Global analysis of the AP2/ERF gene family in rose (Rosa chinensis) genome unveils the role of RcERF099 in Botrytis resistance.

BACKGROUND: The AP2/ERFs belong to a large family of transcription factors in plants. The AP2/ERF gene family has been identified as a key player involved in both biotic and abiotic stress responses in plants, however, no comprehensive study has yet been carried out on the AP2/ERF gene family in rose (Rosa sp.), the most important ornamental crop worldwide. RESULTS: The present study comprises a genome-wide analysis of the AP2/ERF family genes (RcERFs) in the rose, involving their identification, gene structure, phylogenetic relationship, chromosome localization, collinearity analysis, as well as their expression patterns. Throughout the phylogenetic analysis, a total of 131 AP2/ERF genes in the rose genome were divided into 5 subgroups. The RcERFs are distributed over all the seven chromosomes of the rose, and genome duplication may have played a key role in their duplication. Furthermore, Ka/Ks analysis indicated that the duplicated RcERF genes often undergo purification selection with limited functional differentiation. Gene expression analysis revealed that 23 RcERFs were induced by infection of the necrotrophic fungal pathogen Botrytis cinerea. Presumably, these RcERFs are candidate genes which can react to the rose's resistance against Botrytis cinerea infection. By using virus-induced gene silencing, we confirmed that RcERF099 is an important regulator involved in the B.cinerea resistance in the rose petal. CONCLUSION: Overall, our results conclude the necessity for further study of the AP2/ERF gene family in rose, and promote their potential application in improving the rose when subjected to biological stress.

Characterization of black spot resistance in diploid roses with QTL detection, meta-analysis and candidate-gene identification.

KEY MESSAGE: Two environmentally stable QTLs linked to black spot disease resistance in the Rosa wichurana genetic background were detected, in different connected populations, on linkage groups 3 and 5. Co-localization between R-genes and defense response genes was revealed via meta-analysis. The widespread rose black spot disease (BSD) caused by the hemibiotrophic fungus Diplocarpon rosae Wolf. is efficiently controlled with fungicides. However, in the actual context of reducing agrochemical use, the demand for rose bushes with higher levels of resistance has increased. Qualitative resistance conferred by major genes (Rdr genes) has been widely studied but quantitative resistance to BSD requires further investigation. In this study, segregating populations connected through the BSD resistant Rosa wichurana male parent were phenotyped for disease resistance over several years and locations. A pseudo-testcross approach was used, resulting in six parental maps across three populations. A total of 45 individual QTLs with significant effect on BSD resistance were mapped on the male maps (on linkage groups (LG) B3, B4, B5 and B6), and 12 on the female maps (on LG A1, A2, A3, A4 and A5). Two major regions linked to BSD resistance were identified on LG B3 and B5 of the male maps and were integrated into a consensus map built from all three of the male maps. A meta-analysis was used to narrow down the confidence intervals of individual QTLs from three populations by generating meta-QTLs. Two 'hot spots' or meta-QTLs were found per LG, enabling reduction of the confidence interval to 10.42 cM for B3 and 11.47 cM for B5. An expert annotation of NBS-LRR encoding genes of the genome assembly of Hibrand et al. was performed and used to explore potential co-localization with R-genes. Co-localization with defense response genes was also investigated.

RcMYB84 and RcMYB123 mediate jasmonate-induced defense responses against Botrytis cinerea in rose (Rosa chinensis).

Jasmonates (JAs) are important for pathogen resistance in many plants, but the role of these phytohormones in fungal pathogen resistance in rose is unclear. Here, we determined that exogenous application of methyl jasmonate increased resistance to the important fungal pathogen Botrytis cinerea in Rosa chinensis 'Old blush', whereas silencing the JA biosynthetic pathway gene Allene Oxide Synthase (AOS) and JA co-receptor gene CORONATINE INSENSITIVE 1 (COI1) suppressed this response. Transcriptome profiling identified various MYB transcription factor genes that responded to both JA and B. cinerea treatment. Silencing Ri-RcMYB84/Ri-RcMYB123 increased the susceptibility of rose plants to B. cinerea and inhibited the protective effects of JA treatment, confirming the crucial roles of these genes in JA-induced responses to B. cinerea. JAZ1, a key repressor of JA signaling, directly interacts with RcMYB84 and RcMYB123 to deplete their free pools. The JAZ1-RcMYB84 complex binds to the RcMYB123 promoter via the CAACTG motifs to block its transcription. Upon JA treatment, the expression of RcMYB123 is de-repressed, and free forms of RcMYB84 and RcMYB123 are released due to JAZ1 degradation, thereby activating the defense responses of plants to B. cinerea. These findings shed light on the molecular mechanisms underlying JA-induced pathogen resistance in roses.

