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Analysis of Genetic Relatedness of Dieffenbachia Cultivars Using AFLP Markers
Abstract
Dieffenbachia Schott is an important ornamental foliage plant genus. A total of 30 species has been recognized, but most cultivars come from or are related to a single species, D. maculata (Lodd.) G. Don. At least 11 of the cultivars are sports or somaclonal variants. As a result, the potential lack of genetic diversity in cultivated Dieffenbachia has become a concern. However, no research has been conducted to determine the genetic relatedness of the cultivars. This study analyzed the genetic similarity of 42 Dieffenbachia cultivars using amplified fragment length polymorphism (AFLP) markers. Six primer sets, selected from an initial screening of 48, generated a total of 453 scorable AFLP fragments of which 323 (71%) are polymorphic. All cultivars were clearly differentiated by their AFLP fingerprints. A dendrogram was constructed using the unweighted pair-group method of arithmetic averages, and principal coordinated analysis was carried out to show multiple dimensions of the distribution of the cultivars. The 42 cultivars were divided into three clusters; clusters I and II comprise 18 and 23 cultivars, respectively. Jaccard's similarity coefficients for cultivars in the clusters I and II varied from 0.44 to 0.95 and 0.41 to 0.87, respectively. These results indicate that broadening the genetic variability in the Dieffenbachia gene pool is needed, but the genetic similarity of many cultivars is not as close as previously thought. Additionally, Jaccard's similarity coefficients between most sports or somaclonal variants and their parents were 0.73 or lower, suggesting that accumulation of somatic mutations through tissue culture may play a role in the increased variation between some sports or variants and their parents.
... As a result, there is increasing concern over the loss of genetic diversity in caladium via loss of cultivars. Similar concerns have been raised for other ornamental aroids (Chen et al., 2004a(Chen et al., , 2004b(Chen et al., , 2004cHenny, 1977). Reduced genetic diversity could result in increased vulnerability to new diseases and pests and in reduced genetic variation for cultivar development. ...
... Genetic relationships and diversity among cultivars or species have been studied in a number of important ornamental aroids, including aglaonema (Aglaonema Schott) (Chen et al., 2004b), alocasia [Alocasia (Schott) G. Don] (Chen et al., 2004a), dieffenbachia (Dieffenbachia Schott) (Chen et al., 2004c), philodendron (Philodendron Schott) (Devanand et al., 2004), and syngonium (Syngonium Schott) . These studies have established the genetic similarity among cultivars and demonstrated the usefulness in assisting such studies for germplasm identification, preservation, and new cultivar development (Chen et al., 2004a(Chen et al., , 2004b(Chen et al., , 2004cDevanand et al., 2004). ...
... Genetic relationships and diversity among cultivars or species have been studied in a number of important ornamental aroids, including aglaonema (Aglaonema Schott) (Chen et al., 2004b), alocasia [Alocasia (Schott) G. Don] (Chen et al., 2004a), dieffenbachia (Dieffenbachia Schott) (Chen et al., 2004c), philodendron (Philodendron Schott) (Devanand et al., 2004), and syngonium (Syngonium Schott) . These studies have established the genetic similarity among cultivars and demonstrated the usefulness in assisting such studies for germplasm identification, preservation, and new cultivar development (Chen et al., 2004a(Chen et al., , 2004b(Chen et al., , 2004cDevanand et al., 2004). In addition, they have revealed some interesting phenomena in aroids. ...
Caladium (Caladium xhortulanum Birdsey) is an important aroid widely used in the ornamental plant industry. Concerns have been raised about possible loss of genetic diversity due to a drastic decline in the number of cultivars in the last century. This study assessed genetic diversity and relationships among caladium cultivars and species accessions. Forty-five major cultivars and 14 species accessions were analyzed based on 297 DNA fragments produced by the target-region amplification polymorphism marker system. A low level of diversity (44.4% polymorphism) was exhibited in cultivars, while a high level of diversity (96.8% polymorphism) was present among seven accessions of Caladium bicolor (Aiton) Vent., Caladium marmoratum Mathieu, Caladium picturatum C. Koch, and Caladium schomburgkii Schott. A small percentage (7.6%) of DNA fragments was present in cultivars but absent in the seven species accessions, while a high percentage (32.2%) of DNA fragments was present in the seven species accessions but absent in cultivars. Cultivars shared a higher level of similarity at the molecular level with an average Jaccard coefficient at 0.802, formed a large group in cluster analysis, and concentrated in the scatter plot from a principal-coordinate analysis. Two accessions of C. bicolor and C. schomburgkii were very similar to cultivars with Jaccard similarity coefficients from 0.531 to 0.771, while the rest of the species accessions had small similarity coefficients with cultivars (0.060 to 0.386). Caladium steudnirifolium Engler and Caladium lindenii (André) Madison were very dissimilar to C. bicolor, C. marmoratum, C. picturatum, and C. schomburgkii, with Jaccard similarity coefficients from 0.149 to 0.237 (C. steudnirifolium) and from 0.060 to 0.118 (C. lindenii). There is a limited amount of molecular diversity in caladium cultivars, but the great repertoire of unique genes in species accessions could be used to enhance the diversity in future cultivars and reduce potential genetic vulnerability.
... The frequency of somaclonal variants is generally high, and the time required for a new cultivar release can be only 2 to 3 years compared to 7 to 10 years required using traditional breeding methods (Skirvin et al., 1994;Chen et al., 2003a). Chen et al (2004) analyzed genetic relatedness of some cultivated Dieffenbachia using amplified fragment length polymorphism (AFLP) and ...
... Dieffenbachia, a monocot belonging to the family Araceae, noted for its attractive, striking foliage, is one of the most important ornamental plant genera (Chen et al., 2004). Traditional breeding in Dieffenbachia can be hampered by its naturally-occurring dichogamy, long breeding cycle, poor seed production and germination (Henny, 1988). ...
... Camouflage was selected from somaclonal variants of cv. Panther, while Octopus was isolated from somaclonal variants of Camouflage (Chen et al., 2004). ...
... In this chapter, we document somaclonal variation in floriculture crops, discuss underlying genetic mechanisms accounting for the variation, present methods of somaclone identification and characterization, and the use of somaclonal variation for new cultivar development. Chen et al. 20032004a, Henny et al. 2003a Leaf variegation patterns and plant size Chen et al. 2003 Anthurium Enhanced axillary branching, shortened internodes, or changes in leaf shape and variegation Matsumoto and Kuehnle 1997, Chen et al. 2003, Henny et al. 2003b Change in foliar variegation Mujib et al. 2000, Ahmed et al. 2004 Leaf shape and variegation patterns and number of lateral shoots Chen et al. 20032004b Changes in foliar variegation Chen et al. (unpublished data) Philodendron Plant size and overall form Mujib and Jana 1995, Chen et al. 2003, Devenand et al. 2004 Spathiphyllum variants in floriculture crops is much higher than that of natural sport occurrence (Karp 1991, Schum andPreil 1998). This may indicate the action of culture medium induction, where it is assumed that somaclonal variation results from the interaction between pre-existing genetic variation and variation induced by culture media. ...
... 'Sarah' has much narrower green margins and extensive green blotches extending from margin to the inner ivory panel. The three somaclonal cultivars along with their parent are among the most popular Dieffenbachia cultivars produced for interior plantscaping (Chen et al. 2004b). ...
... The genus Dieffenbachia consists of about 30 species and over 100 cultivars with spotted, striped or speckled with cream, white, yellow, gold, silver, or a combination of these colors leaves, 15 to 40 cm or so in length [1, 2]. Dieffenbachia is an ornamental perennial monocot plant native to tropical America [3], belonging to the family Araceae [4]. It is prized among interiorscapers for the attractive variegated foliage, tolerance of interior environments and easy production [5]. ...
... Shen et al. [31] demonstrated the potential for new cultivar development by selecting callus-derived somaclonal variants of Dieffenbachia in 3 -4 years compared to 7 -10 years through traditional breeding methods. Chen et al. [4] analyzed genetic relatedness of some cultivated Dieffenbachia using amplified fragment length polymorphism and found that cultivars selected from somaclonal variants differ genetically from their parents. Shen et al. [32] evaluated the occurrence of somaclonal variation among regenerants derived through indirect shoot organogenesis from leaf explants of three Dieffenbachia cultivars Camouflage, Camille and Star Bright. ...
