ERF. These insights lay the groundwork for further research into the functional roles of these genes, their regulatory networks, and their potential applications in enhancing drought resistance in desert plants and agricultural crops.

Kinetic and thermodynamic analysis of alizarin Red S biosorption by Alhagi maurorum: a sustainable approach for water treatment

Synthetic dyes, such as Alizarin Red S, contribute significantly to environmental pollution. This study investigates the biosorption potential of Alhagi maurorum biosorbent for the removal of Alizarin Red S (ARS) from aqueous solutions. Fourier transform infrared spectroscopy (FTIR) was used to analyze the biosorbent’s adsorption sites. Various parameters were optimized to maximize dye adsorption. An optimal removal efficiency of 82.26% was attained by employing 0.9 g of biosorbent with a 25 ppm dye concentration at pH 6 and 60 °C over 30 min. The data were modeled using various isothermal and kinetic models to understand the adsorption behavior. Thermodynamic parameters indicated that the adsorption process was spontaneous and endothermic. The pseudo-second-order kinetic model best described the data, indicating chemisorption as the rate-limiting step. The data matched best to the Langmuir model, indicating that the adsorption occurs as a monolayer on uniform surfaces with a finite number of binding sites. The model showed a strong correlation (R² = 0.991) and a maximum adsorption capacity (qmax) of 8.203 mg/g. Principal component analysis (PCA) identified temperature as the dominant factor, with the primary component, PC1 capturing 100% of its effect. The mechanisms involved in ARS biosorption on A. maurorum include electrostatic interactions, hydrogen bonding, hydrophobic interactions, dipole-dipole interactions, and π-π stacking. Alhagi maurorum showed promising potential for biosorbing toxic dyes from contaminated water, suggesting further investigation for practical applications.

Alhagi maurorum Ethanolic Extract can halt potential topiramate neurotoxicity on sciatic nerve of adult male albino rats

Alhagi maurorum Ethanolic Extract can halt potential topiramate neurotoxicity on sciatic nerve of adult male albino rats

Morphological and Germination Characteristics of Alhagi maurorum Boiss. and Salsola richteri Kar. Seeds Distributed in the Karakum Desert of Turkmenistan

The Karakum Desert is the largest desert in Turkmenistan and covers a significant part of Central Asia. This desert is home to plant species that can adapt to hot, dry, and barren conditions. During the summer months, the Karakum Desert can get extremely hot, with temperatures reaching up to 50 °C, while in winter, it can drop to -20 °C. Therefore, the desert conditions significant seasonal temperature differences. In winter, rainfall is very low, snow rarely falls, and quickly melts. Afghan winds have an impact on the desert vegetation. The desert vegetation mainly consists of short grasses and woody shrubs. Despite the scarcity of rainfall and high temperatures during the summer months, steppe plants have generally been able to adapt to the harsh conditions of the desert. There are also species that have adapted to the desert climate, such as Alhagi maurorum Boiss., Haloxylon persicum Bunge, Haloxylon aphyllum (Minkw.) Iljin), Solanum nigrum L., and Salsola richteri Kar. These plants are highly valuable in terms of nutrition for animals and are used as winter animal feed in rural areas. In this study, the seed and germination characteristics of Alhagi maurorum Boiss. and Salsola richteri Kar, which are tree and shrub-like plants that can adapt to Karakum Desert conditions, were examined. In laboratory study, the morphological characteristics of the seeds were determined, and vitality and germination tests were conducted. Information was obtained about the germination adaptation of these plants in desert ecosystems. As a result of the study, it was determined that S. richteri had a very low rate of viable seeds and no germination, while the germination process of Alhagi maurorum could take a considerably long time.

Cerium Oxide Nanoparticles Synthesis using Alhagi Maurorum Leaf Extract and Evaluation of Their Cytotoxic Effect on Breast Cancer Cell Lines and Antibacterial Effects.

