Perennial Herbaceous Plant Research Articles

Psophocarpus tetragonolobus: An Underused Species with Multiple Potential Uses.

Natural products, particularly those extracted from plants, have been used as therapy for different diseases for thousands of years. The first written records on the plants used in natural medicine, referred to as “medicinal plants”, go back to about 2600 BC. A thorough and complete understanding of medicinal plants encompasses a multiplex of overlapping and integrated sciences such as botany, pharmacognosy, chemistry, enzymology and genetics. Psophocarpus tetragonolobus, a member of Fabaceae family also called winged bean, is a perennial herbaceous plant characterized by its tuberous roots and its winged pod twinning and a perennial legume rich in proteins, oils, vitamins and carbohydrates. Besides nutrients, winged bean also contains bioactive compounds that have therapeutic activities like anti-oxidant, anti-inflammatory, antinociceptive, antibacterial, antifungal, antiproliferative and cytotoxic activity, a few of which already been reported. This plant can also be used as a medicinal plant for future benefits. With this concept in mind, the present review is designed to shed the light on the interests in the various phytochemicals and pharmacological pharmacognostical aspects of Psophocarpus tetragonolobus.

Helleborus orientalis lam. (Ranunculaceae) Türünün Çiçek, Tohum ve Meyve Yapısının İncelenmesi

Bu çalışmada Helleborus orientalis Lam. (Ranunculacaae) türünün çiçek morfolojisi, tohum ve meyve yapısı incelenmiştir. Helleborus L., düğün çiçeğigiller (Ranunculaceae) familyasından Orta, Güney ve Doğu Avrupa'dan Kafkasya sınırlarına kadar yayılmış yaklaşık 20 türü bulunan zehirli bir bitki cinsidir. Helleborus L. cinsi Türkiye’de iki tür ile temsil edilmektedir. Bunlar H. vesicarius Aucher ve H. orientalis Lam. türleridir. Ülkemizde “çöpleme” olarak bilinen bu tür, Türkiye’de doğal olarak yetişir. H. orientalis halk arasında bohça otu, kara çöpleme, siyah harbak, boynuz otu, danabağırtan, danakıran gibi yöresel isimlerle anılır. Çok yıllık otsu bir bitkidir. Gövdeleri yapraksız ve 60 cm uzunluğunda olup, taban yaprakları her daim yeşildir. Uzun yaprak saplı, yaprakları geniş, digitat biçiminde olup çiçek yapısı simozdur. Sepaller yeşilimsi beyaz, bazen morumsu, ovat, 20-30(-35) mm’ dir. Tek bir çiçeğin tamamen açıldıktan sonraki ortalama ömrü yaklaşık 6 gündür. Çiçek örtüsü iki sıra üzerine dizili, sepale ya da petale benzeyen dışta 5 geniş halkalı, meyvede kalıcı, içteki halkada bulunanlar küçük, tüpsü, 2-dudaklı parçalı (nektaryum) bulunur. Uzun tepecikte kısa anterler, nadiren küttür. Anterler en azından tabanda birleşmiş birkaç tohumlu folikül grubu meyve bulunmaktadır. Tohumlar genellikle 6’ dan daha fazladır. Çiçeklenme genellikle mart-mayıs arasındadır. H. orientalis türü genel olarak Karadeniz Bölgesi’nde yayılış göstermekte olup genel dağılımı Yunanistan ve Kafkasya bölgeleri üzerindedir. Genellikle orman açıklıkları ve orman altlarında, çalılık habitatlarda 1-2200 metre arasında bulunmaktadır. Türkiye’de A2, A3, A4, A5, A6, A7, A8, A9 ve B3 karelerinde yayılış göstermektedir. Çalışma materyali A3 karesi içinde yer alan Bilecik ilinin Çiçekli yayla mevkiinden 23.05.2020 tarihinde toplanarak herbaryum materyali haline getirilmiştir. Morfolojik çalışmalar canlı örnekler üzerinde ölçümler yapılarak incelenmiştir.

