Lateral Root Initiation and the Analysis of Gene Function Using Genome Editing with CRISPR in Arabidopsis

Lateral root initiation is a post-embryonic process that requires the specification of a subset of pericycle cells adjacent to the xylem pole in the primary root into lateral root founder cells. The first visible event of lateral root initiation in Arabidopsis is the simultaneous migration of nuclei in neighbouring founder cells. Coinciding cell cycle activation is essential for founder cells in the pericycle to undergo formative divisions, resulting in the development of a lateral root primordium (LRP). The plant signalling molecule, auxin, is a major regulator of lateral root development; the understanding of the molecular mechanisms controlling lateral root initiation has progressed tremendously by the use of the Arabidopsis model and a continual improvement of molecular methodologies. Here, we provide an overview of the visible events, cell cycle regulators, and auxin signalling cascades related to the initiation of a new LRP. Furthermore, we highlight the potential of genome editing technology to analyse gene function in lateral root initiation, which provides an excellent model to answer fundamental developmental questions such as coordinated cell division, growth axis establishment as well as the specification of cell fate and cell polarity.

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AT-HOOK MOTIF NUCLEAR LOCALISED PROTEIN 18 as a Novel Modulator of Root System Architecture

10.20944/preprints202003.0046.v1 ◽

2020 ◽

Author(s):

Marek Šírl ◽

Tereza Šnajdrová ◽

Dolores Gutiérrez-Alanís ◽

Joseph G. Dubrovsky ◽

Jean Phillipe Vielle-Calzada ◽

...

Keyword(s):

Root System ◽

System Architecture ◽

Lateral Root ◽

Apical Meristem ◽

Lateral Roots ◽

Primary Root ◽

Root System Architecture ◽

Root Apical Meristem ◽

Root Initiation ◽

Lateral Root Initiation

The AT-HOOK MOTIF NUCLEAR LOCALIZED PROTEIN (AHL) gene family encodes embryophyte-specific nuclear proteins with DNA binding activity. They modulate gene expression and affect various developmental processes in plants. We identify AHL18 (At3G60870) as a developmental modulator of root system architecture and growth. AHL18 regulates the length of the proliferation domain and number of dividing cells in the root apical meristem and thereby, cell production. Both primary root growth and lateral root development respond according to AHL18 transcription level. The ahl18 knock-out plants show reduced root systems due to a shorter primary root and a lower number of lateral roots. This change results from a higher number of arrested and non-developing lateral root primordia (LRP) rather than from decreased initiation. Overexpression of AHL18 results in a more extensive root system, longer primary roots, and increased density of lateral root initiation events. Formation of lateral roots is affected during the initiation of LRP and later development. AHL18 regulate root apical meristem activity, lateral root initiation and emergence, which is in accord with localization of its expression.

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A single cell view of the transcriptome during lateral root initiation in Arabidopsis thaliana

10.1101/2020.10.02.324327 ◽

2020 ◽

Author(s):

Hardik P. Gala ◽

Amy Lanctot ◽

Ken Jean-Baptiste ◽

Sarah Guiziou ◽

Jonah C. Chu ◽

...

Keyword(s):

Single Cell ◽

Root Development ◽

Lateral Root ◽

Lateral Roots ◽

Primary Root ◽

Root Initiation ◽

Lateral Root Primordia ◽

Lateral Root Development ◽

Lateral Root Initiation ◽

Molecular Events

AbstractRoot architecture is a major determinant of fitness, and is under constant modification in response to favorable and unfavorable environmental stimuli. Beyond impacts on the primary root, the environment can alter the position, spacing, density and length of secondary or lateral roots. Lateral root development is among the best-studied examples of plant organogenesis, yet there are still many unanswered questions about its earliest steps. Among the challenges faced in capturing these first molecular events is the fact that this process occurs in a small number of cells with unpredictable timing. Single-cell sequencing methods afford the opportunity to isolate the specific transcriptional changes occurring in cells undergoing this fate transition. Using this approach, we successfully captured the transcriptomes of initiating lateral root primordia, and discovered many previously unreported upregulated genes associated with this process. We developed a method to selectively repress target gene transcription in the xylem pole pericycle cells where lateral roots originate, and demonstrated that expression of several of these targets was required for normal root development. We also discovered novel subpopulations of cells in the pericycle and endodermal cell files that respond to lateral root initiation, highlighting the coordination across cell files required for this fate transition.One sentence summarySingle cell RNA sequencing reveals new molecular details about lateral root initiation, including the transcriptional impacts of the primordia on bordering cells.

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Systemic signalling through translationally controlled tumour protein controls lateral root formation in Arabidopsis

Journal of Experimental Botany ◽

10.1093/jxb/erz204 ◽

2019 ◽

Vol 70 (15) ◽

pp. 3927-3940 ◽

Cited By ~ 9

Author(s):

Rémi Branco ◽

Josette Masle

Keyword(s):

Lateral Root ◽

Root Elongation ◽

Developmental Plasticity ◽

Lateral Roots ◽

Primary Root ◽

Dual Function ◽

Growth Promoter ◽

Root Initiation ◽

Long Distance ◽

Lateral Root Initiation

Abstract The plant body plan and primary organs are established during embryogenesis. However, in contrast to animals, plants have the ability to generate new organs throughout their whole life. These give them an extraordinary developmental plasticity to modulate their size and architecture according to environmental constraints and opportunities. How this plasticity is regulated at the whole-organism level is elusive. Here we provide evidence for a role for translationally controlled tumour protein (TCTP) in regulating the iterative formation of lateral roots in Arabidopsis. AtTCTP1 modulates root system architecture through a dual function: as a general constitutive growth promoter enhancing root elongation and as a systemic signalling agent via mobility in the vasculature. AtTCTP1 encodes mRNAs with long-distance mobility between the shoot and roots. Mobile shoot-derived TCTP1 gene products act specifically to enhance the frequency of lateral root initiation and emergence sites along the primary root pericycle, while root elongation is controlled by local constitutive TCTP1 expression and scion size. These findings uncover a novel type for an integrative signal in the control of lateral root initiation and the compromise for roots between branching more profusely or elongating further. They also provide the first evidence in plants of an extracellular function of the vital, highly expressed ubiquitous TCTP1.

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Application of Phytohormones to Pea Roots After Removal of the Apex: Effect on Lateral Root Production

Functional Plant Biology ◽

10.1071/pp9790195 ◽

1979 ◽

Vol 6 (2) ◽

pp. 195 ◽

Cited By ~ 9

Author(s):

PB Goodwin ◽

SC Morris

Keyword(s):

Lateral Root ◽

Lateral Roots ◽

Primary Root ◽

Root Production ◽

Root Tip ◽

Naphthaleneacetic Acid ◽

Root Initiation ◽

Lateral Root Initiation ◽

Lateral Bud ◽

Bud Growth

Removal of 2 mm of the primary root tip of Pisum sativum caused a complete halt to primary root elongation, but did not alter the total number of laterals formed. The auxins indole-3-acetic acid and 1-naphthaleneacetic acid, when applied to the stump in a lanolin emulsion, increased the number of lateral roots. High levels of abscisic acid and low levels of the cytokinins N6-benzylaminopurine and N6-(γ, γ-dimethylallylamino)purine, and of the gibberellins GA3 and GA7, resulted in decreased lateral root production. Kinetin was without effect. There appears to be an inverse relationship between auxins and cytokinins in root/shoot growth coordination. Auxins, which are produced in the shoot tip, inhibit lateral bud growth but promote lateral root initiation. Cytokinins, which are produced in the root tip, inhibit lateral root initiation, but promote lateral stem growth.

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