Green synthesis offers a fast, simple, and economical method for producing metallic nanoparticles.The basis of this method is to obtain nanoparticles using natural materials, such as plants, fungi, and bacteria, instead of harmful and expensive chemical-reducing agents. In this study, CeO2NPs were produced using Alhagi maurorum extract, and their anticancer and antibacterial activities were evaluated. Alhagi maurorum extract was prepared according to a previously described protocol, and CeO2NPs were synthesized from the salt of this extract. The resulting nanoparticles were characterized using Transmission electron microscopy (TEM), scanning electron microscope (SEM), and X-ray diffraction (XRD) techniques. The antibacterial and cytotoxic effects of the nanoparticles were measured by MIC, MBC, and MTT assays, respectively. The results were analyzed using one-way analysis of variance (ANOVA) using Prism software. The MTT assay on breast cancer cell lines showed that the cytotoxic effect of CeO2NPs on cell lines was concentration-dependent. In addition, this nanoparticle was more effective against Gram-positive bacteria. These nanoparticles can be used as cancer drug delivery systems with specific targeting at low concentrations in addition to anticancer treatments. It can also have biological and medicinal applications, such as natural food preservation and wound dressing.

Treatment of functional disorders of gastrointestinal tract of calves with complex phytomineral preparation

Significant economic losses due to diarrhoea morbidity in newborn young animals lead to a decrease in the profitability of the sector in the structure of agricultural production. Therefore, the purpose of this study was to create and investigate the therapeutic effect of application of complex phytomineral preparation using local components: opoka of Taskala deposit and decoction of camelthorn (Alhagi maurorum) for treatment of diarrhoea in calves. The therapeutic effect of the preparation was studied in the conditions of dairy farm in Kyzylorda region on 5-7-day-old calves. The effect of the preparation was studied in a comparative aspect with an analogous preparation of imported production and the results obtained from healthy animals. Clinical, haematological, and biochemical methods of investigation were employed. The findings obtained suggest a high therapeutic effect of the created preparation. Its application allowed reducing the duration of the treatment period by 5-7 days compared to the imported preparation and restoring the positive dynamics of average daily gain as early as on the 5th day after the start of treatment. Haematological studies indicate stimulation of the immune system in animals that was manifested by a 14% increase in the count of lymphocytes and a 37% increase in the proteins of the γ-globulin fraction compared to healthy animals. These indices suggest an increase in the activity of the humoral part of the immune system in animals after treatment with the preparation. Another positive therapeutic effect of the preparation was observed in the protective effect on the liver of diseased animals. Despite the present effect of both preparations, the decrease in the activity of markers characterising hepatocyte destruction was higher with the Kazakh preparation. Thus, the created preparation is characterised by a higher therapeutic effect in comparison with the imported preparation and allow recommending it for treatment and prevention of diarrhoea in young animals

Effects of Alhagi maurorum Medik polysaccharide derived from different regions on the intestinal immune functions of lambs.

Introduction: Plant polysaccharide are widely studied as potential prebiotics because of their potential to protect and enhance the immunity of lambs. Methods: In this study, the polysaccharide content of Alhagi maurorum Medik from Aksu (AK) and Shanshan (SS) at different cutting periods was determined, and the functions of Alhagi maurorum Medik polysaccharide were investigated to useas an immunomodulator. Results: Our results indicated that the content of Alhagi maurorum Medik polysaccharide is the highest at the maturity stage, and the polysaccharide content of Alhagi maurorum Medik produced in Shanshan area is higher as compared to the Aksu area. The serum IgG, duodenum IgA, TNF-α, IL-4, IL-10 contents, jejunum IgA, TNF-α, IL-4, IL-17 contents, ileum IgA, IL-17 contents, duodenum villus height, crypt depth and jejunum crypt depth of lambs were significantly adjusted in the SS group as compared to CK control group and AK groups (p < 0.05). Furthemore, the sequencing results showed that SS polysaccharide promoted the release of large amounts of IgA and enhanced the immunal function of intestine by regulating the IgA production pathway and B-cell receptor signaling to activate B cells in the T-dependent pathway. Discussion: Altogether, Alhagi maurorum Medik polysaccharide from SS group holds a promising potential to be used as a valuable immunopotentiator for optimizing the immune system of intestine in lambs.

