Fresh Plant Samples Research Articles

Transformation of heavy metals from contaminated water to soil, fodder and animals

A serious environmental problem that threatens soil quality, agricultural productivity, and food safety is heavy metal pollution in water sources. Heavy metal pollution is the main problem in tehsil Pasrur, Sialkot, Pakistan. Present study was arranged to notice the heavy metals in water, soil, forages and buffalo milk. There are seven sites that were used for this experiment. Highest malondialdehyde (MDA) contents (3.00 ± 0.01) were noticed in barseem roots at site 7. Ascorbate Peroxidase (APX) was reached at its peak (1.93 ± 0.01) at site 7 in the fresh barseem. Maximum protein contents (0.36 ± 0.01) were observed in fresh plant samples at site 2. Site 3's buffalo milk samples had the highest Ni content (7.22 ± 0.33 ppm), while Site 3's soil samples had the lowest Cr content (8.89 ± 0.56 ppm), Site 1's plant shoots had the lowest Cr content (27.75 ± 1.98 ppm), and Site 3's water had the highest Cr content (40.07 ± 0.49 ppm). The maximum fat content (5.38 ± 2.32%) was found in the milk of the animals at site 7. The highest density (31.88 ± 6.501%), protein content (3.64 ± 0.33%), lactose content (5.54 ± 0.320%), salt content (0.66 ± 0.1673%), and freezing point (− 0.5814 ± 0.1827 °C) were also observed in the milk from animals at site 7, whereas site 5 displayed the highest water content (0.66 ± 0.1673%) and peak pH value (11.64 ± 0.09). In selected samples, the pollution load index for Ni (which ranged from 0.01 to 1.03 mg/kg) was greater than 1. Site 7 has the highest conductivity value (5.48 ± 0.48). Values for the health risk index varied from 0.000151 to 1.00010 mg/kg, suggesting that eating tainted animal feed may pose health concerns. Significant health concerns arise from metal deposition in the food chain from soil to feed, with nickel having the highest health risk index.

DNA EXTRACTION FROM LOCAL SOUVENIR OF SUKU ANAK DALAM JAMBI WITHOUT LIQUID NITROGEN USING CTAB METHOD

One of the local souvenirs from Jambi is accessories made from the seed bark of the sebalik sumpah tree. The sebalik sumpah tree is currently quite rare. Through DNA isolation, sebalik sumpah is one of the efforts of genetic conservation and as a first step in cultivating sebalik sumpah trees. DNA isolation is the initial stage in molecular analysis. The purpose of DNA isolation is to obtain genomic DNA from a sample. Plant DNA isolation generally uses fresh plant samples, with young and soft plant parts. This is recommended to obtain good quality genomic DNA. However, with the limitations of fresh samples, a method is needed that is able to extract DNA from plants that have changed shape and have undergone processing. Each plant requires a different method to obtain DNA. The CTAB (Cethyl Trimethyl Ammonium Bromide) buffer method is a method commonly used in plants because it is able to extract DNA with high polysaccharide and phenol content, so many have modified the method. This study aims to determine the success of the CTAB method without nitrogen in extracting processed materials and the length of incubation time at the precipitation stage. Extraction was carried out by liquid nitrogenless CTAB method with the addition of PVP (Polyvinylpolypyrrolidone). The results show that the CTAB method without liquid nitrogen is able to extract plant samples that have undergone processing and the length of incubation time at the precipitation stage affects the DNA produced.

