Identifying successful combinations by fertility index in old garden roses and hybrid tea roses crosses.

The success of rose breeding programs is low due to poor seed sets and germination rates. Determining fertile parents and cross combinations that show high compatibility could increase the effectiveness of breeding programs. In this study, three rose varieties belonging to Rosa × hybrida (Jumilia, First Red and Magnum), and two old garden rose species (Black Rose and Cabbage Rose) with known ploidy levels were reciprocally crossbred under controlled conditions to determine the successful crosses by checking fertility. The pollen germination rate (PG), crossability rate (CR), seed number per fruit (SNpF), seed production efficiency (SPE), seed germination rate (SGR), fruit weight (FW), seed weight (SW) and stigma number (SiN), etc. were recorded. Comprehensive fertility index value was calculated. Principal component analysis (PCA), correlation matrix, and hierarchical heat map were used to evaluate the data. The findings showed that old garden roses had more viable pollen than hybrid tea roses. The crossing success improved as pollen fertility increased. Also, female parent fertility improved crossing success just as much as pollen fertility. Although the pollen fertility and stigma numbers were low, some combinations had higher CR and SPE. The maximum SPE (from 8.67% to 19.46%) was determined in combinations where Black Rose was the female parent despite the lower stigma number and low pollen fertility. The highest CR was recorded in Black Rose × First Red (94.36%). All combinations in which Black Rose was used as the female parent had a more stable CR. The SNpF of combinations where hybrid rose varieties were female parents and old garden roses were pollen parents was higher than other combinations where hybrid rose varieties were both female and pollen parents. The SPE in intraspecific crosses was lower than that obtained from interspecific crosses. Moreover, the SGR decreased in combinations that produced heavier seeds. The results suggested that SPE is a more accurate parameter than SNpF in demonstrating combination success in breeding programs. Black Rose × First Red, Black Rose × Jumilia, Black Rose × Magnum and Black Rose × Cabbage Rose combinations can be used successfully as the PCA and heat map showed. Black Rose showed better performance as both seed and pollen parents according to the comprehensive fertility index. From the correlation matrix, it is understood that the number of stigmas cannot be an important criterion in parent selection. Old garden roses can be used as parents to increase the success of breeding programs. However, it is necessary to reveal how successful they are in transferring desired characteristics such as scent, petal number, and color.

Modulation of physiological and biochemical traits of two genotypes of Rosa damascena Mill. by SiO2-NPs under In vitro drought stress.

BACKGROUND: Drought is a major abiotic stress that restricts plant growth and efficiency although some nutrients such as silicon improve drought tolerance by regulating the biosynthesis and accumulating some osmolytes. In this regard, a completely randomized factorial design was performed with three factors including two genotypes ('Maragheh' and 'Kashan'), three concentrations of silicon dioxide nanoparticles (SiO2-NPs) (0, 50, and 100 mg L- 1), and five concentrations of PEG (0, 25, 50, 75, and 100 g L- 1) with three replications. RESULTS: The findings showed that drought stress decreased protein content and it was improved by SiO2-NPs, so the genotype of 'Maragheh' treated with 100 mg L- 1 SiO2-NPs had the highest protein content. Under severe drought stress, had a higher membrane stability index (MSI) than 'Kashan', and the 'Maragheh' explants subjected to 100 mg L- 1 SiO2-NPs exhibited the uppermost MSI. The explants supplemented with 100 mg L- 1 SiO2-NPs sustained their photosynthetic parameters more in comparison with other treatments under drought stress conditions and as well as 100 mg L- 1 SiO2-NPs showed higher content of protein and proline of 'Maragheh' than 'Kashan'. Drought stress reduced Fm, Fv/Fm, and Fv, while SiO2-NPs treatment enhanced these parameters. SiO2-NPs also improved water deficit tolerance by enhancing the activity of antioxidant enzymes such as catalase (CAT), peroxidase (POD), guaiacol peroxidase (GPX), and superoxide dismutase (SOD) and reducing lipid peroxidation and H2O2 concentration. CONCLUSIONS: According to the findings, the genotype 'Maragheh' was more tolerance to drought stress than 'Kashan' by improving water balance, antioxidant enzyme activities, and membrane stability as it was obtained from the unpublished previous evaluation in in vivo conditions and we concluded based on these results, in vitro culture can be used for drought screening in Damask rose plants. The results of the current study revealed that the induced drought stress by polyethylene glycol (PEG) in two Damask rose genotypes was ameliorated with SiO2-NPs and the tolerance genotypes were better than the sensitive ones in response to SiO2-NPs treatment.