Variability of the Gender Spectrum in Origanum vulgare L. (Lamiaceae, Magnoliopsida)

Origanum vulgare L. is a gynodioecious species, populations of which include hermaphroditic plants with bisexual flowers and female plants with pistillate flowers. This is a holarctic forest–steppe species belonging to herbaceous perennial plants. The variability of the female-to-hermaphrodite ratio in 26 coenopopulations of O. vulgare from different habitats has been studied. The data on eight populations were obtained in the course of the authors’ studies in southern Siberia, while 18 populations from Eastern Europe were analyzed based on the published data of other researchers. For all habitats, the projective cover of the species was determined according to the standard geobotanical methods. The plants studied represented a long-rhizome life form, so a partial shoot or a partial shrub (ramets) was used as a countable unit. The gender structure of coenopopulations was studied by counting generative plants within transects with an area of 10 to 20 m2. For each population, the total number of plants to be counted was at least 90. A significant positive correlation between the projective cover of O. vulgare in a phytocenosis and the percentage of female plants in coenopopulations was revealed (the Pearson correlation coefficient r = 0.86, p = 0.001). The ANOVA analysis showed no significant difference in the percentage of female plants in six different habitats (Femp = 3.36; Fcrit 5.5 = 5.1; p = 0.05). Taking into account the morphological and reproductive features of different genders of O. vulgare, one can suggest that an increase in the projective cover of the species in a phytocoenosis and an increase in the percentage of female plants in the gender spectrum depend on successful seed regeneration in the phytocenosis.

THE CHEMICAL CONSTITUENTS' OF THE LEAVE ESSENTIAL OIL OF ALTERNANTHERA PUNGENS (KUNTH)

Alternanthera pungens Kunth commonly called khaki weed is from the family Amaranthaceae. It is a herbaceous perennial plant that has stems prostrate, rarely rising and about 10-50 cm long. The work was carried out due to the scarcity of information on the volatile constituents from the plant leaves despite works on the flower and other parts. The extraction of the essential oils from the dried leaves was carried out by the hydro distillation method using an all-glass Clevenger apparatus. The extracted oils were then analyzed using gas chromatography-mass spectrometry (GC-MS). A total of twelve constituents' representing 93.39% of A. Pungens oil with a yield of 0.4% (v/w) was obtained. The analysis of the GC-MS results of the leaf oil showed that it was dominated by â-ionone (42.18%) and hexahydrofarnesyl acetone (15.53%), others in trace amounts include; methyl palmitate (6.13%), 1octadecyne (4.72%), undecane (3.73%), para-mentha-1, 3, 8-triene (3.65%), isophytol (3.21%), ?cadinene (3.06%), 1, 2-dimethyl cyclooctene (3.05%), para-cymene (2.96%), phytol (2.67%) and neophytadiene (2.50%). The common classes of compounds present in the leaves oil are aceto monocyclic monoterpenoid (42.18%), sesquiterpenoids (18.59%), hydrocarbons (11.50%), diterpenoids (8.38%), monoterpenes (6.61%) and fatty acids (6.13%).The constituents and the compositional pattern of essential oil identified from the leaves of Alternanthera pungens grown in Nigeria differ quantitatively and qualitatively from previously reported member of the genus and the presence of sesquiterpenoid as one of the major components of the oils justify the traditional use of the plants in treating pains, headaches and inflammations.

THE CHEMICAL CONSTITUENTS' OF THE LEAVE ESSENTIAL OIL OF ALTERNANTHERA PUNGENS (KUNTH)

Alternanthera pungens Kunth commonly called khaki weed is from the family Amaranthaceae. It is a herbaceous perennial plant that has stems prostrate, rarely rising and about 10-50 cm long. The work was carried out due to the scarcity of information on the volatile constituents from the plant leaves despite works on the flower and other parts. The extraction of the essential oils from the dried leaves was carried out by the hydro distillation method using an all-glass Clevenger apparatus. The extracted oils were then analyzed using gas chromatography-mass spectrometry (GC-MS). A total of twelve constituents' representing 93.39% of A. Pungens oil with a yield of 0.4% (v/w) was obtained. The analysis of the GC-MS results of the leaf oil showed that it was dominated by â-ionone (42.18%) and hexahydrofarnesyl acetone (15.53%), others in trace amounts include; methyl palmitate (6.13%), 1octadecyne (4.72%), undecane (3.73%), para-mentha-1, 3, 8-triene (3.65%), isophytol (3.21%), ?cadinene (3.06%), 1, 2-dimethyl cyclooctene (3.05%), para-cymene (2.96%), phytol (2.67%) and neophytadiene (2.50%). The common classes of compounds present in the leaves oil are aceto monocyclic monoterpenoid (42.18%), sesquiterpenoids (18.59%), hydrocarbons (11.50%), diterpenoids (8.38%), monoterpenes (6.61%) and fatty acids (6.13%).The constituents and the compositional pattern of essential oil identified from the leaves of Alternanthera pungens grown in Nigeria differ quantitatively and qualitatively from previously reported member of the genus and the presence of sesquiterpenoid as one of the major components of the oils justify the traditional use of the plants in treating pains, headaches and inflammations.