Tracking and imaging nano-plastics in fresh plant using cryogenic laser ablation inductively coupled plasma mass spectrometry

Tracking and imaging of nano-plastics are extremely challenging, especially in fresh biological samples. Here, we propose a new strategy in which polystyrene (PS) was doped with the europium chelate Eu (DBM)3bpy to quantify, track, and in situ image nano-plastics in fresh cucumber based on inherent metals using cryogenic laser ablation inductively coupled plasma mass spectrometry (cryo-LA-ICP-MS). The cryogenic conditions provide a stable condition for imaging fresh cucumber, suppressing the evaporation of water in fresh plants, and maintaining the original structure of plants with respect to room temperature imaging in LA-ICP-MS. The plants were cultivated in two types of nano-plastics solutions with low (50 mg/L) and high (200 mg/L) concentrations for 9 days. The results showed that nano-plastics mainly enrich the roots and have negative effects, which decrease the trace elements of Zn, Mn, and Cu in cucumber. Smaller PS particles are able to penetrate the plant more easily and inflict serious damage. Novel imaging method provides a novel insight into the tracking and imaging of nano-plastics in fresh plant samples. The results illustrated that nano-plastics deposition on plants has the potential to have direct ecological effects as well as consequences for potential health.

Comparison of genetic characteristics among species of Suaeda spp. (Amaranthaceae) in the western Anbar plateau - Iraq

Background: Comparison of genetic characteristics among species of Suaeda spp.
 Objective: identifying the genetic characteristics among species Suaeda (Amaranthaceae) in Anbar Governorate. (From August 2020 to September 2021 fresh plant samples were collected in the flowering stage for the studied species).
 Materials and methods: The genetic diversity of the Suaeda species was studied after DNA extraction and using the Inter Simple Sequence Repeats Reaction method to recorded in the genetic aspects. DNA was extracted from young leaves of the studied species. The concentration and purity of the DNA were determined, and the detection of genetic relationships between the studied plant species as well as finding this genetic fingerprint by ISSR Reaction.
 Results: The results showed that the plant samples collected during study belong to species of the genus Suaeda, and these species were: S. aegyptiaca, S. altissima, S. carnosissma, S. fruticosa, S. monoica, S. vera and S. vermiculata symbolized by (S1, S2, S3, S4, S5, S6, S7) respectively for the purpose of brevity. The results showed that there was the highest close between S. aegyptiaca and S. vera which amounted to 0.6319, and the results showed that there was the least closeness between S. aegyptiaca and S. carnosissma which amounted to 0.4231.
 Conclusion: Genetic traits are among the stable traits that can be adopted in separating the studied species. The genetic study, especially at the DNA level, is one of the most important modern taxonomic studies that rely on PCR technology for the accuracy of its results and the speed of obtaining results.There are at least seven species of the genus Suaeda in the western province of Anbar Governorate - Iraq.

Antibacterial Activity of <i>Phyllanthus</i><i> </i><i>Amarus</i> (Schum and Thonn) Extract Against <i>Salmonella </i><i>Typhi</i> Causative Agent of Typhoid Fever

The study was conducted to assess the antibacterial activity of Phyllanthus amarus (Schum and Thonn) extract against Salmonella typhi causative agent of typhoid fever at the laboratories of the Departments of Chemistry and Theoretical and Applied Biology of the College of Science, Kwame Nkrumah University of Science and Technology, Kumasi. The objectives were to determine the highest yield of crude extract of P. amarus using different proportions of water to ethanol and to determine the sensitivity of Salmonella typhi to these. Three different extraction procedures were carried out. In the first procedure, seven extraction setups each containing different proportions of the two extract (water and ethanol) were used with 10g of the plant sample. In the second procedure, eight setups were used for the two solvents. Ten grams of both fresh and dry plant sample were extracted in two different 200ml of water and in another two different 200ml of water; 20g of both fresh and dry plant sample were again extracted. The same procedure was repeated using ethanol as the solvent. In the third procedure, 10g each of fresh plant sample were boiled in 100ml and 200ml of water for 30 minutes. A sensitivity test to determine the zones of inhibition for the various plant extracts was done on Salmonella typhi isolated from human. Results from the crude yield of P. amarus using water only had the highest crude yield of 2.57g, followed by ethanol only which was 2.52g. The sensitivity studies conducted on the fresh P. amarus indicated that aqueous extract of P. amarus inhibited S. typhi to a zone of 5.00mm in 10g/200ml and 7.17mm in 20g/200ml. Ethanol extract also recorded an inhibition zone of 2.67mm and 5.33mm in 10g/200ml and 20g/200ml respectively. Again, sensitivity studies using dry P. amarus samples showed that the aqueous extracts recorded a zone of inhibition of 7.33mm in 10g/200ml and 13.50mm in 20g/200ml. Also ethanol extracts also recorded an inhibition zone of 6.83mm in 10g/200ml and 10.50mm in 20g/200ml. Significant differences were observed among the extracts and the control in both 10g/200ml and 20g/200ml concentrations (P<0.05). Aqueous and ethanol extracts of P. amarus proved inhibitory to S. typhi.

