‘Lord Baltimore’ hardy hibiscus (Hibiscus moscheutos L.) was used in the mass encapsulation protocol, rinsing effects on bulk encapsulation, growth comparisons using K-NAA, acclimatization to greenhouse environment, and leaf anatomy comparisons. Nodal segments, each containing a single axillary bud, were harvested and cut to 4mm prior to mass encapsulation. Results showed that using 2.75% alginate with a concentration of 60 or 80 mM calcium chloride produced an alginate mass, or “cookie”, which was able to hold together during rinsing protocols. The rinsing study resulted with the potential to reduce rinse times from 2 three-minute rinses to 1 one-minute rinse. After 4 weeks under mist, ~58% of root growth was generated when using 1 one-minute rinse. After 8 weeks under mist, the greatest percentage of shoot and root growth occurred when using 1 one-minute rinse. The use of K-NAA in the gel matrix, after four weeks, resulted in no discernable differences. Concentrations of 0 or 0.1 µM will generate a greater percentage of shoot and root growth. Using 1.0 µM K-NAA, and extending the time under mist to 8 weeks, will result in a greater percentage of shoots and roots generated. Acclimatization study investigated the survival rates on mass encapsulated nodal segments, looking at different media, depth of planting and humidity. Increases in shoot lengths can be expected up to 8 weeks under mist and planted either on top or 1 cm deep. A reduction in root growth was seen when placed on top of the medium after 4 weeks. The anatomical study found differences in the leaf cell lengths based on the stage of leaf development. However, there was no indication of a gradual change in anatomy as they adapted from in vitro to greenhouse environment. The best results were found when comparing epidermal, palisade and spongy mesophyll cells on the fourth fully expanded leaf under mist. Hydrangea quercifolia ‘Snow Queen’ was used to test the effects of tissue culture media and plant growth regulators. Research found that using dichloroisocyanuric acid (NaDCC) resulted in a decrease of contamination on explants without a reduction in the number of shoots produced. This would reduce the amount of time and labor on the encapsulation procedure, as no rinsing is required. The best result, looking a nutrient formulation, was found to be WPM with 1 µM BA added.

As an alternative to testing nutrient components separately, four common media formulations were used: Murashige and Skoog (MS), Driver and Kuniyuki (DKW), woody plant medium (WPM) and Anderson's revised medium (AND). Each medium was tested separately. Then high salts formulation (MS and DKW) were mixed with low salts formulation (WPM and AND) at a half and half concentration. In total, eight different media were tested spanning high, intermediate and low nutrient salts concentrations. To test the media two different plant species were chosen, a woody species Hydrangea quercifolia (oakleaf hydrangea) and an herbaceous species Tagetes erecta (African marigold). Nodal explants were harvested from H. quercifolia `Dayspring' stock plants grown in raised beds in a greenhouse. After being disinfested, explants were placed on the eight treatment media, along with 1.0 µM benzyladenine (BA) and 1.0 µM indolebutryic acid (IBA). In addition to the plant growth regulators (PGR's), 10 ppm Orthene insecticide and 150 ppm Benlate fungicide were included in the tested media to prevent contamination from a spider mite infestation. Explants placed on WPM were significantly different than explants on other media. WPM explants initiated growth the most, but elongation was sluggish and leaves showed signs of deficiencies after 8 to 10 weeks in vitro. Intermediate salts media DKW/WPM and DKW/AND had fewer explants initiate growth then explants on WPM, but the few that did elongated. These explants had the maximum lengths of any other explant on all other treatments. Tagetes erecta `Marvel Orange' seeds were sown in polyethylene flats in a lab under cool white fluorescent lamps. After about a month apical shoot tips were harvested, surface disinfested, and placed in vitro on the eight treatment media, with the same PGR's and additives as the hydrangea experiment. Tagetes explants responded substantially faster than the Hydrangea explants, with growth initiating on all treatments within a week. Initially there were significant differences between treatments, but after 3 months in vitro there were no differences between treatments. Although no data was taken, there was observed differences between the treatment media. Explants on low salts media of WPM and AND show signs of nutrient deficiencies, with high salt media showed no deficiencies. Overall the best growth of axillary shoots was seen on high salt media of MS and DKW.

Thesis--Auburn University, 2009. / Abstract. Vita. Includes bibliographical references.