Effect of Valerian Extract on GABRB3 Gene MRNA Expression and Sedation in BALB/C Mice

Background: Valerian, also known as Setwall, Baldrianwurzel, or Phu is an herbaceous perennial plant from Europe and Asia, ubiquitously dispersed in almost all countries with a pH of 6-7. Valeriana officinalis is often used to treat sleep disorders, anxiety, fatigue, seizures, epilepsy, and depression as traditional medicine for 2000 years. The main constituent of Valeriana officinalis is Valerenic acid with hydroxyl and acetone group derivatives. Valerenic acid has anxiolytic, tranquilizing, and sleep-inducing effects that have been demonstrated in both animal studies and clinical trials. Valerenic acid inhibits catabolism-enzyme induced breakdown of Gamma-Aminobutyric Acid (GABA) in the brain resulting in sedation. Gamma-aminobutyric acid is the most important brain inhibitory neurotransmitter with key roles in the regulation and function of the central nervous system. The sedative effect of Valerian extract is facilitated by the GABAA receptor β3 subunit. GABRB3 gene is a ligand-gated ion channel that encodes the GABAA receptor β3 subunit. GABRB3 also functions as a receptor for diazepam and other anesthetic drugs (i.e. phenobarbital). Valerian influences the presynaptic component of GABA-ergic neurons that affect the release of synaptic GABA. Valerian also inhibits GABA reuptake and GABA catabolism by inhibiting the enzyme GABA transaminase. On that basis, this study aims to determine the effect of valerian extract on GABRB3 gene mRNA expression and sedative effect in BALB/c mice. Objective: This is an experimental study using an animal model, with a posttest-only control group design to determine the effect of Valerian extract on GABRB3 gene mRNA expression and sedative effect in BALB/c mice. Methods: Twenty selected BALB/c mice were randomly allocated into four groups; each group consisting of 5 mice. Group A (negative control) was given 5 ml of aqua dest (distilled water). Group B (positive control) was given 0.025 mg/10 g of diazepam. Group C (treatment group 1) was given 2.5 mg/10 g of Valerian extract, and group D (treatment group 2) was given 5 mg/10 g of Valerian extract. The drugs were administered for seven days through a gastric gavage. Rotarod test was performed on the seventh day. A blood sample of 1 ml was taken on the first day before drug administration and after the rotarod test on the seventh day to be analyzed using RT-PCR. Results: GABRB3 gene mRNA expression showed a significant increase in group B, C, and D (p <0.0001). There was a significant difference between group C and D. The examination of motor coordination functions (Rotarod test) showed a significant difference (p<0.05) between group A and group B, between group A and group C, and between group A and group D. There was no significant difference between group B and both group C and D. Conclusions: GABRB3 gene mRNA expression was increased significantly after the administration of Valerian extract. Based on the Rotarod test, Valerian extract and Diazepam had a clinically similar sedation effect. A higher dose of Valerian extract does not yield a higher level of GABRB3 gene mRNA expression nor sedative effects.

