The Decreased apical dominance1/Petunia hybrida CAROTENOID CLEAVAGE DIOXYGENASE8 gene affects branch production and plays a role in leaf senescence, root growth, and flower development

Carotenoids and carotenoid cleavage products play an important and integral role in plant development. The Decreased apical dominance1 (Dad1)/PhCCD8 gene of petunia (Petunia hybrida) encodes a hypothetical carotenoid cleavage dioxygenase (CCD) and ortholog of the MORE AXILLARY GROWTH4 (MAX4)/AtCCD8 gene. The dad1-1 mutant allele was inactivated by insertion of an unusual transposon (Dad-one transposon), and the dad1-3 allele is a revertant allele of dad1-1. Consistent with its role in producing a graft-transmissible compound that can alter branching, the Dad1/PhCCD8 gene is expressed in root and shoot tissue. This expression is upregulated in the stems of the dad1-1, dad2, and dad3 increased branching mutants, indicating feedback regulation of the gene in this tissue. However, this feedback regulation does not affect the root expression of Dad1/PhCCD8. Overexpression of Dad1/PhCCD8 in the dad1-1 mutant complemented the mutant phenotype, and RNA interference in the wild type resulted in an increased branching phenotype. Other differences in phenotype associated with the loss of Dad1/PhCCD8 function included altered timing of axillary meristem development, delayed leaf senescence, smaller flowers, reduced internode length, and reduced root growth. These data indicate that the substrate(s) and/or product(s) of the Dad1/PhCCD8 enzyme are mobile signal molecules with diverse roles in plant development.     

Modifying the acylation of flavonols in Petunia hybrida

The potential for chemically-regulating the acylation of natural products in whole plants has been determined by treating petunia leaves with phenylpropanoid acyl donors supplied as the respective methyl esters. Treatment with derivatives of the naturally-occurring acylating species caffeic acid resulted in a general increase in flavonol derivatives, notably caffeoylated quercetin-3-O-diglucoside (QDG) and kaempferol-3-O-diglucoside (KDG). Similarly, methyl ferulate increased the content of feruloylated KDG 40-fold. Treatment with methyl coumarate resulted in the appearance of a coumaroylated derivative of quercetin-3-O-glucuronyl-glucoside (QGGA). When the feeding studies were repeated with the equivalent phenylpropanoid isosubstituted with fluorine groups a semi-synthetic 4-fluorocinnamoyl ester of QGGA was observed. Our results demonstrate the potential to regulate the acylation of flavonols and potentially other natural products by treating whole plants with methyl esters of natural and unnatural acyl donors.     

Location and variation of the constitutive heterochromatin in Petunia hybrida

The location of heterochromatin in the chromosomes of Petunia hybrida (2n=14) is presented. C-banded mitotic metaphase chromosomes and carmine-stained pachytene bivalents have been studied. It is shown that the heterochromatin is predominantly located near the centromeres and at the secondary constrictions of the satellite chromosomes. The distribution of chromomeres in pachytene bivalents also reveals that heterochromatin is not restricted to distinct blocks, as is the case in tomato, but occurs in smaller chromomeres which gradually decrease in size towards the ends. Conspicuous telomeres have not been observed. Both C-banding technique and pachytene analysis demonstrate large variation of heterochromatin between different lines of Petunia. The study of pachytene morphology has been hampered by a high degree of non-specific stickiness of the bivalents. Both techniques prove to be unsuitable tools for large-scale chromosome identification of Petunia lines.     

The Importance of the Stigma in Flower Senescence in Petunia (Petunia hybrida)

Senescence of flowers of Petunia hybrida Vilm. cv Gypsy is characterizedby colour changes, wilting and abscission. In emasculated detachedflowers the onset of these processes is hastened by any treatmentwhich reduces the vigour of the stigma. Thus pricking it, excisingsegments, or freezing with liquid nitrogen all reduce the timeto morphological changes associated with corolla senescence.Removal of the stigma has the most dramatic effect, reducinglifespan of the flower by about 50 per cent, to 3 d. This reductioncan be lessened if IAA or 2,4-D is applied to the cut surfaceof the style. In intact flowers, the style may usually be implicatedin the production of a stimulus leading to corolla abscission,but abscission will also occur in the absence of the style.Some senescence acceleration takes place not only in the completeabsence of the style, but also when the upper part of the ovaryhas been excised in addition. The speeding up of senescenceand of corolla abscission cannot be due solely to damage perse since when the corolla limb was excised, leaving only thecorolla tube, the tube abscised at about the same time as thecontrols, despite the quite extensive wounding. This also impliesthat the distal parts of the corolla do not play a major rolein the development of the abscission zone at the base of thecorolla tube. A healthy, undamaged stigma appears to be very important incorolla longevity and one of its roles may be to prevent theproduction of an abscission/wilting stimulus by some other componentof the flower. Possibly auxins in the stigma are important inthat either they are mobile and protect the abscission zoneor they create a sink for other substances which are implicatedin flower senescence. Petunia hybrida, abscission, auxins (IAA, 2,4-D), corolla, flower senescence, stigma, style, wilting     

