Later-melting rather than thickening of snowpack enhance the productivity and alter the community composition of temperate grassland.

Snowpack is closely related to vegetation green-up in water-limited ecosystems, and has effects on growing-season ecosystem processes. However, we know little about how changes in snowpack depth and melting timing affect primary productivity and plant community structure during the growing season. Here, we conducted a four-year snow manipulation experiment exploring how snow addition, snowmelt delay and their combination affect aboveground net primary productivity (ANPP), species diversity, community composition and plant reproductive phenology in seasonally snow-covered temperate grassland in northern China. Snow addition alone increased soil moisture and nutrient availability during early spring, while did not change plant community structure and ANPP. Instead, snowmelt delay alone postponed plant reproductive phenology, and increased ANPP, decreased species diversity and altered species composition. Grasses are more sensitive to changes in snowmelt timing than forbs, and early-flowering forbs showed a higher sensitivity compared to late-flowering forbs. The effect of snowmelt delay on ANPP and species diversity was offset by snow addition, probably because the added snow unnecessarily lengthens the snow-covering duration. The disparate effects of changes in snowpack depth and snowmelt timing necessitate their discrimination for more mechanistic understanding on the effects of snowpack changes on ecosystems. Our study suggests that it is essential to incorporate non-growing-season climate change events (in particular, snowfall and snowpack changes) to comprehensively disclose the effects of climate change on community structure and ecosystem functions.

Grassland sensitivity to drought is related to functional composition across East Asia and North America.

Plant traits can be helpful for understanding grassland ecosystem responses to climate extremes, such as severe drought. However, intercontinental comparisons of how drought affects plant functional traits and ecosystem functioning are rare. The Extreme Drought in Grasslands experiment (EDGE) was established across the major grassland types in East Asia and North America (six sites on each continent) to measure variability in grassland ecosystem sensitivity to extreme, prolonged drought. At all sites, we quantified community-weighted mean functional composition and functional diversity of two leaf economic traits, specific leaf area and leaf nitrogen content, in response to drought. We found that experimental drought significantly increased community-weighted means of specific leaf area and leaf nitrogen content at all North American sites and at the wetter East Asian sites, but drought decreased community-weighted means of these traits at moderate to dry East Asian sites. Drought significantly decreased functional richness but increased functional evenness and dispersion at most East Asian and North American sites. Ecosystem drought sensitivity (percentage reduction in aboveground net primary productivity) positively correlated with community-weighted means of specific leaf area and leaf nitrogen content and negatively correlated with functional diversity (i.e., richness) on an intercontinental scale, but results differed within regions. These findings highlight both broad generalities but also unique responses to drought of community-weighted trait means as well as their functional diversity across grassland ecosystems.

Effects of nitrogen inputs and mowing on the abundance and species richness of herbivorous insects in a meadow steppe.

To reveal the effects of nitrogen (N) inputs and mowing on the abundance and richness of insect community in meadow steppe, we investigated the abundance and diversity of herbivorous insects under four treatments in Inner Mongolia meadow steppe in August 2022, including control, N addition, mowing, and combined N addition and mowing. At a long-term control experimental platform, we collected insects using the vacuum sampling method. The results showed that N addition significantly increased the abundance of herbivorous insects, and mowing significantly decreased the abundance of herbivorous insects. Nitrogen addition significantly increased insect abundance in unmown condition but not in the mown condition. The responses of insect abundance at the community level to N addition were mainly driven by the dominant groups, Cicadellidae and Lygaeidae, which was affected by the abundance of Gramineae species. In contrast, their responses to mowing were the opposite. Both N addition and mowing did not affect the diversity of herbivorous insects. Our results indicated that the responses of herbivorous insect abundance to N inputs and mowing were directly regulated by food resources. With increasing food resource availability, the abundance but not the diversity of herbivorous insects increased, with stronger responses of dominant groups than subordinate ones.

First report of Cladosporium cladosporioides causing leaf blight on Sambucus chinensis in China.

