Microsatellite analysis of Damask rose (<Emphasis Type="Italic">Rosa damascena</Emphasis> Mill.) accessions from various regions in Iran reveals multiple genotypes

Background  

Damask roses (Rosa damascena Mill.) are mainly used for essential oil production. Previous studies have indicated that all production material in Bulgaria and Turkey consists of only one genotype. Nine polymorphic microsatellite markers were used to analyze the genetic diversity of 40 accessions of R. damascena collected across major and minor rose oil production areas in Iran.     

Triparental origin of Damask roses

Damask roses are one group of old rose varieties and a key material in old European rose improvement in the 19th century. To clarify the origin of Damask roses, we selected four varieties as the oldest Damask varieties and examined the relationship between the Damask varieties and their putative ancestors at the molecular level. Randomly amplified polymorphic DNA analysis of the Damask varieties proved that they had an identical profile, indicating they were established from a common ancestor. They have never been allowed to reproduce sexually; their reproduction depends entirely on vegetative propagation. We identified three Rosa species, R. moschata, R. gallica and R. fedschenkoana, as parental species of the original hybridization that contributed to forming the four oldest Damask varieties by sequencing the internal transcribed spacer of ribosomal DNA. We also found that all the four oldest Damask varieties had chloroplasts derived only from R. moschata, as judged from psbA-trnH spacer sequences. This triparental origin of the four oldest Damask varieties can explain some morphological characteristics of the four oldest Damask varieties, like fruit shape, leaf color and the 'Moss' character.     

Identification of fungal diseases of Rosa damascena in Kerman province of Iran

The oil rose, Rosa damascena Mill. (Rosaceae) is an agricultural crop cultivated in various countries of the northern hemisphere, such as Turkey, Bulgaria, Morocco, Iran, Egypt, France, China and India. Iran, presently, is the largest producer of rose water in world. The major production areas in Iran are Kashan, Fars, Kerman and Azerbaijan. Kerman province with 2297 hectares ha of rose gardens and 6198 tons of flower production is one of the important rose production regions. The productions of this region are organic and do not use anychemical compounds such as pesticides and fertilisers. The major fungal pathogens were studied during 2008–2010 in oil rose production areas in Kerman province, Iran. Verticillium dahlias, Rosellinia necatrix, Alternaria alternata, Seimatosporium fusisporum and Podosphaera pannosa have been detected in the oil rose from different regions in Kerman province. A. alternata has the most isolates and infected plants per cent in the oil rose. This is the first report fromVerticillium dahliae, R. necatrix, A. alternata, S. fusisporum on oil roses (R.damascena) in the world.     

Changes in abscisic acid and phenols during flower development in two diverse species of rose

Changes in endogenous abscisic acid (ABA) and phenols were determined in petals of two diverse species of rose, viz., Rosa damascena Mill and Rosa bourboniana Desport during flower development. A progressive increase in free ABA was observed during flower development till full bloom in both the species with higher content of free ABA in Rosa damascena. While bound ABA level increased in Rosa damascena till stage 6, in Rosa bourboniana it continued to increase till full bloom. Acidic phenols increased slowly in both the species till stage 4, but sharply afterwards and no significant differences were noticed during full bloom period. Bound phenols content was higher in Rosa damascena during full bloom period. The significance of these changes in relation to flowering in the two diverse species of rose is discussed.     

Composition and structure of lactones from rose flower wax

The presence of seven members of the homologous series of γ-lactones in wax of the Bulgarian oil-bearing rose (Rosa damascena Mill.) was established by means of mass spectrometry of a fraction isolated by chromatography. The lactone of γ-hydroxy hexacosanoic acid was the predominant member.     

Ferricyanide reductase of rose plasma membranes is regulated by nitrogen supply

Summary The ability of suspension-cultured rose (Rosa damascena Mill. cv Gloire de Guilan) cells to reduce ferricyanide is decreased by 50% during an overnight incubation in a low-nutrient (1 mM CaCl₂, 0.1 mM KCl) solution. This loss is not observed when nitrate and/or glutamate is added to the low-nutrient medium, but it occurs in medium containing all the components needed for normal growth except nitrate plus glutamate. Thus, the cells possess both constitutive and inducible enzymes for the reduction of ferricyanide, and nitrate or glutamate is both necessary and sufficient to stimulate the production of the inducible enzyme.     

Development of microsatellite markers in rose

Thirteen polymorphic microsatellite markers were developed in rose (Rosa hybrida L.) from an enriched genomic library. Ten out of 13 microsatellites showed amplification and produced four to 12 polymorphic alleles per locus, with an average of 8.4 in Rosa multiflora. The average values of observed and expected heterozygosities among the 10 loci were 0.62 and 0.82, respectively. Thirteen microsatellites produced three to 22 genotypes with an average value of 11.4 in modern garden roses. The average value of the power of discrimination among the microsatellites was 0.79, ranging from 0.29 to 0.95.     

