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Winged bean (WB), Psophocarpus tetragonolobus, is a tropical legume, the potential of which is not fully understood. We found that 5-week oral administration of a WB seed extract inhibited wrinkle formation induced by repeated tape stripping (TS) as a model of lichenification in human chronic eczematous dermatitis. To elucidate the mechanism of the effect of WB on this model, we applied microarray analysis. Hierarchical clustering revealed that each experimental group formed a distinct cluster, suggesting the presence of a distinct gene expression profile among the three groups of non-TS, TS, and TS with oral administration of WB extract (TS/WB). Gene ontology analysis showed that several gene groups with keratinization and mitosis were significantly upregulated by TS, while other groups with ATP synthesis and glycolysis were significantly downregulated by TS/WB. Moreover, WB extract influenced a number of genes related to epidermal differentiation and inflammation. This suggests that these changes inhibited wrinkle formation by TS. [ABSTRACT FROM AUTHOR]

Toll-like receptors (TLRs) on keratinocytes are important cell surface receptors involved in the innate and acquired immune response to invading microorganisms. In acne vulgaris, TLR2 activation by Propionibacterium acnes (P. acnes) may induce skin inflammation via induction of various proinflammatory molecules that stimulate the invasion of inflammatory cells. Although corticosteroids themselves exert immunosuppressive or anti-inflammatory effects, it is well known clinically that systemic or topical glucocorticoid treatment provokes an acneiform reaction. Nevertheless, the effect of steroids on TLR2 expression in human keratinocytes remains unknown. Here, we found that the addition of glucocorticoids, such as dexamethasone and cortisol, to cultured human keratinocytes increased their TLR2 gene expression. Moreover, these glucocorticoids markedly enhanced TLR2 gene expression, which was further stimulated by P. acnes, tumor necrosis factor-α, and IL-1α. Gene expression of mitogen-activated protein kinase (MAPK) phosphatase-1 was also increased by the addition of dexamethasone. By using several inhibitors and activators, we found that TLR2 gene induction by glucocorticoids was mediated by the suppression of p38 MAPK activity following induction of MAPK phosphatase-1. These findings strongly suggest that steroid-induced TLR2 together with P. acnes existing as normal resident flora plays an important role in the exacerbation of acne vulgaris as well as in possible induction of corticosteroid-induced acne or in that of rosacea-like dermatitis.Journal of Investigative Dermatology (2009) 129, 375–382; doi:10.1038/jid.2008.237; published online 14 August 2008 [ABSTRACT FROM AUTHOR]

Abstract: We used conventional cross-breeding techniques to develop a carnation with low ethylene sensitivity. A time-lapse video recording system for evaluating ethylene sensitivity of flowers, developed in our earlier study, proved useful for this breeding. We selected 10 new ethylene-resistant lines, in addition to the previously selected line 64–54. Lines 902–48a, 234–43S, and 234–36S showed extremely low ethylene sensitivity. The response time to 10μLL−1 ethylene of these three lines exceeded 50h in two independent experiments (2004 and 2005), whereas that of three sensitive control cultivars (‘White Sim’, ‘Excerea’, and ‘Scania’) was 6.2–8.0h. The mean vase life of the ethylene-resistant lines ranged from 7.6 to 12.7 days (1.4–2.3 times that of ‘White Sim’). The application of ethylene to carnation flowers induced autocatalytic ethylene production in both petals and gynoecia of 9 of the 11 ethylene-resistant lines without petal wilting, indicating that the autocatalytic ethylene production was regulated independently of the petal wilting reaction. This result further suggests that these lines have normal ethylene receptors and produce ethylene autocatalytically, but their petal wilting reaction stops at a point beyond the receptor. Two lines (118–64S and 120–69S) showed extremely low ethylene production at flower senescence and very low autocatalytic ethylene production from petals and gynoecia. [Copyright &y& Elsevier]