Transmembrane Adaptor Protein PAG/CBP Is Involved in both Positive and Negative Regulation of Mast Cell Signaling

The transmembrane adaptor protein PAG/CBP (here, PAG) is expressed in multiple cell types. Tyrosine-phosphorylated PAG serves as an anchor for C-terminal SRC kinase, an inhibitor of SRC-family kinases. The role of PAG as a negative regulator of immunoreceptor signaling has been examined in several model systems, but no functions in vivo have been determined. Here, we examined the activation of bone marrow-derived mast cells (BMMCs) with PAG knockout and PAG knockdown and the corresponding controls. Our data show that PAG-deficient BMMCs exhibit impaired antigen-induced degranulation, extracellular calcium uptake, tyrosine phosphorylation of several key signaling proteins (including the high-affinity IgE receptor subunits, spleen tyrosine kinase, and phospholipase C), production of several cytokines and chemokines, and chemotaxis. The enzymatic activities of the LYN and FYN kinases were increased in nonactivated cells, suggesting the involvement of a LYN- and/or a FYN-dependent negative regulatory loop. When BMMCs from PAG-knockout mice were activated via the KIT receptor, enhanced degranulation and tyrosine phosphorylation of the receptor were observed. In vivo experiments showed that PAG is a positive regulator of passive systemic anaphylaxis. The combined data indicate that PAG can function as both a positive and a negative regulator of mast cell signaling, depending upon the signaling pathway involved.     

Photosynthetic carbon fixation in the corollas of Petunia hybrida

Corollas of Petunia hybrida (cv. Hit Parade Rosa) flowers fixed ¹⁴CO₂ under both light and dark conditions. Rates of light fixation were much higher in mature pink corollas than in young, green corollas [57 and 9 nmol (ngchl)¹ min^-1], paralleling the development of chloroplasts in these tissues. Stomatal conductance in corollas was only 12% of that in green leaves, mainly due to the presence of few, and non-functioning stomata in the corolla. The activity and concentration of ribulose bisphosphate carboxylase (EC 4.1.1.39) in corolla extracts were only about 30% (per unit Chi) of those in extracts from green leaves. These results, together with previous results, might indicate a coordinated reduction in activity of systems participating in photosynthesis in corollas. The fixation products following a 6 s pulse with ¹⁴CO₂, were typical of C, plants in both corollas and green leaves, but a higher level of β-carboxylation products was found in the corollas. The activity of phosphoenol-pyruvate carboxylase (EC 4.1.1.31) (per unit protein) was similar in both tissues. Although the total carbon fixed by the corolla constituted only a small part of the metabolites required for flower development, certain photosynthetic metabolites might have a regulatory role in flower development.     

矮牵牛育种研究进展

从常规育种、基因工程育种、体细胞育种和单倍体育种4个方面评述了矮牵牛(Petunia hybrida Vilm.)育种研究进展.国外对矮牵牛育种研究较多,新品种面世推陈出新,国内对其研究则较为薄弱.其常规育种最为成功,不断有新品种推出,基因工程育种也取得了一定成就,已有新品种面世,而体细胞育种及单倍体育种尚无新品种产生,但对这两种方法在育种上应用的可能性和前景作出了一定探索.     

The morphology and anatomy of the stigma of Petunia hybrida

Summary A mature stigma of Petunia hybrida ready for pollination shows 4-6 large shining drops of the exudate along with numerous smaller ones. A developing style and stigma have a columnar tissue that flares at the top. In the stigma there can be distinguished a secretory and a storage zone. In the former, schizogenous cavities are formed which are filled with the exudate. The mode of formation and secretion of the drop has been studied with light and electron microscope. The nature of reserves has been studied histochemically.The exudation takes place in two stages. In the first stage, the epidermal and papillae cells release out the oily exudate upon rupture of the cuticle. The second phase of exudation begins with anthesis. The exudate from the schizogenous cavities is released between the epidermal cells. There are distinct loci on the stigma surface where more exudate is given out than at other places.