Leaf Spot Disease Caused by Alternaria alternata on Rehmannia glutinosa in China

Leaf Spot Disease Caused by <i>Alternaria alternata</i> on <i>Rehmannia glutinosa</i> in China

Molecular Organization of 5S Ribosomal DNA and Taxonomic Status of Avenella flexuosa (L.) Drejer (Poaceae)

The wavy hair-grass, Avenella flexuosa (L.) Drejer (also known as Deschampsia flexuosa (L.) Trin. or Lerchenfeldia flexuosa (L.) Schur), is a species of perennial herbaceous plant of the Poaceae family that is widespread in Eurasia, Africa, North and South America. Whether this species belongs to the Deschampsia genus has long been an open question. Taking into account that the 5S rDNA comparison is a convenient tool in the molecular taxonomy of plants, we cloned and sequenced this region of the A. flexuosa genome and evaluated the genetic distances between this species and other species of the Poeae tribe. It has been found that at least two structural classes of 5S rDNA repeats are present in the A. flexuosa genome. The spacer regions in the 5S rDNA of A. flexuosa and D. antarctica have a low level of similarity that does not exceed the level of similarity with representatives of several other genera of the Poeae tribe. The 5S rDNA clones of A. flexuosa and D. antarctica form two distinct clades on the phyllodendogram. Taking together, our results support the idea that A. flexuosa belongs to a separate genus Avenella.

Эколого-таксономический анализ прибрежной флоры Аулиекольского района (Костанайская область)

The study of flora is an important aspect of biodiversity maintenance and monitoring. Kostanay region is not a sufficiently explored region requiring a detailed analysis of coastal vegetation. During research in 2016–2019 the composition of the coastal flora of 6 geographical points was studied: rivers (Ubagan, Karasu) and natural lakes (Teacher, Solyenoe No. 1, Solyenoe No. 2, Kairankol). As a result of studies conducted on the territory of the Auliekol district of the Kostanay region, 106 species of coastal flora from 84 genera and 37 families were found. The taxonomic composition is characterized by dominance among the families Asteraceae, Poaceae and Fabaceae, among the genera — Artemisia, Plantago, Astragalus. In the Auliekol region, plants of the steppe cenotic group prevail, which is explained by the location of the study area within the Turgai lodge. In the surveyed territory, 7 ecological groups of plants were identified, with the maximum number of species being xero-mesophytes (33 taxa), the minimum — xerophytes (1 species). Among life forms, only 4 groups were noted, of which the maximum falls on perennial herbaceous plants (78 species). Uneven distribution of plants in the surveyed reservoirs was revealed. The maximum amounts falls on the floodplain of the river Ubagan, the minimum — on the lake Teacher.

Field-based individual plant phenotyping of herbaceous species by unmanned aerial vehicle.

Recent advances in Unmanned Aerial Vehicle (UAVs) and image processing have made high‐throughput field phenotyping possible at plot/canopy level in the mass grown experiment. Such techniques are now expected to be used for individual level phenotyping in the single grown experiment.We found two main challenges of phenotyping individual plants in the single grown experiment: plant segmentation from weedy backgrounds and the estimation of complex traits that are difficult to measure manually.In this study, we proposed a methodological framework for field‐based individual plant phenotyping by UAV. Two contributions, which are weed elimination for individual plant segmentation, and complex traits (volume and outline) extraction, have been developed. The framework demonstrated its utility in the phenotyping of Helianthus tuberosus (Jerusalem artichoke), an herbaceous perennial plant species.The proposed framework can be applied to either small and large scale phenotyping experiments.

First Report of Southern Blight on Valeriana officinalis Caused by Sclerotium rolfsii in Brazil