Interaction of roses with a biotrophic and a hemibiotrophic leaf pathogen leads to differences in defense transcriptome activation.

KEY MESSAGE: Transcriptomic analysis resulted in the upregulation of the genes related to common defense mechanisms for black spot and the downregulation of the genes related to photosynthesis and cell wall modification for powdery mildew. Plant pathogenic fungi successfully colonize their hosts by manipulating the host defense mechanisms, which is accompanied by major transcriptome changes in the host. To characterize compatible plant pathogen interactions at early stages of infection by the obligate biotrophic fungus Podosphaera pannosa, which causes powdery mildew, and the hemibiotrophic fungus Diplocarpon rosae, which causes black spot, we analyzed changes in the leaf transcriptome after the inoculation of detached rose leaves with each pathogen. In addition, we analyzed differences in the transcriptomic changes inflicted by both pathogens as a first step to characterize specific infection strategies. Transcriptomic changes were analyzed using next-generation sequencing based on the massive analysis of cDNA ends approach, which was validated using high-throughput qPCR. We identified a large number of differentially regulated genes. A common set of the differentially regulated genes comprised of pathogenesis-related (PR) genes, such as of PR10 homologs, chitinases and defense-related transcription factors, such as various WRKY genes, indicating a conserved but insufficient PTI [pathogen associated molecular pattern (PAMP) triggered immunity] reaction. Surprisingly, most of the differentially regulated genes were specific to the interactions with either P. pannosa or D. rosae. Specific regulation in response to D. rosae was detected for genes from the phenylpropanoid and flavonoid pathways and for individual PR genes, such as paralogs of PR1 and PR5, and other factors of the salicylic acid signaling pathway. Differently, inoculation with P. pannosa leads in addition to the general pathogen response to a downregulation of genes related to photosynthesis and cell wall modification.

Effects of dietary administration of Rose hip and Safflower on growth performance, haematological, biochemical parameters and innate immune response of Beluga, Huso huso (Linnaeus, 1758).

The aim of this study was to investigate effects of two dietary medicinal herbs, Rose hip (Rosa canina) and Safflower (Carthamus tinctorius) supplementation on growth performance, haematological, biochemical parameters and innate immune response of in juvenile beluga, Huso huso. Fish (26.3 ± 0.4 g) were allocated into 15 tanks (20 fish per tank) and triplicate groups were fed a control diet or diets containing 1% and 2% of medicinal herbs, respectively. Feed conversion ratio (FCR), specific growth rate (SGR) and condition factor (CF) did not show significant differences (P > 0.05) in fish given herbal diets. Significant differences were observed in number of white blood cells (WBC) and haemoglobin (Hb) values among the dietary treatments. The serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were significantly lower in supplemented diet groups compared with the control. Innate immune responses (lysozyme activity and ACH50) were significantly higher in 2% Safflower-fed fish compared with other groups (P < 0.05). These results indicate that medicinal herbs in diets can be considered as a beneficial dietary supplement for improving the physiological parameters and enhance the immune response of Persian sturgeon.

Evaluación de la efectividad de un extracto de aceite de rosa mosqueta (ARM) en la prevención de la epitelitis por radioterapia (RT) en pacientes afectos de cáncer de cabeza y cuello (CCC) / Evaluation of the effectiveness of an oil extract of rosehip in the prevention of epithelitis due toradiotherapy in patients with head and neck cancer

Introducción. La epitelitis es un efecto secundario frecuente en el tratamiento con radioterapia (RT). Objetivo. Evaluar la efectividad del aceite de rosa mosqueta (ARM, RepavarR) en la prevencion de epitelitis en pacientes con cancer de cabeza y cuello (CCC) tratados con radioterapia de intensidad modulada (IMRT). Material y métodos. Estudio clinico, intervencionista, prospectivo, exploratorio, controlado y abierto. Entre mayo y octubre de 2013 se incluyeron en el estudio 28 pacientes. Tras firmar el consentimiento informado, se les proporciono ARM para aplicarselo 2 veces/dia desde el inicio del tratamiento. Todo tenian prescritas dosis de IMRT ≥ 60 Gy. Enfermeria los evaluo semanalmente. Se realizo un seguimiento fotografico de la piel que registro el grado de epitelitis y la aparicion de dolor u otros. El grupo control lo constituyeron 34 pacientes consecutivos con CCC tratados con IMRT los meses previos al estudio, cuyo tratamiento topico se realizo con otros productos. Resultados. Ambos grupos son comparables (edad, sexo, localización y dosis RT). En el grupo de estudio (ARM) fue menor el numero de curas por paciente (p = 0.3) y los dias naturales de curas por paciente (p = 0.38); tambien fue menor el numero de pacientes que preciso curas y padecio radiodermitis grado III-IV. Probablemente, el limitado tamano muestral haya influido en la significacion estadistica. Conclusiones. La efectividad del ARM en la prevencion de la epitelitis grave en pacientes con CCC tratados con RT es al menos tan eficaz (probablemente mas) que la de los tratamientos topicos habituales. Seria deseable un estudio leatorizado para confirmar dichos hallazgos (AU)