Differences in ethylene sensitivity among carnation (Dianthus caryophyllus L.) cultivars were evaluated using a time-lapse video recording system. Measurement of time to petal inrolling of ‘White Sim’, ‘Nora’, ‘Chinera’, and line 64-54 flowers subjected to a range of 1–20 μl l−1 ethylene showed that 10 μl l−1 was the optimum concentration for sensitivity evaluation with our video system. With this system we found clear differences in ethylene sensitivity among 10 cultivars and one line. ‘Candy’, ‘Pallas’, ‘Chinera’, and line 64-54 had lower ethylene sensitivities than the other seven cultivars. Line 64-54 had the longest ethylene response time (20.6 h to start of petal inrolling). Video monitoring is a simple and accurate way of evaluating ethylene sensitivity. We have also used the system to study changes in the ethylene sensitivity of carnation flowers after anthesis. We were able to detect a shift in responsiveness to ethylene that was impossible to detect by previous methods. In the Sim-type carnation cultivars tested (‘White Sim’, ‘Scania’, ‘U Conn Sim’, and ‘Nora’), ethylene sensitivity after anthesis decreased significantly with age in both early-cut and late-cut flowers. These results clearly showed that decline of ethylene sensitivity is caused by the increasing physiological age of flowers. Ethylene sensitivity after anthesis decreased with age in normal Sim-type carnations in the same way as in long-vase-life variants such as ‘Sandrosa’. [Copyright &y& Elsevier]

Chrysanthemums are one of the most industrially important cut flowers worldwide. However, their segmental allopolyploidy and self-incompatibility have prevented the application of genetic analysis and modern breeding strategies. We thus developed a model strain, Gojo-0 (Chrysanthemum seticuspe), which is a diploid and self-compatible pure line. Here, we present the 3.05 Gb chromosome-level reference genome sequence, which covered 97% of the C. seticuspe genome. The genome contained more than 80% interspersed repeats, of which retrotransposons accounted for 72%. We identified recent segmental duplication and retrotransposon expansion in C. seticuspe, contributing to arelatively large genome size. Furthermore, we identified a retrotransposon family, SbdRT, which was enriched in gene-dense genome regions and had experienced a very recent transposition burst. We also demonstrated that the chromosome-level genome sequence facilitates positional cloning in C. seticuspe. The genome sequence obtained here can greatly contribute as a reference for chrysanthemum in front-line breeding including genome editing. Nakano et al. sequenced the genome of a model species for hexaploid cultivated chrysanthemum, an important cut flower. While they investigated evolution, duplication events, and a novel retrotransposon in this study, they also demonstrated the translational implication of the genome sequence in breeding the economically important plant. [ABSTRACT FROM AUTHOR]

We report a method for Agrobacterium-mediated transformation of Elatior Begonia (Begonia×hiemalis Fotsch). Young leaf discs were infected with Agrobacterium tumefaciens strains AGL0 and LBA4404. Each strain has a binary vector plasmid, pIG121Hm that includes the β-glucuronidase (GUS) gene with an intron as a reporter gene, and both the neomycin phosphotransferase II and the hygromycin phosphotransferase genes as selection markers. Explants were cultured on modified MS medium supplemented with 1.0 mg/l BA, 0.5 mg/l IAA, 300 mg/l ticarcillin, and either 100 mg/l kanamycin and 5 mg/l hygromycin, or 300 mg/l kanamycin for selection and regeneration. Out of 500 explants infected with AGL0, 16 plantlets were regenerated, and out of 628 explants infected with LBA4404, two plantlets were regenerated after 4 months of culture. Transformation was confirmed by Southern blot analysis of the GUS gene and by histochemical assays of GUS activity in plant tissues. Ten in vitro transgenic plants were obtained from AGL0 infected explants only. [Copyright &y& Elsevier]

