Gibberellin and phytochrome control senescence in alstroemeria leaves independently

In alstroemeria ( Alstroemeria hybrida ), leaf senescence is effectively retarded by the application of gibberellins and by low fluences of red light. In this study we examined the possible interaction of gibberellins and red light in the regulation of senescence. Determination of endogenous gibberellins revealed that leaf senescence is accompanied by significant changes in the concentrations of non‐ 13‐hydroxylated gibberellins, the onset of senescence coinciding with a dramatic drop in GA₄, whereas concentrations of 13‐hydroxylated gibberellins are far less influenced. However, no direct effect of red light on a specific GA‐metabolic step could be determined. When exogenously applied, non‐13‐hydroxylated GAs were more active than the 13‐hydroxylated GAs. It appeared that the effect of red light is additive to that of active GAs. We hypothesise that GA₄ and phytochrome control senescence in alstroemeria mainly through separate mechanisms and have independent effects and that the observed differences in gibberellin concentrations are a consequence of delayed leaf senescence rather than a cause for it.     

Variation in male‐built nest volume with nesting‐support quality,colony, and egg production in whiskered terns

Nest building can represent an energetically costly activity for a variety of animal taxa. Besides, the determinants of within‐species variation in the design of nests, notably with respect to natural and sexual selection, are still insufficiently documented. Based on an observational study, we examined the influence of nesting conditions (nesting‐support quality, colony, laying date, and year) on male‐built nest volume and also its potential role as a postmating sexually selected display in the whiskered tern Chlidonias hybrida. This tern species is a monogamous colonial bird with obligate biparental care breeding on aquatic vegetation. Hence, large nesting platforms are expected to be a selective advantage because they would better withstand adverse environmental conditions and provide a secure structure for eggs. Nest size may also serve as a postmating sexual trait, and variation in egg production would be positively associated with nest size. We found that nest volume was adjusted to different environmental cues. A positive relationship was found between nest volume and nesting‐support quality, indicating that the leaf density of white waterlily is essential for nest stability. Variation in nest volume was not correlated to colony size but varied among colonies and years. Male‐built nest volume was also positively associated with mean egg volume per clutch but not with clutch size. The fitness consequences of building a large nest are yet to be studied, and additional investigations are recommended to better understand whether the activity of males early during breeding season (e.g., nest building and courtship feeding performance) really serves as postmating sexually selected signals.     

Ethylene and flower petal senescence: Interrelationship with membrane lipid catabolism

Accumulated experimental evidence suggests that the decline in the content of membrane components such as phospholipids (PL), is a key event in flower senescence. This loss of membrane integrity can be modulated by ethylene. The aim of this work was to examine the interrelationship between ethylene and one of the products of PL metabolism, diacylglycerol (DAG), during petunia ( Petunia hybrida ) flower senescence. DAG's role was studied using phorbol 12-myristate 13-acetate (PMA), which acts similarly in kinase activation. Our results demonstrate for the first time a senescence-related transient increase in the content of DAG in petunia plasma membranes. The climacteric-like ethylene rise associated with petal wilting appeared in petunia flowers well after PL degradation and DAG increase had commenced. The appearance and peak magnitude of the ethylene rise was enhanced or increased, respectively, by PMA treatment, thereby accelerating appearance and magnitude of all senescence parameters assayed. Conversely, suppression of ethylene action by silver thiosulfate (STS) resulted in retardation of flower wilting, as well as in abolishment of the PMA-enhancing effects on senescence. The results suggest an active role for lipid metabolites like DAG in enhancing flower senescence, through regulation of ethylene production and action, or possible activation of kinases. This sequence of events implies that ethylene is a mediator of flower senescence, rather than a trigger of the process.     