Ornamental industry has applied immensely in vitro propagation approach for large-scale plant multiplication at very high rates of elite superior varieties. As a result, hundreds of plant tissue culture laboratories have come up worldwide. Dieffenbachia species are popular foliage potted plants used in interiorescapes of homes, offices, and malls throughout the world. Most of Dieffenbachia species are now propagated by tissue culture for better utilize of species and expedite plant improvement. This review paper summarizes valuable literature on in vitro techniques including type of explants used, media optimized, ways of propagation and improvement through 45 years of research on Dieffenbachia spp. Which were provide basis for future studies such as genetic transformation for breeding aims, develop new cultivars, develop disease-resistant plants and overcome the environmental obstacles. There is a need for more application of the plant tissue culture techniques on Dieffenbachia to investigate the responses of different cultivars and explants to variable culture media.
... Dieffenbachia, a monocot belonging to the family Araceae noted for its attractive, striking foliage, is one of the most important ornamental plant genera (Chen et al. 2004). Traditional breeding in Dieffenbachia can be hampered by its naturally occurring dichogamy, long breeding cycle, poor seed production and germination (Henny 1988). ...
... Kallak et al. (1997) concluded that callogenesis was not only a result of dedifferentiation of explant tissues but also an essential preparatory stage for in vitro morphogenesis; callus type may be an indicator for shoot organogenic potential. In their analysis of genetic relatedness of Dieffenbachia cultivars using AFLP, Chen et al. (2004) found that cv. Camouflage significantly differed genetically from cvs. ...
The capacity for indirect shoot organogenesis of leaf and root explants of four Dieffenbachia cultivars were examined on a modified Murashige and Skoog (MS; Physiol Plant 15:473–495, 1962) medium supplemented with different plant growth regulators in 112 combinations. Callus formation was only observed from
leaf explants on MS supplemented with 1–10μM thidiazuron (TDZ) and 0.5–1.0μM 2,4-dichlorophenoxyacetic acid (2,4-D) regardless
of cultivars. The combination of 5μM TDZ and 1μM 2,4-D resulted in the greatest callus formation frequency among the four
cultivars tested. Significant differences in callus and shoot formation from leaf explants were also observed among cultivars.
Cultivars Camouflage, Camille, Octopus, and Star Bright produced green nodular, brown nodular, yellow friable, and green compact
calli with corresponding maximum callus formation frequencies of 96%, 62%, 54%, and 52%, respectively. A maximum of 6.7 shoots/callus
was observed in cv. Camouflage, followed by cvs. Camille and Star Bright at 3.7 and 3.5, respectively. Calli of cv. Octopus
displayed no capacity for shoot organogenesis. Regardless of cultivar, callus formation was not observed on root explants.
Regenerated shoots were successfully acclimatized in a shaded greenhouse condition with 100% survival.
... AFLP has been used to study genetic relationships of a wide range of species, including popular ornamentals, such as Aglaonema Schott [10], Alocasia G. Don. [11], Dieffenbachia Schott [12], Caladium Venten. [13,14], Hemerocallis L. [15], and Philodendron Schott [16] and proven to be extremely sensitive to distinguishing closely related cultivars. ...
... Chen et al. [10,12] reported 69% polymorphism in Aglaonema Schott and 71% polymorphism in Dieffenbachia Schott. Devanand et al. [16] identified 64% fragments being polymorphic in cultivated ornamental Philodendron Schott. ...
Calatheas (Calathea Mey.) have been largely produced as ornamental foliage plants for interiorscaping due to their brilliant patterns of leaf color and different leaf textures as well as the ability to tolerate low light levels. Genetic relationships among Calathea species and cultivars, however, have not been documented. This study analyzed the genetic relatedness of 34 commonly grown cultivars across 15 species using amplified fragment length polymorphism (AFLP) markers with near-infrared fluorescence-labeled primers. Six EcoRI + 2 bases/MseI + 3 bases primer set combinations were used in this investigation. Each selected primer set generated 105–136 scorable fragments. A total of 733 AFLP fragments were detected, of which 497 were polymorphic (67.8%). An unweighted pair-group method of the arithmetic averages (UPGMA), principal coordinate analysis (PCOA), and bootstrap analysis were used to analyze the genetic relationships. The 34 cultivars were separated into four clusters. Cluster I contains 19 cultivars that are either from C. roseopicta or C. loesnerii with Jaccard's similarity coefficients ranging from 0.51 to 0.99. Among the 19 cultivars, at least nine are derived from somaclonal variants or sports. Only C. kennedeae ‘Helen’ is positioned in cluster II. Cluster III has 10 cultivars across seven species; Jaccard's similarity coefficients varied from 0.26 to 0.63. Four species are situated in cluster IV with Jaccard's similarity ranging from 0.19 to 0.41. This study establishes the genetic relationships of commonly cultivated calatheas, provides genetic evidence supporting that C. fasciata, C. orbifolia, C. rotundifolia, C. insignis, and C. ornata are independent species, and raises a concern over genetic vulnerability of cultivars in cluster I because of their close genetic similarities.
... 'Sarah' has much narrower green margins and extensive green blotches extending from the margin to the inner ivory panel. The three somaclonal cultivars along with their parent are popular as indoor plants (Chen et al., 2004). ...
Somaclonal variation, where genetic and phenotypic diversity arises among plants regenerated from tissue culture, has significant implications for ornamental horticulture. This variation, induced during in vitro propagation, presents an innovative and cost-effective method for developing new ornamental varieties with desirable traits such as unique color, fragrance, flower shape, and increased resistance to diseases and environmental stressors. Somaclonal variation mechanisms include chromosomal rearrangements, DNA methylation, transposable element activation, and somatic mutations. Although somaclonal variation can sometimes result in undesired traits, careful selection and screening can enhance ornamental plant breeding, leading to the commercialization of novel and improved varieties. Advances in molecular markers and genomic tools allow for early detection and management of beneficial somaclonal variants, improving their stability and reproducibility. Thus, somaclonal variation serves as a vital resource for genetic diversity and innovation in ornamental breeding, contributing to the aesthetic, economic, and ecological value of ornamental plants in landscaping and the floriculture industry. Somaclonal variation is a significant phenomenon in plant biotechnology, offering a valuable tool for enhancing genetic diversity and breeding efficiency in ornamental plants. This explores the mechanisms underlying somaclonal variation, including genetic and epigenetic modifications induced during in vitro culture, such as DNA mutations, chromosomal rearrangements, and changes in DNA methylation patterns. This variation, induced during in vitro propagation, presents an innovative and cost-effective method for developing new ornamental varieties with desirable traits such as unique color, fragrance, flower shape, and increased resistance to diseases and environmental stressors is critically examined, highlighting their importance in ornamental plant improvement. Furthermore, this discusses advanced molecular tools such as marker-assisted selection, next-generation sequencing, and transcriptomic approaches for detecting, characterizing, and stabilizing these variations. By bridging fundamental research with applied breeding, this review underscores the significance of somaclonal variation as an innovative strategy for the rapid development of unique and commercially valuable ornamental plant varieties. Advances in molecular markers and genomic tools allow for early detection and management of beneficial somaclonal variants, improving their stability and reproducibility. Additionally, the challenges associated with somaclonal variation, including genetic instability, undesirable mutations, and limited reproducibility, are addressed, along with strategies to enhance the efficiency and reliability of this approach. Thus, somaclonal variation serves as a vital resource for genetic diversity and innovation in ornamental breeding, contributing to the aesthetic, economic, and ecological value of ornamental plants in landscaping and the floriculture industry. Future prospects for integrating somaclonal variation with modern biotechnological techniques, such as CRISPR-based genome editing and synthetic biology, are also discussed to improve its potential in ornamental plant breeding further.
... To facilitate commercial production, tissue culture methods have been used as a tool for fast and reliable increase of hybridized Dieffenbachia selections [9], [10]. Dieffenbachia is an ornamental perennial monocot plant native to tropical America [11], belonging to the family Araceae [12]. It is prized among interiorscapers for the attractive variegated foliage, tolerance of interior environments and easy production [13]. ...