Transcriptomic profiling of rose flower under treatment of various phytohormones and plant growth regulators.

Rose is one of the most important ornamental plants, accounting for one-third of the world's fresh cut flower market. The vase life refers to the period of a cut flower retaining its appearance in a vase. During this period, the rose was subjected to a variety of abiotic and biotic stresses, resulting in a reduction in the life of cut flowers. Numerous studies have been carried out on cut rose, which proves the effects of various plant hormones on post-harvest dehydration, petal senescence and abscission, disease and vase life of cut rose flowers. In addition, the natural or synthetic hormones or its inhibitor have been successfully used in cut flower preservatives to extend the vase life of rose. However, there is still a lack of systematic and in-depth research on the expression of rose genes related to plant hormone response. Here we analyzed the gene expression changes of the rose flower under treatment of 11 different plant hormones or its inhibitors in order to provide reference for rose studies.

Extraction and identification of chemical and molecular compounds of Rosa damascene Mill) essence in different genotypes cultivated in some provinces of Iran.

The Rosa damascena Mill. is cultivated in different parts of Iran. For evaluation of its essence, the experiment was carried out on 50 accessions during 2014-17 using randomized complete blocks design with three replications. Results showed that the essence of Kermanshah11 (0.412 g), Yazd 1 (0.400 g), Isfahan 4 (0.364 g), Lorestan 1 (0.361 g), Kermanshah 2 (0.350 g) and Isfahan 3 (0.331 g) accessions had a higher amount of essence. Strong positive correlation coefficients (p≤0.01) were found between essence of citronellol- n-eicosan (0.845), citronellol- n-heneicosane (0.879), n-nonadecane-geraniol (0.883), n-heneicosane-geraniol (0.842), n-heneicosane- geraniol (0.850), n-eicosane- geraniol (0.885), n-nonadecane- geraniol (0.930). Comparing the years, the accessions of Isfahan 5, Arak 1, Kordestan 1, West Azarbaijan 1, Tehran 1, Arak1, Fars 1 and Zanjan1 had the highest citronellol, geraniol, geraniol, n-tetradecanal, n-nonadecane, n-eicosane, n-pentacosane, n-heneicosane, respectively.

Characterization of Rosa canina Fruits Collected in Urban Areas of Slovakia. Genome Size, iPBS Profiles and Antioxidant and Antimicrobial Activities.

The studies of plant bacterial endophytes, colonizing the plant tissues without any signs of diseases, are essential for understanding of ecological interactions. The aim of our study is to detect microbiological contamination and to assess the antimicrobial, antioxidant activity, total phenolic, carotenoid content, genome size, and ploidy of non-cultivated Rosa canina sampled from urban areas. Samples of Rosa canina fruits were collected in three locations in Slovakia. The highest total viable count and the Enterobacteriaceae count in fruits were 4.32 log CFU/g and 4.29 log CFU/g, respectively. Counts of the mesophilic anaerobic sporulating bacteria, Pseudomonas spp., and of the microscopic fungi and yeasts were 3.00, 2.15 log CFU/g, 3.65 log CFU/g, and 2.76 log CFU/g, respectively. Regarding the antimicrobial activity, Escherichia coli and Klebsiela oxytoca were the most sensitive species among the assayed microorganisms to the treatment with the ethanolic extracts of Rosa canina fruits. The fruits were rich in bioactive compounds, polyphenols, and carotenoids, that could be related to their antioxidant activity. Genome sizes of analyzed samples ranged from 2.3 to 2.96. DNA-based fingerprinting obtained by iPBS markers of the Rosa canina var. lapidicola Heinr. Braun., was characterized by some distinctive inserted loci. An interdisciplinary study was performed for the dog roses from different parts of Slovakia that resulted in deeper characterization of this species.