Effect of Addition Aqueous Extract of Camel Thorn (Alhagi maurorum) Plant Roots and Some of Immune Enhancers with Drinking Water on some Hematological, Biochemical and Immunological Characteristics of Broilers

Effect of Addition Aqueous Extract of Camel Thorn (Alhagi maurorum) Plant Roots and Some of Immune Enhancers with Drinking Water on some Hematological, Biochemical and Immunological Characteristics of Broilers

Effect of Addition Aqueous Extract of Camel Thorn (Alhagi maurorum) Plant Roots and Some of Immune Enhancers with Drinking Water on Productive Performance of Broilers

Abstract The present study was carried out in a chicken farm owned by the Animal Production Department of the Faculty of Agriculture at the University of Kufa. Utilizing a cohort of 300 broiler chicks (Ross 308) at the age of one day. The chicks were of undetermined sex and had an average starting weight of 37 grams. They were raised in a semi-enclosed home divided into pens of 3 square meters each. The chicks were randomly allocated to five treatments, with three replicates each treatment. Each treatment consisted of 60 chicks, with 20 chicks per replication, for a period of 35 days. The therapies were categorized as follows: Treatment 1 (T1) refers to the control group where no additional substances are added. Treatment 2 (T2) involves adding Immuno-Care to the drinking water at a concentration of 0.4 ml per litre. Treatment 3 (T3) involves adding Immunity-Stim to the drinking water at a concentration of 1 ml per litre. T4: Incorporate a 5 ml/1 litre solution of the aqueous extract derived from the roots of the camel thorn plant into the drinking water. T5: Introduce a 10 ml/1 litre solution of the aqueous extract obtained from the roots of the camel thorn plant into the water. The young birds were provided with a beginning diet from 1 to 10 days old, a grower diet from 11 to 24 days old, and a finisher diet from 25 to 35 days old. The data pertaining to the examined features were evaluated utilizing a totally randomized design. The significance of the disparities between the treatments was assessed by doing the Duncan test at a probability threshold of 0.05. The SAS statistical program was employed for the purpose of conducting the statistical analysis. The key findings were as follows: There were no notable disparities seen in the ultimate live body weight, cumulative weight growth, dressing %, or economic figure. The final feed conversion ratio showed substantial improvement when the aqueous extract of camel thorn plant roots was added at a concentration of 10 ml per liter of drinking water.

Assessing the compost of Prosopis farcta and Alhagi maurorum as organic fertilizer for potato growth and yield (Solanum tuberosum L.)

Growing potatoes requires great care because it needs large nutrients. Proper fertilization is a critical aspect of improving yield. This study comprises a field experiment to assess the impact of several fertilization approaches on the crop performance of potato (cv. Montreal). In this context, we used eight distinct fertilizer treatments: control (T0), standard mineral fertilizer (T1), substance mushroom spent (T2), cow + poultry manure (T3), compost (T4), 50% standard mineral fertilizer comprising substance mushroom spent (T5), 50% standard mineral fertilizer augmented with cow + poultry manure (T6) and 50% standard mineral fertilizer augmented with compost (T7). The results indicate that control treatment gave least days to mean germination time. While the standard mineral fertilizer (T1) had the highest values in chlorophyll content in leaves, single tuber weight, tuber count per plant, and plant yield. On the other hand, (T6) showed significant increment in number of tubers per plant. Meanwhile, (T7) treatment produced higher marketable yield and tuber per plant. Whereas, there are not significant difference among treatments in number of arial stems. on the other hand, these results were the lowest for the control treatment. Hence, the fertilization approach extensively impacts vegetative development and tuber yield.

