72 research outputs found
Regeneration and genetic transformation of rose and evergreen azalea
No full textThe overall goals of this research are to establish tissue culture systems for rose (Rosa hybrida L. and R. chinensis minima) and evergreen azalea (Rhododendron spp.), develop efficient and highly reliable regeneration systems via organogenesis and/or somatic embryogenesis, and develop gene transfer systems for these two important floral crops.Shoot organogenesis was observed on callus tissue upon transfer of rhizogenic explants pre-incubated with 2,4-D to a regeneration medium containing 22.7 M TDZ and 2.9 M GA\sb3. Secondary embryogenesis was observed, and increased numbers of somatic embryos were obtained following transfer of embryogenic calli to a growth regulator-free medium. For 'Carefree Beauty', glucose at 111 mM promoted higher organogenesis and somatic embryogenesis than sucrose at either 59 or 117 mM concentrations; however, for 'Baby Katie', no differences were observed between glucose and sucrose.The best growth regulator combination for adventitious shoot regeneration of azalea 'Fuchsia' and 'Hino Crimson' was 22.74 M TDZ and 22.8 M IAA. In general, incubating leaf explants in the dark for at least 1 week followed by low-light or high-light intensity was enhanced regeneration frequency. Continuous culture under high-light intensity suppresses shoot regeneration; however, a 2-week dark pretreatment promotes shoot organogenesis even when explants were grown under high-light intensity.Several factors for optimizing microprojectile-mediated gene transfer methods for rose and azalea were investigated. Transformation efficiency was increased by elevating the osmotic level of the medium during bombardment. Transformation efficiency was enhanced by combining osmotic treatment, cotyledonary-stage of embryogenic calli, and higher accelerating pressure settings. Comparing GUS (-glucuronidase) transient expression of two particle guns, the PIG (particle inflow gun) and the biolistic PDS-1000/He gene gun, the PIG device resulted in higher GUS expression than the biolistic gene gun. A 10% GUS transient expression was obtained at 1100 psi helium pressure with 6 cm distance from stopping screen-to-leaf sections of azalea using the biolistic gun. A 22.2% GUS transient expression was obtained on shoot tip-derived calli of azalea using the PIG device at 60 psi with an open-chamber accelerating setting.Several factors for optimizing Agrobacterium-mediated gene transfer methods for rose and azalea were also investigated. Highly proliferating tissues, rose somatic embryogenic calli and azalea shoot-tips, have been found to be amenable to Agrobacterium-mediated transformation. The cotyledonary-stage of rose somatic embryos was found to be most amenable for gene transfer. Regeneration and transformation of azalea shoot-tips were better than stem segments or leaf sections. Kanamycin was effective for inhibiting regeneration of azalea leaf sections; however, it was less efficient for selection of rose embryogenic calli. Wounding manipulations were not necessary for rose embryogenic calli; however, for certain Agrobacterium strains, a wounding treatment might increase regeneration and/or transformation frequency. Based on GUS expression, bombardment and fresh cut treatments showed a higher blue color intensity. Adding acetosyringone or a nurse culture of minced tobacco leaf nurse culture to the cocultivation medium enhanced the number and intensity of blue spots; moreover, adding acetosyringone to the bacterial culture probably increased stable transformation in azalea. The susceptibility of various tissue types to different Agrobacterium strains was observed.Made available in DSpace on 2011-05-07T12:32:13Z (GMT). No. of bitstreams: 2
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Regeneration and genetic transformation of rose and evergreen azalea
No full textThe overall goals of this research are to establish tissue culture systems for rose (Rosa hybrida L. and R. chinensis minima) and evergreen azalea (Rhododendron spp.), develop efficient and highly reliable regeneration systems via organogenesis and/or somatic embryogenesis, and develop gene transfer systems for these two important floral crops.Shoot organogenesis was observed on callus tissue upon transfer of rhizogenic explants pre-incubated with 2,4-D to a regeneration medium containing 22.7 M TDZ and 2.9 M GA\sb3. Secondary embryogenesis was observed, and increased numbers of somatic embryos were obtained following transfer of embryogenic calli to a growth regulator-free medium. For 'Carefree Beauty', glucose at 111 mM promoted higher organogenesis and somatic embryogenesis than sucrose at either 59 or 117 mM concentrations; however, for 'Baby