The ornamental plant industry has applied an extremely in vitro propagation approach to large-scale plant propagation with highly selected superior cultivars. As a result, hundreds of plant tissue culture laboratories have emerged worldwide. Dieffenbachia is a very popular ornamental plant which belongs to the Araceae. Dieffenbachia species are popular leafy potted plants used in indoor landscaping of homes, offices and shopping malls worldwide. The plants can be 60 cm to 3 m tall, and have large spotted and/or variegated (white, yellow, green) leaves that may be 30-45 cm long and 15-20 cm wide. They grow well indoors and in some areas outdoor. Most Dieffenbachia species are now propagated by tissue culture to better utilize the species and accelerate plant growth. Further application of plant tissue culture techniques on Dieffenbachia is needed to investigate the response of different cultivars and explants to variable culture media.
... The diverse morphological forms observed in the family Araceae have imposed some complexity and problems in its taxonomy (Ngoka, 1997;Green and Oguzor, 2009) which are being solved by different authors (Onwueme, 1978;Burkill, 1985;Gill, 1988;Hirai et al. 1989;Mayo et al., 1997;Hesse et al., 1999;Bown, 2000;Keating, 2003;Chen et al., 2004;Hesse, 2006;Cabrera et al., 2008;Green and Oguzor, 2009;Osuji and Nwala, 2015;Arogundade and Adedeji, 2016;2017). There is no known work in which numerical methods were applied to members of the family Araceae in order to understand their pattern of relationships. ...
Sixteen taxa belonging to four tribes in the family Araceae were grouped based on numerical data generated from the coding of 16 morphological and 44 anatomical characters with the aim of inferring phenetic relationships among them. The data from these two fields of evidence were subjected to Paired Group Cluster Analysis (PGCA) and Principal Components Analysis (PCA). From the results, two groups emerged. The first group consists of members of genera Alocasia, Colocasia and Xanthosoma which are acaulescent herbs in that their stem(s) are subterranean and are also corm producers while the members of genera Aglaonema and Dieffenbachia which are caulescent herbs exhibiting decumbent character and non-corms producers constitute the second group. Twenty-nine (29) characters comprising of 15 morphological characters and 14 anatomical characters had high component loadings and can be employed in the delimitation of the taxa. The importance of these characters in the delimitation of the Araceae taxa was discussed.
... Although, elliptic shaped stomata are common to all the taxa studied, some of the taxa have Vincent (1989) and Koneman et al. (1997). The genetic relatedness of 42 cultivars of Dieffenbachia was studied using Amplified Fragment Length Polymorphism (AFLP) markers (Chen et al., 2004). Colourless essential oils from fresh leaves and stems of Dieffenbachia picta have been analysed by gas chromatography (Oloyede et al., 2011). ...
This study provides detailed information of the anatomical attributes of the epidermis and the three regions of the petiole of four members of the genus Dieffenbachia Schott. Fresh samples of the leaves of Dieffenbachia picta Schott, Dieffenbachia oerstedii Schott, Dieffenbachia senguine (Jacq) Schott and Dieffenbachia senguine cultivar ‘Candida’ Schott were used. Epidermal peels and transverse sections were made following standard procedures. Generic characters revealed uniform epidermal cell shape on the abaxial surface, wavy to undulating and straight to wavy anticlinal wall patterns on the adaxial and abaxial surfaces respectively, brachyparacytic stomata types, round abaxial petiole outline, the presence of raphides and druses in the petioles of all the taxa. Delimiting features include irregular epidermal cell shape on the adaxial surfaces of D. senguine
and D. senguine cv. ‘Candida’, additional anomocytic stomata types on the abaxial surfaces of D. picta, D. oerstedii and adaxial surface of D. senguine, cuticular striations on the abaxial surfaces of D. oerstedii and D. senguine cv. ‘Candida’, druses and
raphide bundle on the epidermal surface of D. senguine only, flat adaxial petiole outline and slightly concave adaxial petiole outline in the proximal and median regions of D. senguine cv. ‘Candida’ and the presence of lamellar collenchyma cells in the
petiole of D. picta. Data for both quantitative and qualitative characters were subjected to Principal Components Analysis and Single Linkage Cluster Analysis. Interestingly, anomocytic stomata complex, cuticular striations, raphide bundles and druses
and the adaxial petiole outline separated D. senguine and D. senguine cv. ‘Candida’.
... Although, elliptic shaped stomata are common to all the taxa studied, some of the taxa have Vincent (1989) and Koneman et al. (1997). The genetic relatedness of 42 cultivars of Dieffenbachia was studied using Amplified Fragment Length Polymorphism (AFLP) markers (Chen et al., 2004). Colourless essential oils from fresh leaves and stems of Dieffenbachia picta have been analysed by gas chromatography (Oloyede et al., 2011). ...
This study provides detailed information of the anatomical attributes of the epidermis and the three regions of the petiole of four members of the genus Dieffenbachia Schott. Fresh samples of the leaves of Dieffenbachia picta Schott, Dieffenbachia oerstedii Schott, Dieffenbachia senguine (Jacq) Schott and Dieffenbachia senguine cultivar ‘Candida’ Schott were used. Epidermal peels and transverse sections were made following standard procedures. Generic characters revealed uniform epidermal cell shape on the abaxial surface, wavy to undulating and straight to wavy anticlinal wall patterns on the adaxial and abaxial surfaces respectively, brachyparacytic stomata types, round abaxial petiole outline, the presence of raphides and druses in the petioles of all the taxa. Delimiting features include irregular epidermal cell shape on the adaxial surfaces of D. senguine and D. senguine cv. ‘Candida’, additional anomocytic stomata types on the abaxial surfaces of D. picta, D. oerstedii and adaxial surface of D. senguine, cuticular striations on the abaxial surfaces of D. oerstedii and D. senguine cv. ‘Candida’, druses and raphide bundle on the epidermal surface of D. senguine only, flat adaxial petiole outline and slightly concave adaxial petiole outline in the proximal and median regions of D. senguine cv. ‘Candida’ and the presence of lamellar collenchyma cells in the petiole of D. picta. Data for both quantitative and qualitative characters were subjected to Principal Components Analysis and Single Linkage Cluster Analysis. Interestingly, anomocytic stomata complex, cuticular striations, raphide bundles and druses and the adaxial petiole outline separated D. senguine and D. senguine cv. ‘Candida’.
... In this study we could show that levels of similarity with values down to 0.68 are relatively low in dahlias compared to cultivars of other ornamental species from the Asteraceae (CHEN et al. 2004b;GAWENDA 2006). This can be explained by multiple hybridisations between species and species hybrids at the beginning of dahlia breeding about 200 years ago and the common practise of most breeders to select open pollinated seedlings from larger populations of diverse genotypes. ...
Genetic distances between nineteen Dahlia cultivars and three genotypes of wild Dahlia species and a hybrid were analysed with 1432 AFLP-Markers generated by 10 Hind/Mse primer combinations. Markers were transformed into a 0/1-matrix and relative distances were computed by means of the Jaccard-coefficient. From these data a dendrogram was constructed by the UPGMA-method. In the dendrogram the cultivars cluster together into one group, while the wild species and the hybrid branched off at a larger genetic distance. The level of genetic similarity of the cultivars averaged between 0.68 and 0.77. Only two varieties displayed a similarity of 0.92 to each other. This relatively large genetic distance between the cultivars indicates a vast potential for further variety development. Within the cultivars different clusters are formed which do not show any relation to phenotypic characteristics like inflorescence morphology or breeding origin. This lack of correlation indicates that a classification of dahlias into horticultural groups does not reflect genetic relationships between cultivars. Therefore, this classification should not be used to select parents for further breeding apart for the few characters it is built on. With knowledge about the genetic background additional information for the selection of genotypes as parent in breeding is available. Genotypes can be chosen for their low level of relatedness or for their genetically characteristics independent from the horticultural type.
... AFLP analysis was conducted using the GIBCO BRL AFLP system II (Life Technologies, Grand Island, N.Y.) and the products were visualized with a LI-COR 4000-L automated sequencer (LI-COR, Lincoln, Nebr.). The AFLP protocol is described in detail in a previous study (Chen et al., 2004). Thirty-two EcoR I + 2 bases/Mse I + 3 bases AFLP primer sets (E+_ _/M+ _ _ _) were initially screened for markers that were specifi cally linked to ʻCaffi nʼ and ʻFina Sodeaʼ clementine mandarins from Madera, ʻOwariʼ satsuma mandarin from Madera, ʻLane Lateʼ navel orange from Madera, and ʻMinneolaʼ tangelo from Madera, ʻAfourerʼ mandarin from Madera and from Bakersfi eld, navel oranges from , and the distance of the ʻMinneolaʼ tangelo or clementine mandarin pollen transferred within an orchard near Madera, Calif., that found in the pollen parentage study of ʻAfourerʼ mandarin seedlings. ...
