Lectin nucleotide phosphohydrolases (LNPs) are proteins which possess both apyrase catalytic activity (E.C. 3.6.1.5) and specific carbohydrate binding properties, and these are linked. To investigate the structural and functional properties for these proteins, two putative soluble plant LNPs, 4WC and 7WC (from white clover), and a putative soluble plant apyrase 6RG (from ryegrass) were chosen. Rabbit polyclonal antibodies for each plant apyrase were generated using highly purified, overexpressed recombinant 4WC or 7WC. In the case of 6RG, the C-terminal half of the protein constituted the best antigen for generating polyclonal antibodies. These antibodies showed high specificity and sensitivity. Active, recombinant 4WC and 6RG were overexpressed and purified using the baculoviral insect cell expression system (4WCbac-sup and 6RG:Hisbac), while 7WC (7WCcoli) was produced from E. coli inclusion bodies and subsequently refolded to give active enzyme. In course of overexpression, recombinant 4WC was localised in both the cellular fraction (4WCbac) and in the media supernatant (4WCbac-sup), while recombinant 6RG:Hisbac was only found in the cellular fraction (6RG:Hisbac) indicating that it was not secreted during insect cell growth. Secretion of 4WCbac was found to be dependent on N-glycosylation at N313 but not at N85 and elimination of one or both of these sites appeared to have little influence on apyrase activity. In addition, both 4WCbac and 6RG:Hisbac from the cellular fraction were fully functional. These results were compared with similar work performed on the animal ecto-apyrases which have different specific N-glycosylation sites required for secretion and activity. The 4WCbac-sup, 7WCcoli and 6RG:Hisbac proteins all showed apyrase activity, that is they catalysed the hydrolysis of nucleotide tri- and/or di-phosphates to their corresponding nucleotide monophosphates, and released inorganic phosphate in a divalent cation-dependent manner. However, the proteins exhibited different activities, substrate specificities, pH profiles and influence of inhibitors: 4WCbac-sup had a preference for NDPs with a pH optimum ≥9.5; 7WCcoli had a modest preference for NTPs with a pH optimum at 8.5; 6RG:Hisbac was almost exclusively an NTPase with a pH optimum at 6.5. Contrary to predictions based on phylogeny the proteins all bound to sulphated disaccharides and their catalytic activities were influenced both positively and negatively by the binding of specific chitosans. The data indicates that all three soluble plant apyrases investigated here were LNPs, in contrast to predictions from the literature. In order to pinpoint the regions responsible for determining substrate specificity and chitosan binding, chimeras were made using the N- and C-terminal halves of 4WC and 6RG. This resulted in fully functional reciprocal chimeras. Comparison of the apyrase activity for parents and chimeras, substrate specificity, optimal pH, influence of inhibitors on activity and effects of chitosans indicated that the C-terminus was responsible for determining substrate specificity. However, the influence of specific chitosans on the chimeras appeared to be dependent on both the N- and C-terminal portions of the proteins. In addition, chimeras were found to bind to the same sulphated disaccharides as the parent proteins. Preliminary crystal screening experiments were performed with highly purified preparations of 7WCcoli and 6RG:Hisbac. Under specific conditions 7WCcoli was found to form cube-like crystalline arrangements while 6RG:Hisbac formed hexagonal-like crystalline structures. A potential model for carbohydrate binding by LNPs is proposed and the possible biological roles of plant LNPs are discussed.