Katie', no differences were observed between glucose and sucrose.The best growth regulator combination for adventitious shoot regeneration of azalea 'Fuchsia' and 'Hino Crimson' was 22.74 M TDZ and 22.8 M IAA. In general, incubating leaf explants in the dark for at least 1 week followed by low-light or high-light intensity was enhanced regeneration frequency. Continuous culture under high-light intensity suppresses shoot regeneration; however, a 2-week dark pretreatment promotes shoot organogenesis even when explants were grown under high-light intensity.Several factors for optimizing microprojectile-mediated gene transfer methods for rose and azalea were investigated. Transformation efficiency was increased by elevating the osmotic level of the medium during bombardment. Transformation efficiency was enhanced by combining osmotic treatment, cotyledonary-stage of embryogenic calli, and higher accelerating pressure settings. Comparing GUS (-glucuronidase) transient expression of two particle guns, the PIG (particle inflow gun) and the biolistic PDS-1000/He gene gun, the PIG device resulted in higher GUS expression than the biolistic gene gun. A 10% GUS transient expression was obtained at 1100 psi helium pressure with 6 cm distance from stopping screen-to-leaf sections of azalea using the biolistic gun. A 22.2% GUS transient expression was obtained on shoot tip-derived calli of azalea using the PIG device at 60 psi with an open-chamber accelerating setting.Several factors for optimizing Agrobacterium-mediated gene transfer methods for rose and azalea were also investigated. Highly proliferating tissues, rose somatic embryogenic calli and azalea shoot-tips, have been found to be amenable to Agrobacterium-mediated transformation. The cotyledonary-stage of rose somatic embryos was found to be most amenable for gene transfer. Regeneration and transformation of azalea shoot-tips were better than stem segments or leaf sections. Kanamycin was effective for inhibiting regeneration of azalea leaf sections; however, it was less efficient for selection of rose embryogenic calli. Wounding manipulations were not necessary for rose embryogenic calli; however, for certain Agrobacterium strains, a wounding treatment might increase regeneration and/or transformation frequency. Based on GUS expression, bombardment and fresh cut treatments showed a higher blue color intensity. Adding acetosyringone or a nurse culture of minced tobacco leaf nurse culture to the cocultivation medium enhanced the number and intensity of blue spots; moreover, adding acetosyringone to the bacterial culture probably increased stable transformation in azalea. The susceptibility of various tissue types to different Agrobacterium strains was observed.U of I OnlyETDs are only available to UIUC Users without author permissio
eQTL mapping of candidate genes for flower colour as a model for genetical genomics in Azalea
No full textBy its production of approximately 40 million plants per year, azalea (Rhododendron simsii hybrids) is the most important flowering pot plant in Belgium . Flower colour is one of the most important features for azalea breeding. In azalea, flower colour is inherited as a semi-qualitative trait and is mainly determined by differences in anthocyanin and flavonol levels. Its segregation pattern could be explained by a two-gene model (Heursel and Horn 1977). Gene expression of flower colour genes was studied before in an azalea sporting series (De Keyser et al. 2007). Flower colour has served as a model system ever since the beginning of genetics with Mendel and his peas. The multi-disciplinary knowledge on flower colour in azalea made it therefore a perfect model for genetical genomics. Genetical genomics (Jansen and Nap 2001) became available with the introduction of microarrays for gene expression profiling. However, for minor crops this is not an option. cDNA-AFLP was proposed as an alternative profiling method (Vuylsteke et al. 2006) but this technique cannot handle candidate genes directly. The use of RT-qPCR data for eQTL mapping was not reported before in plant genomics, but is reported here as a valuable alternative for eQTL mapping
Azalea (Rhododendron simsii hybrids) germplasm from China assessed by means of fluorescent AFLP