Production of seedless mandarins such as 'Nules' clementine mandarin (Citrus Clementina Hort. Ex Tan.) and 'Afourer' mandarin [C. sinensis (L.) Osbeck x C. reticulata Blanco] is increasing in California as consumers' interest in seedless, easy peeling, and good tasting mandarins increases. The fruit would produce seeds if cross-pollination with compatible pollen source occurred. It is almost impossible to prevent cross-pollination between compatible mandarin cultivars by honeybees (Apis mellifera L.) within the multi-faceted agricultural environment in California. To produce seedless mandarin, growers either plant a single cultivar in a large solid block or try to use pollen-sterile navel oranges (C. sinensis) or satsuma mandarins (C. unshiu Marco.) as buffers to prevent cross-pollination. The question of how many rows of buffer trees or spacing can effectively prevent cross-pollination by honeybees between compatible mandarins is unclear. We initiated a study using fluorescent-labeled AFLP markers to determine the pollen parentages of 'Nules' clementine seedlings and 'Afourer' mandarin seedlings from two orchards in California. The longest distance of pollen flow at an orchard near Madera was 521 m. The pollen of 'Minneola' tangelo (C. reticulata x C. paradisi Macf.) was able to disperse across a minimum of 92 rows of 'Lane Late' navel oranges plus two rows of 'Afourer' mandarins to pollinate 'Afourer' mandarins. We also found that all the seedlings of 'Nules' clementine mandarin at an orchard near Bakersfield had been pollinated by 'Afourer' mandarin pollen. The pollen of 'Afourer' mandarin was able to disperse up to distances between 837 and 960 m to pollinate 'Nules' clementine. The pollen dispersal distance found in this study was at least 16 times longer than previously reported in a citrus orchard. Growers need to consider a much larger space or buffer rows to prevent cross-pollination and produce seedless mandarins in California.
... The patterns of RAPD produced by the primers OPC-02 and OPN-02 are shown in the Figures 1A, B. The genetic variation through molecular markers has been highlighted in a number of ornamental foliage crops (Wen et al. 2001;Chen et al. 2004a, b;Phang et al. 1999). Chen et al. (2004b) indicated that the development of different cultivars of Dieffenbachia were originated from either in sports or somaclonal variants. The present findings show the distant variation among the species and close variation among the varieties by using RAPD markers. ...
The species/varieties of Polyscias and Schefflera are important ornamental foliage pot plants. The germplasm identification and characterization is an important link between the conservation and utilization of plant genetic resources. Investigation were undertaken for identification and determination of genetic variation within 14 species/varieties of Polyscias and one species of Schefflera elegantissima under family Araliaceae through RAPD (random amplified polymorphic DNA) and ISSR (Inter Simple Sequence Repeats) markers. Genetic analysis was made by using 15 selected decamer primers and 9 selected ISSR markers. A total of 164 and 69 distinct DNA fragments ranging from 300 to 2500 bp were amplified by using selected random RAPD and ISSR primers respectively. The genetic similarity was evaluated on the basis of presence or absence of bands. The cluster analysis was made by using similarity coefficient. The cluster analysis indicated that the 14 varieties/species of Polyscias formed one major cluster and Schefflera elegantissima forming another major cluster. There were distant variation among the genus or species and close variation was obtained among the varieties of Polyscias. The correlation matrix indicates that there was significant correlation between ISSR and RAPD markers. Thus, these markers have the potential for identification of species/varieties and variation within the varieties. This is also helpful in breeding programs as well as a major input into conservation biology of foliage crop. Copyright © 2007 The Japanese Society for Plant Cell and Molecular Biology.
... Cytological assay includes chromosome counts and DNA flow cytometry analysis. Among the available molecular marker techniques, amplified fragment length polymorphisms (AFLP) has been widely used for somaclonal and naturally occurring sport evaluation (Chao et al. 2005;Chen et al. 2004). AFLP can be used to detect variation on the DNA level and has proven to be extremely effective in distinguishing closely related genotypes ). ...
... The exact origin of 'Panther' is unknown, but it is the most tolerant cultivar so far identified. 'Camouflage' is a selection of somaclonal variants of 'Panther', and 'Octopus' is a cultivar selected from somaclonal variants of 'Camouflage' (Chen et al., 2004). The chilling-tolerant characteristic of 'Panther' has been largely maintained in 'Camouflage' but moderately lost in 'Octopus'. ...
This study evaluated chilling sensitivity of eight popular Dieffenbachia cultivars. Tissue culture liners were potted in 15-cm diameter pots using Vergro Container Mix A and grown in a shaded greenhouse under maximum photosynthetically active radiation of 285 mu mol center dot m(-2)center dot s(-1) for 5 months. After determining growth indices, the plants were chilled in walk-in coolers at 2, 7, or 12 degrees C for 6,12, or 24 h. Chilled plants were placed back in the shaded greenhouse for chilling injury and growth evaluation. Visible symptoms of injury included chlorosis, necrosis, water-soaked patches on leaves, or complete wilting. In addition to leaf injury, stems of some cultivars chilled at 2 degrees C for 24 h became water-soaked at the base, which resulted in the death of either entire shoots or entire plants depending on cultivars. Leaf injury occurred in all cultivars chilled at 2 degrees C, except for 'Panther'; and the longer the exposure at this temperature, the greater the injury. No visual injury was observed among plants chilled at 7 and 12 degrees C except 'Tropic Honey' that had 26% of leaves injured at 7 degrees C. Based on the percentage of injured leaves 12 days after chilling at 2 degrees C for 24 h, the sensitivity of the eight cultivars ranked as follows: Tropic Honey > Sterling > Carina >= Octopus > Camille > Camouflage > Star Bright > Panther. In addition to visual injury, plant growth was also affected by chilling during the subsequent 3 months of growth. All 'Tropic Honey' chilled at 2 degrees C died regardless of the tested chilling duration. Growth indices of all other cultivars except for 'Panther' chilled at 2 degrees C for 24 h significantly decreased compared with those of controls. 'Camille', 'Camouflage', 'Carina', and 'Sterling' also exhibited significant growth reduction after chilling at 2 degrees C for 12 h. This study showed that genetic variation in chilling sensitivity exists among cultivated Dieffenbachia. The identified chilling-tolerant cultivars could be used for breeding of new chilling-tolerant cultivars. The use of chilling-tolerant cultivars in production may reduce the chance of injury during heating outages and shipment.
... The patterns of RAPD produced by the primers OPC-02 and OPN-02 are shown in the Figures 1A, B. The genetic variation through molecular markers has been highlighted in a number of ornamental foliage crops (Wen et al. 2001;Chen et al. 2004a, b;Phang et al. 1999). Chen et al. (2004b) indicated that the development of different cultivars of Dieffenbachia were originated from either in sports or somaclonal variants. The present findings show the distant variation among the species and close variation among the varieties by using RAPD markers. ...
Abstract The species/varieties of Polyscias and Schefflera are important ornamental foliage pot plants. The germplasm
identification and characterization is an important link between the conservation and utilization of plant genetic resources.
Investigation were undertaken for identification and determination of genetic variation within 14 species/varieties of
Polyscias and one species of Schefflera elegantissima under family Araliaceae through RAPD (random amplified
polymorphic DNA) and ISSR (Inter Simple Sequence Repeats) markers. Genetic analysis was made by using 15 selected
decamer primers and 9 selected ISSR markers. A total of 164 and 69 distinct DNA fragments ranging from 300 to 2500 bp
were amplified by using selected random RAPD and ISSR primers respectively. The genetic similarity was evaluated on the
basis of presence or absence of bands. The cluster analysis was made by using similarity coefficient. The cluster analysis
indicated that the 14 varieties/species of Polyscias formed one major cluster and Schefflera elegantissima forming another
major cluster. There were distant variation among the genus or species and close variation was obtained among the varieties
of Polyscias. The correlation matrix indicates that there was significant correlation between ISSR and RAPD markers. Thus,
these markers have the potential for identification of species/varieties and variation within the varieties. This is also helpful
in breeding programs as well as a major input into conservation biology of foliage crop.
