Levels Of Polyploidy Research Articles

Cell cycle re-entry in the aging Drosophila brain.

The brain is an organ comprised mostly of long-lived, quiescent cells that perform vital functions throughout an animal's life. Due to the brain's limited regenerative ability, these long-lived cells must engage unique mechanisms to cope with accumulated damage over time. We have shown that a subset of differentiated neuronal and glial cells in the fruit fly brain become polyploid during adulthood. Cell cycle re-entry in the brain has previously been associated with neurodegeneration, but there may be a more complex relationship between polyploidy and cell fitness in the brain. Here, we examine how known lifespan modifiers influence the accumulation of polyploidy in the aging fly brain. Flies aged at a low temperature, or with a low protein diet, accumulate polyploid cells in the brain more slowly than expected if this phenotype were solely regulated by lifespan mechanisms. Despite the slower accumulation of polyploid cells, animals under conditions that extend lifespan eventually reach similar levels of polyploidy in the brain as controls. Our work suggests known lifespan modifiers can influence the timing of cell cycle re-entry in the adult brain, indicating there is a flexible window of cell cycle plasticity in the aging brain.

The effect of physical activity of varying intensity on cardiomyocyte hypertrophy and myocardial polyploidy in rats

Regular exercise improves cognitive function, reduces the risk of premature death from cardiovascular disease, stroke, diabetes and improves quality of life.Aim: To study the morphological parameters of cardiomyocytes of the left ventricle of the heart and the proliferative activity of rat cardiomyocytes during physical activity of different intensity.Methodology and Research Methods. Outbred male rats (n = 30) were taken as experimental animals and divided into three series. The first series of rats were given light physical activity – the animals swam in the bath for 15 minutes. The animals of the second series were in the bath for 30 minutes (moderate severity), the third series were in the bath until they began to lose strength and sink (in 55–59 minutes). Animals were taken out of the experiment after 10 sessions of water loading. Some rats (5 animals per series) were slaughtered 30 days after the end of the experiment. After extraction, the heart was stained with hematoxylin and eosin, the amount of DNA in the nuclei of cardiomyocytes was determined, binuclear and Ki-67 positive cells were counted, and the diameter of cardiomyocytes was measured. Statistical analysis was carried out in the program “Statistica 10.0”.Results. Histological examination revealed changes only in the myocardium of the left ventricle of rats of the third series: pronounced dystrophic changes in cardiomyocytes. There were cells with necrobiosis, focal necrosis of groups of cardiomyocytes. The highest level of cardiomyocyte polyploidy occurred in the second and third series (the ratio of diploid and tetraploid cardiomyocytes in the control corresponded to 91.6 ± 7.4%; 8.2±6.3%). In these series, there was also a change in the number of binuclear cells (in the control – 12.7 ± 1.9‰). In all series of Ki-67 positive nuclei were not detected.Conclusion. Severe physical activity leads to structural disorders of the myocardium, persistent hypertrophy of cardiomyocytes and is accompanied by a decrease in the proliferative activity of cardiomyocytes.

Insight into Physiological and Biochemical Determinants of Salt Stress Tolerance in Tetraploid Citrus.

Citrus are classified as salt-sensitive crops. However, a large diversity has been observed regarding the trends of tolerance among citrus. In the present article, physiological and biochemical studies of salt stress tolerance were carried out according to the level of polyploidy of different citrus genotypes. We particularly investigated the impact of tetraploidy in trifoliate orange (Poncirus trifoliata (L.) Raf.) (PO4x) and Cleopatra mandarin (Citrus reshni Hort. Ex Tan.) (CL4x) on the tolerance to salt stress compared to their respective diploids (PO2x and CL2x). Physiological parameters such as gas exchange, ions contents in leaves and roots were analyzed. Roots and leaves samples were collected to measure polyphenol, malondialdehyde (MDA), ascorbate and H2O2 contents but also to measure the activities of enzymes involved in the detoxification of active oxygen species (ROS). Under control conditions, the interaction between genotype and ploidy allowed to discriminate different behavior in terms of photosynthetic and antioxidant capacities. These results were significantly altered when salt stress was applied when salt stress was applied. Contrary to the most sensitive genotype, that is to say the diploid trifoliate orange PO2x, PO4x was able to maintain photosynthetic activity under salt stress and had better antioxidant capacities. The same observation was made regarding the CL4x genotype known to be more tolerant to salt stress. Our results showed that tetraploidy may be a factor that could enhance salt stress tolerance in citrus.