Impact of Drying Methods on Qualitative and Quantitative of Monarda citriodora Cerv. ex Lag. Essential Oil Yield

A genus of Monarda belongs to the family (Lamiaceae), the largest group of angiosperms. The crop has been grown extensively across the world for its flavoring and therapeutic properties as well as for its high-quality essential oil. The present research was designed to study the impact of different drying methods i.e (shed drying, sun-drying, microwave, oven-drying at (40°, 50°, and 60° C), freeze-drying (0°, -10°, and -20° C), and vacuum drying (30°, 45° and 60° C) on the qualitative and quantitative analysis of Monarda citriodora essential oil. Dried and fresh plant samples were hydro-distilled using the clevenger apparatus, and essential oil analyzed by Gas Chromatography (GC) and Gas Chromatography mass spectrophotoscopy (GC/MS). The results showed that different drying methods had significant effects on essential oil yield as shed-drying (2.6 ml) >freeze-drying (2.2 ml at 0°C) >oven drying (2.1 ml) >fresh drying (2.0 ml) >sun drying (2.0 ml) >microwave (0.15 ml). The major compounds obtained by GC/MS were thymol (92.46%) >g-terpinene (10.48%) >o-cymene (9.31%) >carvacrol (8.88%) >cyclohexene (3.51%). The highest essential oil yield was observed under shed dried (2.6 ml) and the major essential oil compound, thymol (92.46%), was observed under microwave-2 (4.0 min, 4.0 min, 4.0 min, and 3.0 min) combination. The results show an inversely proportional correlation between essential oil compounds. Thus, it has been summarized that the M. citriodora plant dry under the shed-drying method was the most appropriate and suitable method for obtaining a high-quality essential oil yield and microwave-2 (MV-2) combination for oil composition.

Evaluating the efficacy of Capparis spinosa Total flavonoids to control Bird Lice (Menacanthus stramineus)

The present study included an in-vitro and in-vivo insecticidal investigation for one of wild Iraqi plant Capparis spinosa total flavonids on locally chicken lice (Menacanthus stramineus). Extraction of total flavonoids from 115g of fresh plant samples by reflex extraction with 600ml distilled water with (10% v/ v) HCl, for 8 hours continuously and the aglycon part was obtained with ethyl acetate. The acetate layer was dried and the residue was weighted and subjected for qualitative and quantitative analysis. Three flavonoids concentrations of Capparis spinosa total Flavonoid (5,3 and 1) mg/ml in 100ml distilled water to treat a collected sample of Bird Lice (Menacanthus stramineus) in vitro and in vivo at different life stages in comparison to a traditional anti lice drug treatment permethrin (0.5mg/ml) as positive control. Distilled water application was considered as negative control. Results of present study showed that the plant was rich with different types of flavonoids and the effect of flavonoid extracts of the Capparis spinosa plant in the decimation of the different stage of chicken body lice, as the concentration factor had non-significant effect on the killing of adult and a significant effect on the killing of eggs/nymph, and this effect increased with the increase in the concentrations of the extract.

RbcL, a potential candidate DNA barcode loci for aconites: conservation of himalayan aconites.