Lectin nucleotide phosphohydrolases (LNPs) are proteins which possess both apyrase catalytic activity (E.C. 3.6.1.5) and specific carbohydrate binding properties, and these are linked. To investigate the structural and functional properties for these proteins, two putative soluble plant LNPs, 4WC and 7WC (from white clover), and a putative soluble plant apyrase 6RG (from ryegrass) were chosen. Rabbit polyclonal antibodies for each plant apyrase were generated using highly purified, overexpressed recombinant 4WC or 7WC. In the case of 6RG, the C-terminal half of the protein constituted the best antigen for generating polyclonal antibodies. These antibodies showed high specificity and sensitivity. Active, recombinant 4WC and 6RG were overexpressed and purified using the baculoviral insect cell expression system (4WCbac-sup and 6RG:Hisbac), while 7WC (7WCcoli) was produced from E. coli inclusion bodies and subsequently refolded to give active enzyme. In course of overexpression, recombinant 4WC was localised in both the cellular fraction (4WCbac) and in the media supernatant (4WCbac-sup), while recombinant 6RG:Hisbac was only found in the cellular fraction (6RG:Hisbac) indicating that it was not secreted during insect cell growth. Secretion of 4WCbac was found to be dependent on N-glycosylation at N313 but not at N85 and elimination of one or both of these sites appeared to have little influence on apyrase activity. In addition, both 4WCbac and 6RG:Hisbac from the cellular fraction were fully functional. These results were compared with similar work performed on the animal ecto-apyrases which have different specific N-glycosylation sites required for secretion and activity. The 4WCbac-sup, 7WCcoli and 6RG:Hisbac proteins all showed apyrase activity, that is they catalysed the hydrolysis of nucleotide tri- and/or di-phosphates to their corresponding nucleotide monophosphates, and released inorganic phosphate in a divalent cation-dependent manner. However, the proteins exhibited different activities, substrate specificities, pH profiles and influence of inhibitors: 4WCbac-sup had a preference for NDPs with a pH optimum ≥9.5; 7WCcoli had a modest preference for NTPs with a pH optimum at 8.5; 6RG:Hisbac was almost exclusively an NTPase with a pH optimum at 6.5. Contrary to predictions based on phylogeny the proteins all bound to sulphated disaccharides and their catalytic activities were influenced both positively and negatively by the binding of specific chitosans. The data indicates that all three soluble plant apyrases investigated here were LNPs, in contrast to predictions from the literature. In order to pinpoint the regions responsible for determining substrate specificity and chitosan binding, chimeras were made using the N- and C-terminal halves of 4WC and 6RG. This resulted in fully functional reciprocal chimeras. Comparison of the apyrase activity for parents and chimeras, substrate specificity, optimal pH, influence of inhibitors on activity and effects of chitosans indicated that the C-terminus was responsible for determining substrate specificity. However, the influence of specific chitosans on the chimeras appeared to be dependent on both the N- and C-terminal portions of the proteins. In addition, chimeras were found to bind to the same sulphated disaccharides as the parent proteins. Preliminary crystal screening experiments were performed with highly purified preparations of 7WCcoli and 6RG:Hisbac. Under specific conditions 7WCcoli was found to form cube-like crystalline arrangements while 6RG:Hisbac formed hexagonal-like crystalline structures. A potential model for carbohydrate binding by LNPs is proposed and the possible biological roles of plant LNPs are discussed.

A universal human dignity, conceived as an inherent and inalienable value or worth in all human beings, which ought to be recognised, respected and protected by others, has become one of the most prominent and widely promoted interpretations of human dignity, especially in international human rights law. Yet, it is also one of the most difficult interpretations of human dignity to justify and ground. The fundamental problem rests on how one can justify bestowing an equal high worth to all human lives, whilst also attributing to all human life a worth that is superior to all non-human animal life. To avoid the speciesist charge it seems necessary to provide further reasons, over and above species membership, for why all humans have a unique worth and dignity. However, intrinsic capacities, such as autonomy, intelligence or language use, are too demanding for many humans (including foetuses or the severely cognitively disabled) to meet the required minimum standard, whilst also being obtainable by some non-human animals, regardless of where the level is set. This thesis offers a solution to this problem by turning instead to the significance of the relational ties between individuals or groups that transcend individual capacities and abilities, and consequently does not require that all individuals in the group need meet the minimum required capacity for full moral status. Rather, it is argued that a universal human dignity could be grounded in our social nature, the interconnectedness and interdependence of human life and the morally considerable relationships that can and do arise from it, especially in regards to our shared vulnerability and dependence, and our ability to engage in caring relationships. Care represents the antithesis to the dehumanizing effects of humiliation, and other degrading and dehumanizing acts, and as a relational concept, human dignity is often best realised through our caring relationships. The way that individuals and groups treat each other has a fundamental role in determining both an individual’s sense of self-worth and well-being, as well as their perceived public value and worth. Thus, whilst species membership is not in itself morally fundamental or basic, it often shapes the nature of our social and moral relations. These relational ties between humans, it is argued, distinguish us most clearly from other non-human animals and accord human relationships a special moral significance or dignity.

