The origins of cultivated chrysanthemums have attracted considerable attention, but they remain poorly known. Here, we reconstructed the phylogeny of representative well-known cultivars and wild species of the genus Chrysanthemum using chloroplast genomes and the nuclear LEAFY gene. Our results suggest that geographic and ecological factors may determine the opportunities for wild species to be involved in the origin of the cultivars. The wild species C. indicum, C. zawadskii, C. dichrum, C. nankingense, C. argyrophyllum, and C. vestitum were likely directly or indirectly involved as paternal species of most of the chrysanthemum cultivars examined in this study. Yet, the maternal species is supported to be a lineage of an extinct wild Chrysanthemum species and its subsequent cultivars, as all accessions of chrysanthemum cultivars sampled formed a strongly supported clade, distinct from all other species of Chrysanthemum in the plastome tree. Thus, the cultivated chrysanthemums originated from multiple hybridizations involving several paternal species rather than only two or a few wild species, with an extinct species and its subsequent cultivars serving as the maternal parents. This finding is consistent with Chrysanthemum having high rates of hybridization and gene flow, which has been demonstrated within previous studies; nevertheless, it is important to unravel the role of an extinct wild Chrysanthemum species as the ultimate maternal parent species for all the chrysanthemum cultivars. Our results also suggest that C. vestitum from Tianzhu and Funiu Mountains in Anhui and Henan Provinces of China represent two distinct cryptic species.

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Origin of Chrysanthemum cultivars-Evidence from nuclear low-copy LFY gene sequences Abstract： The chrysanthemums (Chrysanthemum × morifolium Ramat.) are a well-known group of traditional ornamental flowers in China and have been cultivated all over the world. Yet the origin of chrysanthemum cultivars has been highly debated. In this study we employ the nuclear low-copy LFY gene to study the evolutionary history of chrysanthemum cultivars. The structure of the LFY gene in all Chrysanthemum species examined is highly conserved with three exons and two introns. The length of the LFY gene in Chrysanthemum varied from 2, 887 to 3, 348 bp. The two introns exhibited high levels of variation in length and sequence composition at the intraspecific and interspecific levels. Phylogenetic analysis of the whole LFY sequences of Chrysanthemum resulted in topologies that contained three major clades. The LFY sequences from the same cultivars are present in two or three clades, supporting that hybridization and allopolyploidy were important mechanisms in the origins of different chrysanthemums. Our results suggest that different cultivars had different ancestors. Chrysanthemum indicum, C. zawadskii and C. nankingense were likely the direct ancestors of most chrysanthemum cultivars examined. Chrysanthemum vestitum is a putative ancestor for some cultivars, and may have indirectly involved in the development of the chrysanthemum cultivars. Sequences of the LFY gene are informative to shed insights into the origin of chrysanthemum

During our study on the bioactivities of natural flavonoids, we found that the total flavonoids (TFs) and the main constituent of it, licochalcone A (lico A), activated unfolded protein response (UPR) and induced autophagy and thereby apoptosis in H292 cells. MicroRNAs, such as the tumor repressor miR-144-3p, were reported to be differentially expressed in lung cancer cells and were linked to ER stress, autophagy, and apoptosis. However, the underlying miRNA-based mechanism for lico A modulating proliferation, autophagy and apoptosis in lung cancer cells is elusive. In this study, we found that miR-144-3p was down-regulated in H292 cells comparing to normal embryonic lung cells WI-38, and lico A (10 μM) could increase miR-144-3p level in H292 cells. Knockdown of miR-144-3p significantly abrogated the apoptosis and proliferation-inhibiting effects of lico A, and lico A could enhance the proliferation-inhibiting effect and apoptosis induced by miR-144-3p overexpression. Moreover, overexpression miR-144-3p could induce ER stress by down-regulating Nrf2, and lico A enhanced the Nrf2 down-regulation caused by miR-144-3p overexpression. Co-transfection experiments showed that lico A potentially increased the dicing of pre-miR-144 so as to increase the mature miR-144-3p level. Interestingly, high level of lico A (40 μM) up-regulated CHOP protein, but failed to increase the downstream genes levels of CHOP, including Bim and Bcl-2 in H292 cells. Docking studies indicated that CHOP-mediated pathway was potentially blocked by high dose of lico A. Our results suggested that lico A could cause UPR, autophagy and apoptosis, and the underlying mechanism involved up-regulation of miR-144-3p, and increased lico A level would also increase the potential for lico A inhibiting CHOP-dependent apoptosis in H292 cells.