Aconitum heterophyllum Wall. ex Royle and Aconitum balfourii Stapf, are two highly important, threatened medicinal plants of the Indian Himalayan Region. Root-tubers of Aconites have occupied an important place in Indian pharmacopoeia from very ancient times. India is a hub of the wild-collected medicinal herbs industry in Asia and these two aconites are known to have been heavily traded from the region in illicit manner. Prosecution of these illegal trading crimes is hampered by lack of pharma-forensic expertise and tools. Present study was conducted to evaluate the discriminatory potential of rbcL, a Chloroplast based DNA barcode marker for the authentication of these two Himalayan Aconites. Fresh plant samples were collected from their natural distributional range as well as raw materials were procured from herbal market and a total of 32 sequences were generated for the rbcL region. Analysis demonstrated that rbcL region can successfully be used for authentication and importantly, both the aconites, were successfully discriminated by rbcL locus with high bootstrap support (> 50%). Molecular markers could certainly be relied upon morphological and chemical markers being tissue specific, having a higher discriminatory power and not age dependent. Phylogenetic analysis using Maximum Likelihood Method revealed that the rbcL gene could successfully discriminate Himalayan Aconites to species level and have potential to be used in pharma-forensic applications as well as to curb illicit trade of these invaluable medicinal plants.

Polishing entire stems and roots using sandpaper under water: An alternative method for macroscopic analyses.

PremisePolishing entire stem and root samples is an effective method for studying their anatomy; however, polishing fresh samples to preserve woods with soft tissues or barks is challenging given that soft tissues shrink when dried. We propose sanding fresh or liquid‐preserved samples under water as an alternative, given that it preserves all tissues in an intact and clear state.Methods and ResultsBy manually grinding the surface of the samples under water using three ascending grits of waterproof sandpapers, an excellent polished sanded surface is obtained. The wood swarf goes into the water without clogging the cell lumina, rendering the surfaces adequate for cell visualization and description. We show results in palms, liana stems, roots, and wood blocks.ConclusionsUsing this simple, inexpensive, rapid technique, it is possible to polish either fresh, dry, or liquid‐preserved woody plant samples, preserving the integrity of both the soft and hard tissues and allowing for detailed observations of the stems and roots.

Some regularities in the process of anthocyanin extraction from vegetable sources

This work investigates regularities in the process of extracting anthocyanins from various plant sources in the medium of extractants. For extraction, fresh plant samples were used, including the roots of purple carrots, roots of purple potatoes of the Amethyst variety, fresh fruits of Michurin's aronia, fruits of the garden nightshade, cornelian cherries, blueberries and red cabbage, as well as dried peony petals. The extraction was carried out by infusing the plant material under study in a selected extractant (left overnight). The concentration of anthocyanins was determined spectrophotometrically. It was shown that a 0.1 M aqueous solution of HCl is an effective and environmentally friendly extractant allowing for the extraction of anthocyanins from various sources. Provided that the medium acidity is greater than 1, significant losses of anthocyanins are possible: 5-45% at pH = 2; 33-88% at pH = 3; 41-92% when extracted with distilled water. Acylation does not contribute to an increase in the degree of anthocyanin extraction. The addition of organic solvents (ethanol, acetonitrile and glycerin) can significantly accelerate the anthocyanin extraction in some cases, which is particularly true for cornelian cherries. When using crushed material, the influence of the extractant on the degree of extraction is minimal. Information is presented on the solvatochromic effect, which affects the spectral characteristics of some anthocyanin solutions. It was shown that the shift of the absorption band maximum is the greatest for non-acylated anthocyanins, while decreasing for acylated compounds. The latter is most likely to be associated with the intramolecular copigmentation of acylated anthocyanins in aqueous solutions. When determining the concentration of anthocyanins in solvents with different concentrations of organic additives, the solvatochromic effect should be taken into account. The addition of an organic solvent leads not only to a shift in the absorption band maxima, but also to significant hyperchromic effects. If this effect is ignored, errors in determination of anthocyanins can exceed 70%. This paper proposes a simple and effective approach to considering these effects using a cross-dilution scheme. Therefore, the choice of extractants for efficient anthocyanin extraction from various plant raw materials was experimentally substantiated, and a method for evaluating solvatochromic effects was proposed.