Physicochemical and Rheological Characterization of a Novel Manna Exudate from Alhagi pseudalhagi (Iranian Tarangabin)

Tarangabin manna (TM) is a resinous substance having a yellowish sticky character with a reasonably sweet taste. It is largely collected in Iran and Afghanistan. This study for the first time presents a comprehensive investigation of the techno-functional, rheological, and interfacial characteristics of water-soluble components for TM. The composition analysis revealed protein, moisture, fat, ash, and carbohydrate contents of 1.58, 2.98, 0.51, 2.04, and 92.90%, respectively. The effects of TM concentration on the physicochemical, structural, rheological, interfacial, emulsion, and foaming ability and stability were evaluated. X-ray diffraction analysis showed an amorphous structure for the purified sample and a crystalline structure for the raw sample. TM solutions exhibited Newtonian behavior, with the apparent viscosity decreasing as temperature increased, fitting well with the Arrhenius model. The TM solutions exhibited weak viscoelastic properties, primarily demonstrating a dominant viscous character. The surface tension and interfacial tension of the TM solution prepared at a concentration of 50% were measured at 45.23 mN/m and 7.74 mN/m, respectively. The contact angle of the dry thin layer of TM was determined to be 31.74°. Remarkably, the TM solution at a concentration of 50% exhibited the highest foaming ability (76.80%), foaming stability (91.92%), and emulsifying activity index (24.53%). The findings, coupled with TM appropriate foaming ability and stability, sweetness, and characteristic flavor, suggest that TM holds potential as a special food ingredient.

Structure characterization and intestinal immune promotion effect of polysaccharide purified from Alhagi camelorum Fisch

This study investigated the structure of acid Alhagi camelorum Fischa polysaccharide (aAP) and its impact on intestinal activity in mice. The results showed that aAP comprised of the fucose, arabinose, rhamnose, galactose, glucose, xylose, mannose, galacturonic acid, glucuronic acid with the molar ratio of 0.81:14.97:10.84:11.14:3.26:0.80:0.80:54.92:2.47 with the molecular weight (Mw) of 22.734 kDa. Additionally, the composition of aAP was assessed via FT-IR, methylation, and NMR analyses, indicating that the backbone of the aAP was consisted of →4)-α-D-GalpA-6-OMe-(1 → 4)-α-GalpA-(1 → and →4)-α-D-GalpA-6-OMe-(1 → 2)-α-L-Rhap-(1→, as well as →4)-β-D-Galp- and →5)-α-L-Araf- for the branched chain. Furthermore, ICR mice underwent intragastric administration of different concentrations of aAP for 7 consecutive days. The results showed that aAP enhanced the murine spleen and thymus indices, promoted the secretion of serum lgG antibody, intestinal lgA antibody and intestinal cytokines, improved the morphology of intestinal villi and crypts, enhanced quantity of intestinal IELs and IgA+ cells, and activated T lymphocytes and DC cells in MLNs. In summary, these findings suggest that the utilization of aAP could enhance the immune response of the murine intestinal mucosa.

Vegetation Analysis in the Archaeological Area of Pasargadae WHS (Iran) Enhancing the Naturalistic Value of the Site within the Occurring Environmental Changes

This study provides pioneering research on the vegetation of archaeological areas in Iran to enhance its naturalistic and bioindication values by selecting the Pasargadae World Heritage Site (WHS). Vegetation surveys were carried out in different homogeneous habitats, analyzing the plant communities through statistical elaboration, syntaxonomic role, mapping, and enhancement of plants with conservation interest. In an ecological approach, the study included an analysis of the recent climate changes and human interventions influencing the water resources. Results revealed seven main vegetation types reflecting ecological gradients shaped by environmental, edaphic, and anthropogenic factors. The syntaxonomic analysis showed a primary subdivision in semi-natural grasslands and synanthropic vegetation. Several key species were identified as bioindicators of multiple factors, such as: Launaea acanthodes, Stipa barbata, Alhagi maurorum, Bellevalia saviczii, Glycyrrhiza glabra, Convolvulus arvensis, and Hordeum murinum. The vegetation map showed how the hilly grassland communities hosted the highest number of species with conservation interest and their need to be better protected. Bio-climatic data, such as the construction of dams and the exploitation for irrigation purposes, pointed to the increasing xeric conditions, which make urging conservation efforts for the site’s historical and naturalistic values. The study underscores the importance of preserving places with high plant diversity for effective site management, and enhances the intricate relationship between vegetation and natural features in the occurring environmental changes.