... Aggarwal et al. (2002) identifi ed 501 AFLP markers from basmati rice (Oryza sativa L.) with 65% of them being polymorphic. Chen et al. (2004aChen et al. ( , 2004b reported 69% polymorphism in Aglaonema and 71% polymorphism in Dieffenbachia, two other aroid genera. ...
Philodendrons (Philodendron Schott) are among the most popular tropical ornamental foliage plants used for interior decoration. However, limited information is available on the genetic relationships among popular Philodendron species and cultivars. This study analyzed genetic similarity of 43 cultivars across 15 species using amplified fragment length polymorphism (AFLP) markers with near infrared fluorescence labeled primers. Forty-eight EcoR I + 2 / Mse 1 + 3 primer set combinations were screened, from which six primer sets were selected and used in this investigation. Each selected primer set generated 96 to 130 scorable fragments. A total of 664 AFLP fragments were detected, of which 424 (64%) were polymorphic. All cultivars were clearly differentiated by their AFLP fingerprints, and the relationships were analyzed using the unweighted pair-group method of arithmetic average cluster analysis (UPGMA) and principal coordinated analysis (PCA). The 43 cultivars were divided into five clusters. Cluster I comprises eight cultivars with arborescent growth style. Cluster II has only one cultivar, 'Goeldii'. There are 16 cultivars in cluster III, and most of them are self-heading interspecific hybrids originated from R.H. McColley's breeding program in Apopka, Fla. Cluster IV contains 13 cultivars that exhibit semi-vining growth style. Cluster V has five cultivars that are true vining in morphology, and they have lowest genetic similarity with philodendrons in other clusters. Cultivated philodendrons are generally genetically diverse except the self-heading hybrids in cluster III that were mainly developed using self-heading and semi-vining species as parents. Seven hybrid cultivars have Jaccard's similarity coefficients of 0.88 or higher, suggesting that future hybrid development needs to select parents with diverse genetic backgrounds.
... The frequency of somaclonal variants is generally high, and the time required for a new cultivar release can be only 2-3 years compared to 7-10 years required using traditional breeding methods (Skirvin et al. 1994;Chen et al. 2003;Henny and Chen 2003). Chen et al. (2004) analyzed genetic relatedness of some cultivated Dieffenbachia using amplified fragment length polymorphism and found that cultivars selected from somaclonal variants differ genetically from their parents. ...
A novel protocol for indirect shoot organogenesis of Dieffenbachia cv. Camouflage was established using leaf explants excised from in vitro shoot cultures. The frequency of callus formation
reached 96% for explants cultured on Murashige and Skoog (1962) basal medium supplemented with 5μM thidiazuron and 1μM 2,4-dichlorophenozyacetic acid. The number of shoots regenerated
was high, with up to 7.9 shoots produced per callus cultured on basal medium supplemented with 40μM N
6-(Δ2-isopentenyl)adenine and 2μM indole-3-acetic acid. Regenerated shoots rooted well in a soilless substrate, acclimatized ex
vitro at 100%, and grew vigorously under shaded greenhouse conditions. Somaclonal variations in leaf variegation, color, and
morphology have been observed in regenerated plants.
Ornamental foliage plants have witnessed immense popularity
in recent years because of their captivating array of colours, textures,
and shapes that make them stand out in garden landscapes, indoor
spaces, and commercial settings. This review offers a thorough
summary of the history, origin, objectives, constraints and methods of
breeding of ornamental foliage plants, with a particular emphasis on
important elements such as breeding techniques, and future
prospects.A wide range of characteristics, such as foliage colour,
shape, size, pest and disease resistance, and glowing foliage are sought
after in breeding.The breeding of ornamental foliage plants is a
constantly evolving and diverse area that combines conventional
breeding methods with the latest biotechnological progress. Various
breeding methods, including introduction, selection, mutation
breeding,hybridization, tissue culture, and genetic engineering, are
employed to enhance genetic variability and introduce novel traits.
However, there are certain constraints in ornamental foliage plant
breeding including natural flowering, protandry, protogyny, low seed
quality, and fruit maturity. This can be solved by collaboration between
breeders, researchers, growers, and regulatory authorities leading to
sustainable and responsible breeding practices.The future of breeding
of ornamental foliage plants looks bright, because of the ongoing
research into different methods of breeding of ornamental foliage
plants. By integrating recent breeding technologies with traditional
breeding methods, there is a promise of accelerating trait discovery,
improving breeding efficiency, and developing robust and visually
attractive varieties suitable for various landscapes and climates.This
review aims to provide valuable insights into the current state and
future directions of breeding ornamental foliage plants, offering a
foundation for further research and development in this vital field.
Putri HA, Purwito A, Sudarsono, Sukma D. 2021. Morphological, molecular and resistance responses to soft-rot disease variability among plantlets of Phalaenopsis amabilis regenerated from irradiated protocorms. Biodiversitas 22: 1077-1090. Phalaenopsis amabilis (L.) Blume is a prominent donor for the white petal and sepal trait in Phalaenopsis breeding. However, it has an undesirable character, such as susceptible to soft-rot disease. Therefore, developing soft-rot resistance mutants through gamma irradiation could be explored. This study aimed to evaluate the variability of plantlets regenerated from irradiated and non-irradiated protocorms using morphology, stomatal size, and molecular markers and to test responses of the plantlets against soft-rot disease. The plantlets were regenerated from irradiated (5, 10, 15, or 20 Gy) and non-irradiated protocorms. The results showed that P. amabilis plantlet variants were successfully identified based on their leaf morphology and stomatal size variations. A few plantlets have low stomatal densities, large stomatal size, and high chloroplast numbers, which indicated they were polyploids. Leaf disc assay for soft-rot disease response grouped most of the plantlets into very susceptible or susceptible. Moreover, four soft-rot resistant plantlets regenerated from irradiated and non-irradiated protocorms were successfully identified. The resistant plantlets were identified after three consecutive periods of inoculations with pathogens causing soft-rot disease. The evaluation also confirmed nucleotide variation in the Pto gene isolated from different levels of plantlet variant resistance responses.
Here, we used data generated from amplified fragment length polymorphism (AFLP) analysis to address the biodiversity status and taxonomic relationships among 47 wild accessions representing 9 species of the genus Tulipa in Iran. A high level of genetic diversity within the genus was observed; the most distant taxa were T. humilis and T. schrenkii, while the highest degree of similarity was found between T. montana and T. biflora. Twelve AFLP primer sets amplified 342 fragments, of which 304 were polymorphic (88.1%). The average number of polymorphic bands per AFLP primer pair was 28.5. A hierarchical cluster analysis was carried out on the genetic profile of the taxa, and the results mostly reconfirmed the recognized taxonomy of the genus. However, we found evidence for recognition of a new subgenus for T. biebersteiniana.
RAPD and ISSR analysis was used to evaluate the genetic variation among 13 cultivars of Calathea across 11 species. In total, 262-marker loci were assessed; of which 252 were polymorphic revealing 96.1% polymorphism. Genetic diversity parameter was calculated for RAPD, ISSR and RAPD + ISSR approaches. Nei's similarity index varies from 0.49 to 0.80, 0.44 to 0.85 and 0.51 to 0.82 respectively. Cluster analysis by the unweighted pair group method (UPGMA) of Dice coefficient of similarity generated dendograms with similar topology that gave a better reflection of diversity and affinities between cultivars. The phylogenetic trees divided the 13 Calathea cultivars studied into two groups: group I consisting of only C. bella and the remaining 12 cultivars in group II. This molecular result is comparable to notable morphological characteristics. A mantel test of correlation analysis of the distance matrices was carried out that resulted in significant correlation (r = 0.427 at p = 0.998) between RAPD and ISSR markers. Thus, these markers have the potential for identification of species/varieties and variation within the species that are efficient for germplasm management. This is also helpful in breeding programs as well as a major input into conservation biology of the foliage crop.