Fruit phytochemical composition and color parameters of 21 accessions of five Rosa species grown in North West Iran.

BACKGROUND: The genus Rosa comprises economically important horticultural plants belonging to the family Rosaceae. Recently, the use of different Rosa species has increased owing to their multipurpose properties (ornamental, food and medicinal uses). In this study, 21 accessions of Rosa genotypes were compared for fruit phytochemical composition and color parameters. RESULTS: The highest antioxidant activity (37.86 mg AAE g-1 FW) and total phenolic (8.17 mg GAE g-1 FW), total flavonoid (2.53 mg QUE g-1 FW), total carotenoid (20.21 mg g-1 FW) and ascorbic acid (84.27 mg g-1 FW) contents were observed in G20 (R. canina), G8 (R. canina), G9 (R. canina), G5 (R. damascena) and G10 (R. moschata) respectively. Chlorogenic acid and gallic acid were found as the main phenolic constituents of Rosa fruits. High amounts of apigenin, rutin, quercetin, p-coumaric acid, cinnamic acid, chlorogenic acid, caffeic acid and gallic acid were obtained in fruit extracts of G6, G14, G6, G8, G19, G9, G19 and G12 respectively. Moreover, the level of color parameters also varied among genotypes. The highest values of a*, b*, L* and chroma were obtained in G4 (R. canina). Based on hierarchical clustering analysis with heat-map, five groups of accessions were identified. CONCLUSION: Different Rosa genotypes are rich in certain phytochemical compounds, with significant variations in their levels being observed. Hence evaluation of Rosa genetic resources can supply valuable data for screening accessions containing high levels of individual phenolics, antioxidants and other bioactive compounds for use in breeding programs and food and pharma industries. © 2019 Society of Chemical Industry.

Rosa hybrida RhERF1 and RhERF4 mediate ethylene- and auxin-regulated petal abscission by influencing pectin degradation.

The timing of plant organ abscission is modulated by the balance of two hormones, ethylene and auxin, while the mechanism of organ shedding depends on the loss of middle lamella pectin in the abscission zone (AZ). However, the mechanisms involved in sensing the balance of auxin and ethylene and that affect pectin degradation during abscission are not well understood. In this study, we identified two members of the APETALA2/ethylene-responsive factor (AP2/ERF) transcription factor family in rose (Rosa hybrida), RhERF1 and RhERF4 which play a role in petal abscission. The expression of RhERF1 and RhERF4 was influenced by ethylene and auxin, respectively. Reduced expression of RhERF1 or RhERF4 was observed to accelerate petal abscission. Global expression analysis and real-time PCR assays revealed that RhERF1 and RhERF4 modulate the expression of genes encoding pectin-metabolizing enzymes. A reduction in the abundance of pectin epitopes was detected in the AZs of RhERF1 and RhERF4-silenced plants by immunofluorescence microscopy analysis. In addition, RhERF1 and RhERF4 were shown to bind to the promoter of the pectin-metabolizing gene ß-GALACTOSIDASE 1 (RhBGLA1), and reduced expression of RhBGLA1 delayed petal abscission. We conclude that during petal abscission, RhERF1 and RhERF4 integrate and coordinate ethylene and auxin signals to modulate pectin metabolism, in part by regulating the expression of RhBGLA1.

The strong competitive role of 2n pollen in several polyploidy hybridizations in Rosa hybrida.