Introduction. Epithelitis is a frequent side effect in radiotherapy treatments (RT). Objective. To evaluate the effectiveness of Rosehip Oil (ARM, Repavar) in the prevention of epithelitis in patients with head and neck cancer (HNC) treated with intensity modulated radiotherapy (IMRT). Material and methods. An interventionist, prospective, exploratory, controlled and open clinical study. Between May and October 2013, 28 patients were included in the study. After signing the informed consent form, the patients were provided with ARM to apply this twice daily from the beginning of the treatment. All of them had a prescribed dose of IMRT ≥ 60 Gy. They were evaluated weekly by nurses on the ward. A photographic monitoring of the skin with a registered grade of epithelitis and with signs of pain or other was carried out. The control group was formed of 34 consecutive patients with HNC treated with IMRT months prior to the study, whose skin treatment had been different products. Results. Both groups were comparable (age, sex, region and RT dose). In the study group (ARM), the number of treatments per patient (p = 0.3) and the natural days for treatment per patient (p = 0.38) were less; also, the number of patients who required treatments and who suffered from radiodermitis of III-IV grade, were less. It is probable that the limited specimen size may have influenced in the statistics. Conclusions. The effectiveness of ARM in the prevention of severe epithelitis in patients with HNC treated with RT is, at least, as effective (or more effective) as other and more common skin treatments. A randomized study would be required to confirm these findings (AU)

Astragalin inhibits IL-1ß-induced inflammatory mediators production in human osteoarthritis chondrocyte by inhibiting NF-κB and MAPK activation.

Astragalin, a bioactive component isolated from Rosa agrestis, has been described to exhibit anti-inflammatory activity. The aim of this study was to investigate the anti-inflammatory effects and the underlying mechanisms of astragalin on IL-1ß-stimulated human osteoarthritis chondrocyte. The production of NO and PGE2 was detected by Griess reaction and ELISA. The expression of iNOS and COX-2 was detected by western blotting. The expression of NF-κB and MAPKs was detected by western blot analysis. We found that astragalin dose-dependently inhibited IL-1ß-induced NO and PGE2 production, as well as iNOS and COX-2 expression. Meanwhile, western blot analysis showed that astragalin inhibited IL-1ß-induced NF-κB and MAPK activation in human osteoarthritis chondrocyte. In addition, astragalin was found to activate PPAR-γ. The inhibition of astragalin on IL-1ß-induced NO and PGE2 production can be reversed by PPAR-γ antagonist GW9662. Astragalin suppressed IL-1ß-induced inflammatory mediators via activating PPAR-γ, which subsequently inhibited IL-1ß-induced NF-κB and MAPK activation. Astragalin may be a potential agent in the treatment of osteoarthritis.

Disease resistance breeding in rose: current status and potential of biotechnological tools.

The cultivated rose is a multispecies complex for which a high level of disease protection is needed due to the low tolerance of blemishes in ornamental plants. The most important fungal diseases are black spot, powdery mildew, botrytis and downy mildew. Rose rosette, a lethal viral pathogen, is emerging as a devastating disease in North America. Currently rose breeders use a recurrent phenotypic selection approach and perform selection for disease resistance for most pathogen issues in a 2-3 year field trial. Marker assisted selection could accelerate this breeding process. Thus far markers have been identified for resistance to black spot (Rdrs) and powdery mildew and with the ability of genotyping by sequencing to generate 1000s of markers our ability to identify markers useful in plant improvement should increase exponentially. Transgenic rose lines with various fungal resistance genes inserted have shown limited success and RNAi technology has potential to provide virus resistance. Roses, as do other plants, have sequences homologous to characterized R-genes in their genomes, some which have been related to specific disease resistance. With improving next generation sequencing technology, our ability to do genomic and transcriptomic studies of the resistance related genes in both the rose and the pathogens to reveal novel gene targets to develop resistant roses will accelerate. Finally, the development of designer nucleases opens up a potentially non-GMO approach to directly modify a rose's DNA to create a disease resistant rose. Although there is much potential, at present rose breeders are not using marker assisted breeding primarily because a good suite of marker/trait associations (MTA) that would ensure a path to stable disease resistance is not available. As our genomic analytical tools improve, so will our ability to identify useful genes and linked markers. Once these MTAs are available, it will be the cost savings, both in time and money, that will convince the breeders to use the technology.