RAPD analysis of genetic variation between a group of rose cultivars and selected wild rose species

The genetic variability based on random-amplified polymorphic DNA markers was analysed among 10 cultivated rose varieties and 9 wild species from three different series of the genus Rosa. Using 13 different RAPD primers, 104 polymorphic DNA fragments with a high potential to differentiate rose genotypes could be produced. A dendrogram displaying the relative genetic similarities among the genotypes shows the existence of large genetic diversity among the cultivated roses as compared to the wild species. Furthermore, the main clusters found here are in agreement with known pedigrees and the classical taxonomy. However, the relationships between cultivated roses as inferred by RAPD markers do not correlate with the classical rose classification system. From the present data it is concluded that cultivated roses display a high level of genetic variability despite the fact that single morphological and physiological characters may be less polymorphic within rose groups. This contrasts with the widely accepted opinion of a lack of genetic variability in roses. This is also in accordance with the reported history of rose breeding which makes it highly probable that rose genomes comprise mosaics of different species genomes. As a consequence, it may be possible to utilize the high genetic variability of all genetic traits not under actual selection by breeders for future breeding programmes.     

Composition of rugosa rose (Rosa rugosa thunb.) hydrolate according to the time of distillation

Rugosa rose (Rosa rugosa Thunb.) is one of the most common rose species in Poland. It has mild soil and climate requirements and is resistant to low temperatures. Rugosa rose hips are a valuable raw material used in food and pharmaceutical industries, while flowers and petals may be a source of fragrant products, such as essential oil and hydrolate. The main aim of this study was to verify usefulness of dried R. rugosa petals for essential oil and hydrolate production. We also assessed the use of rugosa rose petals remaining after oil distillation for hydrolate production.The R. rugosa dried petals immersed in water were subjected to simple distillation and five fractions of primary rose hydrolate were obtained. In parallel, essential oil from the second sample of petals was obtained by hydrodistillation in Clevenger-type apparatus. The distillation residue was used for obtaining four fractions of secondary hydrolate. The volatiles from hydrolate fractions were isolated by liquid–liquid extraction with diethyl ether. The essential oil and hydrolate volatiles were analyzed by GC–FID–MS.Hydrolate fractions contained similar amounts of volatiles (20–30 mg/L) with the exception of the first fraction of primary hydrolate (60 mg/L). β-Phenylethanol, citronellol, geraniol, and nerol were the main volatile constituents of primary hydrolate. β-Phenylethanol, citronellic acid, and geranic acid were the main volatile constituents of secondary hydrolate. The content of alcohols decreased, while the content of monoterpene esters (citronellyl, neryl, and geranyl acetate) as well as monoterpene acids (citronellic, neric, and geranic acid) increased in successive fractions of both hydrolates.The scent and composition of essential oil and hydrolate obtained from R. rugosa petals were similar to those of rose oil and rose water produced from damask rose (Rosa damascena Mill.). This proves, that rugosa rose distillation products may become an alternative to fragrant damask rose products.     

In vitro propagation of the Damask rose (Rosa damascena Mill.)

Roses are an important commercial crop available in a wide range of varieties in international markets. Due to its economic value, this study aimed to establish a new and reproducible protocol for the in vitro propagation of Rosa damascena Mill. We developed an efficient and cost-effective method for rapid and high-quality shoot multiplication and in vitro rooting of Damask rose using nodal explants. For each stage of the micropropagation procedure (i.e., explant establishment, shoot multiplication and growth, and rooting), different media and combinations of plant growth regulators were utilized. A new culture medium, termed A₁₉, resulted in significant improvements to shoot proliferation and root induction for this rose cultivar. For optimal explant establishment, shoot growth, and proliferation, a modified Murashige and Skoog medium with higher levels of nitrates, calcium, and iron plus supplementation with 4?mg/l 6-benzylaminopurine and 0.25?mg/l indole-3-acetic acid was utilized. To increase shoot length, 75?d after culture initiation (including two subcultures), shoots were transferred to the same medium additionally supplemented with 0.2?mg/l gibberellic acid. This resulted in vigorous shoot growth, with longer shoots and a greater number of shoots per explant. Shoots were then separated and transferred to various root induction medium for 30?d. The results clearly showed that a liquid ?A₁₉ medium-A (i.e., with half-strength macroelements) supplemented with 0.1?mg/l indole-3-butyric-acid was the most successful medium for in vitro rooting in this cultivar. Shoots were cultured in this medium for 7?d in the dark, before transfer to liquid ?A₁₉ medium-A without hormone supplementation under a 16-h photoperiod. This modified protocol resulted in significant improvement in shoot regeneration and proliferation and obtained stronger shoots over a period of about 20?wk.     

