Carnation Cultivars Research Articles

Esterified carotenoids are synthesized in petals of carnation (Dianthus caryophyllus) and accumulate in differentiated chromoplasts

Although yellow and orange petal colors are derived from carotenoids in many plant species, this has not yet been demonstrated for the order Caryophyllales, which includes carnations. Here, we identified a carnation cultivar with pale yellow flowers that accumulated carotenoids in petals. Additionally, some xanthophyll compounds were esterified, as is the case for yellow flowers in other plant species. Ultrastructural analysis showed that chromoplasts with numerous plastoglobules, in which flower-specific carotenoids accumulate, were present in the pale yellow petals. RNA-seq and RT-qPCR analyses indicated that the expression levels of genes for carotenoid biosynthesis and esterification in pale yellow and pink petals (that accumulate small amounts of carotenoids) were similar or lower than in green petals (that accumulate substantial amounts of carotenoids) and white petals (that accumulate extremely low levels of carotenoids). Pale yellow and pink petals had a considerably lower level of expression of genes for carotenoid degradation than white petals, suggesting that reduced degradation activity caused accumulation of carotenoids. Our results indicate that some carnation cultivars can synthesize and accumulate esterified carotenoids. By manipulating the rate of biosynthesis and esterification of carotenoids in these cultivars, it should be feasible to produce novel carnation cultivars with vivid yellow flowers.

Regeneration of Genetically Stable Plants from in Vitro Vitrified Leaves of Different Carnation Cultivars.

This study was conducted to investigate the efficacy of shoot regeneration from different leaf types (normal leaves and vitrified leaves) from three different carnation cultivars ‘Kumbuyl’, ‘Denev’, and ‘Jinju’ using different combinations of 3-indole butyric acid (IBA) and thidiazuron (TDZ) concentrations. The shoot tips cultured on Murashige and Skoog (MS) basal media (Type 1 media) produced normal leaves, while those cultured-on media supplemented with plant growth regulators and/or vitamin (Type 2 media and Type 3 media) produced vitrified leaves for all cultivars. Culture of normal leaf segments on MS medium containing different combinations of IBA and TDZ concentrations induced callus in all treatments; however, the callus was unable to induce shoots and finally became necrotic. In contrast, no callus induction was observed in the control (hormone-free treatment). When vitrified leaf segments underwent the same treatments, shoots were induced from the vitrified leaves (derived from Type 2 media) but were unhealthy and gradually died, whereas those induced from Type 3 media were vitrified and healthy. The optimal combination for the best shoot regeneration and number of shoots per explants varied depending on the genotypes used. The vitrified shoots induced from the leaves of Type 3 media transformed into normal shoots and survived well under greenhouse conditions. According to the results of random amplified polymorphic DNA (RAPD) analysis, the banding patterns of twelve primers that were detected in vitrified leaf-induced normalized shoots were identical to those of normal in vitro grown plants, indicating that no genetic variation had occurred during the procedure. Taken together, this study indicates that vitrified leaves can be used for shoot regeneration of recalcitrant carnation cultivars, regardless of the genotypes and types of vitrified leaves. However, as the number of shoots per explants was still low, further investigation is warranted to obtain a more efficient shoot regeneration protocol for genetic transformation of the cultivars.

Flower color mutation caused by spontaneous cell layer displacement in carnation (Dianthus caryophyllus)