BACKGROUND: 2n pollen play a strong competitive role in hybridization and breeding of multiploids in Rosa hybrida. The ploidy inheritable characteristic of 'Orange Fire' × 'Old Blush' were analyzed. RESULT: The results of the cytological observations indicated that 2n pollen developed from the defeated cytoplasmic division or nuclear division in the meiosis metaphase II of PMC (pollen mother cell) in 'Old Blush'. The natural generation rate of the 2n pollen in 'Old Blush' (2x) was about 1.39 in percentage of all male gametes, whereas the tetraploids in the F1 offspring possessed a high rate, i.e., 44.00%. The temporal and spatial characteristics of 'Old Blush' pollen germination on the stigma and growth in pistil of 'Orange Fire' and 'DEE' were observed, and the results suggested that the germination rate of 2n pollen on the stigma was not superior to that of 1n pollen, but that the proportion of 2n pollen increased to 30.90 and 37.20%, respectively, while it traversed the stigma and entered into style. The callose plug in the 2n pollen tube was significantly thinner than that of 1n pollen tube. And each trait involved in our experiment probably is very important for F1 morphological phenotypes. CONCLUSION: We conclude that 2n pollen are involved in hybridization and have a competitive advantage while it traversed the stigma and entered into style. The callose plug in the 2n pollen tube was may have strongly influenced the competitive process in R. hybrida.

High penetrance of a pan-canina type rDNA family in intersection Rosa hybrids suggests strong selection of bivalent chromosomes in the section Caninae.

All dogroses (Rosa sect. Caninae) are characterized by the peculiar canina meiosis in which genetic material is unevenly distributed between female and male gametes. The pan-canina rDNA family (termed beta) appears to be conserved in all dogroses analyzed so far. Here, we have studied rDNAs in experimental hybrids obtained from open pollination of F1 plants derived from 2 independent intersectional crosses between the pentaploid dogrose species (2n = 5x = 35) Rosa rubiginosa as female parent (producing 4x egg cells due to the unique asymmetrical canina meiosis) and the tetraploid (2n = 4x = 28) garden rose R. hybrida 'André Brichet' as male parent (producing 2x pollen after normal meiosis). We analyzed the structure of rDNA units by molecular methods [CAPS and extensive sequencing of internal transcribed spacers (ITS)] and determined the number of loci on chromosomes by FISH. FISH showed that R. rubiginosa and 'André Brichet' harbored 5 and 4 highly heteromorphic rDNA loci, respectively. In the second generation of hybrid lines, we observed a reduced number of loci (4 and 5 instead of the expected 6). In R. rubiginosa and 'André Brichet', 2-3 major ITS types were found which is consistent with a weak homogenization pressure maintaining high diversity of ITS types in this genus. In contrast to expectation (the null hypothesis of Mendelian inheritance of ITS families), we observed reduced ITS diversity in some individuals of the second generation which might derive from self-fertilization or from a backcross to R. rubiginosa. In these individuals, the pan-canina beta family appeared to be markedly enriched, while the paternal families were lost or diminished in copies. Although the mechanism of biased meiotic transmission of certain rDNA types is currently unknown, we speculate that the bivalent-forming chromosomes carrying the beta rDNA family exhibit extraordinary pairing efficiency and/or are subjected to strong selection in Caninae polyploids.

Polyploidization mechanisms: temperature environment can induce diploid gamete formation in Rosa sp.

Polyploidy is an important evolutionary phenomenon but the mechanisms by which polyploidy arises still remain underexplored. There may be an environmental component to polyploidization. This study aimed to clarify how temperature may promote diploid gamete formation considered an essential element for sexual polyploidization. First of all, a detailed cytological analysis of microsporogenesis and microgametogenesis was performed to target precisely the key developmental stages which are the most sensitive to temperature. Then, heat-induced modifications in sporad and pollen characteristics were analysed through an exposition of high temperature gradient. Rosa plants are sensitive to high temperatures with a developmental sensitivity window limited to meiosis. Moreover, the range of efficient temperatures is actually narrow. 36 °C at early meiosis led to a decrease in pollen viability, pollen ectexine defects but especially the appearance of numerous diploid pollen grains. They resulted from dyads or triads mainly formed following heat-induced spindle misorientations in telophase II. A high temperature environment has the potential to increase gamete ploidy level. The high frequencies of diplogametes obtained at some extreme temperatures support the hypothesis that polyploidization events could have occurred in adverse conditions and suggest polyploidization facilitating in a global change context.

Botanical origin of mei-gui hua (petal of a Rosa species).