Conserved chamber-specific polyploidy maintains heart function in Drosophila.

Developmentally programmed polyploidy (whole-genome duplication) of cardiomyocytes is common across evolution. Functions of such polyploidy are essentially unknown. Here, in both Drosophila larvae and human organ donors, we reveal distinct polyploidy levels in cardiac organ chambers. In Drosophila, differential growth and cell cycle signal sensitivity leads the heart chamber to reach a higher ploidy/cell size relative to the aorta chamber. Cardiac ploidy-reduced animals exhibit reduced heart chamber size, stroke volume and cardiac output, and acceleration of circulating hemocytes. These Drosophila phenotypes mimic human cardiomyopathies. Our results identify productive and likely conserved roles for polyploidy in cardiac chambers and suggest that precise ploidy levels sculpt many developing tissues. These findings of productive cardiomyocyte polyploidy impact efforts to block developmental polyploidy to improve heart injury recovery.

Genomics for Yield and Yield Components in Durum Wheat.

In recent years, many efforts have been conducted to dissect the genetic basis of yield and yield components in durum wheat thanks to linkage mapping and genome-wide association studies. In this review, starting from the analysis of the genetic bases that regulate the expression of yield for developing new durum wheat varieties, we have highlighted how, currently, the reductionist approach, i.e., dissecting the yield into its individual components, does not seem capable of ensuring significant yield increases due to diminishing resources, land loss, and ongoing climate change. However, despite the identification of genes and/or chromosomal regions, controlling the grain yield in durum wheat is still a challenge, mainly due to the polyploidy level of this species. In the review, we underline that the next-generation sequencing (NGS) technologies coupled with improved wheat genome assembly and high-throughput genotyping platforms, as well as genome editing technology, will revolutionize plant breeding by providing a great opportunity to capture genetic variation that can be used in breeding programs. To date, genomic selection provides a valuable tool for modeling optimal allelic combinations across the whole genome that maximize the phenotypic potential of an individual under a given environment.

Nano-Strainer: A workflow for the identification of single-copy nuclear loci for plant systematic studies, using target capture kits and Oxford Nanopore long reads.

In modern plant systematics, target enrichment enables simultaneous analysis of hundreds of genes. However, when dealing with reticulate or polyploidization histories, few markers may suffice, but often are required to be single-copy, a condition that is not necessarily met with commercial capture kits. Also, large genome sizes can render target capture ineffective, so that amplicon sequencing would be preferable; however, knowledge about suitable loci is often missing. Here, we present a comprehensive workflow for the identification of putative single-copy nuclear markers in a genus of interest, by mining a small dataset from target capture using a few representative taxa. The proposed pipeline assesses sequence variability contained in the data from targeted loci and assigns reads to their respective genes, via a combined BLAST/clustering procedure. Cluster consensus sequences are then examined based on four pre-defined criteria presumably indicative for absence of paralogy. This is done by calculating four specialized indices; loci are ranked according to their performance in these indices, and top-scoring loci are considered putatively single- or low copy. The approach can be applied to any probe set. As it relies on long reads, the present contribution also provides template workflows for processing Nanopore-based target capture data. Obtained markers are further tested and then entered into amplicon sequencing. For the detection of possibly remaining paralogy in these data, which might occur in groups with rampant paralogy, we also employ the long-read assembly tool canu. In diploid representatives of the young Compositae genus Leucanthemum, characterized by high levels of polyploidy, our approach resulted in successful amplification of 13 loci. Modifications to remove traces of paralogy were made in seven of these. A species tree from the markers correctly reproduced main relationships in the genus, however, at low resolution. The presented workflow has the potential to valuably support phylogenetic research, for example in polyploid plant groups.

Optimal protocol for in vitro polyploid induction of Cymbidium aloifolium (L.) Sw

BackgroundCymbidium aloifolium is a popular ornamental flower in Thailand with both economic and medical values. Polyploid induction techniques are used to improve plant quality. This study identified polyploidy levels of C. aloifolium induction by colchicine. Protocorms of C. aloifolium were treated on solid New Dogashima Medium (NDM) with various concentrations of colchicine (0, 0.01, 0.02, 0.03, 0.04 and 0.05% w/v) for 2, 4 and 8 weeks.ResultsColchicine effectively induced plant polyploidy. Tetraploid plants were found after treatment with 0.03% and 0.04% colchicine for 8 weeks, while at increased concentration and duration, survival, response and growth performance decreased. Tetraploid plants showed the lowest growth performance but highest size of stomatal and epidermal cells. Growth performance and leaf surface anatomy data were analyzed by Pearson’s correlation and PCA. Results showed that stomatal and epidermal cell sizes had strongly negative correlations with other variables, while HCA revealed that stomatal and epidermal cell sizes of tetraploid plantlets were larger but stomatal and epidermal cell densities decreased when compared with the diploids.ConclusionColchicine at suitable concentrations and duration produced polyploid plants with alteration of morphological and anatomical traits. This study provides potential information to support orchid quality production as ornamental plants and a source of pharmaceutical raw materials.

