[Show abstract][Hide abstract] ABSTRACT: Cymbidium ensifolium is a Chinese Cymbidium with an elegant shape, beautiful appearance, and a fragrant aroma. C. ensifolium has a long history of cultivation in China and it has excellent commercial value as a potted plant and cut flower. The development of C. ensifolium genomic resources has been delayed because of its large genome size. Taking advantage of technical and cost improvement of RNA-Seq, we extracted total mRNA from flower buds and mature flowers and obtained a total of 9.52 Gb of filtered nucleotides comprising 98,819,349 filtered reads. The filtered reads were assembled into 101,423 isotigs, representing 51,696 genes. Of the 101,423 isotigs, 41,873 were putative homologs of annotated sequences in the public databases, of which 158 were associated with floral development and 119 were associated with flowering. The isotigs were categorized according to their putative functions. In total, 10,212 of the isotigs were assigned into 25 eukaryotic orthologous groups (KOGs), 41,690 into 58 gene ontology (GO) terms, and 9,830 into 126 Arabidopsis Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and 9,539 isotigs into 123 rice pathways. Comparison of the isotigs with those of the two related orchid species P. equestris and C. sinense showed that 17,906 isotigs are unique to C. ensifolium. In addition, a total of 7,936 SSRs and 16,676 putative SNPs were identified. To our knowledge, this transcriptome database is the first major genomic resource for C. ensifolium and the most comprehensive transcriptomic resource for genus Cymbidium. These sequences provide valuable information for understanding the molecular mechanisms of floral development and flowering. Sequences predicted to be unique to C. ensifolium would provide more insights into C. ensifolium gene diversity. The numerous SNPs and SSRs identified in the present study will contribute to marker development for C. ensifolium.
PLoS ONE 12/2013; 8(12):e85480. DOI:10.1371/journal.pone.0085480 · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A total of 85 Chinese Cymbidiums and one Zygopetalum were collected and analysed using 29 simple sequence repeats (SSRs) and 12 intersimple sequence repeats (ISSRs). These Chinese Cymbidiums are some of the most popular cultivars in market. The pairwise genetic distance between these accessions averaged 0.897, ranging from 0.259 to 1.000. A model-based clustering analysis revealed five genetic groups with serious admixture ancestry, as indicated by a fixation index of 0.156 and only 10.67% genetic variation among groups. By analysing the cross-compatibility and pollen morphology, Chinese Cymbidiums are revealed to have high compatibility for intra- and interspecific crosses. Notably, Zygopetalum mackayi had high compatibility with Chinese Cymbidiums and shared similar morphologies in terms of pollinia and pollen grain. This study is the first to report a successful cross between Zygopetalum and Chinese Cymbidiums. The high compatibility offers the potential for introducing Zygopetalum mackayi's superior traits into Chinese Cymbidiums. The similar pollen morphology as well as the low genetic differentiation might be related to the high cross-compatibility among these accessions.
[Show abstract][Hide abstract] ABSTRACT: The FLOWERING LOCUS T (FT) gene plays crucial roles in regulating the vegetative-to-reproductive phase transition. The FT-like gene of spring orchid (Cymbidium goeringii Rchb. f.), CgFT, was isolated and characterized. CgFT mRNA was detected in leaves, pseudobulb, and flowers. In flowers, CgFT was expressed more in young flower buds than in mature flowers, and was predominantly expressed in young ovary and sheath. Seasonal expression analysis in leaves of a three-year-old spring orchid showed that a large increase in transcription, which started on June 20 for CgFT. We propose that the increased transcription in the middle of June marks the beginning of flower induction in this species. The ectopic expression of CgFT in transgenic tobacco plants showed novel phenotypes by flowering earlier than wild-type plants. Further analysis of the flowering time-related genes indicated that the expression of LEAFY, APETALLA1, FRUITFULL and SEPALLATA1 were significantly upregulated in 35S::CgFT transgenic tobacco plants. These results indicated that CgFT is a putative FT homolog in spring orchid that regulates flower transition, similar to its homolog in Arabidopsis. This study provides the first information on the spring orchid floral gene to elucidate the regulation of the flowering transition in spring orchid.