Ornamental peach (Prunus persica (L.) Batsch) is a popular plant for urban landscapes and gardens. However, the genetic relationship among ornamental peach cultivars is unclear. In this report, a group of 51 ornamental peach taxa, originated from P. persica and P. davidiana (Carr.) Franch., has been studied using AFLPs. The samples were collected from China, Japan, and US. A total of 275 useful markers ranging in size from 75 to 500 base pairs were generated using six EcoRI/MseI AFLP primer pairs. Among them, 265 bands were polymorphic. Total markers for each taxon ranged from 90 to 140 with an average of 120. Two clades were apparent on the PAUP-UPGMA tree with P. davidiana forming an outgroup to P. persica, indicates that P. davidiana contributed less to the ornamental peach gene pools. Within P. persica clade, 18 out of 20 upright ornamental peach cultivars formed a clade, which indicated that cultivars with upright growth habit had close genetic relationship. Five dwarf cultivars were grouped to one clade, supported by 81% bootstrap value, indicating that they probably derived from a common gene pool. These results demonstrated that AFLP markers are powerful for determining genetic relationships in ornamental peach. The genetic relationships among ornamental cultivars established in this study could be useful in ornamental peach identification, conservation, and breeding.
Ornamental Ficus L. is a group of lactiferous trees, shrubs, and woody root-climbing vines that are cultivated either as landscape plants in the tropics and subtropics or as foliage plants used worldwide for interiorscaping. With the recent rapid expansion of the ornamental plant industry, more new Ficus species and cultivars have been introduced. However, no study has thus far addressed the genetic relationships of cultivated ornamental Ficus. Using amplified fragment length polymorphism (AFLP) markers with near-infrared fluorescence-labeled primers, this study analyzed the genetic relatedness of 56 commercial cultivars across 12 species. Forty-eight EcoRI + 2/MseI + 3 primer set combinations were initially screened, from which six primer sets were selected and used in this investigation. Most cultivars were differentiated by their AFLP fingerprints, and their relationships were determined using the unweighted pair-group method of arithmetic average cluster analysis. The 56 cultivars were divided into 12 clusters that correspond to 12 species, indicating that no interspecific hybrids of ornamental Ficus are in commercial production. The 12 species are genetically diverse, with Jaccard's similarity coefficients ranging from 0.21 to 0.43. However, cultivars within three species - Ficus benjamina L., Ficus elastica Roxb. Ex Hornem., and Ficus pumila L. - are genetically close. Twenty-seven of the 29 cultivars of F. benjamina and five cultivars of F. pumila had Jaccard's similarity coefficients of 0.98 or higher respectively. Nine cultivars of F. elastica shared Jaccard's coefficients higher than 0.96. These results indicate potential genetic vulnerability of these cultivars within the three species. Because there are increasing reports of invasive pests in the ornamental plant industry, strategies for conserving genetic resources and broadening genetic diversity of cultivated Ficus are discussed.
The aim of this paper is to review progress in phylogenetic research of Araceae during the period since publication of the first major molecular study by French et al. (1995). This, the first cladogram of the whole family inferred from DNA molecular data (Fig 9.1), resulting from research by J.C. French, M. Chung and Y. Hur, was based on chloroplast restriction site data (RFLPs or restriction fragment length polymorphisms). Their paper was highly significant and marked the beginning of the modern era of molecular phylogenetics of Araceae, nowadays based on DNA sequence data (e.g. Cabrera et al., 2008; Cusimano et al., 2011; Nauheimer et al., 2012b). It was innovative for Araceae in other ways as well, being the first family-scale cladistic analysis using computer algorithms and the first published cladogram for the family as a whole using genera as the ultimate operational taxonomic units (OTUs). No attempt has been made in the present chapter to discuss in detail the work of the previous 25 years during which many significant advances in systematic knowledge of the family were made, both in morphological taxonomy, but also in other fields such as cytology, palynology, phytochemistry, anatomy, fossil aroids, pollination biology and seedling morphology. Reviews of this literature have been provided by various authors, including Petersen (1989), Grayum (1990), Mayo et al. (1997) and Keating (2003), and Nicolson (1960) and Croat (1990) summarized classifications published in this earlier period.
This study analysed genetic relationships of 23 Alocasia cultivars across 17 species using amplified fragment length polymorphism (AFLP) markers. Six primer sets, selected from an initial screening of 48, generated a total of 578 scorable AFLP fragments of which 334 (58.4%) were polymorphic. All cultivars were clearly detected by their AFLP fingerprints. A dendrogram was constructed using the unweighted pair-group method of arithmetic averages (UPGMA). Principal coordinated analysis (PCOA) was carried out to show multiple dimensional distributions of cultivars. Both UPGMA and PCOA analyses separated the 23 cultivars into three clusters. Cluster I comprises 16 cultivars, mainly derived from A. crassifolia, A. cuprea, A. longiloba, A. grandis, A. guttata, A. plumbea, A. macrorrhiza, A. micholitziana, and A. villeneuvei or hybrids of A. lowii X A. sanderiana and A. cuprea X A. veitchii. Jaccard's similarity coefficients for these species ranged from 0.43 to 0.77. Cluster II contains six cultivars, which include A. cadieri, A. cucullata, A. gageana, A. odora, and A. portei. Jaccard's similarity coefficients varied from 0.52 to 0.83. There is only one cultivar, 'Hilo Beauty' in the cluster III, whose low similarity (0.21) with the rest of the Alocasia species may suggest that it could actually belong to another genus of Araceae. Based on documented interspecific hybrids, it appears that hybrids were developed from species exclusively within the identified clusters. This may suggest that Alocasia species sharing high Jaccard's similarity coefficients are more likely to be intercrossable. The interspecific relationships detected by the AFLP analysis could provide the genetic basis for selecting parents for future hybrid development.
Japanese barberry (Berberis thunbergii DC.) is a popular ornamental shrub used in garden and urban landscaping. Currently there are over 60 B. thunbergii cultivars in the market. To better distinguish its cultivars, we used the amplified fragment length polymorphism (AFLP) technique to develop DNA marker profiles for 59 cultivars and hybrids. These markers were used to authenticate the trueness-to-name of B. thunbergii cultivars in production and in the market, control for intracultivar genetic variants, and develop a molecular key to identify cultivars approved for im-portation in Canada. Polymorphic markers from seven primer combinations were able to clearly differentiate 57 of 59 cultivars evaluated. Two cultivars, Aurea and Aurea Nana, could not be differentiated because they had identical marker profiles. Among the 274 plants tested, 263 were confirmed to be true-to-name and correctly labeled, whereas 11 plants could not be confirmed true-to-name. Seven of the 20 cultivars evaluated exhibited detectable intracultivar genetic variation. 'Crimson Pygmy' had the highest number of plants exhibiting genetic variability. Overall, nursery producers and retailers do not appear to be mixing or mislabeling cultivars. A molecular key developed from a subset of 25 markers was able to accurately identify and differentiate the 11 B. thunbergii cultivars approved for importation in Canada. This key could be used in a cultivar verification program to facilitate international trade of B. thunbergii cultivars where wheat rust is a concern.
Introduction Production Conditions and Environments Foliage Plant Propagation Foliage Plant Production Postproduction The Future Literature Cited
The occurrence of somaclonal variation among regenerants derived through indirect shoot organogenesis from leaf explants of
three Dieffenbachia cultivars Camouflage, Camille and Star Bright was evaluated. Three types of somaclonal variants (SV1, SV2, and SV3) were
identified from regenerated plants of cv. Camouflage, one type from cv. Camille, but none from cv. Star Bright. The three
variants had novel and distinct foliar variegation patterns compared to cv. Camouflage parental plants. Additionally, SV1
was taller with a larger canopy and longer leaves than parental plants and SV2. SV2 and SV3 did not produce basal shoots (single
stem) but basal shoot numbers between SV1 and parental plants were similar ranging from three to four. The variant type identified
from regenerated cv. Camille had lanceolate leaves compared to the oblong leaves of the parent. This variant type also grew
taller and had a larger canopy than parental plants. The rates of somaclonal variation were up to 40.4% among regenerated
cv. Camouflage plants and 2.6% for regenerated cv. Camille. The duration of callus culture had no effect on somaclonal variation
rates of cv. Camouflage as the rates between plants regenerated from 8months to 16months of callus culture were similar.
The phenotypes of the identified variants were stable as verified by their progenies after cutting propagation. This study
demonstrated the potential for new cultivar development by selecting callus-derived somaclonal variants of Dieffenbachia.