Effects of Alhagi camelorum Fisch polysaccharide from different regions on growth performance and gastrointestinal microbiota of sheep lambs.

Polysaccharides derived from Alhagi camelorum Fisch possess diverse activities, making them a potential prebiotic candidates for enhancing lamb health. This study investigated the immunomodulatory effects of Alhagi camelorum Fisch polysaccharides from Aksu (AK) and Shanshan (SS) regions on sheep lambs. The results showed that sheep lambs in the SS group exhibited significantly increased (p < 0.05) average daily gain, levels of growth hormone (GH), insulin (INS), IgA and IgM, and cytokines IL-4, IL-10, IL-17, TNF-α and IFN-γ compared to those in the control check (CK) group. Moreover, the SS treatment significantly increased the diversity and abundance of beneficial bacteria, while concurrently diminishing the prevalence of harmful bacteria. Additionally, it modulated various metabolic pathways, promoted lamb growth, improved immunity, reduced the risk of gastrointestinal disease and improved the composition of gastrointestinal microbiota. In summary, our findings highlight the potential of SS treatment in enhancing gastrointestinal health of sheep lambs by improving intestinal function, immunity, and gut microbiome. Consequently, these results suggest that Alhagi camelorum Fisch polysaccharides derived from Shanshan regions holds promising potential as a valuable intervention for optimizing growth performance in sheep lambs.

Current Status and Resources of Alhagi pseudalhagi (Fabaceae) in the Atyrau Region, Western Kazakhstan

Alhagi pseudalhagi, which grows in the arid zone of the Atyrau region, and an assessment of the current state of its raw material reserves were studied. Botanical characteristics, ontogenetic spectra, morphological indicators, productivity, areas of thickets, and reserves of raw materials of above-ground organs were assessed. The structural parameters of these populations in the Zhangyr and Coneu Rivers valleys, in the vicinity of Imankara Mountain, and on the Taisoigan sands were studied. It was established that the species composition of the populations includes 63 species from 54 genera and 30 families. The most common species include 49 species, which, according to their occurrence in populations of A. pseudalhagi, are distributed in the following classes: 17 species—class II (0–20%); 7 species—class III (41–60%); and 2 species—classes IV (61–80%) and V (81–100%). The maximum similarity in species composition was noted between populations in the Zhangyr and Coneu Rivers valleys. The highest morphometric indicators were observed among the population of the Coneu river valley, and the lowest are located on the Taisoigan sands. Analysis of the age spectra made it possible to determine that the populations in the Zhangyr and Coneu River valleys are characterized as young and those in the area of Imankara Mountain and on the Taisoigan sands as stable and middle-aged.

Protective Effect of Mesenchymal Stem Cell Active Factor Combined with Alhagi maurorum Extract on Ulcerative Colitis and the Underlying Mechanism.

Ulcerative colitis (UC) is a relapsing and reoccurring inflammatory bowel disease. The treatment effect of Alhagi maurorum and stem cell extracts on UC remains unclear. The aim of the present study was to investigate the protective role of Alhagi maurorum combined with stem cell extract on the intestinal mucosal barrier in an intestinal inflammation mouse model. Sixty mice were randomly divided into a control group, model group, Alhagi group, MSC group, and MSC/Alhagi group. MSC and Alhagi extract were found to reduce the disease activity index (DAI) scores in mice with colitis, alleviate weight loss, improve intestinal inflammation in mice (p < 0.05), preserve the integrity of the ileal wall and increase the number of goblet cells and mucin in colon tissues. Little inflammatory cell infiltration was observed in the Alhagi, MSC, or MSC/Alhagi groups, and the degree of inflammation was significantly alleviated compared with that in the model group. The distribution of PCNA and TNF-alpha in the colonic tissues of the model group was more disperse than that in the normal group (p < 0.05), and the fluorescence intensity was lower. After MSC/Alhagi intervention, PCNA and TNF-alpha were distributed along the cellular membrane in the MSC/Alhagi group (p < 0.05). Compared with that in the normal control group, the intensity was slightly reduced, but it was still stronger than that in the model group. In conclusion, MSC/Alhagi can alleviate inflammatory reactions in mouse colonic tissue, possibly by strengthening the protective effect of the intestinal mucosal barrier.