A change of layer arrangement of shoot apical meristem (SAM) organized by three cell layers (L1, L2 and L3) is thought to be one of the provocations of bud sport, which often induces changes in phenotypic colors in periclinal chimeras. This paper describes a cell layer rearrangement which is the cause of spontaneous flower color mutation by using two carnation (Dianthus caryophyllus L.) cultivars that are presumably periclinal chimeras, ‘Feminine Minami’ (deep pink flower) and its recessive sport ‘Tommy Minami’ (pinkish red flower). The genotype of the acyl-glucose-dependent anthocyanin 5-glucosyltransferase (AA5GT) which is responsible for the color change of red to pink, in each cell layer was deduced by genomic analysis using tissues originated from specific cell layer and investigation of partial petal color mutations. In the results, the genotype of the L1 of ‘Feminine Minami’ was heterozygous for functional AA5GT and non-functional AA5GT carrying retrotransposon Ty1dic1 (AA5GT-Ty1dic1), and its inner cell layer hid red flower genotype, whereas AA5GT-Ty1dic1 of the L1 of ‘Tommy Minami’ became homogenic in absence of the insertion of a new Ty1dic1. Our outcomes concluded that the L1 of ‘Tommy Minami’ harboring the recessive AA5GT alleles are attributed to the inner cell layer of ‘Feminine Minami’ possessing red flower genotype.

Regeneration response of carnation cultivars in response of silver nanoparticles under in vitro conditions

The purpose of this study was to analyze the effect of chemically synthesized silver nanoparticles (AgNPs) on in vitro regeneration of carnation cultivars cv. Noblessa, cv. Antigua and cv. Mariposa. Number of shoots/explant of cv. Noblesse and cv. Antigua significantly increased at 6 mg/L AgNPs (average 7.33 shoots per explant) when supplemented in MS media. While cv. Mariposa showed highest regeneration rate at 8 mg/L (average 10 shoots per explant). High concentration of AgNPs (12 mg/L) in the medium enhanced rooting response, number of roots/plant, and root length as compared with control. The fresh and dry weight of regenerated plants significantly (P < 0.05) increased at 6 mg/L. DPPH based free radical scavenging activity, total antioxidant activity and reducing power potential of regenerated plants varied depending upon concentration of AgNPs in the media. To find non-enzymatic antioxidants to combat oxidative damage, total phenolics and flavonoids were also determined in the regenerated plants. The study concludes that metallic nanoparticles has significant effect on in vitro growth of carnation cultivars however concentration dependent. Furthermore, nanoparticles can be effectively used for increased in vitro shoot multiplication and regeneration of floriculture plants.

Characteristics of flower colour in the bud-mutated cultivars of the carnation Mini Tiara series

Spray carnation cultivars of the Mini Tiara series have unique, narrow sword-shaped petals with various colours, and were bred through bud mutations from pink-flowered 'Mini Tiara Pink' (MP) and pink, creamy-flowered 'Mini Tiara Cream' (MC). MP had pelargonidin glucoside as a major pigment, whereas the pale purple-flowered cultivar 'Mini Tiara Lilac' derived from MC had cyanidin glucoside as a major anthocyanin. Furthermore, orange-flowered 'Mini Tiara Sunny' (MS) had both pelargonidin glucoside and chalcononaringenin 2'-glucoside. Thus, the variation in petal colour in the Mini Tiara series was produced by the difference in the amount and types of flavonoid pigment in the petals. Additionally, MS produced several kinds of flower colour mutation and showed a high frequency of flower colour mutation compared with the other cultivars. Therefore, the Mini Tiara series is not only a highly commercial product for the flower market but also useful material for the study of petal colour mutability in carnations.

Current perspectives on pollen cryopreservation in horticultural species

Cryopreserved pollen collections provide long-term security for wildflora, ornamentals, fruit and vegetable crops, medicinal herbs and endangered species. Technology has been developed for long-term cryopreservation of pollen in the form of nuclear genetic diversity (NGD) of mango, amla, pomegranate, jackfruit, guava, passion fruit, jamun, different species and cultivars of citrus, papaya (different species), grape, tomato, eggplant (Solanum gene pool), onion, capsicum, rose, gladiolus, gerbera, carnation and RET species of medicinal plants. Pollen collecting is usually done on a bright sunny day between 10-11 am. Pollen collections are subjected to viability indexing by germination in vitro by the hanging drop or cellophane techniques. Pollen samples are packed either in gelatin capsules or butter paper packets, sealed air tight in polyethylene aluminium laminated pouches and lowered into a canister of a cryoflask. The fertility of stored pollen is tested under field conditions through controlled pollination. The pollen cryobank is managed by periodic replenishment of the cryogen. With the recent advances in allied bio-scientific areas in which cryopreserved pollen could be used in general crosses, shuttle breeding maintenance of B line in male sterile based hybrid seed production. Security of male parent is also ensured in the cryobank. Pollen cryopreservation protocols developed for different horticultural species will be discussed in details.