CRISPR/Cas9 Induced Duplications, Deletions and Inversions in Mouse Zygotes Lead to Karyotype Instability

CRISPR/Cas9 technology has been widely used for targeted modification of the mammalian genomes. We have analyzed the karyotype of 18 mouse fibroblast cell lines with Cntn6 gene rearrangements introduced by CRISPR/Cas9. We have produced cell lines with 2374 kb Cntn6 gene duplications, 1137 kb deletions and inversions of similar size. In addition, we have performed cytogenetic analysis for five control mouse embryonic fibroblasts with the intact Cntn6 gene alleles. The cell lines heterozygous for Cntn6 gene inversion and homozygous and heterozygous for Cntn6 gene duplication had a high level of polyploidy (20–46%), as well as chromosome 6 monosomy (1–9%) and trisomy (1–8%). No trisomy was detected in the four cell lines with the deletion and duplication of the Cntn6 gene in the compound, and the proportion of polyploid cells was minimal (1.5–5.7%). Thus, we have shown the karyotype destabilization in the cell lines that have undergone genome editing using CRISPR/Cas9 system.

Treated municipal wastewater as option to the use of fresh water for the cultivation of valuable pastoral species Buffel grass (Cenchrus ciliaris L.)

AbstractThe increasing aridity exacerbated by climatic changes is leading to loss of perennial herbaceous plants Cenchrus ciliaris, an apomictic, polyploid grass used as forage in hot and dry areas, and is currently under threat for the increased scarcity of water. In this study, treated municipal wastewater (TWW) was used to irrigate two ploidy levels of C. ciliaris for two consecutive years. The objectives were (1) to assess the possibility of using unconventional water for watering C. ciliaris and (2) to identify at what extent TWW irrigation affected growth and nutritive properties of C. ciliaris that differed for polyploidy level. TWW irrigation positively affected growth and physiological plant parameters. In leaves of TWW watered C. ciliaris, the concentration of Nt, P and K significantly increased. The tetraploid C. ciliaris showed a better growth and quality than the hexaploid ones, appearing the most suitable cultivar to be irrigated with TWW for feed uses. In short, TWW can be strategically used for irrigating forage species with the double output of ameliorating soil properties, recovering degraded area and improving the nutritive values of fodder species contributing to the promotion of a green and sustainable circular economy, more in climatically under pressure developing countries.

The comparison of polymorphism among Avena species revealed by retrotransposon-based DNA markers and soluble carbohydrates in seeds

Here, we compared the polymorphism among 13 Avena species revealed by the iPBS markers and soluble carbohydrate profiles in seeds. The application of seven iPBS markers generated 83 bands, out of which 20.5% were polymorphic. No species-specific bands were scored. Shannon’s information index (I) and expected heterozygosity (He) revealed low genetic diversity, with the highest values observed for A. nuda (I = 0.099; He = 0.068). UPGMA clustering of studied Avena accessions and PCoA results showed that the polyploidy level is the main grouping criterion. High-resolution gas chromatography revealed that the studied Avena accessions share the same composition of soluble carbohydrates, but significant differences in the content of total (5.30–22.38 mg g−1 of dry weight) and particular sugars among studied samples were observed. Sucrose appeared as the most abundant sugar (mean 61.52% of total soluble carbohydrates), followed by raffinose family oligosaccharides (31.23%), myo-inositol and its galactosides (6.16%), and monosaccharides (1.09%). The pattern of interspecific variation in soluble carbohydrates, showed by PCA, was convergent to that revealed by iPBS markers. Thus, both methods appeared as a source of valuable data useful in the characterization of Avena resources or in the discussion on the evolution of this genus.

Navigating the CoGe Online Software Suite for Polyploidy Research.