Valeriana officinalis L. (valerian, garden valerian) is a perennial herbaceous plant in the Valerianaceae family, commonly used as a medicinal herb in the treatment of insomnia and nervousness. In May 2019, at Botucatu city in Sao Paulo state (22°53′25″S, 48°27′19″W), 80% of production of valerian plants from a grower were observed to show symptoms typical of southern blight that included leaf wilt and the presence of sclerotia in rotted tissue. Symptoms on infected plants began with yellowing of older leaves from the base to the apex that was accompanied by a brown rot in which sclerotia were formed. The suspect fungal pathogen was tentatively identified as Sclerotium rolfsii (teleomorph Athelia rolfsii) based on its morphological characters and the signs and symptoms of the fungus. Efforts to isolate the fungus for use in pathogenicity tests were made by surface disinfecting infected tissue for 1 min in 70% alcohol and then 2 min in 2% NaOCl, followed by immersion in sterile distilled water. Tissue pieces were then placed in plates containing potato dextrose agar and maintained at 25°C. White mycelia and sclerotia formed in these plates after 7 to 14 days. Sclerotia were brown, ellipsoid in shape, and ranged from 0.82 to 1.8 cm in diameter (average 1.42 cm, n = 100). These dimensions were similar to those obtained by Erper et al. (2019) from S. rolfsii that infected candyleaf in Turkey. A culture of our valerian isolate was deposited in the collection of the Biological Institute of Sao Paulo under accession number MMBF 04/20. Pathogenicity tests were conducted on healthy 2-month-old valerian plants in a greenhouse maintained at 25°C and 95% relative humidity (RH) for 3 days and at 25°C and 70% RH the following 8 days. Sclerotia were incorporated into the soil near four plant stem repetitions as the method of inoculation, and four noninoculated valerian plants were the control. Disease symptoms began to appear 3 days postinoculation, sclerotia formed after 9 days, and plant death occurred within 11 days. Symptoms on inoculated plants were the same as those observed on the original diseased specimens, and control plants remained healthy. S. rolfsii was reisolated from sclerotia produced on inoculated plants, thus confirming Koch’s postulates. DNA for molecular identification was extracted from our isolate of S. rolfsii using the cetyltrimethylammonium bromide method according to Doyle and Doyle (1987). This was followed by a polymerase chain reaction using primers ITS1/ITS4, which amplify part of the internal transcribed spacer region (White et al. 1990). The amplicon obtained (GenBank accession no. MT023434) was bidirectionally sequenced and shared 97.05% sequence identity with A. rolfsii (593/611 nucleotide identity, MH854711). S. rolfsii is known to infect many economically important plants (Tu and Kimbrough 1978). It has been reported from Japan on Valeriana radix (Morita et al. 2015), in India on V. jatamansi (syn. V. wallichii) (Khosla 2014), and in Korea on V. fauriei (Kwon et al. 2010). In Brazil, other medicinal plants such as Taraxacum officinale (dente de leao) (Severino et al. 2014) and Plantago major (tanchagem) (Dias-Arieira et al. 2014) have been reported to be susceptible to infection by S. rolfsii. To our knowledge, this is the first report of the occurrence of S. rolfsii on valerian in Brazil.

Haplophyllum Buxbaumii Ekstresinin Prostat Kanseri Üzerindeki Apoptotik Etkisi

Prostate cancer is a common type of cancer among men. Recently, studies on new anticancer agents of herbal origin in the treatment of cancer have increased. Haplophyllum buxbaumii (HP) is a perennial herbaceous plant. Studies have reported that Haplophyllum subspecies has antioxidant properties. And the anticancer effect of HP is unknown. This study aims to reveal the apoptotic effect HP on prostate cancer. The cytotoxic effect of HP was evaluated by the MTT method. In addition, apoptotic pathway mediators were analyzed by ELISA method. HP extract application reduced the viability of prostate cancer and activated the apoptotic mediators while decreasing anti-apoptotic mediators. Our study showed that HP extract has an antiproliferative and apoptotic effect on prostate cancer.

The Correlation of Some Nutrient Elements and Antibacterial Activity of the Basil (Ocimum basilicum)

This study aimed to analyze the correlation of some macro and micro nutritional elements of the basil plant (Ocimum basilicum) with the antibacterial activity of the basil extract on two Gram positive and two Gram negative bacteria. The extract of the basil plants grown in laboratory conditions was obtained by solid-liquid extraction with ethanol after drying and burning of the freshly harvested above ground parts. The macro (P, K, Ca, and Mg), micro (Fe, Cu, Zn, Mn, and B) nutrients and a functional element (Al) amounts were detected by ICP-OES. The results revealed that the basil extract was compatible with herbaceous perennial plants in terms of macro nutrient elements P, K, Ca and Mg. Moreover, some micronutrients (Fe, Mn, Zn, B) and the functional element Al was found to be significantly high. The basil extract was significantly effective on inhibiting the Gram negative E. coli, P. aeruginosa, and Gram positive S. aureus and E. faecium. The element B was found to be positively correlated with the antibacterial activity of the basil plant suggesting as the B content increases, the antibacterial activity of basil extract is supposed to increase, too. The P and K contents were found to be positively correlated with antibacterial activity, while Ca and Mg contents were negatively correlated. The correlation analysis of the nutritional elements, this extract can be suggested to be more preferrable against Gram positive bacteria, especially when the Fe, Cu, Zn, Mn levels are high. These results revealed the importance of the macro and micro nutrient contents of medicinal plants which can be used traditionally against various diseases. In basil cultivation the soils low in lime may convert the negative correlation of Ca and Mg into a positive correlation in terms of antibacterial effect. Conscious and proper fertilization therefore is suggested to be effective on antibacterial activity of the basil plant.