Sambucus chinensis, belonging to the Caprifoliaceae family, is an economically large herb plant that is widely cultivated in southern China for its good ornamental characteristics, edible properties, and medicinal values. In July 2021, symptoms of leaf spot were observed on Sambucus chinensis plants in two fields of Chongqing Medicinal Botanical Garden (29º8'26" N, 107º13'23" E) in Nanchuan city, Chongqing, China. Disease incidence was approximately 35 and 50% for each field. The symptoms were initially yellow or black irregular spots on leaves, and then increased to larger dark brown lesions. Finally, the entire infected leaf was blighted, withering, curl and abscission. Ten blight leaves were randomly sampled from fields. Tissues were cut into small pieces and surface sterilized with 75% ethanol for 30 s and sterilized in 2% sodium hypochlorite for 2 min, rinsed thrice with sterile distilled water, plated on potato dextrose agar (PDA) plates, and incubated at 25°C for 7 days in the dark. Later, 20 isolates were obtained from the infected leaves and had similar characteristics. Three isolates were randomly selected (CQ81, CQ82, CQ83) for the further study. Colonies on PDA were olive-green to brown with a velvety texture. Conidia (n=30) were pale- to olive-brown, smooth to verruculose and produced in long, branched chains which were easily disarticulate, single celled, and elliptical to limoniform, and measured as 2.51~4.29 × 1.63~2.14 µm. Conidiophores were solitary, straight or flexous, often unbranched. The DNA of three isolates were extracted and the internal transcribed spacer (ITS) region and translation elongation factor 1-alpha (TEF1-α) were sequenced using primer pairs ITS1/ITS4 (White et al. 1990) and EF1-728F/EF1-986R (Carbone and Kohn 1999), respectively. The sequences of three isolates were 100% identical, and one representative isolate CQ82 were deposited in GenBank (ON387641, ITS; and ON409522, TEF). BLASTn analysis of these sequences showed 99 to 100% nucleotide identity with the sequences of C. cladosporioides CPC 14705 in Korea (Bensch et al. 2010). Phylogenetic analysis using Neighbor-joining method and concatenated sequences (ITS +TEF1) with MEGA7 placed isolate CQ82 in C. cladosporioides with 99% bootstrap support. On the basis of morphological and molecular characteristics, the isolates were identified as C. cladosporioides (Bensch et al. 2010; Nam et al. 2015). A total of sixteen healthy potted plants of S. chinensis were conducted for the pathogenicity test. Eight plants were selected and one shoot of each plant was randomly used for inoculation. Leaves from the shoot of each plant were brushed with 106 conidia/ml suspension of isolate CQ82. Another 8 plants were performed in the same procedure, inoculated with sterile distilled water as control. All plants were covered with plastic bags for two days and then arranged in a greenhouse with 80% relative humidity at 25°C. The pathogenicity test was repeated thrice. After 15 days inoculation, the similar symptoms were observed on the inoculated leaves, whereas controls remained healthy. The pathogen was reisolated from blight tissue and identified as C. cladosporioides by the methods described above. Although this fungus was previously reported to cause leaf disease on many plants (Meneses et al. 2018; Sun et al. 2017), this is the first report of C. cladosporioides causing leaf blight on S. chinensis in China. This study will establish a foundation for controlling the disease.

Extreme drought exacerbates plant nitrogen­phosphorus imbalance in nitrogen enriched grassland.

The nitrogen­phosphorus (N-P) imbalance induced by N enrichment has received increasing concerns, because N:P ratios play a critical role in driving many fundamental ecological processes. Given the simultaneous occurrence of different global change drivers, it is important to understand whether and how would such N-induced N-P imbalance would be mediated by other global change factors. We examined the interactive effects of N addition (10 g N m-2 yr-1) and extreme drought (-66 % rainfall during the growing season) on species- and community-level N:P ratios in both green and senesced leaves in a temperate grassland of northern China. Extreme drought did not alter soil available N:P ratio under ambient N conditions, but increased that under N enriched conditions. Further, extreme drought did not alter the community-level N:P in both green and senesced leaves under ambient N conditions but significantly enhanced that under N enriched conditions. The drought-induced species turnover made a significant positive contribution to the changes in the community-level N:P ratio under N enriched conditions, but not under ambient N conditions. Our results suggest that the N-induced ecosystem N-P imbalance would be exacerbated by extreme drought event, the frequency of which is predicted to increase across global drylands. Such N-P imbalance would have consequences on litter decomposition, nutrient cycling, and the structures of above- and below-ground food webs. Our findings highlighted the complexity in predicting ecosystem N-P imbalance given the interactions between different global change drivers.

Semisynthesis of Matrinic Acid/Alcohol/Ester Derivatives, Their Pesticidal Activities, and Investigation of Mechanisms of Action against Tetranychus cinnabarinus.