The CoGe software suite at genomevolution.org hosts a number of tools that facilitate genomic research on plant and animal whole-genome multiplication-polyploidy. SynMap permits analysis and visualization of two-way syntenic dotplot alignments of genomes, includes many options and data/graphics download possibilities, and even permits three-genome synteny maps and interactive views. FractBias is a tool that operates within SynMap that permits calculation and graphic display of genome fragments (such as chromosomes) of one species mapped to another, displaying both blockwise homology depths and the extent of syntenic gene (syntelog) loss following polyploidy events. SynMap macrosynteny results can segue into the microsynteny tool GEvo, which provides genome-browser-like views of homologous genome blocks. CoGe FeatView allows call-up of given gene features already stored in the CoGe resource, and CoGeBlast permits searches for additional features that can be analyzed or downloadedfurther. Links from these tools can be fed into SynFind, which can find syntenic blocks surrounding a feature across multiple specified genomes while also simultaneously providing overall genome-wide syntenic depth calculations that can be interpreted to reflect polyploidy levels. Here, we describe basic use of these tools on the CoGe software suite.

Development of a 135K SNP genotyping array for Actinidia arguta and its applications for genetic mapping and QTL analysis in kiwifruit.

Kiwifruit (Actinidia spp) is a woody, perennial and deciduous vine. In this genus, there are multiple ploidy levels but the main cultivated cultivars are polyploid. Despite the availability of many genomic resources in kiwifruit, SNP genotyping is still a challenge given these different levels of polyploidy. Recent advances in SNP array technologies have offered a high-throughput genotyping platform for genome-wide DNA polymorphisms. In this study, we developed a high-density SNP genotyping array to facilitate genetic studies and breeding applications in kiwifruit. SNP discovery was performed by genome-wide DNA sequencing of 40 kiwifruit genotypes. The identified SNPs were stringently filtered for sequence quality, predicted conversion performance and distribution over the available Actinidia chinensis genome. A total of 134 729 unique SNPs were put on the array. The array was evaluated by genotyping 400 kiwifruit individuals. We performed a multidimensional scaling analysis to assess the diversity of kiwifruit germplasm, showing that the array was effective to distinguish kiwifruit accessions. Using a tetraploid F1 population, we constructed an integrated linkage map covering 3060.9cM across 29 linkage groups and performed QTL analysis for the sex locus that has been identified on Linkage Group 3 (LG3) in Actinidia arguta. Finally, our dataset presented evidence of tetrasomic inheritance with partial preferential pairing in A. arguta. In conclusion, we developed and evaluated a 135K SNP genotyping array for kiwifruit. It has the advantage of a comprehensive design that can be an effective tool in genetic studies and breeding applications in this high-value crop.

Multiporate Pollen of Poaceae as Bioindicator of Environmental Stress: First Archaeobotanical Evidence from the Early–Middle Holocene Site of Takarkori in the Central Sahara

This paper reports on the most ancient unusual morphological trait of the apertures of Poaceae pollen found in archaeological layers. In Poaceae, high levels of hybridization, polyploidy, apomixis, and multiporate pollen are often related. Multiple genomes in polyploids are critical for the adaptation of plant species to stresses and could be revealed by anomalies in pollen development. Therefore, the paleoenvironmental research can gain great benefits from identifying polyploids in past contexts by observing anomalous pollen morphology during pollen counts. The occurrence of multiporate pollen in Poaceae has also been related to special features of the ecology of the species showing this anomaly, as well as to climatic and environmental stresses experienced by Poaceae living in a given region. Multiporate and bi- or tri-porate instead of monoporate pollen grains have been observed in samples taken from Takarkori rockshelter, an archaeological site in southwestern Libya (central Sahara) that has been occupied between ~10,200 and ~4650 cal BP. Multiporate pollen was found in organic sands and coprolites of ovicaprines. On the basis of archaeobotanical research, this work aims to investigate whether the presence of supernumerary pores in Poaceae pollen may be an effect of both climatic/hydrological changes and continued anthropogenic pressure on the wild grasses living in the region. The presence of multiporate pollen reveals that Poaceae that lived in central Sahara tackled several kinds of stress during the early and middle Holocene. The Takarkori pollen record suggests that climate change could have played a major role in the early Holocene, while human pressure became stronger during the middle Holocene. The change in environmental conditions determined adaptive responses of polyploid grasses even in the form of multiporate pollen.