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Zusammenfassung Bei der Durchsicht der einschlägigen Literatur ergab sich, daß die Kenntnisse über die Entwicklungsgeschichte und den Sekretionsmechanismus des Narben-Schleimes äußerst lückenhaft sind, obgleich dieser im Verlauf des Bestäubungs-Vorganges bei vielen Pflanzen eine bedeutende Rolle spielt. Die vorliegende Untersuchung versuchte daher eine detaillierte Einsicht in die Morphologie und Anatomie der Narbe von Petunia hybrida zu geben, da diese Art als Objekt für zahlreiche physiologische Untersuchungen auf dem Gebiet der Befruchtungs-inkompatibilität dient.Die Blüten von Petunia sind zwittrig, zygomorph und fünfzählig. Sie besitzen fünf epipetale Antheren mit drei verschiedenen Filament-Längen; diese öffnen sich zu verschiedenen Zeitpunkten. Die reife Narbe zeigt zum Zeitpunkt der Bestäubung 4-6 große sowie zahlreiche kleinere, transparente Flüssigkeits-Tropfen.Der sich entwickelnde Griffel mit der Narbe besteht aus einem säulenartigen Gewebe, das sich an der Spitze verbreitert. Bei der Narbe kann eine Sekretions-und eine Reservematerial-Zone unterschieden werden. In der Sekretionszone werden schizogen Hohlräume gebildet, die sich mit dem Exsudat füllen. Die Bildung und Sekretion der Narbenflüssigkeit wurde licht-und elektronenmikroskopisch untersucht. Mittels histochemischer Methoden wurden die Substanzen der Reservematerial-Zone näher charakterisiert.Die Freisetzung der Narben-Flüssigkeit findet in zwei Schritten statt: Während der ersten Phase wird das ölige Exsudat von den epidermalen Zellen und den Papillen der Narbe nach Zerreißen der Narben-Cuticula freigesetzt. In der zweiten Phase jedoch tritt das Exsudat aus den schizogenen Hohlräumen durch die Zwischenräume in der Narben-Epidermis aus. Auf der Narben-Oberfläche gibt es Bezirke, welche offensichtlich in reichlicherem Maße Flüssigkeit produzieren als andere Bezirke.

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Dedicated to the memory of the late Prof. P. Maheshwari FRS, our teacher (R. N. K.) and friend (H. F. L.) who read through the Ms in April at Paris, a month before his death.     

Physiology and biochemistry of the stigmatic fluid of Petunia hybrida

Summary Production of stigma exudate per flower of Petunia hybrida is about 200 g. The effect of light, temperature, metabolic poison and emasculation on the production of the exudate at different ages of the bud has been studied. The presence of a thin film of water below the stigmatic exudate has been demonstrated. Physical properties of the exudate such as relative viscosity and surface tension have also been determined. Chemical analysis of the stigmatic fluid showed that it consists primarily of an oil, sugars and amino acids. No protein could be detected. It also contains no acid phosphatase.Behaviour of the pollen from its deposition on the stigmatic fluid until it germinates on the stigma surface has been studied in vivo and also with the aid of an artificial stigma.The role of the stigmatic fluid in pollination has been determined.

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Zusammenfassung Die Narbenflüssigkeit, welche von zahlreichen Pflanzen ausgeschieden wird zu einem Zeitpunkt, da die Blüte reif ist zur Bestäubung, spielt eine wichtige Rolle bei der Fixierung des Pollens. Um so überraschender ist die Tatsache, daß hinsichtlich der Physiologie und Biochemie des Narbenschleimes so wenig detaillierte Informationen zur Verfügung stehen.Bei Petunia wird je Blüte im Mittel 200 g Narben-Flüssigkeit produziert. Die Intensität der Narbenschleimproduktion ist abhängig von der Temperatur und der Länge der Lichtperiode. Hingegen wird die Ausscheidung der Narbenflüssigkeit durch Hemmung der Atmung und durch partielle Kastration nicht beeinflußt.Die physikalischen Eigenschaften (Dichte, relative Viscosität und Oberflächenspannung) werden bestimmt. Bei der chemischen Analyse des Narbenschleimes ergab sich, daß dieser hauptsächlich aus einem öligen Fett besteht, dessen Fettsäure-Zusammensetzung ermittelt wurde. Hingegen werden nur sehr geringe Spuren von Zuckern und Aminosäuren gefunden. Der Narbenschleim ist frei von Proteinen und zeigt keine enzymatische Aktivität.Das Verhalten des Pollens bei der Landung in einem Tropfen der Narbenflüssigkeit wird in vivo und mit Hilfe einer künstlichen Modell-Narbe untersucht.Die Bedeutung der Narbenflüssigkeit im Zusammenhang mit der Bestäubung und den einleitenden Stadien der Pollenkeimung wird diskutiert.

