Stephen D. Jackson

High-Resolution Temporal Profiling of Transcripts during Arabidopsis Leaf Senescence Reveals a Distinct Chronology of Processes and Regulation

This work presents a high-resolution time-course analysis of gene expression during development of a leaf from expansion through senescence. Enrichment in ontologies, sequence motifs, and transcription factor families within genes showing altered expression over time identified both metabolic pathways and potential regulators active at different stages of leaf development and senescence. Leaf senescence is an essential developmental process that impacts dramatically on crop yields and involves altered regulation of thousands of genes and many metabolic and signaling pathways, resulting in major changes in the leaf. The regulation of senescence is complex, and although senescence regulatory genes have been characterized, there is little information on how these function in the global control of the process. We used microarray analysis to obtain a high-resolution time-course profile of gene expression during development of a single leaf over a 3-week period to senescence. A complex experimental design approach and a combination of methods were used to extract high-quality replicated data and to identify differentially expressed genes. The multiple time points enable the use of highly informative clustering to reveal distinct time points at which signaling and metabolic pathways change. Analysis of motif enrichment, as well as comparison of transcription factor (TF) families showing altered expression over the time course, identify clear groups of TFs active at different stages of leaf development and senescence. These data enable connection of metabolic processes, signaling pathways, and specific TF activity, which will underpin the development of network models to elucidate the process of senescence.

Plant responses to photoperiod.

Photoperiod controls many developmental responses in animals, plants and even fungi. The response to photoperiod has evolved because daylength is a reliable indicator of the time of year, enabling developmental events to be scheduled to coincide with particular environmental conditions. Much progress has been made towards understanding the molecular mechanisms involved in the response to photoperiod in plants. These mechanisms include the detection of the light signal in the leaves, the entrainment of circadian rhythms, and the production of a mobile signal which is transmitted throughout the plant. Flowering, tuberization and bud set are just a few of the many different responses in plants that are under photoperiodic control. Comparison of what is known of the molecular mechanisms controlling these responses shows that, whilst common components exist, significant differences in the regulatory mechanisms have evolved between these responses.

Floral responses to photoperiod are correlated with the timing of rhythmic expression relative to dawn and dusk in Arabidopsis

Daylength, or photoperiod, is perceived as a seasonal signal for the control of flowering of many plants. The measurement of daylength is thought to be mediated through the interaction of phototransduction pathways with a circadian rhythm, so that flowering is induced (in long-day plants) or repressed (in short-day plants) when light coincides with a sensitive phase of the circadian cycle. To test this hypothesis in the facultative long-day plant, Arabidopsis thaliana, we used varying, non-24-hr light/dark cycles to alter the timing of circadian rhythms of gene expression relative to dawn and dusk. Effects on circadian rhythms were correlated with those on flowering times. We show that conditions that displaced subjective night events, such as expression of the flowering time regulator CONSTANS into the light portion of the cycle, were perceived as longer days. This work demonstrates that the perception of daylength in Arabidopsis relies on adjustments of the phase angle of circadian rhythms relative to the light/dark cycle, rather than on the measurement of the absolute duration of light and darkness.

A cis Element within Flowering Locus T mRNA Determines Its Mobility and Facilitates Trafficking of Heterologous Viral RNA

ABSTRACT The Arabidopsis flowering locus T (FT) gene encodes the mobile florigen essential for floral induction. While movement of the FT protein has been shown to occur within plants, systemic spread of FT mRNA remains to be unequivocally demonstrated. Utilizing novel RNA mobility assay vectors based on two distinct movement-defective viruses, Potato virus X and Turnip crinkle virus, and an agroinfiltration assay, we demonstrate that nontranslatable FT mRNA, independent of the FT protein, moves throughout Nicotiana benthamiana and mutant Arabidopsis plants and promotes systemic trafficking of viral and green fluorescence protein RNAs. Viral ectopic expression of FT induced flowering in the short-day N. tabacum Maryland Mammoth tobacco under long-day conditions. Recombinant Potato virus X bearing FT RNA spread and established systemic infection more quickly than the parental virus. The cis-acting element essential for RNA movement was mapped to the nucleotides 1 to 102 of the FT mRNA coding sequence. These data demonstrate that a plant self-mobile RNA molecule can mediate long-distance trafficking of heterologous RNAs and raise the possibility that FT RNA, along with the FT protein, may be involved in the spread of the floral stimulus throughout the plant.