Mei-gui hua has been used as a crude drug in traditional medicine and as herbal tea in China. The scientific name of Mei-gui is Rosa rugosa thunb. However, the morphological characteristics and botanical ecology of Mei-gui were different from those of R. rugosa. Since the botanical origins of Mei-gui cultivated in China have not yet been clarified, we compared Mei-gui and R. rugosa in terms of their morphological characteristics, phylogenetic analysis, and phytochemical studies. Our research suggested that Mei-gui cultivated around Tarim Basin in Xinjiang Province showed homology to Rosa gallica, while those cultivated in the northeastern parts of China are considered to be hybrids of R. rugosa.

The asymmetric meiosis in pentaploid dogroses (Rosa sect. Caninae) is associated with a skewed distribution of rRNA gene families in the gametes.

In pentaploid dogroses, Rosa section Caninae (2n=5x=35), the pollen transmits one basic genome (x=7) derived from the seven segregating bivalents, whereas the egg transmits four basic genomes (4x=28) one set derived from the segregation of seven bivalents and three sets of univalent-forming chromosomes. Chromosomes from all five genomes carry 18-5.8-26S nuclear ribosomal DNA (rDNA) sites. This mode of sexual reproduction, known as permanent odd polyploidy, can potentially lead to the independent evolution of rDNA on bivalent- and univalent-forming chromosomes. To test this hypothesis, we analyzed rRNA gene families in pollen and somatic leaf tissue of R. canina, R. rubiginosa and R. dumalis. Six major rRNA gene families (alpha, beta, beta' gamma, delta and epsilon) were identified based on several highly polymorphic sites in the internal transcribed spacers (ITSs). At least two of the major rRNA gene families were found in each species indicating that rDNAs have not been homogenized across subgenomes. A comparison of ITS1 sequences from leaf and pollen showed differences: the shared beta rRNA gene family was more abundant among pollen clones compared to leaf clones and must constitute a major part of the rDNA loci on bivalent-forming chromosomes. The gamma and delta families were underrepresented in pollen genomes and are probably located predominantly (or solely) on the univalents. The results support the hypothesis that pentaploid dogroses inherited a bivalent-forming genome from a common proto-canina ancestor, a likely donor of the beta rDNA family. Allopolyploidy with distantly related species is likely to have driven evolution of Rosa section Caninae.

The use of bead beating to prepare suspensions of nuclei for flow cytometry from fresh leaves, herbarium leaves, petals and pollen.

"Bead beating" is commonly used to release DNA from cells for genomic studies but it was used here to prepare suspensions of plant nuclei for measurement of DNA amounts by flow cytometry. Plant material was placed in 2-ml screw-capped tubes containing beads of zirconia/silica (2.5 mm diameter) or glass (2.5 or 1.0 mm diameter) and 1 ml of lysis buffer. The tubes were mechanically shaken with an FP120 FastPrep Cell Disrupter to release intact nuclei from plant tissue by the impact of the beads. The nuclei were then stained with propidium iodide (PI) and analyzed by flow cytometry. The method was tested using fresh leaves, fresh petals and herbarium leaves of Rosa canina, leaves and pollen of R. rugosa, and fresh leaves of Petroselinum crispum, Nicotiana tabacum, and Allium cepa. Batches of 12 samples of fresh leaves were prepared, simultaneously, in 45 s by bead beating in the Cell Disrupter. In flow cytometry histograms, nuclei of fresh leaves gave G(1)/G(0) peaks with CVs of less than 3.0% and nuclei from fresh petals and herbarium leaves of R. canina, and pollen of the generative nuclei of R. rugosa gave peaks with coefficients of variation (CVs) of less than 4.0%. DNA amounts estimated from 24-month-old herbarium leaves, using P. crispum as an internal standard, were less than those of fresh leaves by a small but significant amount. Suspensions of nuclei can be prepared rapidly and conveniently from a diversity of tissues by bead beating. Exposure of laboratory workers to harmful substances in the lysis buffer is minimized.

Evolutionary implications of permanent odd polyploidy in the stable sexual, pentaploid of Rosa canina L.