It has long been proposed that white-flowered chrysanthemums ( Chrysanthemum morifolium Ramat.) have a single dominant gene that inhibits carotenoid formation or accumulation in ray petals. However, the precise function of the proposed gene was unknown. We previously isolated a gene encoding carotenoid cleavage dioxygenase 4, designated CmCCD4a, which is specifically expressed in the ray petals of white-flowered chrysanthemums. Because CmCCD4a was a strong candidate for the single dominant gene, we analyzed the relationship between CmCCD4a expression and carotenoid content in two sets of petal color mutants. Here, we show that CmCCD4a represents a small gene family containing at least four members. Two of them, CmCCD4a- 1 and CmCCD4a- 2, were highly expressed in ray petals of two taxa with low carotenoid levels. In petal color mutants derived from these taxa, increases in carotenoid levels accompanied decreases in CmCCD4a expression levels in ray petals. Two different circumstances reduced the levels of CmCCD4a expression in the mutants: either a CmCCD4a gene was lost from the genome or the expression of a CmCCD4a gene was suppressed. In the latter case, suppression may be caused by the loss of a function that normally enhances CmCCD4a transcription. A stepwise decrease in the amount of CmCCD4a expression in either L1 or L2 resulted in a corresponding stepwise increase in the carotenoid content in ray petals. From these results, we propose that CmCCD4a expression is the key factor that controls the carotenoid content in ray petals of chrysanthemum. [ABSTRACT FROM AUTHOR]

Most chrysanthemum ( Chrysanthemum morifolium Ramat.) flowers have a central capitulum, composed of many disc florets that is surrounded by ray petals. CmCCD4a, a gene that encodes a carotenoid cleavage dioxygenase (CCD), is expressed specifically in the ray petals of chrysanthemum cultivars, and its expression leads to white ray petals as a result of carotenoid degradation. Here, we show that wild chrysanthemums with white ray petals have CmCCD4a orthologs, whereas those with yellow ray petals lack these orthologs, as is the case in chrysanthemum cultivars. CmCCD4a orthologs also exist in some lines of Chrysanthemum pacificum and Chrysanthemum shiwogiku, even though these species lack ray petals. Interspecific hybridization between C. shiwogiku and a yellow-flowered chrysanthemum cultivar showed that the CmCCD4a orthologs from C. shiwogiku lead to the development of white ray petals. This indicates that the translation products of the CmCCD4a orthologs maintain enzymatic activity that can degrade carotenoids in chrysanthemums, irrespective of whether or not the ray petals that CmCCD4a expression actually occurred. [ABSTRACT FROM AUTHOR]

Abstract: Ethylene production and the expression of ethylene biosynthetic pathway genes of some carnation cultivars with a long vase life (‘Miracle Rouge’, ‘Miracle Symphony’ and ‘Sandrosa’) and a cultivar with a normal vase life (‘White Sim’) were investigated. None of the long-life cultivars exhibited normal climacteric-like ethylene production, or petal in-rolling during senescence, but they did exhibit slow petal desiccation. Ethylene production rates from senescing flowers were very low in all long-life cultivars, although slightly higher in ‘Sandrosa’ than in the two other long-life cultivars. Aminoethoxyvinylglycine (AVG) treatment prolonged flower life in ‘Sandrosa’, but did not affect flower life in ‘Miracle Rouge’ and ‘Miracle Symphony’, which indicated a role of 1-aminocyclopropane-1-carboxylate synthase (ACS) only in ‘Sandrosa’. 1-Aminocyclopropane-1-carboxylate (ACC) treatment markedly accelerated senescence of ‘Sandrosa’ but had only a small effect on ‘Miracle Rouge’ and ‘Miracle Symphony’, suggesting that ACC oxidase (ACO) was inhibited or downregulated in the two ‘Miracle’ cultivars. The levels of DC-ACS1, DC-ACS2 and DC-ACO1 transcripts in the gynoecium and petals were high at 5 and 6 days after harvest in ‘White Sim’ but were below the detection levels in ‘Miracle Rouge’ and ‘Miracle Symphony’. In ‘Sandrosa’, DC-ACS1 was below the detection limit, whereas the expression of DC-ACS2 and DC-ACO1 was very low. The results show that long vase life in these carnation cultivars, ‘Miracle Rouge’, ‘Miracle Symphony’ and ‘Sandrosa’, is correlated with low DC-ACS1, DC-ACS2 and DC-ACO1 expression in the gynoecium and petals. [Copyright &y& Elsevier]

