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Tetrastigma hemsleyanum Diels et Gilg (Sanyeqing, SYQ) is a perennial climbing liana and an endemic plant to southern China. Its tuberous roots (TRs) are used in traditional Chinese medicine for treating some diseases such as high fever, pneumonia, asthma, hepatitis, and cancers. However, the mechanisms underlying the development of TR and the content of flavonoids and phenylpropanoids (FPs) are not well-understood. In this study, we performed a transcriptomic analysis of 12 fully developed TR (FD-TR) samples harvested in four seasons [spring (Sp), summer (Su), autumn (Au), and winter (Wi)] using the RNA-Sequencing (RNA-Seq). We obtained a total of 78.54 Gb raw data and 65,578 unigenes. Then, the unigenes were annotated by using six databases such as non-redundant protein database (NR), Pfam, eggNOG, SWISSProt, Kyoto Encyclopedia of Genes and Genomes (KEGG), and gene ontology (GO). The transcriptomic profiling showed closer relationships between the samples obtained in Su and Au than those obtained in Sp and Wi based on the results of both total unigenes and differentially expressed genes (DEGs). Three pathways, including the biosynthesis of FPs, metabolism of starch and sucrose, and signaling of phytohormones, were highly enriched, suggesting a gene-level seasonal variation. Based on the numbers of DEGs, brassinosteroid (BR) signal transduction factors appeared to play a key role in modulating the development of TRs while most of the auxin signaling genes were mainly activated in Wi and Sp FD-TRs. Most genes in the biosynthesis and biodegradation of starch and biodegradation of cellulose were activated in Wi FD-TRs. As determined by the high performance liquid chromatography (HPLC) and aluminum nitrate colorimetric method, the contents of total flavonoids and most detected FP components increased from Sp to Au but decreased in Wi. Enhanced expression levels of some genes in the biosynthetic pathways of FPs were detected in Su and Au samples, which corroborated well with metabolite content. Our findings provide the first transcriptomic and biochemical data on a seasonal variation in the composition of medically important metabolites in SYQ FD-TRs.

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Understanding the relationship between intraspecific phenotypic variation and habitat is fundamentally important to ecology and evolution. However, it is largely unknown whether the environment drives the morphological variation in riverine fish otoliths, which are potential phenotypic markers. In this study, we investigated morphological variations in the otoliths of an endemic Chinese fish (Schizothorax nukiangensis Tsao) collected from seven sites with varying environmental gradients along the Nu‐Salween River. We compared morphological characters of S. nukiangensis otoliths among sampling sites and identified environmental sources of otolith morphological variation using multivariate regression trees and multiple factor analysis. Results showed that S. nukiangensis otoliths collected from different habitats were significantly different in morphology, mainly at the rostrum, excisura and posterior rim. Variations in otolith morphology (specifically the increase in otolith length) were predominantly governed by average river gradient; this might be a functional response to hydrogeomorphic conditions. Other environmental gradients (i.e., altitude, latitude and average annual temperature) played a minor role in otolith shape. This study highlighted the role of environmental factors in determining the otolith shape in riverine fish; thus, species and population discriminations based on otolith morphology should consider intraspecific variability.