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Le virus des nervures jaunes et nécrotiques de la betterave (Beet necrotic yellow vein virus, BNYVV) est l’agent infectieux responsable de la rhizomanie de la betterave sucrière, une maladie caractérisée par une prolifération anarchique du chevelu racinaire. Le Beet soil-borne mosaic virus (BSBMV) appartient également au genre Benyvirus mais n’est retrouvé qu’en Amérique du Nord. Ce virus, identifié pour la première fois au Texas, est morphologiquement et génétiquement semblable au BNYVV mais sérologiquement éloigné. Compte tenu des différences moléculaires existant, le BSBMV et BNYVV correspondent à deux espèces virales distinctes. Mon projet de thèse a consisté à étudier les interactions moléculaires entre le BNYVV et le BSBMV et rechercher les mécanismes impliqués dans la pathogénicité de ces deux virus. Des clones complets cDNA infectieux du BNYVV étaient disponibles, tout comme ceux de BSBMV. Compte tenu de l’aspect versatile de l’obtention de transcrits infectieux de ces différents clones, j’ai entrepris de produire des clones cDNA de chacun des ARN viraux sous contrôle d’un promoteur constitutive végétal pour initier l’infection par agroinfiltration. Les plantes hôtes Chenopodium quinoa et Nicotiana benthamiana ont été inoculées par des transcrits et agroinfiltrées pour initier l’infection virale et étudier l’interaction entre les ARN génomiques 1 et 2 des deux virus et étudier les propriétés de constructions chimères. En parallèle à ce travail, j’ai réalisé la caractérisation du suppresseur de RNA silencing du BSBMV en le comparant à celui du BNYVV. / The genus Benyvirus includes the most important and widespread sugar beet viruses transmitted through the soil by the plasmodiophorid Polymyxa betae. In particular Beet necrotic yellow vein virus (BNYVV), the leading infectious agent that affects sugar beet, causes an abnormal rootlet proliferation known as rhizomania. Beet soil-borne mosaic virus (BSBMV) is widely distributed in the United States and, up to date has not been reported in others countries. My PhD project aims to investigate molecular interactions between BNYVV and BSBMV and the mechanisms involved in the pathogenesis of these viruses.BNYVV full-length infectious cDNA clones were available as well as full-length cDNA clones of BSBMV RNA-1, -2, -3 and -4. Handling of these cDNA clones in order to produce in vitro infectious transcripts need sensitive and expensive steps, so Ideveloped agroclones of BNYVV and BSBMV RNAs, as well as viral replicons allowing the expression of different proteins.Chenopodium quinoa and Nicotiana benthamiana plants have been infected with in vitro transcripts and agroclones to investigate the interaction between BNYVV and BSBMV RNA-1 and -2 and the behavior of artificial viral chimeras. Simultaneously I characterized BSBMV p14 and demonstrated that it is a suppressor of posttranscriptional gene silencing sharing common features with BNYVV p14.

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Conhecemos o papel dos leucócitos no controle do Trypanosoma cruzi, mas ignoramos qual a contribuição das populações estruturais (miócitos, hepatócitos, etc). Estas populações poderiam sinalizar a presença do parasita e/ou ter ação efetora que poderia aumentar em resposta a citocinas como o IFN-g. Para verificar se as células estruturais respondem ao IFN-g contribuindo à destruição do T. cruzi, analisamos a infecção em quimeras WT/IFNgR-KO (WT/KO) geradas em camundongos IFNgR-KO reconstituídos com medula óssea de camundongos selvagens (WT), nas quais parte dos leucócitos é IFNgR+, mas as células não leucocitárias são deficientes. Quimeras WT/KO e quimeras controle WT/WT foram estudadas em relação à carga parasitária e inflamação. O parasitismo sistêmico e tissular é maior nas quimeras WT/KO do que nas quimeras WT/WT, mas inferior à dos controles IFNgR-KO. Nos animais WT/KO o aumento de ninhos no coração e músculo não se acompanha de aumento no infiltrado leucocitário. Os resultados sugerem que as células não leucocitárias contribuem à defesa frente ao T. cruzi. / Although we know the role played by leukocytes in Trypanosoma cruzi control, we ignore the contribution of structural cells (myocytes, hepatocytes, etc). These cells could signal the presence of the parasite and/or mediate an effector activity which could increase in response to cytokines, as IFN-g. To evaluate if non-leukocytes respond to IFNg, contributing to T. cruzi destruction, we analysed the infection in WT/IFNgR-KO (WT/KO) chimaeras, generated in IFNgR-KO mice reconstituted with bone marrow of wild-type mice (WT) so that most leukocytes are IFNgR+ but structural cells are deficient. WT/KO chimaeras and control WT/WT chimaeras were infected by T. cruzi and the parasite load and inflammation analyzed in various organs. Systemic and tissue parasite loads were higher in WT/KO chimeras than in WT/WT chimeras, but lower than in control IFNgR-KO mice. In WT/KO chimaeras the increased parasite load at the heart and skeletal muscle was not followed by an increase of inflammatory infiltrates. Our results suggest that structural cells contribute to T. cruzi control.