Phytoremediation of pollutants in oil-contaminated soils by Alhagi camelorum: evaluation and modeling

Phytoremediation is a cost-effective and environmentally friendly method, offering a suitable alternative to chemical and physical approaches for the removal of pollutants from soil. This research explored the phytoremediation potential of Alhagi camelorum, a plant species, for total petroleum hydrocarbons (TPHs) and heavy metals (HMs), specifically lead (Pb), chromium (Cr), nickel (Ni), and cadmium (Cd), in oil-contaminated soil. A field-scale study spanning six months was conducted, involving the cultivation of A. camelorum seeds in a nursery and subsequent transplantation of seedlings onto prepared soil plots. Control plots, devoid of any plants, were also incorporated for comparison. Soil samples were analyzed throughout the study period using inductively coupled plasma-optical emission spectroscopy (ICP‒OES) for HMs and gas chromatography‒mass spectrometry (GC‒MS) for TPHs. The results showed that after six months, the average removal percentage was 53.6 ± 2.8% for TPHs and varying percentages observed for the HMs (Pb: 50 ± 2.1%, Cr: 47.6 ± 2.5%, Ni: 48.1 ± 1.6%, and Cd: 45.4 ± 3.5%). The upward trajectory in the population of heterotrophic bacteria and the level of microbial respiration, in contrast to the control plots, suggests that the presence of the plant plays a significant role in promoting soil microbial growth (P < 0.05). Moreover, kinetic rate models were examined to assess the rate of pollutant removal. The coefficient of determination consistently aligned with the first-order kinetic rate model for all the mentioned pollutants (R2 > 0.8). These results collectively suggest that phytoremediation employing A. camelorum can effectively reduce pollutants in oil-contaminated soils.

Studying the Response of Anethum graveolens L. to the Stress of Salinity and Foliar Spray with Alhagi Maurorum Aqueous Extract

Background Weed infestation and salinity are both biotic and abiotic stresses that are highly present in the Basrah governorate in Iraq and widely influence crop growth Methods The present work was conducted to evaluate the effect of salinity at (0, 6, 12 dS/m) and aqueous extract of Alhagi maurorum at (0, 3 g/L) on growth and biochemical parameters as well as phytocomponents of Dill plant Anethum graveolens L. Results Results revealed that growth parameters and photopigment content increased with salinity at 12 dS/m and reduced with Alhagi maurorum aqueous extract. Besides that, carbohydrate content was reduced with both stresses, and proline content increased with Alhagi maurorum aqueous extract. GC-MS chromatogram of Anethum graveolens methanolic extract displayed high alteration in phytocomponents quantity post salinity and camelthorn extract treatments. Conclusion Current work inferred that the Dill plant has high tolerance properties to salinity contrary to weed infestation stress. Stresses combination declined growth parameters and reduced metabolite content of Dill plants.

Joint action of malathion with herbicides against Alhagi pseudalhagi (Bieb.) Desv.

Joint action of malathion with herbicides against Alhagi pseudalhagi (Bieb.) Desv.

Analysis of the amount of flavonoids contained in the surface part of the Alhagi maurorum by chromatographic method

The article provides information about the plant Alhagi maurorum and its chemical structure, pharmacological properties, place in folk medicine, and the high-performance liquid chromatography (HPLC) method used in its analysis. When the leaves, stems, and seeds of Alhagi maurorum were analyzed by chromatography, dehydroquercetin, rutin, quercetin, lutein, and seneroside substances belonging to the class of flavonoids were determined. These flavonoids are natural antioxidants. Their beneficial properties in medicinal plants have been studied in the literature review. Alhagi maurorum above-ground parts of leaves, stems, and seeds were recommended to be used in a generalized state.