BIOCHEMICAL CHARACTERIZATION OF Ferula rutbaensis MEDICINAL PLANT IN IRAQI WESTERN DESERT

Plants used in folk medicine not only represent rich sources for therapeutic materials, but it also plays a crucial role in developing completely or partially novel synthesized drugs. Mharut plant (Ferula rutbaensis) is an integral part of Bedouin therapeutic practices in the western desert of Anbar province-Iraq. Still, to date, this is the first study describing its phytochemical constituents. The plant was growing near the Iraq-Saudi Arabia borders and adapted to a wide range of soils. Traditionally, F. rutbaensis has been widely used to treat acne, stomach and bowel disorders, food poisoning and respiratory problems. Fresh plant samples were collected and morphologically characterized. Likewise, the ITS-based DNA barcoding technique was efficiently used to approve the morphological identification of F. rutbaensis. The GC-MS spectrum was adopted in the phytochemical characterization of aqueous and methanol extracts of fresh and dry plant parts. The aqueous extract of dry roots was the richest source for bioactive compounds than fresh or methanolic extracts of either fresh or dry plant parts. In general, the detected phytochemicals falling into fatty acids, terpenes, hydrocarbon alkanes, and esters. Notably, fatty acids in Oleic and Palmitic acids were the two most abundant bioactive compounds in both aqueous and methanolic extracts of plant fresh and dry roots. The detected unsaturated fatty acids and/or other bioactive components are laying behind the therapeutic properties of F. rutbaensis that can be useful ingredients to prepare Mharut-based cosmetics such as medical soaps, body lotions, skin conditioners and sunscreens. Additionally, some other components were found to have anti-inflammatory, antioxidants, and antimicrobial properties. Further investigations will be necessary to confirm the antimicrobial activity of F. rutbaensis extracts.

Restoration of Water Storage Potential in a Degraded Dry Dipterocarp Forest with Enrichment Planting of Three Needle Pine (Pinus kesiya Royle ex Gordon), Northern Thailand

The research assessed water storage in a dry dipterocarp forest (DDF) with enriched 34-year-old pine planting and the role of pine. Plant surveys were carried out using 10, 40×40 m2 plots, and data were obtained by measuring tree stem girths and heights. Plant features, biomass, and stored water amounts were measured. Fresh plant samples of abundant species were taken one time per month from January to December 2018. Three soil pits were made in three plots, and soil samples along 100 cm depth were taken on the same days of collecting plant samples for studying fied capacity, water content and water amount. The DDF was divided into three stands based on the most dominant tree species; Shorea obtusa, Dipterocarpus tuberculatus, and Dipterocarpus obtusifolius. The forest was composed of 86 species with biomass at 101.62 Mg/ha and contained an average water amount of 88.01 m3/ha. The water amount in biomass varied with sampling times from 58.74 to 111.83 m3/ha. The average MWHC of 100 cm soil was estimated to be 5,113.74 m3/ha. The water amount in soil also varied with sampling times from 3,651.50 to 4,481.06 m3/ha. As a result, the total water amount in plant biomass and soil (ecosystem) of the DDF varied in a range from 3,735.0 to 4,558.67 m3/ha. The pine contributed to 30.87 m3/ha (35.07% of the total) and could increase by 64.92% the water storage potential of the forest, and thus these results support the concept of pine enrichment planting in the poor DDF.