Evaluation of Carnation Cultivars; and Nitrogen and Indole Acetic Acid for Their Growth, Yield and Quality in Khumaltar, Nepal

Carnation is an important cut flower in Nepal. Several cultivars imported from abroad are being adopted for their commercial production. However, the performance of these cultivars in the specific agro-climate has not yet determined. Thus, study aimed to evaluate the performance of common cultivars, including mineral nutrition and IAA hormone on the plant growth and flowering. The study was carried out in Horticulture Research Division, Khumaltar, Nepal during 2012 and 2013. Four cultivars: King Lion, Delson, Eskimo and White Liberty including nitrogen and foliar application of IAA were evaluated for plant growth and floral characteristics. The experiment was held in randomized complete block design (RCBD) with four replications.Based on the results, cultivar King Lion had significantly highest plant height (20.66 cm), while cultivar Eskimo had the highest flower stem length (53.59 cm) and stem diameter (5.63 cm) followed by Delson (51.66 cm). The lower flower stems lengths of cultivars: White Liberty (42.48 cm) and King Lion (43.87 cm) were at par. The effect of mineral nutrition and hormone on plant height, flower stem length and flower head diameter was non-significant, however, IAA 25 ppm foliar spray was found superior over N 40 g/m2 and IAA 50 ppm applications. Thus, Eskimo is recommended as the excellent cultivar for its bigger flower stem, straightness and attractive flower looking, while cultivars; Delson and White Liberty as promising for higher flower yield. Similarly, soil application of nitrogen @ 40 g/m2 and foliar spray of auxin @ 25 ppm are recommended for higher flower yield.

Evaluation of genetic diversity of carnation cultivars using CDDP and DAMD markers and morphological traits

This study was aimed to evaluate genetic diversity using molecular markers and morphological traits in carnation, Dianthus caryophyllus (fam: Cryophyllaceae) widely grown as an ornamental. Fifteen cultivars were examined employing seven Conserved DNA-derived Polymorphism (CDDP) and seven Directed Amplification of Minisatellite DNA (DAMD) markers. These varieties were planted in a randomized complete block design in three replicates at the experimental farm of the National Institute of Ornamental Plants, Mahallat, Iran. Analysis of variance for morphological traits showed significant differences among the examined cultivars. Morphological traits such as flower longevity, flower diameter and number of branches were statistically significant at 1 percent probability level. CDDP and DAMD markers produced an average polymorphic index content of 0.38 and 0.29 for CDDP and DAMD markers, respectively. Genetic distance matrix ranged from 0.02 to 0.84 in CDDP and from 0.09 to 0.78 in DAMD marker analysis. Cluster analysis for the two markers revealed that cultivars were grouped in three clusters. Results of molecular evaluation showed that CDDP and DAMD markers have high ability to differentiate carnation cultivars through assessment of the genetic diversity.

Fusarium oxysporum f. sp. dianthi virus 1 Accumulation Is Correlated with Changes in Virulence and Other Phenotypic Traits of Its Fungal Host.

