Chinese raspberry (Rubus chingii Hu) is a fruit valued for it's health benefits, which is indigenous to China. It is a great source of antioxidants. However, the fruit phytochemicals are poorly understood. Phenolics and carotenoids attract much attention for their antioxidant capability, and they dramatically change during fruit ripening, leading to the difference in color, flavor and medicinal components. In this study, we investigated the change of carotenoids, phenolics and antioxidant activity using spectrophotometry during four different ripening phases i.e. mature green (MG), green yellow (GY), yellow orange (YO) and red (RE). The major components of carotenoids, anthocyanins, ellagitannins and flavonols were identified and quantified by LC-MS/MS. As a result, five carotenoids (mainly β-Citraurin and its esters), six anthocyanins (mainly anthocyanins covalently linked to another flavonoid unit), methyl (S)-flavogallonate and rourinoside were first identified in Rubus. In contrast to other known raspberries, R. chingii had a continuous decrease in total phenolics during fruit ripening, which was due to a continuous decrease in flavonoids (including anthocyanin). Total anthocyanin and flavonoid respectively declined from 19.5 to 6.9 mg/100 g FW, and 646.2 to 128.5 mg/100 g FW during fruit maturation and coloration. Accordingly, the components of anthocyanins, ellagitannins and flavonols also declined, thus resulting in a decrease in antioxidant activity (from 41.2 to 10.1 TEAC/100 g FW in ABTS and from 35.3 to 7.7 mmol TEAC/100 g FW in FRAP). In contrast, total carotenoid increased from 184.2 to 305.4 mg/100 g FW. Accordingly, the components of carotenoids also increased, with the exception of lutein. Additionally, kaempferol and quercetin were the main flavonoid aglycones, which were linked to a variety of glycosides. These kaempferol- and quercetin-glycosides mainly accumulated in epidermal hair and placentae. Notably, carotenoids (i.e. β-citraurin esters), instead of anthocyanins, gradually accumulated during fruit ripening, imparting the reddish color to ripe fruit.

Cymbidium ensifolium is one of the most important ornamental flowers in China, with an elegant shape, beautiful appearance, and a fragrant aroma. Its unique flower shape has long attracted scientists. MicroRNAs (miRNAs) are critical regulators in plant development and physiology, including floral development. However, to date, few studies have examined miRNAs in C. ensifolium. In this study, we employed Solexa technology to sequence four small RNA libraries from two flowering phases to identify miRNAs related to floral development. We identified 48 mature conserved miRNA and 71 precursors. These conserved miRNA belonged to 20 families. We also identified 45 novel miRNA which includes 21 putative novel miRNAs*, and 28 hairpin forming precursors. Two trans-acting small interfering RNAs (ta-siRNAs) were identified, one of which was homologous to TAS3a1. TAS3a1 belongs to the TAS3 family, which has been previously reported to target auxin response factors (ARF) and be involved in plant growth and floral development. Moreover, we built a C. ensifolium transctriptome database to identify genes targeted by miRNA, which resulted in 790 transcriptomic target unigenes. The target unigenes were annotated with information from the non-redundant (Nr), gene ontology database (GO), eukaryotic orthologous groups (KOGs) and Kyoto encyclopedia of genes and genomes (KEGG) database. The unigenes included MADS-box transcription factors targeted by miR156, miR172 and miR5179, and various hormone responding factors targeted by miR159. The MADS-box transcription factors are well known to determine the identity of flower organs and hormone responding factors involved in floral development. In expression analysis, three novel and four conserved miRNA were differentially expressed between two phases of flowering. The results were confirmed by RNA-seq and qRT-PCR. The differential expression of two miRNA, miR160 and miR396, targeted ARFs and growth regulating factor (GRF), respectively. However, most of these small RNA were clustered in the uncharacterized group, which suggests there may be many novel small non-coding RNAs yet to be discovered. Our study provides a diverse set of miRNAs related to cymbidium floral development and serves as a useful resource for investigating miRNA-mediated regulatory mechanisms of floral development.