Zwei Populus-Pfropfchimären (MA und AI), die aus P. x canadensis ‘Marilandica’ (M), P. maximowizcii x P. trichocarpa ‘Androscoggin’ (A) and P. nigra L. ’Italica’ (I) aufgebaut sind, wurden für Untersuchungen zur Laub- und Blütenblattentwicklung genutzt. In MA bildet M die äußere Lage (L1) und ihr Derivat, die Epidermis, während die inneren Lagen (L2, L3 etc.) von A gebildet werden. Bei AI stammt die L1 von A und L2, L3 etc. werden von I gebildet. Die genotypisch andersartige Epidermis bedingt bei Periklinal-Chimären morphologische Effekte wie zum Beispiel einen Fruchtknoten in einigen MA-Blüten. Morphologische Besonderheiten verschiedener Gewebe sowohl von M und A als auch von MA wurden verglichen, um festzustellen, wie sie durch die Gewebetransplantation verändert oder beeinflusst wurden und, um mögliche Genotyp- Interaktionen oder -Wechselwirkungen in einem Gewebe ausfindig zu machen. Für die Genotypidentifizierung in verschiedenen Organen wurde die RAPD-PCR getestet. Einer von 20 getesteten 10mer Zufallsprimern (GGAGTGGACA) ermöglichte bei der Verwendung von DNA aus Blattmaterial die Erzeugung verschiedener Bandenmuster für M und A. Bei der Verwendung von MA-Blattmaterial zeigte sich eine Kombination der Muster von M und A, sodass ein Chimärennachweis für das MA-Blattmaterial erbracht wurde. Für ein übertragbares System wurde die spezifische PCR getestet. Unter Verwendung spezifischer Primer für die 16s-rDNA zeigten die PCR-Produkte einheitliche Banden und nach anschließender Sequenzierung eine weitgehende Übereinstimmung der phylogenetischen Verwandtschaft von I, M und A. Weiterhin wurden die kernkodierten rDNA Bereiche ITS 1 und ITS 2 zwischen 18S und 25S getestet. Für I, M und A konnten jeweils zwei Banden von unterschiedlicher Größe und Sequenz ermittelt werden, die vermutlich auf funktionierende rDNA aber auch auf Pseudogene (beschnitten) in niedriger Kopienzahl hinweisen. Die ITS-Regionen von I, M und A wurden charakterisiert, um einen Einblick in die Struktur und Phylogenie der Salicacaee-Familie zu erhalten. Aus den Sequenzunterschieden konnten für I und A spezifische Primerpaare abgeleitet werden, die für die Identifizierung von I und A in AI und MA verwendet werden können. Mittels A-Marker konnte nachgewiesen werden, dass Fruchtknoten aus MA-Blüten neben M-Gewebe auch den A-Genotyp enthalten. / Two Populus graft chimeras (MA and AI) produced of P. x canadensis ‘Marilandica’ (M), P. maximowizcii x P. trichocarpa ‘Androscoggin’ (A) and P. nigra L. ’Italica’ (I) were used for investigations of leaf and flower development. In MA the exogenous layer (L1) forms the epidermis and is derived from M while inner layers (L2, L3 etc.) descend from A whereas in AI L1 is formed by A while L2, L3 etc. descend from I. The exogenous epidermis of the periclinal chimeras imposes morphological effects such as an extra female sex in some of the MA flowers. The morphological characteristics of different plant tissues of parents and chimera were compared to determine how they were modified or altered by the tissue transplantation and possibly identify co-existing or interacting genotypes in one tissue. RAPD-PCR was tested for its usefulness to amplify polymorphic fingerprints including donor specific DNA fragments. One random 10mer primer (GGAGTGGACA) out of 20 tested revealed the amplification of patterns including donor specific DNA bands using extracts from leaf tissues of the M and A parents that were combined using extract from leaf tissue of the MA chimera. This indicates that the leaves of the MA chimera are formed by tissues of M and A. However, the inherent disadvantage of RAPD-PCR is the reproducibility of PCR product generation. Therefore the discriminative potential of the ITS region located between the rRNA genes was investigated. The application of specific 16S ribosomal DNA (rDNA) primers for amplification and sequencing of PCR products revealed a closely phylogenetic relationship between I, M and A. Consequently the ITS1 and ITS2 of nuclear rDNA between 18S and 25S were used. The amplified fragments were purified, cloned in E. coli and sequenced. Analyses of multiple clones demonstrated extensive paralogy within and between I, M and A ITS operons. For each parent were at least two rDNA operons as well as multiple paralogous sequences within operons identified. The use of PCR and sequence analyses showed that one of the operons encodes a putative expressed (functional) rDNA whereas the second encodes a pseudogen (truncated) in low copy number. We also characterized the ITS regions of I, M and A to gain insights into structure and phylogeny of the Salicacaee family. Based on sequence divergence primers were designed for A and I and used for the identification of A in MA carpels.

