Clonal sector analysis and cell ablation confirm a function for DORNROESCHEN-LIKE in founder cells and the vasculature in Arabidopsis

Abstract

Main conclusion Inducible lineage analysis and cell ablation via conditional toxin expression in cells expressing the DORNRÖSCHEN-LIKE transcription factor represent an effective and complementary adjunct to conventional methods of functional gene analysis. Abstract Classical methods of functional gene analysis via mutational and expression studies possess inherent limitations, and therefore, the function of a large proportion of transcription factors remains unknown. We have employed two complementary, indirect methods to obtain functional information for the AP2/ERF transcription factor DORNRÖSCHEN-LIKE (DRNL), which is dynamically expressed in flowers and marks lateral organ founder cells. An inducible, two-component Cre–Lox system was used to express beta-glucuronidase GUS in cells expressing DRNL, to perform a sector analysis that reveals lineages of cells that transiently expressed DRNL throughout plant development. In a complementary approach, an inducible system was used to ablate cells expressing DRNL using diphtheria toxin A chain, to visualise the phenotypic consequences. These complementary analyses demonstrate that DRNL functionally marks founder cells of leaves and floral organs. Clonal sectors also included the vasculature of the leaves and petals, implicating a previously unidentified role for DRNL in provasculature development, which was confirmed in cotyledons by closer analysis of drnl mutants. Our findings demonstrate that inducible gene-specific lineage analysis and cell ablation via conditional toxin expression represent an effective and informative adjunct to conventional methods of functional gene analysis.

Genetic architecture behind developmental and seasonal control of tree growth and wood properties in Norway spruce

“…As an example of an association specific to JW, an SNP in the gene model MA_77420g0010, encoding a putative member of the ethylene response factor family, was associated with the multi-traits composed of the number of cells, ring width, and tangential ring width ( Table 3 ). The homologous Arabidopsis thaliana gene AT1G24590 controls organogenesis and is also linked to provasculature development (Glowa et al, 2021 ). An SNP in another gene model MA_14038g0010, encoding a member of the GATA transcription factor family, was associated with latewood density in JW ( Table 3 ).…”

Post-Embryonic Lateral Organ Development and Adaxial—Abaxial Polarity Are Regulated by the Combined Effect of ENHANCER OF SHOOT REGENERATION 1 and WUSCHEL in Arabidopsis Shoots

“…Using this line, we confirmed the consistent expression pattern of ESR1 in the upper layers in the CZ and PZ of the SAM ( Figure 4 D), as reported previously [ 17 ]. On the other hand, the expression of ESR2 is predominantly enriched in the founder cells of leaf primordia in the early vegetative phase [ 13 , 28 ]. The expression pattern of ESR1, ESR2, WUS, STM, and REV in the vegetative shoot has been reported previously [ 13 , 29 , 30 , 31 , 32 ] and their protein distribution is schematically represented ( Supplementary Figure S3 ).…”

A versatile CRISPR-based system for lineage tracing in living plants

“…Controlled activation of recombinases can excise or flip matching target sequences in reporter genes and thereby label a cell lineage (e.g., Efroni et al 2015, Shi et al 2019, Suzuki et al 2020, or eliminate lineages, ablating precursor cells by expressing diphtheria toxin (Glowa et al 2021). Targeting a key gene in pigment biosynthesis with CRISPR-Cas9 created sharp sectors lacking colour in the petals of Japanese morning glory and allowed to study periclinal chimaera (Watanabe et al 2017).…”

“…Their excision can be induced by trans-activation, e.g., by heat activation of the autonomous Ac leading to excision of a non-autonomous Ds element between a promoter and a reporter gene or similar combinations (e.g., Barkoulas et al 2008, Kurup et al 2005, Torres-Martínez et al 2020). Controlled activation of recombinases can excise or flip matching target sequences in reporter genes and thereby label a cell lineage (e.g., Efroni et al 2015, Shi et al 2019, Suzuki et al 2020), or eliminate lineages, ablating precursor cells by expressing diphtheria toxin (Glowa et al 2021). Targeting a key gene in pigment biosynthesis with CRISPR-Cas9 created sharp sectors lacking colour in the petals of Japanese morning glory and allowed to study periclinal chimaera (Watanabe et al 2017).…”