Fusarium oxysporum f. sp. dianthi virus 1 (FodV1) was detected in isolate 116 (116V+) of Fusarium oxysporum f. sp. dianthi, reaching such a high accumulation level that it was clearly visible after agarose gel electrophoresis of total DNA extracts. FodV1 consists of four double-stranded RNA segments that correspond to a new mycovirus in the Chrysoviridae family. We obtained an isolate of F. oxysporum f. sp. dianthi 116 (116V-) with only a residual level of FodV1 RNA accumulation by single-conidia selection. Compared with 116V-, isolate 116V+ showed significant phenotypic alterations in vegetative growth and virulence. Thus, the presence of a high titer of mycovirus FodV1 was associated with a modified morphology and a reduced growth of the colonies on solid medium, and with a diminished conidiation in liquid medium. Inoculation of four susceptible carnation cultivars with either 116V- or 116V+ showed that the presence of a high titer of FodV1 was also correlated with a significantly reduced virulence of its fungal host. All of the results suggest that FodV1 could be associated with hypovirulence, identifying it as a potential biocontrol agent for Fusarium wilt of carnation. This is the first report of a mycovirus potentially associated with the induction of hypovirulence in the species F. oxysporum.

Petal color characteristics of carnation mutative cultivars MINAMI series

Petal color of 10 carnation (Dianthus caryophyllus L.) cultivars MINAMI series is divided into two color groups, the acyanic group and the cyanic group. The acyanic group consists of yellow-flowered cultivars 'Poly Minami' and 'Lemon Minami' and off-white-flowered cultivar 'Vanilla Minami'. The cyanic group consists of orange-flowered cultivars 'Orange Minami' and 'Minami', pinkish-flowered cultivars 'Pippi Minami', 'Passion Minami', 'Feminine Minami' and 'Sakura Minami', and reddish-flowered cultivar 'Tommy Minami'. MINAMI series produced from bud mutation has a unique direction of flower color changes. Therefore, the usage of these plant materials might contribute to the elucidation of the mechanism of flower color mutation.

A comprehensive phylogeny of auxin homeostasis genes involved in adventitious root formation in carnation stem cuttings.

Understanding the functional basis of auxin homeostasis requires knowledge about auxin biosynthesis, auxin transport and auxin catabolism genes, which is not always directly available despite the recent whole-genome sequencing of many plant species. Through sequence homology searches and phylogenetic analyses on a selection of 11 plant species with high-quality genome annotation, we identified the putative gene homologs involved in auxin biosynthesis, auxin catabolism and auxin transport pathways in carnation (Dianthus caryophyllus L.). To deepen our knowledge of the regulatory events underlying auxin-mediated adventitious root formation in carnation stem cuttings, we used RNA-sequencing data to confirm the expression profiles of some auxin homeostasis genes during the rooting of two carnation cultivars with different rooting behaviors. We also confirmed the presence of several auxin-related metabolites in the stem cutting tissues. Our findings offer a comprehensive overview of auxin homeostasis genes in carnation and provide a solid foundation for further experiments investigating the role of auxin homeostasis in the regulation of adventitious root formation in carnation.

Enhanced Conjugation of Auxin by GH3 Enzymes Leads to Poor Adventitious Rooting in Carnation Stem Cuttings.

Commercial carnation (Dianthus caryophyllus) cultivars are vegetatively propagated from axillary stem cuttings through adventitious rooting; a process which is affected by complex interactions between nutrient and hormone levels and is strongly genotype-dependent. To deepen our understanding of the regulatory events controlling this process, we performed a comparative study of adventitious root (AR) formation in two carnation cultivars with contrasting rooting performance, “2101–02 MFR” and “2003 R 8”, as well as in the reference cultivar “Master”. We provided molecular evidence that localized auxin response in the stem cutting base was required for efficient adventitious rooting in this species, which was dynamically established by polar auxin transport from the leaves. In turn, the bad-rooting behavior of the “2003 R 8” cultivar was correlated with enhanced synthesis of indole-3-acetic acid conjugated to aspartic acid by GH3 proteins in the stem cutting base. Treatment of stem cuttings with a competitive inhibitor of GH3 enzyme activity significantly improved rooting of “2003 R 8”. Our results allowed us to propose a working model where endogenous auxin homeostasis regulated by GH3 proteins accounts for the cultivar dependency of AR formation in carnation stem cuttings.