An Updated Overview on Insights into Sugarcane Genome Editing via CRISPR/Cas9 for Sustainable Production

Sugarcane crop constitutes one of the most vital sources of sugar and bioenergy globally; however, higher level of polyploidy makes its genome editing an intricate task. Recently, genome editing has become easier with CRISPR/Cas9 system that uses Cas9 to target sequence-specific regions and introduce double-strand breaks into the target region. This technique has been successfully employed to develop new varieties of sugarcane having desired phenotypic and physiological traits. Several genes can be fused with the CRISPR/Cas9 system leading to successful metabolic engineering and biological improvement for ensuring sustainable enhancement in sugarcane production. This simple RNA-guided genome editing technique has become a revolutionary tool and innovative application in biology that might be effectively employed for inducing specified genomic modifications in plant tissues. This review attempts to synthesize fundamentals of genome editing techniques with an ultimate aim to find out opportunities and challenges of sugarcane genome editing. It also tends to summarize the advances and achievements of gene editing via CRISPR-based genome editing technique for numerous field crops. Lastly, the enormous potential of CRISPR/Cas9 for gene editing in sugarcane, major challenges and future perspectives have been objectively elaborated.

Integrated Approach in Genomic Selection to Accelerate Genetic Gain in Sugarcane.

Marker-assisted selection (MAS) has been widely used in the last few decades in plant breeding programs for the mapping and introgression of genes for economically important traits, which has enabled the development of a number of superior cultivars in different crops. In sugarcane, which is the most important source for sugar and bioethanol, marker development work was initiated long ago; however, marker-assisted breeding in sugarcane has been lagging, mainly due to its large complex genome, high levels of polyploidy and heterozygosity, varied number of chromosomes, and use of low/medium-density markers. Genomic selection (GS) is a proven technology in animal breeding and has recently been incorporated in plant breeding programs. GS is a potential tool for the rapid selection of superior genotypes and accelerating breeding cycle. However, its full potential could be realized by an integrated approach combining high-throughput phenotyping, genotyping, machine learning, and speed breeding with genomic selection. For better understanding of GS integration, we comprehensively discuss the concept of genetic gain through the breeder’s equation, GS methodology, prediction models, current status of GS in sugarcane, challenges of prediction accuracy, challenges of GS in sugarcane, integrated GS, high-throughput phenotyping (HTP), high-throughput genotyping (HTG), machine learning, and speed breeding followed by its prospective applications in sugarcane improvement.

Soil water leaf gas exchange and biomass production of Buffelgrass (Cenchrus ciliaris L.) with two ploidy levels under arid zone

Cenchrus ciliaris corresponds to a perennial grass that is widespread in arid environments. This study aimed to assess growth and leaf exchange under water deficit stress of tetraploid (P1) and hexaploid (P2) individuals of C. ciliaris. The experiment was conducted under four treatments (Tc, T1, T2, and T3). Growth phenology, photosynthesis (A), stomatal conductance (Gs), CO2 concentration, and transpiration (Tr) were observed during the stress cycle. The obtained results revealed that the highest growth potentialities were recorded for P2 population in both treatments. Both populations responded differently to water deficit stress displaying various values for tiller and the number of spikes. A significant effect of soil moisture treatments on C. ciliaris plant biomass was recorded (P < 0.05). Moreover, there were different responses to drought stress between both contrasting populations. The results also demonstrated that polyploidy level affected only the phenological parameters. Photosynthetic parameters, such as (A) and (Gs) significantly decreased under severe stress between populations and treatments. It could be inferred that C. ciliaris accessions respond to drought through showing significant changes in their physiological and phenological behavior.

New Chromosomal Data of Dianthus Section Leiopetali (Caryophyllaceae, Sileneae)

In section Leiopetali, the diploid chromosome numbers were reported from only six of total 25 taxa. There is no karyological data relating to other 19 taxa. In this study, it is intended to determine the diploid chromosome numbers of taxa of section Leiopetali. The diploid number of 13 species were detected, four of which were recorded for the first time and nine numbers compatible with the previous reports. Twelve species were diploid with 2n = 2x =30. D. leptopetalus was diploid and polyploid, which reveals only one polyploidy level of tetraploidy (2n = 4x = 60). Polyploidy might have played a role in the karyotype evolution of the genus. One of the most important reasons for this situation is that although polyploidy is observed in the genus Dianthus, the dysploidy mechanism that causes a change in the basic chromosome number has not been found until now. The basic number is x = 15 according to the all chromosomal reports. In conclusion, this study reports new data into the karyological characteristics of section Leiopetali (genus Dianthus) that can be useful for interpreting or understanding relationships among sections.