Multifaceted plant responses to circumvent Phe hyperaccumulation by downregulation of flux through the shikimate pathway and by vacuolar Phe sequestration

Detrimental effects of hyperaccumulation of the aromatic amino acid phenylalanine (Phe) in animals, known as phenylketonuria, are mitigated by excretion of Phe derivatives; however, how plants endure Phe accumulating conditions in the absence of an excretion system is currently unknown. To achieve Phe hyperaccumulation in a plant system, we simultaneously decreased in petunia flowers expression of all three Phe ammonia lyase (PAL) isoforms that catalyze the non‐oxidative deamination of Phe to trans‐cinnamic acid, the committed step for the major pathway of Phe metabolism. A total decrease in PAL activity by 81–94% led to an 18‐fold expansion of the internal Phe pool. Phe accumulation had multifaceted intercompartmental effects on aromatic amino acid metabolism. It resulted in a decrease in the overall flux through the shikimate pathway, and a redirection of carbon flux toward the shikimate‐derived aromatic amino acids tyrosine and tryptophan. Accumulation of Phe did not lead to an increase in flux toward phenylacetaldehyde, for which Phe is a direct precursor. Metabolic flux analysis revealed this to be due to the presence of a distinct metabolically inactive pool of Phe, likely localized in the vacuole. We have identified a vacuolar cationic amino acid transporter (PhCAT2) that contributes to sequestering excess of Phe in the vacuole. In vitro assays confirmed PhCAT2 can transport Phe, and decreased PhCAT2 expression in PAL‐RNAi transgenic plants resulted in 1.6‐fold increase in phenylacetaldehyde emission. These results demonstrate mechanisms by which plants maintain intercompartmental aromatic amino acid homeostasis, and provide critical insight for future phenylpropanoid metabolic engineering strategies.     

S-RNase-based self-incompatibility in Petunia inflata

Background

For the Solanaceae-type self-incompatibility, also possessed by Rosaceae and Plantaginaceae, the specificity of self/non-self interactions between pollen and pistil is controlled by two polymorphic genes at the S-locus: the S-locus F-box gene (SLF or SFB) controls pollen specificity and the S-RNase gene controls pistil specificity.

Scope

This review focuses on the work from the authors'' laboratory using Petunia inflata (Solanaceae) as a model. Here, recent results on the identification and functional studies of S-RNase and SLF are summarized and a protein-degradation model is proposed to explain the biochemical mechanism for specific rejection of self-pollen tubes by the pistil.

Conclusions

The protein-degradation model invokes specific degradation of non-self S-RNases in the pollen tube mediated by an SLF, and can explain compatible versus incompatible pollination and the phenomenon of competitive interaction, where SI breaks down in pollen carrying two different S-alleles. In Solanaceae, Plantaginaceae and subfamily Maloideae of Rosaceae, there also exist multiple S-locus-linked SLF/SFB-like genes that potentially function as the pollen S-gene. To date, only three such genes, all in P. inflata, have been examined, and they do not function as the pollen S-gene in the S-genotype backgrounds tested. Interestingly, subfamily Prunoideae of Rosaceae appears to possess only a single SLF/SFB gene, and competitive interaction, observed in Solanaceae, Plantaginaceae and subfamily Maloideae, has not been observed. Thus, although the cytotoxic function of S-RNase is an integral part of SI in Solanaceae, Plantaginaceae and Rosaceae, the function of SLF/SFB may have diverged. This highlights the complexity of the S-RNase-based SI mechanism. The review concludes by discussing some key experiments that will further advance our understanding of this self/non-self discrimination mechanism.     

The chalcone synthase multigene family of Petunia hybrida (V30): differential,light-regulated expression during flower development and UV light induction