To discover new natural-product-based potential pesticides, 85 matrinic acid/alcohol/ester derivatives were synthesized by structural modifications of a quinolizidine alkaloid matrine. N-(4-Methyl)benzylmatrinyl n-decylate (76) and N-(2-chloro)benzylmatrinyl n-undecylate (86) exhibited greater than seven-fold more pronounced acaricidal activity than matrine against Tetranychus cinnabarinus; N-(2-chloro)benzylmatrinyl benzoate (80) showed the most promising insecticidal activity against Mythimna separata. The carboxyl group of matrinic acids and introduction of n-decyl/ n-undecylcarbonyl into matrinic alcohols were important for the acaricidal activity; introduction of alkyloxy into the carboxyl of matrinic acids and introduction of the electron-withdrawing groups on the N-benzyl of matrinic esters were necessary for the insecticidal activity. Through RT-PCR and qRT-PCR analysis, it was shown that the lactam ring of matrine was vital for action on VGSC; opening the lactam ring of matrine and the alkylcarbonyl of side-chain were two important factors for acting with α1, α2, and α4 nAChR subunits; α1, α2, α4, and ß3 subunits may be the target of action of compound 86 against T. cinnabarinus.

Synthesis of andrographolide-related esters as insecticidal and acaricidal agents.

In continuation of our program aimed at the development of natural product-based pesticidal agents, a series of andrographolide-related esters, such as 3,19-dialkyl(aryl)carbonyloxy andrographolide (3a-g), 3-alkyl(aryl)carbonyloxyandrographolide (4a-g), and 19-alkyl(aryl)carbonyloxyandrographolide (5a-g), were prepared. Their structures were well characterized by 1H NMR, IR, optical rotation, HRMS and mp. Especially three-dimensional structures of compounds 3a, 4g, and 5g were unambiguously confirmed by single-crystal X-ray diffraction. Compounds 3a and 5a exhibited good insecticidal and acaricidal activities against Mythimna separata and Tetranychus cinnabarinus. Their structure-activity relationships were also discussed.

Synthesis of some monosaccharide-related ester derivatives as insecticidal and acaricidal agents.

To develop natural-product-based pesticidal agents, a series of monosaccharide-related ester derivatives (17a-q and 18a-f), glucose (xylose)-piperic acid/piperic acid-like conjugates, were synthesized. Three-dimensional structures of compounds 17b, 17g, 17h, and 17n were unambiguously determined by single-crystal X-ray diffraction. Especially compounds 18e and 18f exhibited the most potent insecticidal and acaricidal activities against Mythimna separata and Tetranychus cinnabarinus. Their structure-activity relationships were also discussed.

3D-QSAR and Molecular Docking Studies on the TcPMCA1-Mediated Detoxification of Scopoletin and Coumarin Derivatives.

The carmine spider mite, Tetranychus cinnabarinus (Boisduval), is an economically important agricultural pest that is difficult to prevent and control. Scopoletin is a botanical coumarin derivative that targets Ca2+-ATPase to exert a strong acaricidal effect on carmine spider mites. In this study, the full-length cDNA sequence of a plasma membrane Ca2+-ATPase 1 gene (TcPMCA1) was cloned. The sequence contains an open reading frame of 3750 bp and encodes a putative protein of 1249 amino acids. The effects of scopoletin on TcPMCA1 expression were investigated. TcPMCA1 was significantly upregulated after it was exposed to 10%, 30%, and 50% of the lethal concentration of scopoletin. Homology modeling, molecular docking, and three-dimensional quantitative structure-activity relationships were then studied to explore the relationship between scopoletin structure and TcPMCA1-inhibiting activity of scopoletin and other 30 coumarin derivatives. Results showed that scopoletin inserts into the binding cavity and interacts with amino acid residues at the binding site of the TcPMCA1 protein through the driving forces of hydrogen bonds. Furthermore, CoMFA (comparative molecular field analysis)- and CoMSIA (comparative molecular similarity index analysis)-derived models showed that the steric and H-bond fields of these compounds exert important influences on the activities of the coumarin compounds.Notably, the C3, C6, and C7 positions in the skeletal structure of the coumarins are the most suitable active sites. This work provides insights into the mechanism underlying the interaction of scopoletin with TcPMCA1. The present results can improve the understanding on plasma membrane Ca2+-ATPase-mediated (PMCA-mediated) detoxification of scopoletin and coumarin derivatives in T. cinnabarinus, as well as provide valuable information for the design of novel PMCA-inhibiting acaricides.