La valorisation paysagère et horticole des espèces du genre Hydrangea est basée sur l'exploitation de leur diversité phénotypique s'exprimant notamment dans les formes architecturales et les coloris. La collection de ressources génétiques, située à Angers, comprend treize espèces introduites en Europe à partir du 18ème siècle. L'analyse de la composition chimique des inflorescences, réalisée pour la première fois sur l'ensemble du genre, permettra de compléter la caractérisation génétique des espèces et des clones. L'objectif de cette étude était ainsi d'analyser les profils phénoliques d'un ensemble de clones ainsi que la diversité intra et interspécifique existant dans la collection. La caractérisation structurale et la quantification de 80 composés phénoliques présents dans les inflorescences de 99 génotypes répartis dans 11 espèces du genre Hydrangea ont été réalisées. Ces composés se répartissent parmi 4 grandes classes chimiques : les flavonoïdes, les anthocyanes, les acides cinnamiques et les amines polyacylées. Une diversité importante existe entre les profils aussi bien des espèces que des sous-espèces. La structuration de cette diversité inter et intrapécifique est analysée. Ces connaissances nouvelles devraient permettre d'orienter les stratégies de conservation et de sélection.

Thesis--Auburn University, 2009. / Abstract. Vita. Includes bibliographical references.

Made available in DSpace on 2014-06-11T19:28:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-12-17Bitstream added on 2014-06-13T20:37:44Z : No. of bitstreams: 1 doria_kmabvs_me_botfca.pdf: 1765521 bytes, checksum: 3c3876d40aef109f0fbe73859a7a9972 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A hortênsia é um arbusto semilenhoso muito apreciado como ornamental no Brasil. No Brasil podemos ressaltar a “Região das Hortênsias” no Sul do país, onde esta ornamental é utilizada em projetos de jardinagem em casas e rodovias. A cidade de Gramado têm a hortênsia como sua flor símbolo. No Estado de São Paulo, ela é comumente encontrada na Região de Campos do Jordão. Plantas de hortênsia apresentando anéis cloróticos e necróticos foram observadas por Yuki et al. (2005) em material proveniente de Arujá, estado de São Paulo. Transmissões por extrato vegetal permitiram a observação de lesões locais cloróticas em Chenopodium quinoa e Gomphrena globosa, indicando infecção causada por vírus. Desta forma, o presente trabalho teve como objetivo a identificação e caracterização da espécie viral presente nestas amostras. Inicialmente as amostras foram analisadas por microscopia eletrônica, onde puderam ser observadas partículas alongadas filamentosas, medindo cerca de 490 nm, indicando a provável presença de um potexvirus. Oligonucleotídeos específicos Hyd_senso e Hyd_anti_senso foram desenhados para o Hydrangea ringspot virus (HdRSV), um potexvirus encontrado comumente em países Europeus e nos Estados Unidos. O RNA total foi extraído pelo método de Bertheau et al. (1998), para posterior análise por RT-PCR utilizando-se estes oligonucleotídeos. Dois fragmentos, um em torno de 550 e outro de 250 nucleotídeos foram amplificados e purificados para realização do sequenciamento genético. Uma identidade de nucleotídeos de 96% e 88% para o fragmento maior e menor respectivamente foi observada para HdRSV (número de acesso AJ 707100.1), indicando tratar-se desta espécie viral. O HdRSV até então era uma praga exótica no Brasil, de forma que foi realizada comunicação ao Ministério da Agricultura, Pecuária e Abastecimento, que emitiu parecer... / The hydrangea is an ornamental plant very appreciated in Brazil. In South of Brazil, this plant is used in projects for gardening in houses and highways. Hydrangea is the symbol of Gramado´s city. In State of São Paulo this ornamental plant is commonly found in Campos do Jordão. Hydrangea plants showing leaves with chlorotic and necrotic rings were observed by Yuki (2005) in material proceeding from Arujá, State of São Paulo. Chlorotic local lesions were observed on Chenopodium quinoa and Gomphrena globosa, after sap transmission, indicating infection caused by virus. On electron microscope analysis, virus particles with 490 nm could be 4 visualized indicating infectin by a potexvirus. In order to identify the species of virus infecting these plants, specifics primers (Hyd_senso and Hyd_anti_senso) were design for Hydrangea ringspot virus (HdRSV), a potexvirus commonly found infecting hydrangea in Europe and United States. Total RNA was extracted following Bertheau et al., 1998 protocol’s and the primers were used in RT-PCR. Two fragments, one around 550bp and another one of 250 nucleotides were amplified and sequenced. An identity of nucleotide of 96% and 88%, respectively, was observed for HdRSV (number of access AJ 707100.1), indicating that both fragments amplified were from the virus. As the HdRSV is an exotic pest in Brazil, the occurrence was notified to the Ministry of Agriculture (MAPA) that gave us the permission for publication this data (process 21052.015361/2007-08). To evaluate the dissemination of this virus in the matrices of hydrangea used in the commercial production in Brazil, 17 samples of the region of Arujá – SP were analysed for the presence of the virus. Eight of them were infected by virus, and the RT-PCR fragment from the varieties Azul Rendado, Azul LZR, Renat Blue, Rosa Japonesa, Rosita and Vermelho Comum were sequenced for analysis... (Complete abstract click electronic access below)