The genus Rheum L., commonly known as rhubarb, is composed of approximately equal to 60 species, primarily distributed throughout northern and central Asia. Rhubarb species have been used for medicinal purposes for thousands of years; however, it was not until the 18th century that the culinary use of petioles was first reported. Although the origin(s) of culinary rhubarb is not clear, it is thought that they originated from hybridization of rhubarb species originally brought to Europe for medicinal purposes. Most rhubarb cultivars lack pedigree information, and the genetic relationship among cultivars is largely unknown. Amplified fragment length polymorphism (AFLP) markers were generated for fingerprint analysis of 37 cultivars and four putative Rheum species accessions. Ten EcoRI and MseI primer combinations were analyzed for a total of 1400 scored polymorphisms, with an average of 140 polymorphisms per primer combination. Results show at least two clusters of related cultivars, as well as distantly related accessions. This study provides an estimate of rhubarb cultivar genetic diversity using AFLP analysis.
Aglaonema is an important ornamental foliage plant genus, but genetic relationships among its species and cultivars have not been reported. This study analysed genetic relatedness of 54 cultivars derived from nine species using amplified fragment length polymorphism (AFLP) markers.
Initially, 48 EcoRI + 2/MseI + 3 primer set combinations were screened, from which six primer sets that showed clear scoreable and highly polymorphic fragments were selected and used for AFLP reactions. AFLP fragments were scored and entered into a binary data matrix as discrete variables. Jaccard's coefficient of similarity was calculated for all pair-wise comparisons among the 54 cultivars, and a dendrogram was constructed by the unweighted pair-group method using the arithmetic average (UPGMA).
The number of AFLP fragments generated per primer set ranged from 59 to 112 with fragment sizes varying from 50 to 565 bp. A total of 449 AFLP fragments was detected, of which 314 were polymorphic (70 %). All cultivars were clearly differentiated by their AFLP fingerprints. The 54 cultivars were divided into seven clusters; cultivars within each cluster generally share similar morphological characteristics. Cluster I contains 35 cultivars, most of them are interspecific hybrids developed mainly from A. commutatum, A. crispum or A. nitidum. However, Jaccard's similarity coefficients among these hybrids are 0.84 or higher, suggesting that these popular hybrid cultivars are genetically much closer than previously thought. This genetic similarity may imply that A. nitidum and A. crispum are likely progenitors of A. commutatum.
Results of this study demonstrate the efficiency and ease of using AFLP markers for investigating genetic relationships of ornamental foliage plants, a group usually propagated vegetatively. The AFLP markers developed will help future Aglaonema cultivar identification, germplasm conservation and new cultivar development.
Although Dieffenbachia have been important ornamental tropical foliage plants for many years, relatively few cultivars have become widely grown. Most new cultivars have originated from private plant collections or mutations of commercial cultivars, with breeding playing a small role. Due to the natural diversity of this genus, a breeding program involving Dieffenbachia was initiated in 1976 at the Central Florida Research and Education Center, Apopka. ‘Tropic Star’, herein described, is the third hybrid Dieffenbachia to be released to Florida foliage growers from that program (1, 2).
Pelargoniums are of considerable commercial importance, mainly because they are highly decorative and relatively easy to cultivate. As the numbers of pelargonium cultivars are continuously increasing, the adoption of reliable and discriminant molecular tools capable of identifying new cultivars and determining their diversity with respect to already registered ones promises to be the prime requirement for a valuable decorative plant market, as well as a genetic guarantee of the value of plant materials. A judicial trial offered a unique opportunity to develop an innovative line of research for molecularly characterizing Pelargonium peltatum genotypes on the basis of RAPD and AFLP fingerprinting. The DNA polymorphism of a pelargonium flower recovered at the site of a crime was compared with those of nine plants seized in the house of the suspected murderer. Once an efficient protocol for isolating genomic DNA in pelargonium had been optimized, a total of 162 RAPD and 568 AFLp markers were detected using respectively twenty 10-mer primers and twelve Eco-RI/Mse-I primer combinations. RAPD fingerprints were not reproducible using the recovered flower template. AFLP fingerprints detected an average of 47 markers and furnished evidence that the recovered flower was genetically different from each of the seized plants. This new type of molecular marker proved to be a powerful tool for discriminating genetic differences even between phenotypically similar pelargonium individuals. AFLP markers could, therefore, be adopted for the granting of Plant Breeders' Rights with the aim of making the protection of new pelargonium cultivars more specific and effective. Because they can detect reliable genetic polymorphisms and supply effective cultivar descriptors, AFLP fingerprinting could be crucial for the future of pelargonium breeding and new cultivar testing.
Sports from two cut rose varieties, as well as a garden rose variety, were analysed with molecular markers. Between 695 and 752 random amplified polymorphic DNA and AFLP fragments were used to infer genetic differences between the sports, the original variety and seedlings of these varieties. Whereas no polymorphisms between the sports of the cut rose varieties and the original variety were observed, five polymorphisms could be detected between the garden rose variety and its sports. In contrast, a large number of polymorphisms occurred between all varieties and their seedlings. Therefore molecular markers can be used to verify the origin of vegetatively propagated rose plants of doubtful origin, thus enabling breeders in the future to claim plant breeders rights on sports of varieties already registered.
In the European Alps, Rhododendron ferrugineum can constitute dense populations with almost 100% of cover. The developmental pattern by layering and the resulting complexity of population structure make it difficult to identify distinct clones even by excavation. Therefore genotypic structure of a R. ferrugineum population, in the French Alps, was inferred from AFLP markers. In a first step, we analysed 400 samples using AFLP profiles generated by one selective primer pair. Seventeen bands out of 25 were polymorphic (68%). We identified a total of 32 multilocus genotypes. In a second step, the 32 genotypes were verified by applying two additional primer pairs to the two most distant samples from each genotype. The mean similarity (proportion of band sharing) between pairs of clones was 0.85 (range from 0.52 to 0.94). The spatial distribution of clones showed that vegetative spreading mainly occurred down a slope. Based on an annual shoot mean growth of 2.6 cm/year and the size of the widest clone, we estimated the age of the oldest individual to be at least 300 years. A single genotype can occupy a large surface and sometimes form a dense patch, suggesting that this species adopts a phalanx growth form with limited intermingling of some genets.
The AFLP− (Keygene, Wageningen, The Netherlands) technology allows the simultaneous amplification of many restriction fragments in
a single polymerase chain reaction (PCR). Thus, the technology can be used to very efficiently detect restriction fragment
polymorphisms. As an example of the vast numbers of markers that can be detected, the author describes here a protocol to
construct a high-density genetic map mArabldopsis. The map may be used to identify primer combinations that will target fragments mapped to a specific area of the genome.
RAPD analysis was performed to assess DNA variation among rye plants regenerated from immature embryos and inflorescences.
From the studied plants, 40% showed at least one variation, and the number of mutations per plant was quite high, ranging
from 1 up to 12. On some occasions (2.9% of the scored bands) the modified band was observed in only one plant or in several
but originated from the same callus (variable band). In other cases (5.25%) the same band varied in several plants obtained
from different calli. We call these hypervariable bands and they could vary between plants belonging to different cultivars
and/or with different origins, inflorescences or embryos. Thus, they must originate through independent mutational events.
We assume that these bands represent hypervariable regions of the rye genome and so detect hot spots of DNA instability. Some
of these bands proved to be unique sequences, others were present in a low copy number while the remaining ones were moderately
or highly repetitive.
Since amplified fragment length polymorphism (AFLP) analysis has proved useful in distinguishing cultivars of Caladium, it was used to assess the status of species of Caladium vs. Xanthosoma, both in tribe the Caladieae, and to reassess the position of Hapaline in the same tribe. AFLP analysis using three primer combinations was carried out on four species of Caladium(C. bicolor, C. humboldtii, C. lindenii and C. schomburgkii). Results showed that AFLP can distinguish between the different species by their unique and different banding patterns.