[A case of anaphylaxis due to rose-flavored soft-serve ice cream with pollen food allergy syndrome].

We experienced a 10-year-old boy who had anaphylaxis after eating rose-flavored soft-serve ice cream. The patient felt a sense of discomfort in his throat when eating apple, peach, loquat, Japanese pear, and kiwi fruit. Therefore, we measured specific IgE antibodies to allergen components by ImmunoCAP ISAC. Consequently, the patient gave positive results for all PR-10 proteins from birch, alder, hazel, apple, peach, peanut, hazelnut, and soybean, so we diagnosed him with Pollen Food Allergy Syndrome (PFAS) induced by cross reactivity with pollens of birch family and fruits of rose family. When we conducted the skin prick test as is for red rose syrup because of the belief that anaphylaxis was caused by the rose ingredient contained in rose-flavored soft-serve ice cream, the patient gave a strong positive result. However, the results were negative for rose essence and Food Red No. 2 contained. Subsequently, it was found that red rose syrup contained apple juice. Therefore, we conducted the prick-prick test for apple, and the patient was confirmed to be strongly positive to apple. We thus identified apple as the cause of anaphylaxis. Since there is no legal obligation of labeling specific raw materials when directly selling manufactured and processed food products to general consumers, it is possible for general consumers to mistakenly take them in without knowing the containment of allergic substances. It is believed that the labeling method should be improved in the future.

The photorespiratory pathway is involved in the defense response to powdery mildew infection in chestnut rose.

Chestnut rose (Rosa roxburghii Tratt), a non-domesticated potential fruit crop, is susceptible to infection by powdery mildew. A small-scale customized macro-array was assembled with cDNA clones from a suppression subtractive hybridization library enriched for defense transcripts and used to screen for differentially expressed genes induced by powdery mildew infection. Three photorespiratory genes, encoding the small subunit of RuBisCO (ribulose-1,5-bisphosphate carboxylase/oxygenase), RuBisCO activase and serine-glyoxylate aminotransferase, were identified in this screen. Quantitative RT-PCR analysis of these three genes showed that they were significantly up-regulated 16 h after the host was inoculated with powdery mildew, and assays at the enzyme level confirmed high levels of enzymatic activity 24 h after infection. Of six phytohormones tested, salicylic acid (SA) noticeably induced their expression, and HPLC analysis showed that SA accumulated in the leaves after fungal infection.

Rdr3, a novel locus conferring black spot disease resistance in tetraploid rose: genetic analysis, LRR profiling, and SCAR marker development.

Black spot disease of rose, incited by the fungus Diplocarpon rosae, is found worldwide and is the most important disease of garden roses. A gene-for-gene interaction in this pathosystem is evidenced by the presence of pathogenic races of D. rosae and the previous discovery of a dominant resistance allele at the Rdr1 locus in the diploid Rosa multiflora. The objective of the present study was to genetically analyze resistances to North American black spot races 3, 8, and 9 previously reported in tetraploid roses. Resistance to North American races 3 and 8 segregated 1:1 in multiple F(1) populations, indicating that both are conferred by dominant alleles at single loci and are present in simplex (Rrrr) configuration. Gene pyramiding was demonstrated by combining both resistances into single genotypes. Resistance to race 9 was partial and segregated in a quantitative fashion. Analysis of these populations with microsatellite markers previously developed for Rdr1 revealed that the gene conferring race 3 resistance resides within the same R gene cluster as Rdr1. Race 8 resistance segregated independently and is, therefore, a novel locus for black spot resistance in rose which we have named Rdr3. NBS and LRR profiling were used in a bulked segregant analysis to identify a marker 9.1 cM from Rdr3, which was converted to a SCAR marker form for marker-assisted breeding.

Occupational asthma, eosinophil and skin prick tests and serum total IgE values of the workers in a plant manufacturing rose oil.

