Germplasm resources are the source of herbal medicine production. The cultivation of superior germplasm resources helps to resolve the conflict between long-term population persistence and growing market demand by consistently producing materials with high quality. The fern speciesCibotium barometzis the original plant of cibotii rhizoma (“Gouji”), a traditional Chinese medicine used in the therapy of pain, weakness, and numbness in the lower extremities. Long-history medicinal use has caused serious wild population decline in China. Without sufficient understanding of the species and lineage diversity ofCibotium, it is difficult to propose a targeted conservation scheme at present, let alone select high-quality germplasm resources. In order to fill such a knowledge gap, this study sampledC. barometzand relative species throughout their distribution in China, performed genome skimming to obtain plastome data, and conducted phylogenomic analyses. We constructed a well-supported plastome phylogeny of ChineseCibotium, which showed that three species with significant genetic differences are distributed in China, namelyC. barometz,C. cumingii, andC. sino-burmaensesp. nov., a cryptic species endemic to NW Yunnan and adjacent regions of NE Myanmar. Moreover, our results revealed two differentiated lineages ofC. barometzdistributed on the east and west sides of a classic phylogeographic boundary that was probably shaped by monsoons and landforms. We also evaluated the resolution of nine traditional barcode loci and designed five new DNA barcodes based on the plastome sequence that can distinguish all these species and lineages of ChineseCibotiumaccurately. These novel findings on a genetic basis will guide conservation planners and medicinal plant breeders to build systematic conservation plans and exploit the germplasm resources ofCibotiumin China.

Camphor tree [Cinnamomum camphora (L.) J. Presl], a species in the magnoliid family Lauraceae, is known for its rich volatile oils and is used as a medical cardiotonic and as a scent in many perfumed hygiene products. Here, we present a high-quality chromosome-scale genome of C. camphora with a scaffold N50 of 64.34 Mb and an assembled genome size of 755.41 Mb. Phylogenetic inference revealed that the magnoliids are a sister group to the clade of eudicots and monocots. Comparative genomic analyses identified two rounds of ancient whole-genome duplication (WGD). Tandem duplicated genes exhibited a higher evolutionary rate, a more recent evolutionary history and a more clustered distribution on chromosomes, contributing to the production of secondary metabolites, especially monoterpenes and sesquiterpenes, which are the principal essential oil components. Three-dimensional analyses of the volatile metabolites, gene expression and climate data of samples with the same genotype grown in different locations showed that low temperature and low precipitation during the cold season modulate the expression of genes in the terpenoid biosynthesis pathways, especially TPS genes, which facilitates the accumulation of volatile compounds. Our study lays a theoretical foundation for policy-making regarding the agroforestry applications of camphor tree.

Jasmonic acid (JA) is an important hormone that functions in plant defense. cam1 and wrky53 mutants were more resistant to Spodoptera littoralis than in the wild-type (WT) Arabidopsis group. In addition, JA concentration in cam1 and wrky53 mutants was higher compared with the WT group. To explore how these two proteins affect the resistance of Arabidopsis plants, we used a yeast two-hybrid assay, firefly luciferase complementation imaging assay and in vitro pull-down assay confirming that calmodulin 1 (CAM1) interacted with WRKY53. However, these two proteins separate when calcium concentration increases in Arabidopsis leaf cells. Then, electrophoretic mobility shift assay and luciferase activation assay were used to verify that WRKY53 could bind to lipoxygenases 3 (LOX3) and lipoxygenases 4 (LOX4) gene promoters and negatively regulate gene expression. This study reveals that CAM1 and WRKY53 negatively regulate plant resistance to herbivory by regulating the JA biosynthesis pathway via the dissociation of CAM1-WRKY53, then the released WRKY53 binds to the LOXs promoters to negatively regulate LOXs gene expression. This study reveals WRKY53′s mechanism in insect resistance, a new light on the function of WRKY53.

Gasterophilosis is a significant threat to equids in the desert steppe of X injiang, C hina, where G asterophilus pecorum ( F abricius) ( D iptera: G asterophilidae) is the dominant botfly species. A population analysis was conducted on 195 individual G . pecorum larvae from three host species, P rzewalski's horse, the domestic horse and the A siatic wild ass. The distribution of haplotypes of the maternally inherited mitochondrial cytochrome oxidase subunit I ( COI ) gene was analysed to assess the population differentiation of G . pecorum . High haplotype diversity was observed among G . pecorum populations from all host species, indicating that the G . pecorum infecting one host had multiple maternal ancestors. A phylogenetic tree showed six clades, suggesting a high degree of genetic differentiation. A constructed haplotype network described both the origin of the haplotypes and the population structure. The findings indicated that G . pecorum infections within P rzewalski's horses were mainly transmitted from A siatic wild asses. Clade 1 was found to be the most primitive group and to have evolved to be highly adaptable to the desert steppe. Clade 2 originated from C lade 1, potentially as a result of the annual migration of domestic horses. Revealing the differentiation of the G . pecorum population is important for elucidating the aetiology of G asterophilus infection in X injiang and for planning appropriate control measures.

Cinnamomum camphora is an evergreen tree that contains essential oils and the leaf is the main organ used to extract essential oils. Isoprenoid is a key component contributing to the essential oils of C. camphora. Still, the molecular mechanisms and regulatory pathways underlying isoprenoid biosynthesis remain to be explored. In our study, we found that the content of linalool was higher in 65-day-old leaves than that in 15-day-old leaves of linalool-type C. camphora. The leaf proteome of these two materials was then analyzed based on data-independent acquisition, respectively, and 11,503 proteins were identified, of which 11,076 were annotated. Analysis of differentially expressed proteins revealed that the expression levels of MCT, MDS, and AACT increased significantly in 65-day-old leaves. Further analysis of the protein interaction network indicated that 15 differentially expressed proteins related to isoprenoid biosynthesis were co-expressed, and also suggested that the transcription factor families of BES1, C3H, MYB, NF-YC, Nin-like, WRKY, ZF-HD, and bHLH may act as candidate regulators of leaf development in C. camphora. Our study verified that the process of isoprenoid biosynthesis in C. camphora is regulated by a complicated network consisting of conserved synthetases, and provides proteomic information for further investigation of isoprenoid metabolic.