Nutritional Composition of ‘Aju Mbaise’ Herbal Cocktail

This study was carried out to evaluate the nutritional content of ‘Aju Mbaise’ herbal mixture. The experiment involves the collection of fresh plant samples that make up ‘Aju Mbaise’ cocktail. The study was done in the Research Laboratory of the Department of Biochemistry, Faculty of Science, University of Port Harcourt, Nigeria. The plants samples were collected from Mbaise locality in Imo State, Nigeria, and were identified as Cnestis ferruginea, Xylopia aethiopica, Uvaria chamae, Palisota hirsuta, Scleria sp., Napoleona imperialis, Dialium guineense, Combretum racemosun, and Heterotis rotundifolia respectively. The dietary, mineral, and vitamin compositions were determined accordingly. The proximate, vitamin, and mineral composition of the herbal cocktail were investigated in line with their standard methods of analysis. The proximate analysis revealed that the cocktail extract contains carbohydrate (69.51%), crude protein (10.05%), moisture (8.89%), crude fat (5.17%), fibre (3.745), and ash content (2.65%). The vitamin analysis revealed reasonable concentration of vitamins A, B1, B2, B3, B6, B12, C, D, and K. The mineral evaluation showed high concentration of calcium, magnesium, potassium, sodium, zinc, and iron. This study revealed high nutritional value of ‘Aju Mbaise’ herbal cocktail which is responsible for the plant’s usefulness in the management, treatment, as well as maintenance of good health.

Assessment of Proximate, Phytochemical and Selected Mineral Content of Acanthus montanus Leaf

Acanthus montanus (Nees) T. Anders belongs to the family Acanthaceae and is one of the most threatened and underutilized species of vegetables in Africa. However the leaves of this plant are part of a consortium infusion (agabada nkwu) given to post-natal mothers within and around Mbaise in Imo State, South Eastern Nigeria, to ensure health and vitality of both nursing mother and child. The proximate, phytochemical and selected mineral contents of the Acanthus montanus leaf were investigated to ascertain the various components. The fresh plant sample was obtained from Ife, Ezinihitte Mbaise Local Government Area of Imo state, Nigeria. The sample was air-dried at room temperature for 72 hours prior to the various analyses, which were done using standard methods. The results of the phytochemical analysis showed 1.60 % flavonoids, 6.67 % tannins, 5.60 % alkaloids, 6.45 % saponins, 0.26 % phytate, 5.41% oxalate, 0.49 % phenol and 0.019 % Hydrogen Cyanide (HCN). The percentage proximate evaluation for moisture content, carbohydrate, protein, fibre, ash content and fat were: 11.85, 37.86, 17.72, 16.70, 10.56 and 5.31 respectively, while the percentage quantities of selected minerals; iron, magnesium, potassium, calcium, phosphorus, sodium, manganese, zinc and copper were: 0.014, 0.569, 3.152, 0.909, 0.089, 0.202, 0.009, 0.010 and 0.001 respectively. Acanthus montanus is reported for its nutritional and medicinal values throughout Africa. The result of this work suggests further exploitation of the parts of Acanthus montanus to unveil more of its potential uses for the treatment of diseases.

Phytochemical constituents and free radical scavenging activity of fresh and dried samples of Curculigo pilosa (Schum. & Thonn.) Engl. (Hypoxidaceae)

Curculigo pilosa (Schum. & Thonn) Engl. is a tropical African flowering plant belonging to the family Hypoxidaceae. It is a highly valued medicinal plant used in traditional medicine to treat constipation, impotence, limb limpness, arthritis, knee joints, and watery diarrhea. It is also used as a potent immunomodulator and aphrodisiac. The aim of this study is to evaluate the phytochemical constituents and free radical scavenging activity of the fresh and dried samples of C. pilosa. Fresh plant samples were purchased from herbal market at Iyana-Iba market in Ojo local government area. Some of the samples were oven dried at 30 oC to get the dried sample. Phytochemical analysis was carried out using standard laboratory procedures while ferric reducing power, nitric oxide and DPPH scavenging assays were used to evaluate the antioxidant activity of the samples. Both dried and fresh samples contained phenols, flavonoids, saponins, alkaloids and cardiac glycosides. Tannins was present in the extracts except the aqueous extract of the dried sample while terpenoids was present only in the acetone extract of the fresh and dried sample but absent in the aqueous extract of both samples. Fresh sample contained the highest amount of phytochemicals in both extracts. The amount of phenols in aqueous and acetone extracts of the fresh sample was 68.51 mg g-1 and 80.94 mg g-1 respectively. Fresh samples exhibited higher scavenging activity than the dried sample. Fresh sample of C. pilosa contained more phytochemicals and exhibited higher free radical scavenging activity than the dried sample.