Ceratopteris chunii and Ceratopteris chingii (Pteridaceae), two new diploid species from China, based on morphological, cytological, and molecular data

Understanding how natural hybridization and polyploidizations originate in plants requires identifying potential diploid ancestors. However, cryptic plant species are widespread, particularly in Ceratopteris (Pteridaceae). Identifying Ceratopteris cryptic species with different polyploidy levels is a challenge because Ceratopteris spp. exhibit high degrees of phenotypic plasticity. Here, two new cryptic species of Ceratopteris, Ceratopteris chunii and Ceratopteris chingii, are described and illustrated. Phylogenetic analyses reveal that each of the new species form a well-supported clade. C. chunii and C. chingii are similar to Ceratopteris gaudichaudii var. vulgaris and C. pteridoides, respectively, but distinct from their relatives in the stipe, basal pinna of the sterile leaf or subelliptic shape of the fertile leaf, as well as the spore surface. In addition, chromosome studies indicate that C. chunii and C. chingii are both diploid. These findings will help us further understand the origin of Ceratopteris polyploids in Asia.

Targeting glioma cells by antineoplastic activity of reversine.

Gliomas are the most common type of primary central nervous system tumors and despite great advances in understanding the molecular basis of the disease very few new therapies have been developed. Reversine, a synthetic purine analog, is a multikinase inhibitor that targets aurora kinase A (AURKA) and aurora kinase B (AURKB). In gliomas, a high expression of AURKA or AURKB is associated with a malignant phenotype and a poor prognosis. The present study investigated reversine-related cellular and molecular antiglioma effects in HOG, T98G and U251MG cell lines. Gene and protein expression were assessed by reverse transcription-quantitative PCR and western blotting, respectively. For functional assays, human glioma cell lines (HOG, T98G and U251MG) were exposed to increasing concentrations of reversine (0.4–50 µM) and subjected to various cellular and molecular assays. Reversine reduced the viability and clonogenicity in a dose- and/or time-dependent manner in all glioma cells, with HOG (high AURKB-expression) and T98G (high AURKA-expression) cells being more sensitive compared with U251MG cells (low AURKA- and AURKB-expression). Notably, HOG cells presented higher levels of polyploidy, while T98G presented multiple mitotic spindles, which is consistent with the main regulatory functions of AURKB and AURKA, respectively. In molecular assays, reversine reduced AURKA and/or AURKB expression/activity and increased DNA damage and apoptosis markers, but autophagy-related proteins were not modulated. In conclusion, reversine potently induced mitotic catastrophe and apoptosis in glioma cells and higher basal levels of aurora kinases and genes responsive to DNA damage and may predict improved antiglioma responses to the drug. Reversine may be a potential novel drug in the antineoplastic arsenal against gliomas.

In vivo polyploidy induction of Phalaenopsis amabilis in a bubble bioreactor system using colchicine

Abstract Phalaenopsis amabilis Blume var. grandiflora Bateman is economically important as cut and pot flower. Polyploidy is considered as a valuable tool in improvement and evolution of ornamental plants. Protocorm-like bodies (PLBs) of P. amabilis were cultured on Murashige and Skoog medium containing 0.20 mg L-1 IBA together with 2.00 mg L-1 KIN and 1.00 g L-1 activated charcoal and grown for a period of five months. Fully-developed plantlets from in vitro grown PLBs were immersed in a bubble reactor filled with half-strength Hoagland solution containing the antimitotic agent colchicine (0.05%, 0.10% and 0.15%, w/v) for 72 h with a few drops (1 mL of 0.1%) of octylphenoxypolyethoxyethanol or Nonidet (P-40) as a surfactant. Plantlets were aerated to prevent hypoxia. Colchicine-treated and untreated plantlets were transferred to pots for a period of 60 days. Tetraploidy was successfully induced by 0.15% colchicine. Polyploidy levels were firstly detected using flow cytometry and then confirmed by cytological and morphological observations. The chromosome number was 2n = 2x = 38 in diploids and 2n = 4x = 76 in tetraploid. Incubation of plantlets in liquid medium containing 0.15% colchicine induced the maximum recovered tetraploids with minimum frequency of survival (50%). The tetraploid plants were more compact and exhibited round and thick leaves with darker green color than diploids. Stomata size in tetraploids were larger with less density than diploids. Chloroplast number in guard cells of tetraploids was about two times more than that of control. These results indicate that induction of tetraploids are a reliable and powerful tool for generation of novel phenotypes with ornamental and horticultural value for genetic improvement and breeding. Produced tetraploids in current study have potential in the ornamental/floriculture trade.