No full textThe current assortment of pot azalea (Rhododendron simsii hybrids) has been created from a relatively narrow basis of collectors material (botanical gardens, private collections) brought from the far east. R. simsii, the main ancestor, originates from hilly areas in China (Chang Jiang valley), Thailand, Laos and Burma. However, several other species from the Tsutsusi subgenus, from South-Asia and Japan, might have contributed: e.g. R. indicum, R. mucronatum and R. scabrum. From the Kunming Institute of Botany (China) 33 seed lots from natural populations in mountain area's with an altitude ranging from 250 to 3500 In were obtained. The majority of these Rhododendron species belong to the Tsutsusi subgenus; 8 are R. simsii. per population 10 plants were analysed by AFLP using 3 primer combinations. They were compared to the breeders pot azalea genepool (70 plants, some with common pedigree and bud sports). AFLP was performed using the commercially available kit (ABI-Perkin-Elmer) for fluorescent fragment detection on an ABI Prism 377 DNA Sequencer. The genetic diversity of the different gene pools was analysed by comparison of the marker frequencies, by calculating similarity indices, by multivariate analysis and AMOVA. Small differences within populations were observed. Large variation was observed within the R. simsii species and between the different species from the Tsutsusi subgenus. The pot azalea genepool was clearly distinguishable from the Chinese accessions. On dendrograms it was more closely clustered to R, simsii and X. mucronatum than to less related species
Curry County fairyland of flowers
No full textTitle from PDF caption (viewed on June 13, 2018)."March 30, 1944"--Page 9.This archived document is maintained by the State Library of Oregon as part of the Oregon Documents Depository Program. It is for informational purposes and may not be suitable for legal purposes.Mode of access: Internet from the Oregon Government Publications Collection.Text in English
Street Improvement Photographs -- Box 21, Folder 36 (Bramble Avenue) -- print, 1929-01-10
No full textPhotographer's notes written on front of print: Jan. 10, 1929 / Bramble Ave. west from Azalea Ave. after imp.GeoCoordinates: 39.151987,-84.39588
“Azalea” and “Wildflowers of the Mountains”: An Analysis of Four Song Settings on Two Poems by So-wol Kim
No full textabstract: This research project will focus on two poems by the Korean poet So-wol Kim (1902-1934). His poems are admired throughout Korea and are often set by Korean art song composers. This paper will examine four art song settings by composers Sung-tae Kim (1910-2012) and Soon-nam Kim (1917-1986) of two poems by So-wol Kim: “Azalea” and “Wildflowers of the Mountains.” The discussion will examine in detail the varied interpretations and expressions of the texts by each composer. To be clear, the translations of the poems investigated in this paper are poetic renderings and are not meant for performance purposes.Dissertation/ThesisDoctoral Dissertation Music 202
Comparative Physiology of Natural Deacclimation in Ten Azalea Cultivars
No full textSeasonal deacclimation was investigated during Jan. to Mar. 2014 in leaves of 10 azalea cultivars (Rhododendron section Tsutsusi) under natural conditions in eastern China. Based on the midwinter leaf freezing tolerance (LFT), these cultivars were grouped as “more-hardy” vs. “less-hardy.” Eight of the 10 cultivars first showed deacclimation when daily mean temperature over 2-week period preceding the LFT measurement was ≈9.5 °C. Deacclimation for other two cultivars was somewhat delayed and might have involved deacclimation–reacclimation cycling before eventual deacclimation. Our data indicate that the “more-hardy” group deacclimated slower than the “less-hardy” ones over the first half of the deacclimation period. This trend reversed during the second half of the deacclimation period. Accordingly, “more-hardy” and “less-hardy” cultivars depicted a “curvilinear” and “reverse curvilinear/linear” deacclimation kinetics. “More-hardy” cultivars generally had higher total soluble sugars (TSS) than “less-hardy” ones at acclimated state. TSS declined during deacclimation in all cultivars, and the loss was positively correlated with the loss in LFT. Leaf starch content generally followed opposite trend to that of TSS, i.e., it was at lowest during acclimated state and increased during deacclimation.</jats:p
Multipoint-likelihood maximization mapping on 4 segregating populations to achieve an integrated framework map for QTL analysis in pot azalea (Rhododendron simsii hybrids)
No full textBackground: Azalea (Rhododendron simsii hybrids) is the most important flowering pot plant produced in Belgium, being exported world-wide. In the breeding program, flower color is the main feature for selection, only in later stages cultivation related plant quality traits are evaluated. As a result, plants with attractive flowering are kept too long in the breeding cycle. The inheritance of flower color has been well studied; information on the heritability of cultivation related quality traits is lacking. For this purpose, QTL mapping in diverse genetic backgrounds appeared to be a must and therefore 4 mapping populations were made and analyzed.