Resumo: A hortênsia é um arbusto semilenhoso muito apreciado como ornamental no Brasil. No Brasil podemos ressaltar a "Região das Hortênsias" no Sul do país, onde esta ornamental é utilizada em projetos de jardinagem em casas e rodovias. A cidade de Gramado têm a hortênsia como sua flor símbolo. No Estado de São Paulo, ela é comumente encontrada na Região de Campos do Jordão. Plantas de hortênsia apresentando anéis cloróticos e necróticos foram observadas por Yuki et al. (2005) em material proveniente de Arujá, estado de São Paulo. Transmissões por extrato vegetal permitiram a observação de lesões locais cloróticas em Chenopodium quinoa e Gomphrena globosa, indicando infecção causada por vírus. Desta forma, o presente trabalho teve como objetivo a identificação e caracterização da espécie viral presente nestas amostras. Inicialmente as amostras foram analisadas por microscopia eletrônica, onde puderam ser observadas partículas alongadas filamentosas, medindo cerca de 490 nm, indicando a provável presença de um potexvirus. Oligonucleotídeos específicos Hyd_senso e Hyd_anti_senso foram desenhados para o Hydrangea ringspot virus (HdRSV), um potexvirus encontrado comumente em países Europeus e nos Estados Unidos. O RNA total foi extraído pelo método de Bertheau et al. (1998), para posterior análise por RT-PCR utilizando-se estes oligonucleotídeos. Dois fragmentos, um em torno de 550 e outro de 250 nucleotídeos foram amplificados e purificados para realização do sequenciamento genético. Uma identidade de nucleotídeos de 96% e 88% para o fragmento maior e menor respectivamente foi observada para HdRSV (número de acesso AJ 707100.1), indicando tratar-se desta espécie viral. O HdRSV até então era uma praga exótica no Brasil, de forma que foi realizada comunicação ao Ministério da Agricultura, Pecuária e Abastecimento, que emitiu parecer... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The hydrangea is an ornamental plant very appreciated in Brazil. In South of Brazil, this plant is used in projects for gardening in houses and highways. Hydrangea is the symbol of Gramado's city. In State of São Paulo this ornamental plant is commonly found in Campos do Jordão. Hydrangea plants showing leaves with chlorotic and necrotic rings were observed by Yuki (2005) in material proceeding from Arujá, State of São Paulo. Chlorotic local lesions were observed on Chenopodium quinoa and Gomphrena globosa, after sap transmission, indicating infection caused by virus. On electron microscope analysis, virus particles with 490 nm could be 4 visualized indicating infectin by a potexvirus. In order to identify the species of virus infecting these plants, specifics primers (Hyd_senso and Hyd_anti_senso) were design for Hydrangea ringspot virus (HdRSV), a potexvirus commonly found infecting hydrangea in Europe and United States. Total RNA was extracted following Bertheau et al., 1998 protocol's and the primers were used in RT-PCR. Two fragments, one around 550bp and another one of 250 nucleotides were amplified and sequenced. An identity of nucleotide of 96% and 88%, respectively, was observed for HdRSV (number of access AJ 707100.1), indicating that both fragments amplified were from the virus. As the HdRSV is an exotic pest in Brazil, the occurrence was notified to the Ministry of Agriculture (MAPA) that gave us the permission for publication this data (process 21052.015361/2007-08). To evaluate the dissemination of this virus in the matrices of hydrangea used in the commercial production in Brazil, 17 samples of the region of Arujá - SP were analysed for the presence of the virus. Eight of them were infected by virus, and the RT-PCR fragment from the varieties Azul Rendado, Azul LZR, Renat Blue, Rosa Japonesa, Rosita and Vermelho Comum were sequenced for analysis... (Complete abstract click electronic access below) / Orientador: Renate Krause Sakate / Coorientador: Marcelo Agenor Pavan / Banca: Jorge Alberto Marques Rezende / Banca: Valdir Atsushi Yuki / Mestre

