Adaptation Insights from Comparative Transcriptome Analysis of Two Opisthopappus Species in the Taihang Mountains

Opisthopappus is a major wild source of Asteraceae with good cold and drought resistance. Two species of this genus (Opisthopappus taihangensis and Opisthopappus longilobus) have been employed as model systems to address the evolutionary history of perennial herb biomes in the Taihang Mountains of China. However, further studies on the adaptive divergence processes of these two species are currently impeded by the lack of genomic resources. To elucidate the molecular mechanisms involved, a comparative analysis of these two species was conducted. Among the identified transcription factors, the bHLH members were most prevalent, which exhibited significantly different expression levels in the terpenoid metabolic pathway. O. longilobus revealed a higher expression than did O. taihangensis in terms of terpenes biosynthesis and metabolism, particularly regarding monoterpenoids and diterpenoids. Analyses of the positive selection genes (PSGs) identified from O. taihangensis and O. longilobus, 1203 genes were found that related to adaptative divergence, which were under rapid evolution and/or have signs of positive selection. Different PSG expressions occurred primarily in the mitochondrial electron transport, starch degradation, secondary metabolism, as well as nucleotide synthesis and S-metabolism pathway processes. Two PSGs were obviously differentially expressed in terpenes biosynthesis that might result in the fragrances divergence between O. longilobus and O. taihangensis, which would provide insights as to how the two species adapted to different environments, characterized by sub-humid warm temperate and temperate continental monsoon climates. The comparative analysis for these two species of Opisthopappus not only revealed how the divergence occurred from molecular perspective, but also provided novel insights into how differential adaptations occurred in Taihang Mountains.

Identification of the Volatile Compounds and Observation of the Glandular Trichomes in Opisthopappus taihangensis and Four Species of Chrysanthemum

Opisthopappus taihangensis (Ling) Shih, a wild relative germplasm of chrysanthemum, releases a completely different fragrance from chrysanthemum species. We aimed to identify the volatile compounds of the leaves of O. taihangensis and four other Chrysanthemum species using headspace solid-phase micro-extraction combined with gas chromatography-mass spectrometry (HS-SPME-GC/MS). In total, 70 compounds were detected, and terpenoids accounted for the largest percentage in these five species. Many specific compounds were only emitted from O. taihangensis and not from the other four species. In particular, 1,8-cineole could be responsible for the special leaf fragrance of O. taihangensis as it accounted for the largest proportion of the compounds in O. taihangensis but a small or no proportion at all in other species. The glandular trichomes (GTs) in the leaves are the main organs responsible for the emission of volatiles. To explore the relationship between the emissions and the density of the GTs on the leaf epidermis, the shape and density of the GTs were observed and calculated, respectively. The results showed that the trichomes have two shapes in these leaves: T-shaped non-glandular trichomes and capitate trichomes. Histochemical staining analyses indicated that terpenoids are mainly emitted from capitate glandular trichomes. Correlation analysis showed that the volatile amount of terpenoids is highly related to the density of capitate trichomes. In O. taihangensis, the terpenoids content and density of capitate trichomes are the highest. We identified the diversity of leaf volatiles from O. taihangensis and four other Chrysanthemum species and found a possible relationship between the content of volatile compounds and the density of capitate trichomes, which explained the cause of the fragrance of O. taihangensis leaves.