Results: An integrated framework map on four individual linkage maps in Rhododendron simsii hybrids was constructed. For genotyping, mainly dominant scored AFLP (on average 364 per population) and MYB-based markers (15) were combined with co-dominant SSR (23) and EST markers (12). Linkage groups were estimated in JoinMap. A consensus grouping for the 4 mapping populations was made and applied in each individual mapping population. Finally, 16 stable linkage groups were set for the 4 populations; the azalea chromosome number being 13. A combination of regression mapping (JoinMap) and multipoint-likelihood maximization (Carthagene) enabled the construction of 4 maps and their alignment. A large portion of loci (43%) was common to at least two populations and could therefore serve as bridging markers. The different steps taken for map optimization and integration into a reference framework map for QTL mapping are discussed.
Conclusions: This is the first map of azalea up to our knowledge. AFLP and SSR markers are used as a reference backbone and functional markers (EST and MYB) were added as candidate genes for QTL analysis. The alignment of the 4 maps on the basis of framework markers will facilitate in turn the alignment of QTL regions detected in each of the populations. The approach we took is thoroughly different than the recently published integrated maps and well-suited for mapping in a non-model crop
How to perform RT-qPCR accurately in plant species?: a case study on flower colour gene expression in an azalea (Rhododendron simsii hybrids) mapping population
No full textBackground: Flower colour variation is one of the most crucial selection criteria in the breeding of a flowering pot plant, as is also the case for azalea (Rhododendron simsii hybrids). Flavonoid biosynthesis was studied intensively in several species. In azalea, flower colour can be described by means of a 3-gene model. However, this model does not clarify pink-coloration. The last decade gene expression studies have been implemented widely for studying flower colour. However, the methods used were often only semi-quantitative or quantification was not done according to the MIQE-guidelines. We aimed to develop an accurate protocol for RT-qPCR and to validate the protocol to study flower colour in an azalea mapping population.
Results: An accurate RT-qPCR protocol had to be established. RNA quality was evaluated in a combined approach by means of different techniques e.g. SPUD-assay and Experion-analysis. We demonstrated the importance of testing noRT-samples for all genes under study to detect contaminating DNA. In spite of the limited sequence information available, we prepared a set of 11 reference genes which was validated in flower petals; a combination of three reference genes was most optimal. Finally we also used plasmids for the construction of standard curves. This allowed us to calculate gene-specific PCR efficiencies for every gene to assure an accurate quantification. The validity of the protocol was demonstrated by means of the study of six genes of the flavonoid biosynthesis pathway. No correlations were found between flower colour and the individual expression profiles. However, the combination of early pathway genes (CHS, F3H, F3'H and FLS) is clearly related to co-pigmentation with flavonols. The late pathway genes DFR and ANS are to a minor extent involved in differentiating between coloured and white flowers. Concerning pink coloration, we could demonstrate that the lower intensity in this type of flowers is correlated to the expression of F3'H.
Conclusions: Currently in plant research, validated and qualitative RT-qPCR protocols are still rare. The protocol in this study can be implemented on all plant species to assure accurate quantification of gene expression. We have been able to correlate flower colour to the combined regulation of structural genes, both in the early and late branch of the pathway. This allowed us to differentiate between flower colours in a broader genetic background as was done so far in flower colour studies. These data will now be used for eQTL mapping to comprehend even more the regulation of this pathway
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