Bacterial wilt caused by Burkholderia caryophylli is one of the most important and damaging diseases of carnations (Dianthus caryophyllus) in Japan. We aimed to identify random amplified polymorphic DNA (RAPD) markers associated with the genes controlling bacterial wilt resistance in a resistance-segregating population of 134 progeny plants derived from a cross between ‘Carnation Nou No. 1’ (a carnation breeding line resistant to bacterial wilt) and ‘Pretty Favvare’ (a susceptible cultivar). We screened a total of 505 primers to obtain RAPD markers useful for selecting resistant carnation lines: 8 RAPD markers identified by bulked segregant analysis were linked to a major resistance gene; of these, WG44-1050 had the greatest effect on resistance to bacterial wilt. A locus with large effect on bacterial resistance was mapped around WG44-1050 through QTL analysis. The RAPD marker WG44-1050 was successfully converted to a sequence-tagged site (STS) marker suitable for marker-assisted selection (MAS). Five combinations of primers were designed for specific amplification of WG44-1050. In addition, the STS marker we developed was useful and reliable as a selection marker for breeding for resistance to bacterial wilt, using a highly resistant wild species, D. capitatus ssp. andrzejowskianus and a resistant line, ‘Carnation Nou No. 1’, as breeding materials. [ABSTRACT FROM AUTHOR]

In vegetatively propagated plants, branches of periclinal chimera give rise to bud mutants, or `sports', which have been used to breed new cultivars. Here, we have examined DNA methylation profiles in the cells of the L1 and L2+3 layers from single plants of vegetatively propagated carnations. The band patterns of methylation-sensitive amplified fragment polymorphism (M-AFLP) of the genomic DNAs in the L1 and L2+3 layers of the carnation cultivar, `White Sim', vegetatively propagated over the past 50 years, were completely different. The bud mutant, `White Mind', obtained from `White Sim' about 25 years ago, also had very different M-AFLP patterns to the parental `White Sim' line. The cultivar, `Red', which was separated from `Satisfaction' as a bud mutant two years ago, showed similar patterns to `Satisfaction' in each corresponding layer. However, we were able to detect minor, but significantly different M-AFLP patterns between the cultivars, `Red' and `Satisfaction'. The number of bands that differed between these cultivars was larger in the L2+3 layers than in the L1 layers. The results indicate that the DNA methylation profiles differ between each cell layer derived from distinct cell lineages of vegetatively propagated plants, and that changes in these profiles occur frequently and accumulate in the cells of the L2+3 layers, rather than in the L1 layer. [ABSTRACT FROM AUTHOR]

Four cDNA clones were isolated from Vigna mungo seedlings by the screening with cDNA encoding UDP-glu-cose:flavonoid 3-0-glucosyltransferase (UF3GT) of Antirrhinum majus as a probe; the product of the gene corresponding to one cDNA was more highly expressed in the first simple leaves than in stems. Nucleotide sequence analysis revealed 1,691 bp (including 326 bp non-reading) containing an open reading frame of 455 amino acids. The deduced amino acid sequence showed 42% and 23% identity with those of A. majus UDP-glucose:flavonoid 3-O-glucosyltransferase (UF3GT) and Petunia hybrida UDP-rhamnose:anthocyanidin 3-0-glucoside rhamnosyltrans-ferase (RT), respectively. One region of the cDNA (amino acids 325 to 387) showed similarity to ceramide UDP-galac-tosyltransferases of mice, rats and humans. A crude extract from Escherichia coli, in which the protein was expressed from the cDNA, showed high UF3GaT activity but low UF3GT activity, and was similar in Km, optimal pH and substrate specificity to UF3GaT from V. mungo. We conclude that we have obtained UDP-galactose:flavonoid 3-0-galactosyltransferase (UF3GaT) cDNA from V. mungo. [ABSTRACT FROM AUTHOR]