Environmental contamination issues related to phosphorus (P) in surface waters substantiates the need to identify minimally-sufficient P fertilization amounts for production of containerized nursery crops and better understand the effect of routine amendments (i.e., dolomite [DL] and micronutrient fertilizer [MF]) added to pine bark substrates on chemical fate of P fertilizer. Four studies were conducted to accomplish two overarching objectives: 1) determine the minimum P fertilization amount and corresponding pore-water P concentration needed to achieve maximal growth of common containerized nursery crops and 2) determine the effect of DL and MF amendments in pine bark on P retention during irrigation and P fractions in substrate pore-water. In a fertigation, greenhouse study, calculated lowest P-fertilizer concentration that sustained maximal growth in Hydrangea paniculata ‘Limelight’ (panicle hydrangea) and Rhododendron ‘Karen’ (azalea) was 4.7 and 2.9 mg·L⁻¹ , respectively, and shoot growth Ilex crenata ‘Helleri’ (holly) was the same when fertilized with 0.5 to 6.0 mg·L⁻¹ P. Porewater P concentrations corresponding with treatments that sustained maximal growth of panicle hydrangea, azalea and holly were as low as 0.6, 2.2 and 0.08 mg·L⁻¹ P, respectively. In a separate study, utilizing low-P controlled-release fertilizers (CRFs), shoot growth of Hydrangea macrophylla ‘P11HM-11’ (bigleaf hydrangea) produced in two ecoregions was maximal when fertilized with as little as 0.3 g CRF-P per 3.8-L container, a 50% P reduction from the industrystandard CRF. Holly required 0.2 or 0.4 g CRF-P depending on ecoregion. Mean pore-water P concentrations that corresponded with highest SDW were 0.8 and 1.2 mg·L⁻¹ for hydrangea and holly, respectively. When irrigating fallow pine bark columns containing CRF for 48 d, amending pine bark with DL and MF reduced orthophosphate-P (OP-P) leachate concentrations by ≈ 70%, most of which was retained within the substrate. In a greenhouse study, containerized Lagerstroemia ‘Natchez’ (crape myrtle) were grown for 91 d in pine bark containing CRF. In pine bark amended with DL and MF, pore-water OP-P and total P concentrations, measured approximately weekly, were reduced by, on average, 64% and 58%, respectively. Total dry weight values of plants grown with DL plus MF or MF-only were 40% higher than those grown with no amendments; however, tissue P amounts and relative P uptake efficiency were the same among plants in these three treatments. Therefore, sorption of OP-P by DL and MF reduced water-extractable OP-P but did not limit P uptake by plants. / Ph. D. / Phosphorus (P) pollution of surface waters is a global issue that has led to widespread fish kills, drinking water contamination and disruption of aquatic ecosystems. Nutrient runoff from agricultural sites is among the leading contributors to P loads in impaired waters. Optimizing P fertilization for containerized nursery crop production is particularly challenging since the primary soilless substrates used to grow containerized crops retain P poorly. Consequently, much of the applied P leaches from containers during irrigation. Reducing amounts of applied P fertilizer and amending substrates (e.g., pine bark) with P-sorbing materials are two methods previously shown to reduce P leaching and increase the proportion of applied P that is absorbed by containerized plants. Four studies were conducted to accomplish two overarching objectives: 1) determine the minimum P fertilization amount necessary for maximal growth of common containerized nursery crops and 2) determine the effect of dolomite (DL) and micronutrient fertilizer (MF) amendments in pine bark on P retention during irrigation. Our findings indicated that P fertilization requirements of woody ornamental crops is species-dependent. When using liquid fertilizer, Japanese holly and evergreen azalea achieved maximal growth when P fertilizer concentrations were reduced by 90% and 40%, respectively, compared to current recommendations. In contrast, the current minimum fertilizer recommendation of 5 ppm P was optimal for panicle hydrangea. In a subsequent study in which containerized woody ornamentals were grown using low-P controlled-release fertilizers (CRFs), bigleaf hydrangea reached maximal growth when given CRFs containing 50% less P than amounts in conventional CRFs. Considering hydrangea and azalea are among the top woody ornamental shrubs produced in the US, using fertilizers with minimally sufficient P amounts for these species could greatly reduce P runoff from nursery sites. Results of two studies conducted to achieve the second aforementioned objective indicated that amending CRF-fertilized pine bark with DL and MF can reduce water-extractable total P concentrations by > 50%. Despite lower levels of plant-available P in the substrate, P uptake by crape myrtle was unaffected by the amendments. The DL was primarily responsible for P retention in pine bark; however, the addition of MF was needed for maximal growth and P uptake of crape myrtle. According to this research, amending pine bark with DL and MF could be considered a best management practice for reducing P leaching from containerized crops.

