Humulus scandens-Derived Biochars for the Effective Removal of Heavy Metal Ions: Isotherm/Kinetic Study, Column Adsorption and Mechanism Investigation

Humulus scandens was first adopted as a biomass precursor to prepare biochars by means of a facile molten salt method. The optimized biochar exhibits a high specific surface area of ~450 m2/g−1, a rich porous structure and abundant oxygen functional groups, which demonstrate excellent adsorption performance for heavy metal ions. The isotherm curves fit well with the Langmuir models, indicating that the process is governed by the chemical adsorption, and that the maximum adsorption capacity can reach 748 and 221 mg/g−1 for Pb2+ and Cu2+, respectively. In addition, the optimized biochar demonstrates good anti-interference ability and outstanding removal efficiency for Cu2+ and Pb2+ in simulated wastewater. The mechanism investigation and DFT calculation suggest that the oxygen functional groups play dominant roles in the adsorption process by enhancing the binding energy towards the heavy metal ions. Meanwhile, ion exchange also serves as the main reason for the effective removal.

Novelties in the vascular flora of Croatia

We report on and discuss records of five species new to the vascular flora of Croatia. Calamagrostis canescens was found near Đurđevac in remnants of fen vegetation, and we consider it a critically endangered species in the flora of Croatia. Agrimonia procera is native or archaeophyte in central Croatia and its discovery is in accordance with its earlier use as a medicinal plant. Amaranthus emarginatus is an exotic newcomer that was previously found in urban habitats on the north Adriatic coast. Carex vulpinoidea and Oenothera suaveolens were shown to be locally established species, in meadows along the Kupa River, and on the bars of the Drava River, respectively. In addition, we present new distributional and ecological data on two neophytes (Cardamine occulta, Humulus scandens) that were only briefly mentioned in earlier sources from Croatia.

Colletotrichum fioriniae infecting invasive Japanese hop (Humulus scandens) in the United States

Japanese hop (Humulus scandens) is a non-native, invasive plant that colonizes disturbed riparian areas throughout the eastern United States and Canada, forming dense, monocultural stands that displace native plant communities due to a high reproductive rate, rapid growth, climbing bines, and dense shading (Balogh and Dancza 2008). It is capable of serving as a reservoir for agronomically important plant pathogens, such as the Tomato spotted wilt virus and powdery mildew species that infect commercial hemp and hop fields (Yoon et al. 2018; Weldon et al. 2020). In the spring of 2016, diseased populations of H. scandens were observed along the Monocacy River in Frederick County, Maryland with severe chlorotic and necrotic leaf lesions. Symptomatic leaves were surface sterilized and placed in moist chambers at 25°C for sporulation. Sporulating acervuli, lacking setae, developed on irregular, tan necrotic leaf lesions following 7 to 12 days in a moist chamber (Figure 1). Conidia were hyaline, aseptate, smooth-walled, fusiform to cylindrical with both ends acute (Figure 1B). Conidia measured (n = 100) [L x W; Average (+ Std. Err), range]: 12.42 µm (± 0.10), 8.41 – 14.48 µm; x 3.91 µm (±0.03), 3.03 – 4.91 µm. Monoconidial fungal cultures were obtained by transferring conidia with a sterile glass needle to acidified potato dextrose agar and incubated at 25°C for 2 to 3 days. Based on phenotypic characteristics and conidial morphology and size, the pathogen appeared to belong to the Colletotrichum acutatum complex (Damm et al. 2012). Therefore, six loci (ITS, GADPH, CHS1, HIS3, ACT, and TUB2) were amplified and sequenced from a representative isolate, 16-008, for species characterization (GenBank accessions MW023070 to MW023075) (Damm et al. 2012). For the ITS region and ACT, GADPH, and CHS1 loci, isolate 16-008 was 100% identical to C. fioriniae and shared 99% similarity to TUB2 and HIS3 for multiple accessions of C. fioriniae in GenBank. Gene sequences were aligned, trimmed, concatenated, and analyzed against 32 reference strains, within the C. acutatum complex (Damm et al. 2012). Concatenated loci were used to generate a maximum likelihood phylogeny using W-IQ-TREE (Trifinopoulos et al. 2016). Results from the phylogenetic analysis demonstrated that isolate 16-008 was most genetically similar to C. fioriniae with a bootstrap support of 100% (Figure 2). Based on phenotypic and sequence analyses, isolate 16-008 was identified as C. fioriniae. Humulus scandens seedlings from Maryland (n = 3) were inoculated with a conidia suspension (107 conidia mL-1) with 0.125% Tween 20® and applied with an atomizer until runoff. Inoculated plants were placed in a dew chamber at 25°C for 2 days. Experimental plants were distributed in a mist tent at 25°C with 14 h of light and monitored for 2 weeks. Negative control plants (n = 2) were sprayed with a sterile 0.125% Tween 20® water solution. All inoculated plants were symptomatic by 12 days post inoculation. No symptoms were observed on the mock-inoculated plants. Symptoms were identical to disease field samples. Inoculations were repeated with the same results. Colletotrichum fioriniae was reisolated and confirmed from excised leaf lesions via ITS and ACT sequencing. To our knowledge, this is the first report of C. fioriniae naturally infecting H. scandens within the United States (Farr and Rossman 2020). Future studies will evaluate the host range of this isolate due to the species broad host range and the weed’s extensive distribution.

Identify potential allelochemicals from Humulus scandens (Lour.) Merr. root extracts that induce allelopathy on Alternanthera philoxeroides (Mart.) Griseb.

Although it is well-documented that invasion of invasive plants is promoted with allelopathic effects by inhibiting the growth and phenotypic performance of native plants, little is known conversely. In this study, the allelopathy effects of a native plant, Humulus scandens (Lour.) Merr., on a typical invasive species Alternanthera philoxeroides (Mart.) Griseb., was investigated by exposing A. philoxeroides seedlings to three chemical solvent extracts (i.e., petroleum ether extract (PE), ethyl acetate extract (EE), and n-butanol extract (NE) of H. scandens root (HR). The three chemical extracts inhibited the growth, stem length, node number, leaf number, leaf area, and root number, and increased malondialdehyde (MDA) content of A. philoxeroides seedlings, which indicated that the extracts inhibited the plant growth by damaging the membrane system of leaves. And the synthetical effect of allelopathy (SE) index indicated that EE had the greatest inhibition on the growth of A. philoxeroides. Fifty compounds were identified from the three extracts of HR using GC–MS analysis, among which 5 compounds (dibutyl phthalate, stigmasta-3,5-diene, 2,6-Di-tert-butylphenol campesterol, and neophytadiene) were identified from H. scandens root extracts for the first time. And n-hexadecanoic acid exists in all three extracts. The findings of the present study provide a novel method to potentially control the invasion of A. philoxeroides. However, field monitoring under natural conditions would be necessary to confirm in practice the results obtained with the bioassays.