Gateway-compatible tissue-specific vectors for plant transformation

Michniewicz, Marta; Frick, Elizabeth M.; Strader, Lucia C.

doi:10.1186/s13104-015-1010-6

Cited by 43 publications

(36 citation statements)

References 43 publications

(50 reference statements)

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“…bZIP binding sites mutations were introduced into pENTR3C - gGLFY by Ω-PCR to generate gGLFYm3 . Both constructs were shuffled into pMCS:GW 20 using LR reaction. To create gLFY : GUS , the 4,929-bp genomic LFY clone minus stop codon and GUS fragments were PCR amplified and inserted into linearized pENTR3C by Gibson Assembly.…”

Section: Methodsmentioning

confidence: 99%

“…23 ). The amiRFT fragments were introduced into EcoRI-HF (NEB, R3101S) digested pENTR3C (Thermo Fisher Scientific, A10464) by Gibson Assembly (NEB, E5510S) and shuffled into binary vector pMCS:GW 20 using LR reaction, which resulted in pMCS:amiRFT. The previously described 3994-bp FT promoter was amplified and inserted to XhoI (NEB, R0146S) digested pMCS:amiRFT by Gibson Assembly.…”

Section: Methodsmentioning

confidence: 99%

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Florigen family chromatin recruitment, competition and target genes

Zhu

Klasfeld

et al. 2020

Preprint

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17Plants monitor seasonal cues, such as day-length, to optimize life history traits including 18 onset of reproduction and inflorescence architecture 1-3 . Florigen family transcriptional 19 co-regulators TERMINAL FLOWER 1 (TFL1) and FLOWERING LOCUS T (FT) 20 antagonistically regulate these vital processes 4-6 yet how TFL1 and FT execute their 21 roles and what the mechanism is for their antagonism remains poorly understood. We 22 Main 35Plant development occurs after embryogenesis and is plastic, this allows modulation of 36 the final body plan in response to environmental cues to enhance growth and 37 reproductive success 7,8 . Of particular importance for species survival is the timing of the 38 formation of flowers that give rise to seeds which occurs in response to the seasonal 39 cues photoperiod and temperature 1,3,9 . For example, in plants that flower only once, like 40Arabidopsis and most crops, an early switch to flower formation allows rapid completion 41 of the life-cycle in a short growing season, but reduces total seed set or yield 10-12 . By 42 contrast, delaying flower formation supports formation of more seeds, but extends the 43 time to seed set. In many plant species, these alternative developmental trajectories are 44 tuned in response to daylength in antagonistic fashion by two members of the florigen 45 family of proteins 12,13 . FT promotes onset of the reproductive phase and flower 46 formation (determinacy), while TFL1 promotes vegetative development and branch fate 47 (indeterminacy) 4-6,13 . In Arabidopsis, which flowers in the spring, FT accumulates only 48 when the daylength exceeds a critical threshold, while TFL1 is present in both short-day 49 and long-day conditions 1,3,14 . A key unanswered question is how FT and TFL1 50 modulate plant form -what are the downstream processes they set in motion and what 51 is molecular basis for their antagonism? 52 53Accumulating evidence points to roles of FT and TFL1, which lack ability to bind DNA, 54 in transcriptional activation and repression, respectively, by forming complexes with a 55 bZIP transcription factor, FLOWERING LOCUS D (FD) 12,15-19 , although non-nuclear 56 functions for both proteins have also been described 20,21 . Mechanistic insight into 57 florigen activity has been hampered by their low protein abundance. To overcome this 58 limitation and to test the role of TFL1 in the nucleus, we conducted TFL1 chromatin 59 immunoprecipitation followed by sequencing (ChIP-seq). Towards this end, we first 60 generated a biologically active, genomic GFP-tagged version of TFL1 (gTFL1-GFP tfl1-61 1) (Supplementary Fig. 1a, b). Next, we enriched for TFL1 expressing cells by isolating 62 shoot apices from 42-day-old short-day-grown inflorescences just prior to onset of 63 flower formation (Fig. 1a). Finally, we optimized low abundance ChIP by combining 64 eight individual ChIP reactions per ChIP-seq replicate. We conducted FD ChIP-seq in 65 4 analogous fashion using a published 22 , biologically active ( Supplementary Fig. 1c), 66 genomic...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Florigen family chromatin recruitment, competition and target genes

Zhu

Klasfeld

et al. 2020

Preprint

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“…To generate the pSCR: C4 construct, the coding sequence of C4 was cloned into pENTR/D-TOPO (Invitrogen, USA), and subsequently Gateway-cloned into the pSCR:GW vector 13 through an LR reaction (Invitrogen, USA). A .…”

Section: Methodsmentioning

confidence: 99%

The receptor-like kinases BAM1 and BAM2 promote the cell-to-cell movement of miRNA in the root stele to regulate xylem patterning

Fan

Wang

Hou

et al. 2019

Preprint

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Xylem patterning in the root is established through the creation of opposing gradients of miRNAs and their targets, enabled by the cell-to-cell spread of the former. The miRNAs involved in xylem patterning, miR165/6, move through plasmodesmata, but how their trafficking is regulated remains elusive. Here, we describe that the receptor-like kinases BAM1/2 are required for the intercellular movement of miR165/6 in the stele and hence proper xylem patterning in the root.

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“…pENTR-MSL2 (Haswell and Meyerowitz, 2006) was used in a Gateway technology (Life Technologies) recombination reaction with pSCR:GW (Michniewicz et al, 2015) to create pSCR:MSL2.…”

Section: Subcloning and Transgenic Linesmentioning

confidence: 99%

Plastid osmotic stress influences cell differentiation at the plant shoot apex

Wilson¹,

Mixdorf²,

Berg³

et al. 2016

Journal of Cell Science

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The balance between proliferation and differentiation in the plant shoot apical meristem is controlled by regulatory loops involving the phytohormone cytokinin and stem cell identity genes. Concurrently, cellular differentiation in the developing shoot is coordinated with the environmental and developmental status of plastids within those cells. Here, we employ an Arabidopsis thaliana mutant exhibiting constitutive plastid osmotic stress to investigate the molecular and genetic pathways connecting plastid osmotic stress with cell differentiation at the shoot apex. msl2 msl3 mutants exhibit dramatically enlarged and deformed plastids in the shoot apical meristem, and develop a mass of callus tissue at the shoot apex. Callus production in this mutant requires the cytokinin receptor AHK2 and is characterized by increased cytokinin levels, downregulation of cytokinin signaling inhibitors ARR7 and ARR15, and induction of the stem cell identity gene WUSCHEL. Furthermore, plastid stressinduced apical callus production requires elevated plastidic reactive oxygen species, ABA biosynthesis, the retrograde signaling protein GUN1, and ABI4. These results are consistent with a model wherein the cytokinin/WUS pathway and retrograde signaling control cell differentiation at the shoot apex.

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Gateway-compatible tissue-specific vectors for plant transformation

Cited by 43 publications

References 43 publications

Florigen family chromatin recruitment, competition and target genes

Florigen family chromatin recruitment, competition and target genes

The receptor-like kinases BAM1 and BAM2 promote the cell-to-cell movement of miRNA in the root stele to regulate xylem patterning

Plastid osmotic stress influences cell differentiation at the plant shoot apex

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