The role of microRNAs in the control of flowering time

The onset of flowering in plants is regulated by complex gene networks that integrate multiple environmental and endogenous cues to ensure that flowering occurs at the appropriate time. This is achieved by precise control of the expression of key flowering genes at both the transcriptional and post-transcriptional level. In recent years, a class of small non-coding RNAs, called microRNAs (miRNAs), has been shown to regulate gene expression in a number of plant developmental processes and stress responses. MiRNA-based biotechnology, which harnesses the regulatory functions of such endogenous or artificial miRNAs, therefore represents a highly promising area of research. In this review, the process of plant miRNA biogenesis, their mode of action, and multiple regulatory functions are summarized. The roles of the miR156, miR172, miR159/319, miR390, and miR399 families in the flowering time regulatory network in Arabidopsis thaliana are discussed in depth.

Mobile FT mRNA contributes to the systemic florigen signalling in floral induction.

In inducing photoperiodic conditions, plants produce a signal dubbed “florigen” in leaves. Florigen moves through the phloem to the shoot apical meristem (SAM) where it induces flowering. In Arabidopsis, the FLOWERING LOCUS T (FT) protein acts as a component of this phloem-mobile signal. However whether the transportable FT mRNA also contributes to systemic florigen signalling remains to be elucidated. Using non-conventional approaches that exploit virus-induced RNA silencing and meristem exclusion of virus infection, we demonstrated that the Arabidopsis FT mRNA, independent of the FT protein, can move into the SAM. Viral ectopic expression of a non-translatable FT mRNA promoted earlier flowering in the short-day (SD) Nicotiana tabacum Maryland Mammoth tobacco in SD. These data suggest a possible role for FT mRNA in systemic floral signalling, and also demonstrate that cis-transportation of cellular mRNA into SAM and meristem exclusion of pathogenic RNAs are two mechanistically distinct processes.

Expression of the Arabidopsis gai gene under its own promoter causes a reduction in plant height in chrysanthemum by attenuation of the gibberellin response

Several transgenic chrysanthemum (Chrysanthemum morifolium Ramat.) lines have been produced that express the Arabidopsis thaliana gai (gibberellic acid insensitive) gene under its own promoter. These transformants exhibit a range of dwarf phenotypes, the extent of dwarfing being related to the reduction in the response to gibberellin in each transgenic line. Physiological measurements of growth, chlorophyll content and flowering time demonstrate that the extent of the transgene effects correlate with the level of transgene expression. Production of many ornamental crops relies heavily on the use of expensive and harmful agrochemicals in order to produce a blemish free and uniform product. We demonstrate the feasibility of producing a dwarf (pot) chrysanthemum without the need for growth retardant chemicals through heterologous expression of the mutant Arabidopsis gai gene driven from its own promoter.

Floral induction and flower formation--the role and potential applications of miRNAs.

The multiple regulatory pathways controlling flowering and flower development are varied and complex, and they require tight control of gene expression and protein levels. MicroRNAs (miRNAs) act at both the transcriptional and post-transcriptional level to regulate key genes involved in flowering-related processes such as the juvenile-adult transition, the induction of floral competence and flower development. Many different miRNA families are involved in these processes and their roles are summarized in this review, along with potential biotechnological applications for miRNAs in controlling processes related to flowering and flower development.

DAY NEUTRAL FLOWERING Represses CONSTANS to Prevent Arabidopsis Flowering Early in Short Days

This work describes the flowering time gene DAY NEUTRAL FLOWERING (DNF), which acts in the same flowering pathway as CONSTANS (CO). DNF is a membrane-bound E3 ligase that represses CO expression and plays an important role in maintaining low levels of CO expression in short days; it is thus essential for the ability of the Arabidopsis plant to have a different flowering response in long and short days. The photoperiodic response in Arabidopsis thaliana requires the precise regulation of CONSTANS (CO) expression in relation to the light period during the day. In short days (SDs) levels of CO expression are normally low during the light period, and this results in delayed flowering compared with long days (LDs) when CO expression rises to high levels before the end of the light period. We identified a novel flowering time gene called DAY NEUTRAL FLOWERING (DNF) that acts in the same flowering pathway as CO. DNF is a membrane-bound E3 ligase that represses CO expression and plays an important role in maintaining low levels of CO expression in SDs. The effect of DNF on the rhythm of CO expression is essential for the photoperiodic response of Arabidopsis, enabling it to have a different flowering response in LDs and SDs.

Jasmonic acid spraying does not induce tuberisation in short-day-requiring potato species kept in non-inducing conditions

The potato speciesSolanum andigena (Juz. and Buk.) andSolanum demissum (Lindl.) that both require short days for tuberisation were kept in either long days (16 h light), or short days (8 h light) with a 30-min night break mid-way through the dark period. Tuberisation of these species was inhibited under both conditions. Repeated spraying of these plants with up to 100 μM jasmonic acid did not induce them to tuberise even though jasmonic acid was shown to be taken up and transported within the plant. This result argues against jasmonic acid itself being the transported tuber-inducing signal, although it does not exclude a role for jasmonic acid later in tuber formation and development once induction has taken place.