Zanthoxylum bungeanum is one of the most important medicinal and edible homologous plants because of its potential health benefits and unique flavors. The chemical components in compositions and contents vary with plant genotype variations and various environmental stress conditions. Fatty acids participate in various important metabolic pathways in organisms to resist biotic and abiotic stresses. To determine the variations in metabolic profiling and genotypes, the fatty acid profiling and key differential genes under low temperature stress in two Z. bungeanum varieties, cold-tolerant (FG) and sensitive (FX), were investigated. Twelve main fatty acids were found in two Z. bungeanum varieties under cold stress. Results showed that the contents of total fatty acids and unsaturated fatty acids in FG were higher than those in FX, which made FG more resistant to low temperature. Based on the result of orthogonal partial least squares discriminant analysis, palmitic acid, isostearic acid, linolenic acid and eicosenoic acid were the important differential fatty acids in FG under cold stress, while isomyristic acid, palmitic acid, isostearic acid, stearic acid, oleic acid, linolenic acid and eicosenoic acid were the important differential fatty acids in FX. Furthermore, fatty acid synthesis pathway genes fatty acyl-ACP thioesterase A (FATA), Delta (8)-fatty-acid desaturase 2 (SLD2), protein ECERIFERUM 3 (CER3), fatty acid desaturase 3 (FAD3) and fatty acid desaturase 5 (FAD5) played key roles in FG, and SLD2, FAD5, 3-oxoacyl-[acyl-carrier-protein] synthase I (KAS I), fatty acyl-ACP thioesterase B (FATB) and acetyl-CoA carboxylase (ACC) were the key genes responding to low temperature in FX. The variation and strategies of fatty acids in two varieties of Z. bungeanum were revealed at the metabolic and molecular level. This work provides a reference for the study of chemical components in plant stress resistance.

Zanthoxylum bungeanum Maxim. is an economically important tree species that is resistant to drought and infertility, and has potential medicinal and edible value. However, comprehensive genomic data are not yet available for this species, limiting its potential utility for medicinal use, breeding programs, and cultivation. Transcriptome sequencing provides an effective approach to remedying this shortcoming. Herein, single-molecule long-read sequencing and next-generation sequencing approaches were used in parallel to obtain transcript isoform structure and gene functional information in Z. bungeanum. In total, 282,101 reads of inserts (ROIs) were identified, including 134,074 full-length non-chimeric reads, among which 65,711 open reading frames (ORFs), 50,135 simple sequence repeats (SSRs), and 1492 long non-coding RNAs (lncRNAs) were detected. Functional annotation revealed metabolic pathways related to aroma components and color characteristics in Z. bungeanum. Unexpectedly, 30 transcripts were annotated as genes involved in regulating the pathogenesis of breast and colorectal cancers. This work provides a comprehensive transcriptome resource for Z. bungeanum, and lays a foundation for the further investigation and utilization of Zanthoxylum resources.

Because of extensive cultivation areas, various cultivars, nonstandard naming notations, and morphology similarity among relative cultivars, adulteration and associated business fraud may happen in the marketplaces of prickly ash pericarps due to higher financial gain and high-frequency trading. This study presents variations in the chemical components and contents of different prickly ash species from different plantations. Alkylamide profiling of pericarps derived from Zanthoxylum armatum, Z. bungeanum, and some relative Zanthoxylum species from 72 plantations across China were tested using ultra-performance liquid chromatography. Then, several chemometrics were applied to classify the prickly ash pericarps to reveal potential indicators that distinguish prickly ash pericarps and to identify the key factors that affect pericarp alkylamide profiling. The dominating alkylamides in the prickly ash pericarps were Z. piperitum (ZP)-amide C (0–20.64 mg/g) and ZP-amide D (0–30.43 mg/g). Alkylamide profiling of prickly ash pericarps varied significantly across species and geographical variations. ZP-amide D in prickly ash pericarps was identified as a potential indicator to distinguish prickly ash species. Longitude and aluminum content in soils were identified as key factors that affected alkylamide profiling of prickly ash pericarps. This study provides a useful tool to classify prickly ash species based on pericarp alkylamide profiling and to determine the key influence factors on pericarp alkylamide variations.