Studies on rose mosaic disease in field-grown roses produced in the United Kingdom

Arabis mosaic virus (AMV) and prunus necrotic ringspot virus (PNRSV), separately or together, caused in field-grown roses the range of symptoms recognised as rose mosaic disease. PNRSV infection alone generally induced chlorotic line patterns, ring-spots or mottles in the leaves at some time during the growing season; AMV plus PNRSV normally caused chlorotic vein-banding. However, during prolonged periods of high temperatures (c. 21 °C or more) vein banding occurred in some roses infected only with PNRSV. Isolates of PNRSV from rose had particles which were similar in shape, protein mol. wt, density and sedimentation coefficients to previously described isolates of PNRSV from cherry, plum and rose; all were cherry serotypes. In graft-inoculated roses, apple serotypes of PNRSV induced stunting and chlorosis, puckering and distortion of leaves, which closely resembled symptoms associated with rose mosaic in the USA and chlorotic mottle rose mosaic in New Zealand. To avoid possible confusion in using the name rose mosaic it is suggested that the virus(es) present in roses should be named.     

Changes in endogenous polyamines during flower development in two diverse species of rose

Changes in the concentrations of endogenous free, conjugated and bound polyamine were determined in petals of two different species of rose, viz. Rosa damascena and Rosa bourboniana, from small bud (stage 1) till full bloom (stage 8). High free putrescine and spermidine concentrations were associated with early stages of flower development and then decreased in R. damascena. At full bloom, the concentration of free putrescine was higher than rest of the polyamines measured. A steady increase in conjugated putrescine, spermidine and spermine was observed during entire period of flower development with predominance of conjugated putrescine at full bloom. In R. damascena the bound spermine was higher than rest of the polyamines during full bloom. In R. bourboniana, during the early stages of flower development, similar situation was observed, however, at full bloom, free spermidine concentration was higher than rest of the polyamines. In this species, the concentration of conjugated and bound spermine was higher than rest of the polyamines during full bloom. Polyamine concentrations were generally lower in the petals of R. bourboniana than R. damascena which may be due to genotypic differences. The possible roles of the observed polyamines are discussed in relation to flower development.IHBT Communication no, 0345.     

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 F₁ 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.     

Tocopherol,Carotene, Phenolic Contents and Antibacterial Properties of Rose Essential Oil,Hydrosol and Absolute

The antioxidant and antibacterial activities, and total phenolic contents of Rosa damascena Mill. flower extracts (absolute, essential oil and hydrosol) were investigated. The chemical compositions of these extracts were analysed by GC-MS. Phenylethyl alcohol (78.38%) was found to be the main constituent of rose absolute, while citrenellol and geraniol were the major compounds (>55%) of rose essential oil and hydrosol. Tocopherol and carotene levels were determined by high performance liquid chromatography (HPLC) analysis. The levels of beta carotene (422.3±35.6 ppm), alpha tocopherol (2397.1±72.5 ppm) and gamma tocopherol (343.1±28.4 ppm) of rose absolute were found to be higher than that of essential oil and hydrosol. Their total phenolic contents were also evaluated. The total phenolic content of the tested extracts varied from 5.2 to 2134.3 GAE/mg L⁻¹. Rose absolute and essential oil contained high levels of phenolics and demonstrated strong antibacterial activity against Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), Bacillus subtilis (ATCC 6633), Staphylococcus aureus (ATCC 6538), Chromobacterium violaceum (ATCC 12472) and Erwinia carotovora (ATCC 39048) strains.     

Post-harvest alterations in polyamines and ethylene in two diverse rose species

Changes in the concentrations of endogenous polyamines and ethylene were determined in two diverse species of rose viz. Rosa damascena and Rosa bourboniana during post-harvest periods. At full bloom, the concentrations of free putrescine was significantly higher than rest of the polyamines, i.e. spermine and spermidine in both the species. The concentrations of all the polyamines decreased during subsequent periods upto 48 h after full bloom. Similar situation was also observed in conjugated fraction but in bound fraction, during full bloom, the concentration of spermine was higher than rest of the polyamines. In both the species, ethylene showed higher levels during full bloom with maximum in R. damascena, which increased, during post-harvest periods. The possible significance of polyamines, ethylene and their interactions is discussed during post-harvest periods in flowers.     