AFLP analysis confirmed that C. humboldtii is a species distinct from C. bicolor and that C. lindenii is a true Caladium species and does not belong to Xanthosoma. UPGMA cluster analysis showed that C. bicolor and C. schomburgkii are most similar and that C. humboldtii is closer to the C. bicolor / C. schomburgkii cluster compared with C. lindenii. Genetic relationships between Caladium, Xanthosoma, Hapaline, Alocasia and Protarum were also examined by AFLP analysis using eight primer combinations. Several useful molecular markers were specific either
to Caladium orXanthosoma , so that AFLP can be used to distinguish species of these two genera. Genetic analysis of the genera examined confirms that
the Caladieae and Colocasieae tribes are distinct and that Hapaline falls within the tribe Caladieae and that Protarum is most distant from all the genera examined. Copyright 2000 Annals of Botany Company
A novel DNA fingerprinting technique called AFLP is described. The AFLP technique is based on the selective PCR amplification
of restriction fragments from a total digest of genomic DNA. The technique involves three steps: (i) restriction of the DNA
and ligation of oligonucleotide adapters, (ii) selective amplification of sets of restriction fragments, and (iii) gel analysis
of the amplified fragments. PCR amplification of restriction fragments is achieved by using the adapter and restriction site
sequence as target sites for primer annealing. The selective amplification is achieved by the use of primers that extend into
the restriction fragments, amplifying only those fragments in which the primer extensions match the nucleotides flanking the
restriction sites. Using this method, sets of restriction fragments may be visualized by PCR without knowledge of nucleotide
sequence. The method allows the specific co-amplification of high numbers of restriction fragments. The number of fragments
that can be analyzed simultaneously, however, is dependent on the resolution of the detection system. Typically 50—100 restriction
fragments are amplified and detected on denaturing polyacrylamide gels. The AFLP technique provides a novel and very powerful
DNA fingerprinting technique for DNAs of any origin or complexity.
When genetic diversity among organisms was measured with molecular markers, the question of genome coverage was currently stressed out. In order to check if well-distributed, mapped AFLP markers were more efficient in assessing varietal identification of carrot accessions than randomly chosen markers, nine closely related genotypes were analysed. A software was developed to realise 1,000 random choices of 20 to 70 mapped or unmapped markers, offering numerous genome coverages. We statistically showed that taking into account marker position does not provide a better estimation of genetic distances. Moreover, in the case of carrot, we concluded that 60 AFLP markers offer the best compromise between the level of precision and minimal expense.
New Guinea impatiens ( Impatiens hawkeri) is an economically important floral crop, however, little work has been conducted to further our understanding of the genetics of this crop. In this study, we used amplified fragment length polymorphism (AFLP) technology to investigate the level of polymorphism present among 41 commercial cultivars of New Guinea impatiens, study their genetic relatedness, and assess the genetic diversity in this material. An efficient DNA extraction protocol was developed, and a total of 48 EcoRI and MseI primer combinations were used for PCR amplification. Amplification products were then subjected to polyacrylamide gel electrophoresis. The AFLP analysis showed that all 41 cultivars generated between 73 and 130 scoreable polymorphic bands per primer combination. Gower's Genetic Dissimilarity estimates for the entire set of cultivars ranged between 0.940 and 0.488. A dendogram was generated from these dissimilarity data that revealed four groupings among these 41 cultivars. The implications of these results on genotypic variation, genetic relationships, and genetic diversity in New Guinea impatiens will be discussed.
Caladiums are popular ornamental plants that have not been well studied at the molecular level. Identification of species
within the genus Caladium (Araceae) has been based primarily on morphology. However, the lack of comprehensive references makes identification of Caladium cultivars extremely difficult. Amplified fragment length polymorphism (AFLP) analysis using 17 primer combinations was carried
out on two species of Caladium (C. bicolor and C. schomburgkii), including six cultivars of C. bicolor. Results showed that AFLP can be used to distinguish these two species by their unique and different banding patterns. Unweighted
Pair Group Method using Arithmetic Averages (UPGMA) permitted cluster analysis of data from 17 selected primer combinations
on six cultivars of C. bicolor and one cultivar ofC. schomburgkii . It showed that closely related species can clearly be differentiated and that genetic difference between cultivars can
also be established. Unique AFLP molecular markers were detected for all the C. bicolor cultivars used. The use of AFLP has potential for precisely characterizing and identifying particular caladium cultivars
as well as for the registration of new cultivars. It will also be useful in future breeding programmes and systematics studies.
Copyright 1999 Annals of Botany Company
The generation of AFLPs in spring barley cultivars provided genetic information relating to the development of the crop in the UK since 1953. Principal co-ordinate (PCO) analysis of genetic similarities (gs) confirmed the marked contrast in the cultivars used in the 1970s and 1980s. The earliest cultivars, many derived from Proctor, were succeeded by tall-strawed, disease-resistant types with high yield but poor malting potential. In the 1980s they were in turn replaced by short-strawed cultivars with excellent yield and good malting quality, which originated from Triumph. A PCO plot of gs provided insight into the effects of selection for disease resistance and the antagonism between malting quality and particular resistance genes. The analysis of gs was more useful than pedigrees and estimates of kinship in revealing the genetic relationship between cultivars. Theoretical considerations for maximising the efficiency of an AFLP genotyping programme are discussed in the context of the number of primer pairs required to distinguish genotypes at varying levels of similarity.
The daylily (Hemerocallis spp.) is one of the most economically important ornamental plant species in commerce. Interestingly, it is also one of the
most heavily bred crops during the past 60 years. Since the American Hemerocallis Society began acting as the official registry
of daylily cultivars in 1947, more than 40 000 registrations have been processed. In order to determine the effects of intensive
breeding on cultivar development, and to study relationships among different species, genetic variation in the daylily was
estimated using AFLP markers. Nineteen primary genotypes (species and early cultivars) and 100 modern cultivars from different
time periods were evaluated using 152 unambiguous bands (average 79% polymorphism rate) derived from three AFLP primer combinations.
Overall, pairwise similarity estimates between entries ranged between 0.618 and 0.926 (average=0.800). When comparing cultivar
groups from different time periods (1940–1998), genetic similarity was initially increased, compared to the primary diploid
genotypes, remained constant from 1940 to 1980, and then steadily increased as breeding efforts intensified and hybridizers
began focusing on a limited tetraploid germplasm pool derived by colchicine conversion. Among modern (1991–1998) daylily cultivars,
genetic similarity has increased by approximately 10% compared to the primary genotypes. These data were also used to evaluate
recent taxonomic classifications among daylily species which, with a few minor exceptions, were generally supported by the
AFLP data.
An unstable, dominant mutation for purple-red pigmentation in rice (Oryza sativa L.) has been identified in seed hulls of the tissue culture regenerated somaclone SC86-20973. Genetic analyses suggested that the purple red hull character is conditioned by a single dominant nuclear gene. The purple red hull is dominant over the wild-type green hull in the F1, but a reversal of the purple-red hull phenotype in the F2 reciprocal crosses was observed. Distorted segregation ratios were detected among 81 F3 families that were derived from purple-red hulled F2 plants. All green-hulled F2 and F3 individuals bred true. A stable, dominant purple apiculus mutation was also observed in the original somaclone, and it was strongly linked to the purple-red hull trait. It is postulated that the purple-red hull mutation arose as a dominant character that is stable when selfed in the homozygous condition, but which undergoes unidirectional meiotic gene conversion as a heterozygote.
DNA amplification fingerprinting (DAF) using a high primer-to-template ratio and single, very short arbitrary primers, was used to generate amplified fragment length polymorphic markers (AFLP) in soybean (Glycine max (L.) Merr.). The inheritance of AFLPs was studied using a cross between the ancestral Glycine soja PI468.397 and Glycine max (L.) Merr. line nts382, F1 and F2 progeny. The amplification reaction was carried out with soybean genomic DNA and 8 base long oligonucleotide primers. Silver-stained 5% polyacrylamide gels containing 7 M urea detected from 11 to 28 DAF products with primers of varying GC content (ranging from 50 to 100% GC). Depending on their intensity, AFLPs were classified into three classes. DAF profiles were reproducible for different DNA extractions and gels. Forty AFLPs were detected by 26 primers when comparing G. soja and G. max. Most AFLPs were inherited as dominant Mendelian markers in F1 and F2 populations. However, abnormal inheritance occurred with about 25% of polymorphisms. One marker was inherited as a maternal marker, presumably originating from organelle DNA while another showed apparent paternal inheritance. To confirm the nuclear origin and utility of dominant Mendelian markers, three DAF polymorphisms were mapped using a F11 mapping population of recombinant inbred lines from soybean cultivars Minsoy x Noir 1. The study showed that DAF-generated polymorphic markers occur frequently and reliably, that they are inherited as Mendelian dominant loci and that they can be used in genome mapping.
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