     

矮牵牛育种研究进展

从常规育种、基因工程育种、体细胞育种和单倍体育种4个方面评述了矮牵牛(Petunia hybrida Vilm.)育种研究进展。国外对矮牵牛育种研究较多,新品种面世推陈出新,国内对其研究则较为薄弱。其常规育种最为成功,不断有新品种推出,基因工程育种也取得了一定成就,已有新品种面世,而体细胞育种及单倍体育种尚无新品种产生,但对这两种方法在育种上应用的可能性和前景作出了一定探索。     

The Cell Adhesion Molecule L1 Is Developmentally Regulated in the Renal Epithelium and Is Involved in Kidney Branching Morphogenesis

We immunopurified a surface antigen specific for the collecting duct (CD) epithelium. Microsequencing of three polypeptides identified the antigen as the neuronal cell adhesion molecule L1, a member of the immunoglobulin superfamily. The kidney isoform showed a deletion of exon 3. L1 was expressed in the mesonephric duct and the metanephros throughout CD development. In the adult CD examined by electron microscopy, L1 was not expressed on intercalated cells but was restricted to CD principal cells and to the papilla tall cells. By contrast, L1 appeared late in the distal portion of the elongating nephron in the mesenchymally derived epithelium and decreased during postnatal development. Immunoblot analysis showed that expression, proteolytic cleavage, and the glycosylation pattern of L1 protein were regulated during renal development. L1 was not detected in epithelia of other organs developing by branching morphogenesis. Addition of anti-L1 antibody to kidney or lung organotypic cultures induced dysmorphogenesis of the ureteric bud epithelium but not of the lung. These results suggest a functional role for L1 in CD development in vitro. We further postulate that L1 may be involved in the guidance of developing distal tubule and in generation and maintenance of specialized cell phenotypes in CD.     

Stamens and gibberellin in the regulation of corolla pigmentation and growth in Petunia hybrida

Removal of stamens, or even of only the anthers, at an early stage of corolla development, before the start of main anthocyanin production, inhibited both growth and pigmentation of attached corollas of Petunia. When only one or two stamens were removed from one side, the inhibition was restricted to the corolla side adjacent to the detached stamens. Application of gibberellic acid (GA₃) substituted for the stamens in its effect on both growth and pigmentation. In detached corollas, isolated at the early-green stage and grown in vitro in sucrose medium, GA₃ promoted growth and was essential for anthocyanin synthesis. A marked enhancement of anthocyanin production was observed 48 h before the increase in corolla growth rate. Corollas detached at later stages were able to continue their growth and pigmentation in sucrose without GA₃. When Paclobutrazol (-[(4-chlorophenyl)-ethyl]-(1,1-dimethylethyl)-H-1,2,4-triazol-1-ethanol), an inhibitor of gibberellin biosynthesis, was added to the growth medium of in-vitro-grown corollas, pigmentation was inhibited but there was no effect on corolla growth. Low levels of GA₃ counteracted the Paclobutrazol effect on pigmentation but did not affect growth. The above results indicate that the effect of GA₃ (and probably that of the stamens) on corolla growth is independent of its effect on pigmentation. Gibberellic acid and paclobutrazol had no effect on [¹⁴C]sucrose uptake by in-vitro-grown corollas. The activity of phenylalanine ammonialyase was correlated with the effect of stamens and GA₃ on pigmentation in corollas grown in vivo and in vitro.Abbreviations GA gibberellin - GA₃ gibberellic acid - PAC Paclobutrazol - PAL phenylalanine ammonia-lyase     

Regulation of anthocyanin and lignin synthesis in Petunia hybrida cell suspensions

