Developmentally and transgene regulated nuclear processing of primary transcripts of chalcone synthase A in petunia

The introduction of chalcone synthase A transgenes into petunia plants can result in degradation of chalcone synthase A RNAs and loss of chalcone synthase, a process called cosuppression or post-transcriptional gene silencing. Here we show that the RNA degradation is associated with changes in premRNA processing, i.e. loss of tissue specificity in transcript cleavage patterns, accumulation of unspliced molecules, and use of template-specific secondary poly(A) sites. These changes can also be observed at a lower level in leaves but not flowers of nontransgenic petunias. Based on this, a model is presented of how transgenes may disturb the carefully evolved, developmentally controlled post-transcriptional regulation of chalcone synthase gene expression by influencing the survival rate of the endogenous and their own mRNA.

Epigenetic inactivation of chalcone synthase-A transgene transcription in petunia leads to a reversion of the post-transcriptional gene silencing phenotype

Petunia plants that exhibit a white-flowering phenotype as a consequence of chalcone synthase transgene-induced silencing occasionally give rise to revertant branches that produce flowers with wild-type pigmentation. Transcription run-on assays confirmed that the production of white flowers is caused by post-transcriptional gene silencing (PTGS), and indicated that transgene transcription is repressed in the revertant plants, providing evidence that induction of PTGS depends on the transcription rate. Transcriptional repression of the transgene was associated with cytosine methylation at CpG, CpNpG and CpNpN sites, and the expression was restored by treatment with either 5-azacytidine or trichostatin A. These results demonstrate that epigenetic changes occurred in the PTGS line, and these changes interfere with the initiation of transgene transcription, leading to a reversion of the PTGS phenotype.

Failure to launch: The self-regulating Md -MYB10R6 gene from apple is active in flowers but not leaves of Petunia

Coloured foliage due to anthocyanin pigments (bronze/red/black) is an attractive trait that is often lacking in many bedding, ornamental and horticultural plants. Apples (Malus × domestica) containing an allelic variant of the anthocyanin regulator, Md-MYB10R6, are highly pigmented throughout the plant, due to autoregulation by MYB10 upon its own promoter. We investigated whether Md-MYB10R6 from apple is capable of functioning within the heterologous host Petunia hybrida to generate plants with novel pigmentation patterns. The Md-MYB10R6 transgene (MYB10–R6pro:MYB10:MYB10term) activated anthocyanin synthesis when transiently expressed in Antirrhinumroseadorsea petals and petunia leaf discs. Stable transgenic petunias containing Md-MYB10R6 lacked foliar pigmentation but had coloured flowers, complementing the an2 phenotype of ‘Mitchell’ petunia. The absence of foliar pigmentation was due to the failure of the Md-MYB10R6 gene to self-activate in vegetative tissues, suggesting that additional protein partners are required for Md-MYB10 to activate target genes in this heterologous system. In petunia flowers, where endogenous components including MYB-bHLH-WDR (MBW) proteins were present, expression of the Md-MYB10R6 promoter was initiated, allowing auto-regulation to occur and activating anthocyanin production. Md-MYB10 is capable of operating within the petunia MBW gene regulation network that controls the expression of the anthocyanin biosynthesis genes, AN1 (bHLH) and MYBx (R3-MYB repressor) in petals.

Growth and flowering of petunia and impatiens: effects of competition and reduced water content within a container

The primary objective of this research was to determine how the presence of more than one plant and more than one species in a container influence plant quality, particularly when the volume of water given to the container is reduced. Petunia xhybrida 'Hurrah White' and Impatiens 'Cajun Violet' were chosen as typical bedding plant species. Plants were grown in 2 1 containers either under "100% ETp" (i.e., replacing all the water lost by evapotranspiration in the previous 24 h) or under a moisture-restrictive regime of "25% ETp," in which plants received 25% of the "100% ETp" value. An ancillary experiment investigated whether low watering resulted in floral buds being aborted. Results demonstrated that watering requirements of Petunia under "100% ETp" (i.e., replacing all the water lost by evapotranspiration in the previous 24 h) were on average 30% greater than those of Impatiens. However, when two Petunia plants were growing in the same container, the volume of water required to maintain soil moisture content at container capacity was on average only 10% greater than for a single plant. Under a "25% ETp" regime in which plants received 25% of the "100% ETp" value, flower number, plant height, and flower size were reduced by 50%,33%, and 13%,respectively,in Petunia compared with "100% ETp." For example, flower numbers decreased from an average of 71 to 33 flowers per plant in "100% ETp" and "25% ETp," respectively. Petunia plants in the "25% ETp" regime, however, were more efficient at producing both biomass and flowers in relation to the volume of water applied. Petunia plants that experienced both competition from other plants in the container and lower irrigation rates had enhanced efficiency of flower production (i.e., more flowers per unit biomass). For Impatiens, however, the growing of single plants at "25% ETp" was plausible, but the addition of a Petunia plant at "25% ETp" was detrimental to plant quality (Impatiens flower numbers reduced by 75%).