Enzyme-Treated Zizania latifolia Ethanol Extract Protects from UVA Irradiation-Induced Wrinkle Formation via Inhibition of Lysosome Exocytosis and Reactive Oxygen Species Generation.

Ultraviolet A (UVA) is a risk factor for photoaging and wrinkle formation. Zizania latifolia is an herbaceous perennial plant. It contains many bioactive compounds such as tricin that show antioxidative and anti-inflammatory effects. The aim of this study was to investigate the antiwrinkle effect of a mixture of hydrolytic enzyme (cellulase, hemicellulase and pectinase)-treated Z. latifolia extract (ZLE) and tricin on UVA-irradiated human dermal fibroblasts (HDFs) and SKH-1 hairless mice. Treatment of UVA-irradiated HDF cells with ZLE and tricin significantly decreased UVA induced-plasma membrane rupture, generation of ROS, expression levels of total and secreted lysosomal associated membrane protein (LAMP-1), cathepsin B and metalloproteinases (MMPs) and inhibited NF-κB activation. In the animal study, UVA-damaged epidermal and dermal tissues were repaired by the ZLE and tricin treatments. Administration of ZLE or tricin to UVA-irradiated animals recovered skin surface moisture and collagen fiber in dermal tissue. Treatment of ZLE or tricin decreased wrinkle formation, secretion of MMPs and expression levels of vascular endothelial growth factor (VEGF) and cathepsin B, and increased the expression level of collagen-1 in UVA-irradiated animals. Overall, the ZLE and tricin treatments decreased the skin damage induced by UVA irradiation via inhibition of lysosomal exocytosis and ROS generation. Therefore, ZLE and tricin are promising as antiwrinkle and antiphotoaging agents.

Ethnopharmacological Potential of Tithonia diversifolia (Hemsl) A. Gray

Tithonia diversifolia (Hemsl) A. Gray, commonly known as “Mexican Sunflower”, is a perennial herbaceous plant of the Asteraceae family, native to Mexico and Central America. At present, however, this species is widely found in many countries outside of these areas, including Brazil. It has been the subject of many scientific studies which by highlighting the chemical constitution of its main structures such as leaves, inflorescences, stalk and roots, consequently bring to light this species’ nutritional and therapeutic potential in current literature. The aim of this study is to produce a literature review on the ethnobotanical aspects and possible pharmacological applications of T. diversifolia, with emphasis on its chemical composition, morphological characteristics and pharmacotherapeutic applications. The present study is a bibliographical, exploratory and qualitative research, carried out due to the need to elucidate the proposed theme and fulfills the role of reviewing the specific bibliography on the addressed topic. Based on the produced bibliography, it is noticeable that Tithonia diversifolia (Hemsl) A. Gray is a plant of potential nutritional and pharmacological interest. In general, the studies reviewed here strengthen and highlight the evidence of the diverse applications of this species by traditional medicine, which, in turn, contribute to the development of studies that guarantee the identification of active constituents and their respective biotechnological applications.

Ecological significance of hellebore growth (Epipactis helleborine) in artificial tree plantations of the Eastern Forest-Steppe forest reclamation area

Population growth features of Epipactis helleborine (L.) Crantz, a rare plant species listed in the Red Book of Ukraine in three new habitats on the eastern foreststeppe forest meleorative area, are presented. Epipactis helleborine is a perennial herbaceous plant, 30–100 cm tall, hemicryptophyte, mesophyte, sciophyte. It blooms in June-September, bears fruit in July-October, propagated by seeds and rhizomes. This species has a fairly wide ecological amplitude. Plants can withstand moderate anthropogenic load. It has the ability to grow in disturbed or even artificially created forest ecosystems. At two location there were artificial forest plantations, in one case with a predominance of Fraxinus excelsior and Acer platanoides, in the other Quercus robur and Acer campestre. The third studied group, which was formed on the site of Pinus sylvestris plantation, is dominated by Quercus robur and Acer platanoides. In general,we can state the increase participation of a tree number and shrub species, and a wide range of herbaceous plants in all locations. «Forest weeds» reduce the participation and species characteristic of deciduous forests increase its participation. This became the basis for classifying the studied groups as different syntaxons of the union Querco roboris-Tilion cordatae of the order Carpinetalia betuli of the class Carpino-Fagetea sylvaticae. The identified new habitats supplement the information on the distribution of the rare species Epipactis helleborine in the eastern part of the forest-steppe zone of Ukraine.

