Phalaenopsis has a zygomorphic floral structure, including three outer tepals, two lateral inner tepals and a highly modified inner median tepal called labellum or lip; however, the regulation of its organ development remains unelucidated. We generated RNA-seq reads with the Illumina platform for floral organs of the Phalaenopsis wild-type and peloric mutant with a lip-like petal. A total of 43,552 contigs were obtained after de novo assembly. We used differentially expressed gene profiling to compare the transcriptional changes in floral organs for both the wild-type and peloric mutant. Pair-wise comparison of sepals, petals and labellum between peloric mutant and its wild-type revealed 1,838, 758 and 1,147 contigs, respectively, with significant differential expression. PhAGL6a (CUFF.17763), PhAGL6b (CUFF.17763.1), PhMADS1 (CUFF.36625.1), PhMADS4 (CUFF.25909) and PhMADS5 (CUFF.39479.1) were significantly upregulated in the lip-like petal of the peloric mutant. We used real-time PCR analysis of lip-like petals, lip-like sepals and the big lip of peloric mutants to confirm the five genes’ expression patterns. PhAGL6a, PhAGL6b and PhMADS4 were strongly expressed in the labellum and significantly upregulated in lip-like petals and lip-like sepals of peloric-mutant flowers. In addition, PhAGL6b was significantly downregulated in the labellum of the big lip mutant, with no change in expression of PhAGL6a. We provide a comprehensive transcript profile and functional analysis of Phalaenopsis floral organs. PhAGL6a PhAGL6b, and PhMADS4 might play crucial roles in the development of the labellum in Phalaenopsis. Our study provides new insights into how the orchid labellum differs and why the petal or sepal converts to a labellum in Phalaenopsis floral mutants.

The Phalaenopsis orchid is an important potted flower of high economic value around the world. We report the 3.1 Gb draft genome assembly of an important winter flowering Phalaenopsis ‘KHM190’ cultivar. We generated 89.5 Gb RNA-seq and 113 million sRNA-seq reads to use these data to identify 41,153 protein-coding genes and 188 miRNA families. We also generated a draft genome for Phalaenopsis pulcherrima ‘B8802,’ a summer flowering species, via resequencing. Comparison of genome data between the two Phalaenopsis cultivars allowed the identification of 691,532 single-nucleotide polymorphisms. In this study, we reveal that the key role of PhAGL6b in the regulation of labellum organ development involves alternative splicing in the big lip mutant. Petal or sepal overexpressing PhAGL6b leads to the conversion into a lip-like structure. We also discovered that the gibberellin pathway that regulates the expression of flowering time genes during the reproductive phase change is induced by cool temperature. Our work thus depicted a valuable resource for the flowering control, flower architecture development, and breeding of the Phalaenopsis orchids.

BackgroundIn a breeding program, usually only superior parents are chosen for cross hybridization. Pollens of elite cultivars may not be available at hand. Properly stored pollens provide an opportunity for cross hybridization at unavailable time.ResultsPollen of a Phalaenopsis hybrid was evaluated for the storage ability at different temperatures, including room temperature, 4, − 20, and − 80 °C for up to 96 weeks. The viability of pollen was assessed by TTC staining, in vitro germination and hand pollination during and after storage. Pollen stored at all temperatures for 4 weeks remained viable and capable of successful pollination. Pollen lost its viability after 4 weeks at room temperature. Pollen remains viable after 40 weeks at 4 °C, and after 96 weeks at both − 20 and − 80 °C of storage. Viable pollen could be successfully pollinated to the female parent at all effective storage conditions and produced seeds.ConclusionsOur results indicate that Phalaenopsis pollen can be stored at 4 °C up to 40 weeks for short-term purpose. For long-term storage, pollen can be kept at both − 20 and −80 °C.

ABSTRACT. In vitro grown cabbage (Brassica oleracea var. capitata) seedlings exposed to excess molybdenum (Mo) ions exhibited severely reduced plant growth at the cotyledonary stage. Adding 80 mM proline (Pro) to the Mo-treated medium could help 50% seedlings to overcome the toxicity and grow true leaves. Under excess Mo stress, seedlings accumulated blue/purple anthocyanin in their cotyledons and hypocotyls. The anthocyanin content under Mo with 40 mM Pro was 4-fold higher than the control medium, MS with 40 mM Pro. The presence of Pro in the excess-Mo condition reduced chlorophyll a, whereas the chlorophyll b content was much higher than the control media of MS with and without Pro. Moreover, supplementing various concentrations of Pro into the Mo-stressed condition promoted the seedlings with higher antioxidant enzyme activities of superoxide dismutase, ascorbate peroxidate, and catalase. In addition, genes in the anthocyanin biosynthesis and accumulation pathways, phenylalanine ammonia lyase (PAL), chalcone synthase (CHS), flavonone 3-hydroxylase (F3H), leucoanthocyanidin dioxygenase (LDOX), and glutathione-S-transferase (GST), were all upregulated. Our study indicated that, under excess Mo stress, the antioxidant activity of cabbage seedlings was induced in an attempt to protect plants from the Mo-induced toxicity and exacerbated growth. Pro, on the other hand, functioned in producing higher antioxidant enzyme activity to partially help recover plant growth.

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