This study was aimed to determine the rate of occupational asthma (OA) in workers at a rose extracting plant. Specific clinical tests of 52 workers, randomly chosen from four local rose extracting plants, were statistically compared with the test results of 30 local control subjects of similar age and sex as the plant workers, but who had never worked in such a plant. There were no significant differences in pulmonary function tests (FVC, FEV1, FEV1/FVC, PEFR) between the control and test groups. Significantly higher serum total IgE values (p < 0.0001) were observed for the test subjects (239.08+/-240 IU/ml) compared to the control subjects (81.33+/-61.45 IU/ml). There were also significant differences (p < 0.0001) in the number of eosinophils between the control and test groups, with corresponding mean values of 2.28+/-2.75% and 0.73+/-1.72%, respectively. A specifically prepared skin prick test using a rose allergen (Rosa domescena) was positive for 53.84% in the test subjects whereas only 5.33% positive test results were seen in the control group. We have demonstrated the involvement of Rosa domescena pollen in occupational allergy, through IgE-mediated hypersensitivity. It was concluded that the workers of a rose oil extracting plant are more susceptible to the rose pollens.

Isolation and identification of eight races of powdery mildew of roses (Podosphaera pannosa) (Wallr.: Fr.) de Bary and the genetic analysis of the resistance gene Rpp1.

Powdery mildew, caused by Podosphaera pannosa, is one of the most-severe diseases of roses grown under glass. The differentiation into physiological races and the genetic analysis of resistance in a segregating host population was investigated using single conidial isolates of the pathogen. Using ten rose genotypes, all eight isolates of the pathogen could be ascribed to different races. Five races were isolated from one location, which indicates that populations of P. pannosa exhibit a high racial diversity. Infection experiments in a backcross-population of 114 rose plants resulted in a 1:1 segregation, suggesting control by a single dominant gene. Rpp1 is the first resistance gene against rose powdery mildew to be described.

Allergy symptoms and IgE immune response to rose: an occupational and an environmental disease.

BACKGROUND: Turkey is one of the four major producers of rose. Cultivation of rose is the main economic activity of many villagers in the Lakes region. Rose allergy has not been reported before. We investigated the prevalence of allergy symptoms and specific-IgE immune response due to rose in villagers who had been cultivating rose in Güneykent village in the Lakes region. METHODS: A screening questionnaire including respiratory and allergic symptoms was administered to 290 villagers. The investigation team visited and studied 75 randomly selected villagers with an interviewer-administered questionnaire, pulmonary function testing, skin prick testing and serum total IgE. Specific IgE against Rosa rugosa was measured in 41 villagers. RESULTS: Villagers reported asthma/allergy symptoms outside the rose season (17.6%), during the rose season (6.2%), and both during the rose season and outside the rose season (whole the year) (17.6%). Atopy and specific IgE against Rosa rugosa were detected in 12 (19%), and 8 (19.5%) of the villagers tested. Villagers who had symptoms whole the year reported wheeze more frequently than those who reported symptoms only outside the rose season (41.2% vs 11.1%). CONCLUSIONS: IgE-mediated allergy could occur due to rose and/or its pollen. Symptoms mainly affect the upper airways. Further studies in the rose handlers, florists and workers of the rose industry would help elucidate the occurrence of rose allergy.

Allergic rhinitis in Rosa domescena cultivators: a novel type of occupational allergy?

After the diagnosis of allergic rhinitis due to Rosa domescena was suspected in one subject with skin prick and nasal provocation tests, we recruited all other workers employed in rose cultivation in Yakaören village, Isparta. From May 2000 to July 2000 (exposure period 'time of rose handling'), we studied 600 individuals employed in rose cultivation. A questionnaire was administered by a physician, and skin testing was done by a nurse on 75 individuals suffering from asthma and/or rhinitis. Sera from these 75 subjects were available for immunologic testing. The diagnosis of atopy against rose was based on the presence of work-related symptoms, positive skin prick test (SPT) with rose extracts, and positive RAST. While no subject reported asthmatic symptoms, twenty of them reported either rhino-conjunctivitis, rhinitis or both on exposure to Rosa domescena. Fourteen of these subjects had increased specific IgE levels to Rosa domescena. Eleven (78.5%) also had a positive skin reaction to Rosa domescena. Out of the six negative sera with normal specific IgE levels to rose, only two (33.3%) had positive skin reactivity. No subject had significant daily peak expiratoy flow rate (PEFR) variations. We conclude that exposure to Rosa domescena may represent a risk for allergic rhinitis. The possibility of an occupational rose allergy should therefore be taken into consideration in the subjects working in rose cultivation.