Proximate, Mineral and Phytochemical Composition of Dioscorea dumetorum (Bitter Yam) Commonly Cultivated in South Eastern Nigeria

Standard procedures were used to investigate proximate, mineral, amylose content, gelatinization temperature and phytochemical composition of Dioscorea dumetorum (Both cooked and fresh samples). Significant increase (p< 0.05) of protein, fats and oil, moisture and crude fibre contents was observed in the fresh sample as against cooked sample. A value of (43.80%) was recorded in the cooked sample while in the fresh sample the carbohydrate value was 23.15%. There is a significant increase (p< 0.05) of Fe, Mg, Na and P concentration of the fresh sample compared with the cooked sample. Inversely, significant decrease (p> 0.05) of, Mn, Ca. and K contents in the cooked sample was recorded in comparison with the fresh plant sample. Gelatinization temperature increased significantly (p< 0.05) in the cooked sample as against fresh. Non- significant increase (p> 0.05) of the amylose content was documented in the fresh sample compared with cooked sample. Phytochemical screening revealed alkaloid, saponin glycosides were steadily present, flavonoid, tannin and phenol were moderately present in the cooked sample. In the same vein, similar observation was made in the fresh sample. Steroids were not detected in both samples. Medicinal use of the tuber is hereby justified by the results and suggests it could contribute to the nutrition of man and animals. It could be recommended to diabetic patients and obedience owning to its low starch contents.

Simultaneous Determination of Multiclass Phytohormones in Submilligram Plant Samples by One-Pot Multifunctional Derivatization-Assisted Liquid Chromatography-Tandem Mass Spectrometry.

Phytohormones play crucial roles in every aspect of plant life, and their regulatory functions rely on complex crosstalk networks among the different classes of phytohormones in an antagonistic or synergistic manner. Therefore, the simultaneous determination of multiclass phytohormones is important for studies of phytohormone functions and networks. However, due to the heterogeneity of the sensitivity resulting from structural diversity and the low content in the plant samples, simultaneous determination of multiclass phytohormones is challenging, especially in very tiny plant tissues or organs. Here, we describe a novel method for the simultaneous determination of 31 phytohormones from different classes in a single run. This uses a one-pot multifunctional derivatization coupled to liquid chromatography-tandem mass spectrometry (LC-MS/MS). N, N-Diethyl ethylenediamine (DEED) and 2-methyl-4-phenylaminomethyl-benzeneboronic acid (2-methy-4-PAMBA) were used for derivatization of carboxylated phytohormones and brassinosteroids (BRs), respectively, to simultaneously improve detection sensitivities of carboxylated phytohormones and BRs. The method was fully validated for the 31 targeted phytohormones and could quantify multiclass endogenous phytohormones in small amounts of fresh plant samples (0.02-5 mg, FW). Finally, we used this method to investigate the spatial-temporal distribution of multiple phytohormones in reproductive organs of a single flower of Arabidopsis thaliana for the first time. This method could be a powerful auxiliary tool for studies of phytohormone functions and regulatory networks.

Quantification of lignin oxidation products as vegetation biomarkers in speleothems and cave drip water