Die Arbeit ist in drei Teile untergliedert: Die Struktur "Hypodermales Wasserspeichergewebe" wird unter anatomischen, ökomorphologischen und evolutionsbiologischen Gesichtspunkten betrachtet. Die Anwesenheit eines farblosen Hypoderms erschwert bei der Musteranalyse variegater Periklinalchimären die Bestimmung der Konstitution der L2. Variegate Periklinalchimären mit Hypodermbildung wurden auf die Möglichkeiten der Bestimmung der L2 hin untersucht. Es werden verschiedene Entstehungsformen von maskierenden Mustern und die noch nicht beschriebenen Ringzellen vorgestellt, die den Idiotyp der L2-bürtigen Schicht anzeigen können. Ringzellen sind die Zellen, die im Bereich der Schließzellen an den substomatären Interzellularraum grenzen. Sie bilden dabei einen Ring um die Schließzellen, der im Flächenschnitt zu erkennen ist. Hypodermale Wasserspeichergewebe sind hauptsächlich bei tropischen Arten verbreitet. Die xeromorphe Struktur kommt sowohl bei den epiphytischen Bromelien als auch bei den hygromorphen Schattenpflanzen des tropischen Regenwaldes vor. Die beiden Selektionsfaktoren Trockenheit und Lichtintensität werden als mögliche Einflussfaktoren auf die Hypodermbildung diskutiert. Beispiele dafür, dass der Faktor Licht auch einen modifikativen Einfluss auf die Differenzierung der Hypodermzellen zu haben scheint, werden vorgestellt. Die Struktur "Hypodermales Wasserspeichergewebe" ist sowohl bei Monokotylen als auch Dikotylen gleichermaßen verbreitet. Es wird daher vermutet, dass es sich um eine analoge Struktur handelt, die mehrmals voneinander unabhängig zu verschiedenen Zeiten bei verschiedenen Arten entstanden ist. Innerhalb einer Gruppe verwandter Arten konnte sie mithilfe der Homologiekriterien als homolog eingestuft werden. / This paper contains three different issues: The structure "hypodermal water storage tissue" is considered from the anatomical, the ecomorphological and evolutionary aspect. Because hypodermal layers are non-green, it is difficult to make a pattern analysis of variegated periclinal chimeras and to determine the constitution of L2. Variegated periclinal chimeras with hypodermal layers were examined to the possibilities of determining L2. Different origins of masking patterns and the non-yet described ring cells are presented. Both structures are able to show the L2-genotype. Ring cells are those cells bordering the intercellular space near the stomata. In a cut parallel to the surface the ring built by ring cells is seen. The hypodermal water storage tissue is mainly distributed among tropical species. The xeromorphic structure occurs both to the epiphytic bromeliads and to the hygromorphic shadow plants of the tropical rainforest. The environmental factors humidity and solar radiation are discussed as possible influences on the development of hypodermal layers. Examples for the apparent modifying influence of solar radiation on the development of hypodermal cells are presented. The structure "hypodermal water storage tissue" occurs both to monocots and dicots. That indicates that it is an analogues structure and that it evolved several times independent of each other in different species. Among a group of nearly related species it could be classified by the aid of the criteria of homology as a homologues structure.

