Increased CG, CHG and CHH methylation at the cycloidea gene in the "Peloria" mutant of Linaria vulgaris.

Heritable DNA methylation variation is frequently observed in natural populations of plants, but is thought mostly to be functionally inconsequential. An exception to this is the "Peloria" mutant of Linaria vulgaris, which was originally described by Carl von Linné in 1744. A study in 1999 found that the Peloria phenotype is caused by an epiallele of the L. vulgaris cycloidea homolog Lcyc that showed increased levels of DNA methylation compared to wild-type. The DNA methylation results in silencing of Lcyc, which causes radial flower symmetry in the peloric mutant, whereas wild-type plants have flowers with bilateral symmetry. However, a detailed view of DNA methylation at Lcyc at the single-nucleotide level has not been available. In this study, we investigated DNA methylation at Lcyc and, as a control, at the LvHIRZ gene in wild-type and peloric plants of L. vulgaris using DNA bisulfite treatment coupled to next-generation sequencing. We found strong increases in CHG and CHH methylation at Lcyc, but not LvHIRZ, in Peloria. CG methylation was also increased, but wild-type Lcyc also showed moderate levels of CG methylation. Our results suggest that DNA methylation in all three sequence contexts has been maintained, and potentially transgenerationally inherited, in the peloric L. vulgaris population over decades or even centuries.

Comparative transcriptomic analyses of normal and peloric mutant flowers in Cymbidium goeringii Rchb.f identifies differentially expressed genes associated with floral development.

Cymbidium geringii has high ornamental and economic importance. Its traits, including flower shape, size, and color, are highly sought by orchid breeders. Gaining insights into the molecular basis of C. geringi flower development would accelerate genetic improvement of other orchids. Methods and Results: Here, C. goeringii RNA was purified from normal and peloric mutant flowers, and cDNA libraries constructed for Illumina sequencing. We generated 329,156,782 clean reads, integrated them, and then assembled into 236,811 unigenes averaging 595 bp long. A total of 11,992 differentially expressed genes s, of which 6119 were upregulated and 5873 downregulated, were uncovered in peloric mutant flower buds relative to normal flower buds. Kyoto Encyclopedia of Genes and Genomes enrichment assessments posited that these differentially expressed genes are associated with "Photosynthesis", "Linoleic acid metabolism", as well as "Plant hormone signal transduction" cascades. The DEGs were designated to 12 remarkably enriched GO terms, and 16 cell wall associated GO terms. The expression level of 16 determined genes were verified using RT-qPCR. Conclusions: Our gene expression data may be used to study the regulatory mechanism of flower organ development in C. geringi.

Aphyllorchisperiactinantha (Orchidaceae, Neottieae), a new mycoheterotrophic species from peninsular Thailand.

A new orchid species from southern Thailand, Aphyllorchisperiactinantha, is described and illustrated. The novelty is characterized by the subactinomophic flowers, the concave labellum, not divided into hypochile and epichile, the reduced staminodes, the shallowly bilobed stigma and the semicircular rostellum. A key to the species of Aphyllorchis in Thailand is updated.

Molecular systematics and the evolution of mycoheterotrophy of tribe Neottieae (Orchidaceae, Epidendroideae).

Neottieae comprise about 150-200 species and are distributed mainly in temperate and subtropical zones of the northern hemisphere. Mycoheterotrophy is common in Neottieae. Based on three DNA markers and a broad sampling of Neottieae, these results indicate that Neottieae is strongly supported as monophyletic and Palmorchis is sister to the remaining genera of Neottieae. Holopogon and Neottia s.s. are deeply nested within Listera. The habit of leafless mycotrophy has independently evolved at least three times in Neottieae, one in Cephalanthera, another in Neottia s.l. and the third in the clade formed by Limodorum and Aphyllorchis.

Perspectives on MADS-box expression during orchid flower evolution and development.

The diverse morphology of orchid flowers and their complex, often deceptive strategies to become pollinated have fascinated researchers for a long time. However, it was not until the 20th century that the ontogeny of orchid flowers, the genetic basis of their morphology and the complex phylogeny of Orchidaceae were investigated. In parallel, the improvement of techniques for in vitro seed germination and tissue culture, together with studies on biochemistry, physiology, and cytology supported the progress of what is now a highly productive industry of orchid breeding and propagation. In the present century both basic research in orchid flower evo-devo and the interest for generating novel horticultural varieties have driven the characterization of many members of the MADS-box family encoding key regulators of flower development. This perspective summarizes the picture emerging from these studies and discusses the advantages and limitations of the comparative strategy employed so far. I address the growing role of natural and horticultural mutants in these studies and the emergence of several model species in orchid evo-devo and genomics. In this context, I make a plea for an increasingly integrative approach.