Induction of proteases secreted by fusarium oxysporum F. SP. Dianthi in the presence of carnation root cell walls. Biochemical characterization of a serine protease

We determined that protease activity is secreted by Fusarium oxysporum f. sp. dianthi race 2 (Fod) in vitro. The cell wall extracted from resistant and susceptible cultivars of carnation (Dianthus caryophyllus L.) was used as an inducer. The activity increased in the presence of the two cultivars, and higher levels were detected with the susceptible cultivar. One of the three proteases secreted by Fod was purified using ion exchange (DEAE-cellulose), gel filtration (Sephadex G-75), and affinity chromatographies (Benzamidine Sepharose 6B). Using protease inhibitors, the purified enzyme was classified as a trypsin-like serine protease that was inhibited by phenylmethanesulfonyl fluoride, diisopropylfluorophosphate and the soybean trypsin inhibitor. In addition, the enzyme was biochemically characterized and determined to have an optimal pH of 8.0, an optimal temperature of 50°C, stability from pH 6 to 8, temperature stability between 40 to 60°C, a molecular weight of 54 kDa, and an isoelectric point of 4.4. The values of the Km and Vmax kinetic constants were found to be 0.31 mg/ml and 24.7 µmol/min, respectively.

Cloning and Expression of Omission of Second Division-like Genes from Carnation

The omission of second division gene (OSD1) gene plays a fundamental role in meiosis and is associated with 2n gamete formation in Arabidopsis thaliana. The objective of this work was to unravel the mechanisms leading to 2n pollen production, and isolate and analyze the expression patterns of OSD-like (OSDL) genes in carnation (Dianthus caryophyllus). We found an absence of the second meiotic division caused the formation of 2n pollen. Three homoeologous genes were cloned and labeled as OSDLa, OSDLb, and OSDLc in a diploid carnation. The cDNAs were 1180 bp for OSDLa, 1288 bp for OSDLb, and 971 bp for OSDLc. A strong similarity was found between the amino sequences of OSDLb and OSDLc. An evident feature of OSDLs proteins is the presence of D-box and MR-tail domains; however, the GxEN/KEN-box domain, which is distinct among the other plant proteins was absent. Quantitative real time polymerase chain reaction (qRT-PCR) analysis showed that OSDL genes maintain continuous expression in buds and other tissues. OSDLa has the highest expression in buds of 1.1–1.2 cm long (stage 2), and OSDLb has a high level of expression in buds of 0.9–1.0 cm long (stage 1) and stage 2 buds and ovary tissues in three carnation cultivars. The expression level of OSDLc was highest in ovaries. These expression patterns strongly suggest that OSDLs in carnation involve male meiosis and ovary development. These findings can have potential applications in fundamental polyploidization research and plant breeding programs in carnation.

Characterization of the role of sodium nitroprusside (SNP) involved in long vase life of different carnation cultivars

BackgroundSodium nitroprusside (SNP) has been previously shown to extend the vase life of various cut flowers; however, its positive effect on extending vase life of carnations has not been well documented. Moreover, the role of SNP in the mechanisms underlying determination of vase life of cut carnations has also not been well addressed.ResultsSNP increased vase life of Tico Viola carnations along with their relative fresh weight (RFW). Among the treatments, the flowers treated with 10 mg L−1 SNP had the longest vase life and maximum relative fresh weight (RFW). This was achieved through significant suppression of ethylene production via downregulation of ethylene biosynthesis and petal senescence-related genes, and through an increase in the scavenging mechanism of reactive oxygen species (ROS) by antioxidant activity during flower vase life. In addition, the positive efficacy of SNP could also be confirmed using 1-aminocyclopropane-1-carboxylic acid (ACC) and different cultivars, resulting in similar trends for both experiments.ConclusionTaken together, these results suggest that SNP plays a crucial role in multiple modes of action that are associated with the longevity of cut carnation flowers.