Far-red light filtering by plastic film, greenhouse-cladding materials: effects on growth and flowering in Petunia and Impatiens

Photoselective plastic films with low transmission to far-red (FR) light (700-800 nm) are now available so that plants grown in greenhouses clad with such plastics exhibit reduced stem extension and, consequently, plant height. Here we compare the action of three FR-absorbing polythene films on extension growth of Petunia (Petunia X hybrida) cv. 'Express Blue' and Impatiens walleriana cv. 'Accent Deep Pink' with plants grown under a control polythene film (standard UVI/EVA film). Half of the plants under the control film were treated with a chemical plant growth regulator (PGR; diaminozide, B-Nine) and half were sprayed with water alone. Possible negative effects of such film plastics on flowering, and on fresh and dry weight accumulation, were also quantified. Plants were harvested destructively when all plants in each treatment had reached the first open flower stage. In Petunia, plant height was reduced by all three FR-filtering films and by PGR-treatment. The FR-filtering films giving the highest R:FR ratios also reduced plant height in Impatiens. Leaf number, leaf area and total dry Weight in both species. were greatest in the controls and smallest under films with the lowest PAR transmission. The film giving the highest R:FR ratio and PAR transmission also produced the most compact Petunia plants;, while the film. with. the lowest PAR transmission produced the least compact plants in both species. There was no significant effect of treatments on time to first flower in Impatiens. However, Petunia plants under low PAR transmission films took longer to flower. Plastic-films which filter out FR light to increase the R:FR ratio, combined With high PAR transmission, can therefore be used as an alternative to conventional PGRs.

Sub-irrigation of Petunia: benefits in dry summers

With the increasing frequency and magnitude of warmer days during the summer in the UK, bedding plants which were a traditional part of the urban green landscape are perceived as unsustainable and water-demanding. During recent summers when bans on irrigation have been imposed, use and sales of bedding plants have dropped dramatically having a negative financial impact on the nursery industry. Retaining bedding species as a feature in public and even private spaces in future may be conditional on them being managed in a manner that minimises their water use. Using Petunia x hybrida ‘Hurrah White’ we aimed to discover which irrigation approach was the most efficient for maintaining plants’ ornamental quality (flower numbers, size and longevity), shoot and root growth under water deficit and periods of complete water withdrawal. Plants were grown from plugs for 51 days in wooden rhizotrons (0.35 m (h) x 0.1 m (w) x 0.065 m (d)); the rhizotrons’ front comprised clear Perspex which enabled us to monitor root growth closely. Irrigation treatments were: 1. watering with the amount which constitutes 50% of container capacity by conventional surface drip-irrigation (‘50% TOP’); 2. 50% as sub-irrigation at 10 cm depth (‘50% SUB’); 3. ‘split’ irrigation: 25% as surface drip- and 25% as sub-irrigation at 15 cm depth (‘25/25 SPLIT’); 4. 25% as conventional surface drip-irrigation (‘25% TOP’). Plants were irrigated daily at 18:00 apart from days 34-36 (inclusive) when water was withdrawn for all the treatments. Plants in ‘50% SUB’ had the most flowers and their size was comparable to that of ‘50% TOP’. Differences between treatments in other ‘quality’ parameters (height, shoot number) were biologically small. There was less root growth at deeper soil surface levels for ‘50% TOP’ which indicated that irrigation methods like ‘50% SUB’ and ‘25/25 SPLIT’ and stronger water deficits encouraged deeper root growth. It is suggested that sub-irrigation at 10 cm depth with water amounts of 50% container capacity would result in the most root growth with the maximum flowering for Petunia. Leaf stomatal conductance appeared to be most sensitive to the changes in substrate moisture content in the deepest part of the soil profile, where most roots were situated.

Leaf Structure and Taxonomy of Petunia and Calibrachoa (Solanaceae)

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