In Petunia hybrida cv. Violet 30 cell suspensions the phenylpropanoid pathway can be induced to produce lignin and anthocyanins. Orthovanadate addition leads to lignin accumulation, subculturing the cells using small inoculum sizes (<2 g fresh weight l^-1) gives rise to both anthocyanin and lignin production. Orthovanadate has a negative effect on cell growth. By replacing the medium, one day after orthovanadate addition, by medium without elicitor, we were able to restore growth without disturbing the lignin accumulation. The activity of phenylalanine ammonia-lyase (PAL) increased immediately after orthovanadate addition; this increase stopped upon medium replacement without affecting the lignin production. Reduction of the NAA concentration from 2 mg l^-1 to 0.1 mg l^-1, subsequent to the elicitation by orthovanadate or dilution stress, gave rise to a further increase in the production of lignin and anthocyanins respectively. Decreasing the NAA concentration without a prior elicitation, didn't have any effect on either PAL activity or product formation.Abbreviations 2,4-D 2,4 dichlorophenoxyacetic acid - BSA bovine serum albumine - FW fresh weight - NAA naphthaleneacetic acid - PAL phenylalanine ammonia-lyase - PPP phenyl propanoid pathway     

Influence of glyphosate on flower morphogenesis and pigmentation in Petunia hybrida

Glyphosate showed a remarkable effect inducing the change of flower symmetry from the actinomorphic to the zygomorphic type in Petunia hybrida. Glyphosate [N-(phosphonomethyl)glycine] reduced the anthocyanin content and showed a weak inhibitory effect against phenylalanine ammonia-lyase (PAL) activity. L-2-Aminooxy-3-phenylpropionic acid (APA), an inhibitor of PAL activity, reduced the anthocyanin content but had no effect on flower shape. Additional phenylalanine or trans-cinnamic acid, the intermediates of glyphosate inhibition against PAL activity, could not recover the change of flower shape induced by glyphosate. These results suggested that the reduction of PAL activity alone could not account for the two characteristic changes of flower symmetry and pigmentation induced by glyphosate. On the other hand, the results of application of glyphosate-related compounds suggested that the structure of glyphosate contributed to induce the morphological change of Petunia flower. Glyphosate may thus be a very useful agent in the elucidation of unresolved questions of flower morphogenesis and the related metabolism.     

PhGRL2 Protein,Interacting with PhACO1, Is Involved in Flower Senescence in the Petunia

Dear Editor, The pathways of ethylene biosynthesis and signaling have been studied in detail （Ji and Guo, 2013）. Arabidopsis REVERSION-TO-ETHYLENE SENSITIVITY1 （RTE1）, interacting with ETR1, and its homologs tomato GREEN RIPE （SlGR） and SlGRL1, and rice OsRTH1 negatively regulate the ethylene signaling （Barry and Giovannoni, 2006, Resnick et al., 2006; Dong et al., 2010; Zhang et al., 2012）. A newly published study suggested that a cytochrome b5 and RTE1 are functional partners in promoting ETRl-mediated repression of ethylene signaling in Arabidopsis （Chang et al., 2014）. However, AtRTH, RTE1 homolog in Arabidopsis, and its closest homolog in the tomato, SlGRL2 （GR-like2）, do not play a role in ethylene signaling （Dong et al., 2010）, and the function of the homologs of these members is not well known.     

Patterns of cell division and expansion in developing petals of Petunia hybrida

The definition of the patterns of cell division and expansion in plant development is of fundamental importance in understanding the mechanics of morphogenesis. By studying cell division and expansion patterns, we have assembled a developmental map of Petunia hybrida petals. Cycling cells were labelled with in situ markers of the cell cycle, whereas cell expansion was followed by assessing cell size in representative regions of developing petals. The outlined cell division and expansion patterns were related to organ asymmetry. Initially, cell divisions are uniformly distributed throughout the petal and decline gradually, starting from the basal part, to form a striking gradient of acropetal polarity. Cell areas, in contrast, increased first in the basal portion and then gradually towards the petal tip. This growth strategy highlighted a cell size control model based on cell-cycle departure time. The dorso-ventral asymmetry can be explained in terms of differential regulation of cell expansion. Cells of the abaxial epidermis enlarged earlier to a higher final extent than those of the adaxial epidermis. Epidermal appendage differentiation contributed to the remaining asymmetry. On the whole our study provides a sound basis for mutant analyses and to investigate the impact of specific (environmental) factors on petal growth.