A screening test for the determination of cut flower longevity and ethylene sensitivity of carnation

Strategies to prevent postharvest losses include the use of genotypes that have a longer life. The objective of this study was to develop a screening test for the estimation of cut flower postharvest life and the response to exogenous ethylene of different carnation cultivars at an early stage of plant growth. Ethylene sensitivity and production in different cut flower cultivars was evaluated, and a similar response in the vegetative stage was studied. Also, the possible relationship between the morphological parameters of cuttings and flower postharvest life was studied. Ethylene production of cuttings may be a useful tool for estimating ethylene production of cut flowers. There is a strong relationship between cut flower vase life and the root length of cuttings, as well as cut flower ethylene sensitivity and the number of internodes the cuttings have. Applications of exogenous ethylene to cutting cultivars have an effect on the growth parameters of the cuttings, but the response to ethylene in cut flowers does not behave in the same way in the vegetative stage.

Blue and red light-emitting diodes improve the growth and physiology of in vitro-grown carnations ‘Green Beauty’ and ‘Purple Beauty’

The objective of this study was to determine the effect of light quality on the growth, physiology, and antioxidant enzyme activity of two important domestic carnation (Dianthus caryophyllus) cultivars: ‘Green Beauty’ and ‘Purple Beauty’. In vitro-grown shoot tip explants were cultured on the plant growth regulator (PGR)-free Murashige and Skoog (MS) medium under a conventional cool white fluorescent lamp (control), blue light-emitting diode (LED), or red LED. Growth traits, photosynthetic and biochemical parameters, activity of antioxidant enzymes, and nutrient content were measured after 8 weeks. Interestingly, the blue and red LED treatments resulted in a significant increase in growth, photosynthetic parameters, and nutrient content in comparison to the conventional cool white florescent lamp treatment. In addition, red LED treatment increased the activities of antioxidant enzymes and elemental contents in both cultivars. Thus, incorporating blue or red LED treatments enhances the quality of ‘Green Beauty’ and ‘Purple Beauty’ carnations propagated in vitro.

Reproductive ability of three Japanese populations of &lt;i&gt;Heterodera trifolii&lt;/i&gt; on five crops and three carnation cultivars

Reproductive ability of three Japanese populations of &lt;i&gt;Heterodera trifolii&lt;/i&gt; on five crops and three carnation cultivars

Multiple factors influence adventitious rooting in carnation (Dianthus caryophyllus L.) stem cuttings

Efficient propagation of uniform starting material is a critical requirement for mass production of most ornamental plants, including carnation. For some elite cultivars, the production of young plantlets is limited by poor adventitious root formation from stem cuttings. We previously characterized the molecular signature during adventitious rooting in two carnation cultivars, 2101-02 MFR and 2003 R 8, which were selected because of their contrasting rooting performance. To determine additional factors that contribute to the differences observed in adventitious rooting during the commercial scaling-up of this species, we characterized rooting performance and endogenous hormone levels in stem cuttings of these two cultivars during one production season. We found that stem cutting production declined during the harvest season in a cultivar-dependent manner. In addition, the initiation of adventitious roots in the stem cutting base depended on its endogenous auxin and cytokinin levels at harvest time, while their subsequent growth and development was mainly influenced by the physiological status of the mother plant at harvest time and of the stem cutting during the rooting process.

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Plant growth regulators have significant effect on the delaying senescence in buds and flowers. Present study was conducted to find out the effects of Salicylic acid and Benzyl adenine spraying on various biochemical traits such as membrane stability index, chlorophyll a, and b content, total chlorophyll content, sugar content, catalase, and peroxidase enzymes activity, and proline content in Carnation. Study was conducted in a split plot design with three replications and various concentrations of hormones, cultivars of carnation and measurement time was used as a factor of interaction. Results of study showed that all the traits were significantly affected by spraying of these tow hormones. Cultivar factor was most effective in chlorophyll a, b, and total chlorophyll content, catalase enzyme activity, proline content and sugar content. Among these two hormones, spraying of Salicylic acid @ 100 μM was found most effective treatment. Further, time of measurements and cultivar factors were also found effective in all of traits. Anita Ramtin 1 , Sepideh Kalatejari 1,* , Roohangiz Naderi 2 and Mohammad Matinizadeh 3