CO expression was used as a measure of activity of the photoperiodic flowering pathway to determine whether inactivity of the pathway could be the cause of reduced levels of FT expression during juvenility (Fig. Molecules involved in vegetative phase change in a range of species have been identified, including microRNA156 (miR156) and miR172. Leaf material was harvested from antirrhinum when plants were 12 days (juvenile) and 24 days (adult) old and pooled. No. miR172, which promotes competence to flower, is repressed by SPL genes that are direct targets of miR156 and consequently shows a reciprocal pattern of increasing expression with age36. (a) Real-time PCR analysis of developmental expression of AtFT, AtCO, AtTEM1 and AtTEM2 in aerial parts of Col-0 plants grown under LD harvested at ZT15. Thomas, B. Leaf material was harvested at ZT15 from seven replicate plants to generate each antirrhinum sample and 10 replicate seedlings to generate each Arabidopsis sample. 52, 655–662 (2001). Induction of FT expression in wild-type plants grown under LDs occurred just prior to the end of the JVP (Fig. English ivy (Hedera helix) Juvenility is referred to the period during which a plant cannot be induced to flowers whereas maturity is occurred if a plant gives flowers. S1), in a total volume of 10 μl made up with SDW. RNA AmTEM complemented both the floral repressor and JVP regulator functions of the ArabidopsisTEM1 gene when overexpressed in the tem1 mutant. Attainment of floral competence is the most distinct and consistent manifestation of phase change and hence could be regarded as the most robust indicator of the end of juvenility1. ACTIN (HQ853640) and ELONGATION FACTOR 1 α (AJ805055) were used as reference genes for the expression analysis in antirrhinum and ACTIN2 (BE038458) and β-TUBULIN (AY040074) were used as reference genes in Arabidopsis analysis (Supplementary Tab. Massiah, A. J. and A.J. AVP: Col-0 vs. tem1 p = 0.042; Col-0 vs. RNAi- tem1/2 p = 0.0021; tem1 vs. RNAi- tem1/2 p = 0.0072. Nat. Analyses were conducted according to MIQE guidelines38. Two plant viral suppressors of silencing require the ethylene-inducible host transcription factor RAV2 to block RNA silencing. Plants were grown under LD conditions and aerial parts harvested at ZT15. In the juvenile phase it grows as a creeping vine, with palmate, lobed leaves. 3, http://dx.doi.org/10.1038/ncomms1810 (2012). Trends Plant Sci. S11). Arabidopsis plants transformed with AmTEM gene were sown, stratified and grown under SD conditions, while transgenic plants transformed with OeTEM and AtTEM1 were grown under LD conditions. AmFT shared 78.3% identity at the amino acid level to FT and was shown to functionally complement the Arabidopsis ft-1 late-flowering mutant to restore early flowering (Supplementary Fig. This work is licensed under a Creative Commons Attribution 3.0 Unported License. The genes were then cloned, using Gateway® LR Clonase® II (Invitrogen, USA), into a pB2GW7 binary vector (Invitrogen Ltd., USA) to produce the pBAmTEM, pBOeTEM and pBAtTEM1 vectors. In studies utilising AmTEM we show that TEM regulates juvenile phase length through a mechanism involving repression of both FT and CO genes. 18064-14) following the manufacturers' guidelines using 3 μg total RNA for each sample. Scientia Horticulturae 124, 74–77 (2010). 5a, b). Juvenility in Plants During their early stages of growth and devlopment, many plants will not flower in response to environmental conditions that will induce flowering when they are older. GIGANTEA directly activates Flowering Locus T in Arabidopsis thaliana. JVP: juvenile phase, AVP: adult vegetative phase. Juvenility may be induced in mature plants by treating shoots that develop from lateral buds with GA3 (discussed later in the course). As lower TEM expression levels lead to shortening of the JVP we hypothesised that over-expression of TEM should lead to an increase in JVP length and that this should also affect patterns of CO and FT expression. (b) Phylogenetic analysis of the deduced amino acid sequence of AmTEM and other RAV sub-family class I member homologs. Olea europaea L. (olive) leaf samples were collected from trees of cv Leccino grown in the agricultural farm of Tuscia University (Italy) on the 25th of June 2010 at 20:00 (sun rise was at 5:34, sun set was at 20:52), frozen in liquid nitrogen and stored at −80°C until their utilization.

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