A method for the production of somatic embryos and subsequent plant regeneration for Anthurium andraeanum Linden ex André (Monocotyledonae) hybrids is described. Whole leaf blade explants, derived from plantlets grown in vitro, formed translucent embryogénic calli at their basal ends within one month of culture in the dark. Secondary somatic embryos formed frequently and without an intervening callus on surfaces of primary embryos. Embryogenesis was induced with three genotypes using a modified half-strength Murashige and Skoog (MS) medium supplemented with 1.0 to 4.0 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.33 to 1.0 mg l(-1) kinetin. A combination of 2% sucrose with 1% glucose in the medium favored embryogenesis over 3% sucrose alone. Whole leaf blades on medium solidified with 0.18% Gelrite produced more somatic embryos than leaves on medium with 0.7% Bacto-agar. Within two to three months after culture initiation, embryos were transferred to modified MS medium containing 0.2 mg l(-1) 6-benzyladenine (BA) and 2% sucrose and placed in the light for conversion into plantlets. Rooted plantlets were recovered and transferred into pots with tree fern fiber medium and grown in the greenhouse.

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.

Orchids comprise one of the largest, most highly evolved angiosperm families, and form an extremely peculiar group of plants. Various orchids are available through traditional breeding and micro-propagation since they are valuable as potted plants and/or cut flowers in horticultural markets. The flowering of orchids is generally influenced by environmental signals such as temperature and endogenous developmental programs controlled by genetic factors as is usual in many flowering plant species. The process of floral transition is connected to the flower developmental programs that include floral meristem maintenance and floral organ specification. Thanks to advances in molecular and genetic technologies, the understanding of the molecular mechanisms underlying orchid floral transition and flower developmental processes have been widened, especially in several commercially important orchids such as Phalaenopsis, Dendrobium and Oncidium. In this review, we consolidate recent progress in research on the floral transition and flower development of orchids emphasizing representative genes and genetic networks, and also introduce a few successful cases of manipulation of orchid flowering/flower development through the application of molecular breeding or biotechnology tools.

Several procedures were tested in development of a gene transfer protocol for anthurium. Etiolated internode segments of anthurium cultivars `Rudolph' and `UH1060' were co-cultured with Agrobacterium tumefaciens LBA4404 carrying the chimeric genes neo, for antibiotic resistance, and att encoding antibacterial attacin. Assays of genomic DNA and RNA from kanamycin-resistant `Rudolph' and DNA from `UH1060' plantlets, recovered as soon as 1 year after culture on selection media, indicated the presence of introduced genes, including neo and att, and transcription of att. Western analysis confirmed the expression of attacin protein in calli induced from laminae of regenerated kanamycinresistant `Rudolph' plantlets. Use of tobacco nurse cells during co-cultivation of internodes with Agrobacterium did not increase recovery of shoots under the regeneration conditions used. Improvements in culture and antibiotic selection conditions during plant development are suggested.

An efficient micropropagation procedure via adventitious shoot proliferation was developed for Aglaonema using the popular red cultivar 'Lady Valentine. ' Aseptic culture was initiated by culturing stem nodal segments on Murashige and Skoog (MS) medium supplemented with 32 mg·l −1 gentamicin, 8 mg·l −1 tetracycline and 4 mg·l −1 chloramphenicol. The growth of the axillary buds performed the best when 10 mg·l −1 6-benzyladenine (BA) was incorporated into the medium, and neither gibberellic acid (GA 3 ) nor dark exposure could improve the elongation of the axillary shoots. The single stem nodal segments excised from the elongated shoots were treated with different combinations of α-naphthaleneacetic acid (NAA) and thidiazuron (TDZ) and an average of 10.9 adventitious shoots per stem segment was produced with 0.5 mg·l −1 NAA and 2 mg·l −1 TDZ. Small shoot clusters were subsequently incubated with different concentrations of BA and GA 3 and results showed that 0.5-5 mg·l −1 BA treatments were more effective for shoot proliferation and elongation than 0.5-1 mg·l −1 GA 3 treatments. The longest shoots (reaching 2.69 cm after three months) were obtained on medium containing 5 mg·l −1 BA. Up to 80% of the elongated shoots successfully rooted ex vitro with the application of 1 and 2 mg·l −1 indole-3-butyric acid (IBA) and 92.5% of these rooted shoots survived following transfer to the greenhouse.

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.

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