Discrete element modeling of switchgrass particles under compression and rotational shear

Switchgrass is a perennial herbaceous plant regarded as a biomass energy crop in the United States for its high adaptability and yield potential. Processing and handling of switchgrass particles are challenging due to the erratic mechanical and flow behavior originating from their intrinsic particulate properties. In this work, we present a bonded-sphere discrete element model designed specifically for switchgrass particles. The model simultaneously captures three key particulate features, i.e., fibrous particle shapes, a wide range of particle sizes, and particle deformability. Realistic yet computationally efficient particle shape templates are created based on the image analysis data of switchgrass specimens. A fitting procedure is proposed to ensure both the particle width and length distributions are captured, a unique requirement for fibrous particles. Two full-scale numerical models, i.e., a uniaxial compression model and a Schulze ring shear model, are developed using information from physical experiments. The model is calibrated using experimental data of chopped-small switchgrass specimens, and then, is validated using data of chopped-large specimens in both compression and ring-shear tests. Numerical results show that the numerical models capture bulk densities accurately (with an error of 3%) while slightly underestimate the bulk friction angle. Furthermore, an extensive sensitivity analysis reveals that (1) switchgrass particles with rougher edges (due to different processing techniques) exhibit a higher shear strength and a lower flowability; (2) stiffer particles yield a lower bulk density (up to 21% lower) compared to more deformable particles, indicating particle deformability should be incorporated when modeling biomass flow in a preprocessing system.

Seasonal storage of carbon, nitrogen, and phosphorus in two perennial forest herbaceous plants

Herbaceous perennials in a forest ecosystem play a role in seasonal nutrient cycling. Asarum canadense L. (wild ginger) and Hydrophyllum virginianum L. (Virginia waterleaf) are perennial herbaceous plants with different seasonal growth patterns native to mesic hardwood forests. Both flower early, but A. canadense leaves persist after flowering while H. virginianum senesces and produces a second flush of leaves by late summer. Our specific question of interest was whether there is a difference in nutrient storage in leaves and roots seasonally in these two species. We excavated plants in spring, summer, and fall to determine carbon (C), nitrogen (N), and phosphorus (P) concentration and content in above and below ground tissues, as well as C:N and N:P ratios. Our findings indicate complex patterns in the location of nutrients in A. canadense and H. virginianum across the growing season. Asarum canadense exhibits relatively consistent above ground growth, with seasonal nutrient accumulation focused below ground for C, N, and P. Patterns are more varied in H. virginianum as a result of the second flush of leaves, notably, the C:N ratio of H. virginianum decreases across the growing season while the N:P ratio increases, but to different degrees in above/below ground tissues. We suggest these patterns are linked to changing photosynthetic capacity across the seasons and a prioritization of vegetative growth over reproductive investment.

Ixeris Chinensis is a New Host for Peanut Witches' Broom Phytoplasma, a 16SrII-V Subgroup Strain in Taiwan.