Background Identifying orthologous molecular markers that potentially resolve relationships at and below species level has been a major challenge in molecular phylogenetics over the past decade. Non-coding regions of nuclear low- or single-copy markers are a vast and promising source of data providing information for shallow-scale phylogenetics. Taking advantage of public transcriptome data from the One Thousand Plant Project (1KP), we developed a genome-scale mining strategy for recovering potentially orthologous single-copy markers to address low-scale phylogenetics. Our marker design targeted the amplification of intron-rich nuclear single-copy regions from genomic DNA. As a case study we used Hydrangea section Cornidia, one of the most recently diverged lineages within Hydrangeaceae (Cornales), for comparing the performance of three of these nuclear markers to other "fast" evolving plastid markers. Results Our data mining and filtering process retrieved 73 putative nuclear single-copy genes which are potentially useful for resolving phylogenetic relationships at a range of divergence depths within Cornales. The three assessed nuclear markers showed considerably more phylogenetic signal for shallow evolutionary depths than conventional plastid markers. Phylogenetic signal in plastid markers increased less markedly towards deeper evolutionary divergences. Potential phylogenetic noise introduced by nuclear markers was lower than their respective phylogenetic signal across all evolutionary depths. In contrast, plastid markers showed higher probabilities for introducing phylogenetic noise than signal at the deepest evolutionary divergences within the tribe Hydrangeeae (Hydrangeaceae). Conclusions While nuclear single-copy markers are highly informative for shallow evolutionary depths without introducing phylogenetic noise, plastid markers might be more appropriate for resolving deeper-level divergences such as the backbone relationships of the Hydrangeaceae family and deeper, at which non-coding parts of nuclear markers could potentially introduce noise due to elevated rates of evolution. The herein developed and demonstrated transcriptome based mining strategy has a great potential for the design of novel and highly informative nuclear markers for a range of plant groups and evolutionary scales.

Background Identifying orthologous molecular markers that potentially resolve relationships at and below species level has been a major challenge in molecular phylogenetics over the past decade. Non-coding regions of nuclear low- or single-copy markers are a vast and promising source of data providing information for shallow-scale phylogenetics. Taking advantage of public transcriptome data from the One Thousand Plant Project (1KP), we developed a genome-scale mining strategy for recovering potentially orthologous single-copy markers to address low-scale phylogenetics. Our marker design targeted the amplification of intron-rich nuclear single-copy regions from genomic DNA. As a case study we used Hydrangea section Cornidia, one of the most recently diverged lineages within Hydrangeaceae (Cornales), for comparing the performance of three of these nuclear markers to other 'fast' evolving plastid markers. Results Our data mining and filtering process retrieved 73 putative nuclear single-copy genes which are potentially useful for resolving phylogenetic relationships at a range of divergence depths within Cornales. The three assessed nuclear markers showed considerably more phylogenetic signal for shallow evolutionary depths than conventional plastid markers. Phylogenetic signal in plastid markers increased less markedly towards deeper evolutionary divergences. Potential phylogenetic noise introduced by nuclear markers was lower than their respective phylogenetic signal across all evolutionary depths. In contrast, plastid markers showed higher probabilities for introducing phylogenetic noise than signal at the deepest evolutionary divergences within the tribe Hydrangeeae (Hydrangeaceae). Conclusions While nuclear single-copy markers are highly informative for shallow evolutionary depths without introducing phylogenetic noise, plastid markers might be more appropriate for resolving deeper-level divergences such as the backbone relationships of the Hydrangeaceae family and deeper, at which non-coding parts of nuclear markers could potentially introduce noise due to elevated rates of evolution. The herein developed and demonstrated transcriptome based mining strategy has a great potential for the design of novel and highly informative nuclear markers for a range of plant groups and evolutionary scales.