The effect of polyamines on leaf senescence in two diverse rose species

Polyamines (PAs) retarded the senescence of leaf discs of two diverse speciesof rose viz., Rosa bourboniana andRosa damascena, while polyamine biosynthetic inhibitorsdifluoromethylornithine (DFMO), difluoromethylarginine (DFMA), methylglyoxal-bis(guanylhydrazone) (MGBG) and abscisic acid (ABA) promoted senescence. Sperminewas significantly the most effective polyamine in retarding senescence inR. bourboniana while MGBG and DFMA were more prominent inaccelerating senescence in R. damascena and R.bourboniana respectively. Protein and RNA content were significantlyhigher in polyamine treated leaf discs compared to those treated with polyaminebiosynthetic inhibitors and ABA. Total and reducing sugars decreased under alltreatments while the starch content increased significantly only in polyaminetreated leaf discs. Peroxidase and cellulase activities were retarded bypolyamine treatments and accelerated by polyamine biosynthetic inhibitors andABA. The role of PAs is discussed in relation to senescence.     

基于过氧化物同工酶分析月季种质资源的亲缘关系及杂种真实性

为了提高种质资源利用率,加速月季育种进程,对27份月季种质及3个杂交组合的8个杂交后代,采用过氧化物酶(POD)同工酶方法分析其亲缘关系并进行杂种真实性鉴定。结果表明:月季种质采用POD同工酶分析具有一定的可行性。酶谱分析中,在相对迁移率为0.264~0.858的位点处共获得7条酶带,其中共有酶带3条,特征酶带4条,表明不同月季种质间遗传多样性丰富,但又存在一定同源性。基于酶带特征进行聚类分析,在相似系数为0.57处,可将27份供试材料分为3个大组。合柱组与月季组材料聚在一个大组中,两个组在形态上的相似性再次得到确认。金樱子与硕苞蔷薇分别聚在两个不同的组中,表明二者之间的亲缘关系较远。供试的古老月季品种被聚在两个不同的大组中,与野生种聚成的一组呈平行关系,表明古老月季在起源上的差异较大,可利用其作为杂交亲本进行广泛杂交以选育具有丰富遗传多样性的杂交后代。根据有无父本特征酶带对杂种后代真实性进行鉴定,初步确定2个杂交组合的6个杂交后代中5个为真实杂种,1个为自交种。该研究结果为进一步开展月季遗传育种奠定了基础。     

Xylem and Phloem Derived Polyamines during Flowering in Two Diverse Rose Species

Polyamine contents in xylem (root) and phloem (leaf) exudates in two diverse species of rose, viz. Rosa damascena Mill and Rosa bourboniana Desport, were analyzed before, during, and after flowering in the main flowering season, that is, April–May. Only free putrescine (Put) was detected in the xylem and phloem exudates at these time points, and it was high during the peak flowering period. In phloem, Put content was significantly higher in R. bourboniana than in R. damascena at all three stages; whereas in the xylem exudate it was relatively higher in R. damascena at the peak flowering period. A spray of α-difluoromethylornithine (DFMO), an irreversible inhibitor of the putrescine biosynthetic inhibitor ornithine decarboxylase (ODC), markedly decreased the flowering. This effect was reversed by application of Put alone or in combination with DFMO. The significance of this finding is discussed in light of polyamine translocation during flowering. *IHBT Communication: 0354     

Epidemiology of three viruses infecting the rose in the United Kingdom

Studies on the epidemiology of arabis mosaic (AMV), prunus necrotic ringspot (PNRSV) and strawberry latent ringspot (SLRV) viruses were made in relation to commercial production of standard and bush roses. AMV or SLRV apparently induced either symptomless infection in rose cultivars and Rosa spp., or leaf symptoms ranging from small chlorotic flecks to severe chlorotic mosaic and, occasionally, plant death. Infection of R. canina ‘inermis’ or R. corymbifera by an isolate of SLRV from R. corymbifera also severely depressed flowering and hip formation. In addition, whereas this isolate could be graft-transmitted to all Rosa spp. tested, isolates from R. rugosa and R. multiflora failed to be graft-transmitted to R. canina ‘inermis’ or R. corymbifera. No difference was detected in graft-transmission tests of Rosa spp. with several isolates of AMV or PNRSV. In plantings of up to 7 yr none of the viruses was transmitted through pollen to healthy roses grown in nematode-free soil, and only SLRV was readily seed-transmitted, particularly in R. rugosa. Nevertheless, in soil containing viruliferous nematodes, AMV and/or SLRV were transmitted to c. 80% of healthy plants. AMV and particularly SLRV were each damaging to field-grown maiden rose bushes cv. Fragrant Cloud. SLRV delayed the onset of flowering, and reduced the number and size of blooms. Diseased bushes were less vigorous, and half or none of the AMV- or SLRV- infected bushes respectively, conformed to the British Standards Institution specifications for maiden bush roses. These results are discussed in relation to the commercial production of field-grown roses in the UK.