In Rosa canina (2n = 5x = 35), the pollen and ovular parents contribute, respectively, seven and 28 chromosomes to the zygote. At meiosis I, 14 chromosomes form seven bivalents and 21 chromosomes remain as univalents. Fluorescent in situ hybridization to mitotic and pollen mother cells (PMC) of R. canina showed that 10 chromosomes (two per genome) carry ribosomal DNA (rDNA) loci. Five chromosomes carry terminal 18S-5.8S-26S rDNA loci; three of these also carry paracentric 5S rDNA loci and were designated as marker chromosomes 1. Five chromosomes carry only 5S rDNA loci and three of these were designated as marker chromosomes 2. The remaining four of the 10 chromosomes with rDNA loci were individually identifiable by the type and relative sizes of their rDNA loci and were numbered separately. At PMC meiosis, two marker chromosomes 1 and two marker chromosomes 2 formed bivalents, whereas the others were unpaired. In a gynogenetic haploid of R. canina (n = 4x = 28), obtained after pollination with gamma-irradiated pollen, chromosomes at meiosis I in PMC remained predominantly unpaired. The data indicate only one pair of truly homologous genomes in R. canina. The 21 unpaired chromosomes probably remain as univalents through multiple generations and do not recombine. The long-term evolutionary consequence for the univalents is likely to be genetic degradation through accumulated mutational change as in the mammalian Y chromosome and chromosomes of asexual species. But there is no indication that univalents carry degenerate 5S rDNA families. This may point to a recent evolution of the R. canina meiotic system.

O-methyltransferases involved in the biosynthesis of volatile phenolic derivatives in rose petals.

Rose (Rosa hybrida) flowers produce and emit a diverse array of volatiles, characteristic to their unique scent. One of the most prominent compounds in the floral volatiles of many rose varieties is the methoxylated phenolic derivative 3,5-dimethoxytoluene (orcinol dimethyl ether). Cell-free extracts derived from developing rose petals displayed O-methyltransferase (OMT) activities toward several phenolic substrates, including 3,5-dihydroxytoluene (orcinol), 3-methoxy,5-hydroxytoluene (orcinol monomethyl ether), 1-methoxy, 2-hydroxy benezene (guaiacol), and eugenol. The activity was most prominent in rose cv Golden Gate, a variety that produces relatively high levels of orcinol dimethyl ether, as compared with rose cv Fragrant Cloud, an otherwise scented variety but which emits almost no orcinol dimethyl ether. Using a functional genomics approach, we have identified and characterized two closely related cDNAs from a rose petal library that each encode a protein capable of methylating the penultimate and immediate precursors (orcinol and orcinol monomethyl ether, respectively) to give the final orcinol dimethyl ether product. The enzymes, designated orcinol OMTs (OOMT1 and OOMT2), are closely related to other plant methyltransferases whose substrates range from isoflavones to phenylpropenes. The peak in the levels of OOMT1 and OOMT2 transcripts in the flowers coincides with peak OMT activity and with the emission of orcinol dimethyl ether.

Pollen mother cells of Tradescantia clone 4430 and Tradescantia pallida var. purpurea are equally sensitive to the clastogenic effects of X-rays.

The Tradescantia micronucleus test is a sensitive bioassay for mutagenesis that may be employed both under field and laboratory conditions. This test has been standardized mostly on the basis of the results obtained with clone 4430. However, this clone is not well adapted to tropical weather, frequently showing problems with growth and flowering. In addition, it is attacked by parasites and insects, a fact that limits its use in field studies aiming at the biomonitoring of air pollution. In the city of São Paulo, Tradescantia pallida (Rose) Hunt. var. purpurea Boom is widely distributed as an ornamental plant in gardens and along roadsides and streets, mostly because of its natural resistance and its easy propagation. In this report, we present dose-response curves indicating that the sensitivity of T. pallida and clone 4430 to X-radiation (1, 10, 25 and 50 cGy) is similar. The results confirm our previous suggestion that T. pallida represents a good alternative for in situ mutagenesis testing in tropical regions, especially biomonitoring studies in which the exposure conditions may not be fully controllable.