Ixeris chinensis (Thunb. ex Thunb.) Nakai, a perennial herbaceous plant that belongs to the family of Asteraceae, is widely distributed at mid-low altitude regions in Taiwan. I. chinensis is commonly used as traditional herbal medicine for the treatment of inflammation, bronchitis, pneumonia, and diarrhea. In March 2020, disease symptoms such as shoot proliferation, phyllody, virescence, purple top, and witches' broom were observed on I. chinensis at the sansheng community park in Mailiao, Yunlin County, Taiwan. Totally, eight I. chinensis plants were checked and half of them were symptomatic. These disease symptoms are similar to those associated with peanut witches' broom (PnWB) disease identified in the same area (Liu et al. 2015). Three samples mixed with leaf, stem, and flower were tested including one healthy and two symptomatic I. chinensis. The total DNA of each sample was extracted and examined by nested PCR for the amplification of 16S rDNA with the phytoplasma universal primer pairs P1/P7 followed by R16F2n/R16R2 (Lee et al. 1993). A specific signal of expected size (1.2 kb) for 16S rDNA was only detected in the symptomatic I. chinensis, but not in healthy I. chinensis. The nucleotide sequence (accession no. MT416114) of the amplified DNA fragment using primer pairs P1/P7 from symptomatic I. chinensis is identical to that of GenBank accession NZ_AMWZ01000008 (complement [31109 to 32640]) of phytoplasma associated with PnWB disease (Chung et al. 2013). Analysis of the virtual RFLP pattern of MT416114 generated by iPhyClassifier revealed that the phytoplasma detected in symptomatic I. chinensis belongs to a 16SrII-V subgroup. The total protein of each sample was also extracted and examined by western blotting using the polyclonal antibody raised against Imp protein of purple coneflower witches' broom phytoplasma (Chien et al. 2020), which is identical with that (accession no. ADD59806) of PnWB phytoplasma. An expected signal of 19 kDa specific for Imp was detected in symptomatic I. chinensis, but not in healthy I. chinensis. Subsequent PCR, DNA sequencing and western blotting assays further confirmed that the gene encoding a SAP11-like protein was only detected in symptomatic I. chinensis, and shares 100% identity with that (accession no. EMR14684) of PnWB phytoplasma. Our results indicate that PnWB phytoplasma causes disease in I. chinensis, a common weed, which may act as an alternative natural host and facilitate the spreading of phytoplasma disease in Taiwan.

Threeflower Tickclover (Desmodium triflorum) is a New Host for Peanut Witches' Broom Phytoplasma, a 16SrII-V Subgroup Strain in Taiwan.

Three-flower Tick-clover (Desmodium triflorum) is a perennial herbaceous plant that belongs to the family of Leguminosae. Threeflower tickclover widely grows at mid-low altitude regions in Taiwan and is commonly used as a traditional herbal medicine for the treatment of dysmenorrheal, muscle spasm, cough, pain and poisoning. In March 2020, disease symptoms such as little leaf, phyllody, virescence, and witches' broom were observed on threeflower tickclover at the sansheng community park in Mailiao, Yunlin County, Taiwan. Similar disease symptoms were observed on peanut infected with peanut witches' broom (PnWB) phytoplasma grown in the same area (Liu et al. 2015). Leaf samples collected from the healthy and symptomatic threeflower tickclover were used to extract total DNA and protein for PCR and western blotting assays, respectively. Nested PCR was performed with the phytoplasma universal primer pairs P1/P7 followed by R16F2n/R16R2 for the amplification of 16S ribosomal RNA (rRNA) gene (Lee et al. 1993). A specific DNA fragment of expected size (1.2 kb) for 16S rRNA was only amplified from leaf samples of symptomatic threeflower tickclover. The nucleotide sequence of the amplified DNA fragment using primer pairs P1/P7 was deposited into the GenBank (accession no. MT452308). Blast analysis revealed that MT452308 shares 100% identity with that of GenBank accession NZ_AMWZ01000008 (complement [31109 to 32640]) of phytoplasma associated with PnWB disease (Chung et al. 2013). Based on the virtual RFLP pattern of MT452308 generated by iPhyClassifier, the phytoplasma detected in symptomatic threeflower tickclover could be classified into the 16SrII-V subgroup. For western blotting, the polyclonal antibody raised against Imp protein of purple coneflower witches' broom phytoplasma (Chien et al. 2020), which is identical with that (accession no. ADD59806) of PnWB phytoplasma, was used. An expected signal of 19 kDa specific for Imp was only detected in threeflower tickclover exhibiting disease symptoms. Subsequent assays including PCR, DNA sequencing and western blotting further confirmed that the gene encoding a SAP11-like protein (accession no. EMR14684) identified in PnWB phytoplasma was also found in samples of symptomatic threeflower tickclover, and shares 100% identity with each other. Our results indicate that threeflower tickclover, a common weed in Taiwan, may act as an alternative natural host for PnWB phytoplasma, and contributes to the spreading of phytoplasma disease.