We have analysed the expression of the 8–10 members of the gene family encoding the flavonoid biosynthetic enzyme chalcone synthase (CHS) from Petunia hybrida. During normal plant development only two members of the gene family (CHS-A and CHS-J) are expressed. Their expression is restricted to floral tissues mainly. About 90% of the total CHS mRNA pool is transcribed from CHS-A, wheares CHS-J delivers about 10% in flower corolla, tube and anthers. Expression of CHS-A and CHS-J during flower development is coordinated and (red) light-dependent. In young seedlings and cell suspension cultures expression of CHS-A and CHS-J can be induced with UV light. In addition to CHS-A and CHS-J, expression of another two CHS genes (CHS-B and CHS-G) is induced in young seedlings by UV light, albeit at a low level. In contrast to CHS genes from Leguminoseae, Petunia CHS genes are not inducible by phytopathogen-derived elicitors. Expression of CHS-A and CHS-J is reduced to a similar extent in a regulatory CHS mutant, Petunia hybrida Red Star, suggesting that both genes are regulated by the same trans-acting factors. Comparison of the promoter sequences of CHS-A and CHS-J reveals some striking homologies, which might represent cis-acting regulatory sequences.     

Growth kinetics of glucose-limited petunia hybrida cells in chemostat cultures: Determination of experimental values for growth and maintenance parameters

With glucose-limited continuous cultures of Petunia hybrida six steady states were obtained at specific growth rates varying from 0.0035 to 0.012 h(-1) (corresponding with culture residence times varying from 285 to 85 h). The macromolecular and the elemental biomass composition which were determined in four steady states showed no major differences over the range of growth rates examined. During all six steady states specific subtrate and oxygen consumption as well as biomass and extracellular product formation rates were monitored. Moreover the specific activities of the mitochondrial cytochrome and alternative pathway were determined and used to estimate specific adenosine triphosphate (ATP) production rates. Data thus obtained were used in the determination of maintenance and true growth yield parameters. For the maintenance on glucose and ATP values of 0.0070 C-mol/C-mol/h and 0.034 mol/C-mol/h were obtained, respectively. True yields of biomass on glucose and ATP were 0.50 C-mol/C-mol and 0.28 C-mol/mol, respectively. (c) 1995 John Wiley & Sons, Inc.     

Somatic hybridization of sexually incompatible petunias: Petunia parodii,Petunia parviflora

Summary Somatic hybrid plants were regenerated following the fusion of leaf mesophyll protoplasts of P. parodii with those isolated from a nuclear-albino mutant of P. parviflora. Attempts at sexual hybridization of these two species repeatedly failed thus confirming their previously established cross-incompatibility. Selection of somatic hybrid plants was possible since protoplasts of P. parodii would not develop beyond the cell colony stage, whilst those of the somatic hybrid and albino P. parviflora produced calluses. Green somatic hybrid calluses were visible against a background of albino cells/calluses, and upon transfer to regeneration media gave rise to shoots. Shoots and the resultant flowering plants were confirmed as somatic hybrids based on their growth habit, floral pigmentation and morphology, leaf hair structure, chromosome number and Fraction 1 protein profiles. The relevance of such hybrid material for the development of new, and extensively modified cultivars, is discussed.     

Inheritance of two foreign genes co-introduced intoPetunia hybrida by direct gene transfer

In previous work, transformedPetunia hybrida plants were obtained by direct gene transfer, using two different genes on separate plasmids (NPT II gene and a cDNA of PEPC from green sorghum leaves). In this study, we have analysed the sexual transmission of the acquired genes by genetic crossing analysis of 2 of the transgenic petunias. The ploïdies of the two clones were determined by flow cytometric analysis showing that one was 2n and the other 4n. Self and back crosses show that the kanamycin character was inherited as a single dominant trait, and that the two clones were heterozygotes for this character. Therefore, the 4n clone probably arises from an endoploidization followed by a transformation event. Southern blot analyses show that all of the resistant progenies which were analysed harboured the kanamycin gene, and expressed the phosphorylation activity in vitro. The DNA of several progenies were also tested for the presence of co-transformed PEPC cDNA sequence. All of the kanamycin-resistant progenies tested contained the second coding sequence, indicating that the two foreign genes might be genetically co-inherited in the transgenic plants. The way in which the two genes are integrated into the genome is discussed.Abbreviations NPT neomycin phosphotransferase - PEG polyethyleneglycol - PEPC phosphoenolpyruvate carboxylase