The color of Zanthoxylum bungeanum Maxim. (Z. bungeanum) is a key quality indicator and a factor limiting the development of its industry. However, the underlying mechanisms governing color formation remain largely unexplored. In this study, an integrative analysis of transcriptome and metabolome profiles was conducted across four developmental stages to elucidate the color formation mechanism in Z. bungeanum. A total of 137 flavonoids were identified as the fruits ripened, with high levels of differentially accumulated metabolites (DAMs), including tricetin and (-)-epigallocatechin, which were strongly associated with color formation. This suggests their significant contribution to the pigmentation process. Nine differentially expressed genes (DEGs) were identified as candidate genes involved in color development. Additionally, 15 transcription factors (TFs) (12 MYB and 3 bHLH) exhibited expression patterns similar to those of structural genes in the flavonoid biosynthetic pathway, indicating their role in regulating flavonoid synthesis. The bioinformatics analysis of three key flavonoid synthesis genes—ZbCHI, ZbFLS, and ZbANR—revealed that all three proteins exhibit hydrophobic structures without transmembrane domains. Among them, ZbANR possesses signal peptide regions, whereas ZbCHI and ZbFLS do not. Subcellular localization predictions suggest that ZbCHI is most likely localized in the chloroplast, ZbFLS in the cytoplasm, and ZbANR in the membrane. Functional analyses revealed that their transient expression in Nicotiana benthamiana (N. benthamiana) increased the flavonoid content, with ZbANR overexpression producing a distinct white phenotype in the plants. This study enriches transcriptomic data and provides a comprehensive understanding of flavonoid metabolism and the molecular basis of color formation in Z. bungeanum, offering a valuable theoretical foundation for future breeding programs.

Chinese pepper, referring to Zanthoxylum bungeanum Maxim. and Zanthoxylum armatum DC. species, is an important spice crop that has long attracted people’s interest due to its extensive application in Asian cuisine to improve taste. Numerous cultivars have been developed during the long history of domestication and cultivation. However, little to no information is available on the genetic diversity and evolutionary relationships of Chinese pepper cultivars and their historical diversification has not been clarified. Herein, we sequenced two nrDNA non-coding region markers, the external transcribed spacer (ETS) and the internal transcribed spacer 2 (ITS2), to assess genetic diversity and phylogenetic relationships among 39 cultivated and wild populations of Chinese pepper from eight provinces and to address the question of ancient demographic trends which were probably influenced by changing climate during evolutionary history. In total, 31 haplotypes were identified based on 101 polymorphism sites. Our results revealed relatively high level of genetic variation despite long-term cultivation of this crop. AMOVA revealed that genetic variation existed predominantly within provinces rather than among provinces. The genetic structure result based on haplotype network analysis largely reflected historical records, which suggested a Gansu origin for Chinese pepper and an ancient west-to-east spread of Chinese pepper circulating in China. We also provided evidence that changing Pleistocene climates had shaped the demographic trends of Chinese pepper. Taken together, our findings not only suggest that Chinese pepper is a dynamic genetic system that responds to evolutionary forces, but it also provides a fundamental genetic profile for the conservation and responsible exploitation of the extant germplasm of Chinese pepper and for improving the genetic basis for breeding the cultivars.

Zanthoxylum, belonging to the Rutaceae family, is widely distributed in tropical and subtropical regions. The genus has high economic value as spices, oils, medicinal plants, and culinary applications. Zanthoxylum has a long history of domestication and cultivation in China. However, the phylogenetic relationships and origin of wild and cultivated Zanthoxylum species in China remain largely unknown. Moreover, there is still no clear molecular phylogenetic system for Zanthoxylum species. Herein, 373 Zanthoxylum samples were collected from all presently known provenances of Zanthoxylum in China. In this study, four chloroplast DNA (cpDNA) markers (matK, ndhH, psbB, rbcL) were used to comprehensively analyze the genetic diversity, relatedness, and geographical origin of Chinese Zanthoxylum species. The results were as follows: (1) The aligned length of the four pairs of cpDNA sequences was 3836 bp, and 68 haplotypes were identified according to 219 variable polymorphic sites, including 90 singleton variable sites, 129 parsimony informative sites, 3 Indels (insertions and deletions). (2) Phylogenetic tree and haplotype network strongly supported the division of Zanthoxylum species consistent with the taxonomic recognition of five species: Z. bungeanum, Z. piasezkii, Z. piperitum, Z. armatum, and Z. micranthum. (3) Divergence time estimation suggested that Zanthoxylum genus originated from the Late Eocene, and most Zanthoxylum species diverged after the Middle Miocene. (4) Haplotype 16 (H16) was at the bottom of the phylogenetic tree, had higher haplotype diversity (Hd) and nucleotide polymorphism (Pi) than other haplotypes, and was located in the center of the network figure. Therefore, we deduced that the cultivated Zanthoxylum species may originate in Zhouqu County, Gansu Province, China. Meanwhile, our research provided a scientific basis for the identification and breeding programs of Chinese Zanthoxylum species.