Abstract. Here we present a sensitive method to analyze lignin oxidation products (LOPs) in speleothems and cave drip water to provide a new tool for paleo-vegetation reconstruction. Speleothems are valuable climate archives. However, compared to other terrestrial climate archives, such as lake sediments, speleothems contain very little organic matter. Therefore, very few studies on organic biomarkers in speleothems are available. Our new sensitive method allows us to use LOPs as vegetation biomarkers in speleothems. Our method consists of acid digestion of the speleothem sample followed by solid-phase extraction (SPE) of the organic matter. The extracted polymeric lignin is degraded in a microwave-assisted alkaline CuO oxidation step to yield monomeric LOPs. The LOPs are extracted via SPE and finally analyzed via ultrahigh-performance liquid chromatography (UHPLC) coupled to electrospray ionization (ESI) and high-resolution Orbitrap mass spectrometry (HRMS). The method was applied to stalagmite samples with a sample size of 3–5 g and cave drip water samples with a sample size of 100–200 mL from the Herbstlabyrinth-Advent Cave in Germany. In addition, fresh plant samples, soil water, and powdered lignin samples were analyzed for comparison. The concentration of the sum of eight LOPs (Σ8) was in the range of 20–84 ng g−1 for the stalagmite samples and 230–440 ng L−1 for the cave drip water samples. The limits of quantification for the individual LOPs ranged from 0.3–8.2 ng per sample or 1.5–41.0 ng mL−1 of the final sample solution. Our method represents a new and powerful analytical tool for paleo-vegetation studies and has great potential to identify the pathways of lignin incorporation into speleothems.

Medicinal plants used in the treatment of diabetes in karo ethnic, north sumatra, indonesia

Medicinal plants derived from traditional medicines have played an important role in managing a variety of healthcare and diseases in Karo ethnic in North Sumatra, Indonesia. The study aimed to document the ethnobotanical information on medicinal plants used by traditional healers of Karo ethnic in the treatment of diabetes and to assess the crude extract of phytochemical constituents qualitatively from medicinal plant organ. The study was conducted on two Karo sub-ethnic living at the highland (Karo Gugung) in Karo Regency and the lowland (Karo Jahe) in Langkat Regency with the length of the study was eight months of observation. The survey was conducted using open-ended interviews among four traditional healers those who were selected by snowball sampling method; quantitative analysis of ethnobotanical data was performed by calculating the familiarity index (Fi). Fresh plant samples which were used for phytochemical analysis were collected using participatory method. The results showed that 15 plants were used to treat diabetes by Karo traditional healers. The plants are Blumea balsamifera, Nypa fruticans, Bischofia javanica, Eleutherine americana, Allium cepa, A. sativum, Eugenia polyantha, Piper betle, P. nigrum, Citrus aurantiifolia, Boesenbergia pandurata, Curcuma longa, Kaempferia galanga, Zingiber montanum, and Z. officinale. Familiarity index (Fi) value of each plant was 25 which explained that each of medicinal plant was used by only one traditional healer. The phytochemical screening showed that the crude plant extracts contained phenolic, terpenoid, steroid, and saponin.

DNA barcoding of selected UAE medicinal plant species: a comparative assessment of herbarium and fresh samples.

It is commonly difficult to extract and amplify DNA from herbarium samples as they are old and preserved using different compounds. In addition, such samples are subjected to the accumulation of intrinsically produced plant substances over long periods (up to hundreds of years). DNA extraction from desert flora may pause added difficulties as many contain high levels of secondary metabolites. Herbarium samples from the Biology Department (UAE University) plant collection and fresh plant samples, collected from around Al-Ain (UAE), were used in this study. The three barcode loci for the coding genes matK, rbcL and rpoC1-were amplified. Our results showed that T. terresteris, H. robustum,T. pentandrus and Z. qatarense were amplified using all three primers for both fresh and herbaium samples. Both fresh and herbarium samples of C. comosum, however, were not amplified at all, using the three primers. Herbarium samples from A. javanica, C. imbricatum, T. aucherana and Z. simplex were not amplified with any of the three primers. For fresh samples 90, 90 and 80% of the samples were amplified using matK, rbcL and rpoC1, respectively. In short, fresh samples were significantly better amplified than those from herbarium sources, using the three primers. Both fresh and herbarium samples from one species (C. comosum), however, were not successfully amplified. It is also concluded that the rbcL regions showed real potentials to distinguish the UAE species under investigation into the appropriate family and genus.