Der Ursprung, die Entwicklung und die Formierung von Laubblatt-Mustern konnten bei ausgewählten variegaten Formen der Araceae, Asteraceae, Ericaceae, Maranthaceae und Rosaceae erklärt werden. Die Pflanzen wurden je nach Problematik untersucht und in drei verschiedene Gruppen verteilt: In der ersten Gruppe, Blattmuster mit unregelmäßiger makulater Musterung, Monstera deliciosa, Syngonium podophyllum und die Sorten 'Pirol' und 'Luyona' von Dendranthema grandiflorum zeigen ein unregelmäßiges Laubblattmuster und keine weiß-randigen bzw. weißkernigen Periklinalchimären. Mischzellen wurden durch direkte (mikroskopisch) und indirekte (In-vitro-Kultur und Selbstungen) Nach der Plastidenentmischung in den Schichten des Sprossmeristems wurde bei Syngonium, Monstera und den zwei Sorten von Dendranthema die GW-Form als einzige stabile Periklinalchimäre nachgewiesen. In der zweiten Gruppe, Immerspaltende Periklinalchimären, die chimärische Konstitution GA bei Spiraea bumalda 'Goldflame' konnte durch Wurzelaustriebe (BATESON-Test) und die Adventivsprossinduktion aus Kallus nachgewiesen werden. Darüber hinaus konnten mehrfach perikline Aufspaltungen der ersten Sprossscheitelschicht (Reduplikation von L1) nachgewiesen werden, die zur Entstehung der Panaschierung von 'Goldflame' zur Entmischung führten. Bei den Untersuchungen der In-vitro-Regenerate aus der Kalluskultur und der Wurzelaustriebe an S. bumalda 'Shirobana' wurde festgestellt, dass diese Pflanze keine Chimäre ist und das auftretende Muster der Blüten genetisch kontrolliert ist. In der dritten Gruppe, Hypoderm und Beeinflussung der Musterbildung: Die unmaskierten Binnenfelder bei Ctenanthe lubbersiana 'Variegata' und der Rhododendron-Hybride 'Goldflimmer' sind durch die Existenz eines Hypoderms zu erklären. Bei Ctenanthe lubbersiana 'Variegata' befindet sich ein Hypoderm an der Blattoberseite und der Blattunterseite. Bei 'Goldflimmer' liegt nur unter der oberen Epidermis ein einschichtiges Hypoderm vor. Infolgedessen fehlt an der Blattoberseite die Maskierung Das gelbe Binnenfeld des Blattmusters ist durch eine grüne Mesophyllschicht unterlagert. / The origin, development and formation of foliage-leaf-patterns could be explained with selected variegaten forms of the Araceae, Asteraceae, Ericaceae, Marantaceae and Rosaceae. In order to prove this the plants were examined according to the problem and classified in three different groups: In the first group, leaf-patterns with irregular maculated patterns, Monstera deliciosa, Syngonium podophyllum and the sorts 'Pirol' and 'Luyona' of Dendranthema grandiflorum showed an irregular foliage-leaf-pattern, thought neither to show periclinal chimeras with white edges nor with green edges. Mixed cells were detected by direct (microscopic) and indirect (In vitro culture and self pollination) test. After the plastid sorting out in the layers of the meristems, the green over white Form was proven with Syngonium, Monstera and the two sorts of Dendranthema as a single stable periclinal chimera. In the second group, eversporting periclinal chimeras, the green over aurea chimeral constitution with Spiraea bumalda 'Goldflame' was proved by the regeneration of adventitious shoots from their roots (BATESON-Test) and also by the induction of adventitious bud from callus. Periclinal divisions of the first layer of meristems (reduplication of L1), which are responsible for the appearing of green pattern of the leafs was proved many times. Examination of the regenerated shoots from callus and from the adventitious shoots from roots of S. bumalda 'Shirobana' showed that this plant is not a chimera and that the appearing pattern of the blooms is genetically controlled. In the third group, hypoderm and influence of the pattern-formation, the unmasked inner-fields with Ctenanthe lubbersiana 'Variegata' and the Rhododendron-hybrid 'Goldflimmer' were explained through the existence of one layered hypoderm under the upper Epidermis as well as over the lower Epidermis of C. lubbersiana 'Variegata', thought in 'Goldflimmer' it is only found a one layer Hypoderm under the upper epidermis. Subsequently, the masking is missing at the upper-leaf side the yellow inner-field of the leaf-pattern is through a green Mesophyllslayer masked.