Long-Term In Vivo Imaging of Luciferase-Based Reporter Gene Expression in Arabidopsis Roots

Xuan, Wei; Opdenacker, Davy; Vanneste, Steffen; Beeckman, Tom

doi:10.1007/978-1-4939-7747-5_13

Cited by 18 publications

(20 citation statements)

References 15 publications

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“…A Lumazone imaging system equipped with a charge‐coupled device camera (iKon‐M from Andor, Model No, DU934P‐BV) was applied to image the luciferase signal in the vertical growing Arabidopsis root tip as described previously (Xuan, Opdenacker, Vanneste, & Beeckman, ). To perform time‐lapse imaging of the DR5:Luciferase expression in the OZ, square plates contain 1/2 MS medium with or without 30 μM Cd were sprayed with 1 mM D‐Luciferin solution (0.01% Tween 80) and left to dry in the dark (about 10 min), then 3‐day‐old DR5:Luciferase transgenic seedlings were transferred to the plates supplemented with or without 30 μM Cd, and imaged immediately with a macro lens every 10 min (exposure time) for indicated time period.…”

Section: Methodsmentioning

confidence: 99%

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Cadmium stress suppresses lateral root formation by interfering with the root clock

Xie

Wang

Zheng

et al. 2019

Plant Cell & Environment

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A biological clock activated by oscillating signals, known as root clock, has been linked to lateral root (LR) formation and is essential for regular LR spacing along the primary root. However, it remains unclear how this internal mechanism is influenced by environmental factors known to affect the LR pattern. Here, we report that excessive cadmium (Cd) inhibits LR formation by disrupting the lateral root cap (LRC)-programmed cell death (PCD)-regulated root clock. Cd restricts the frequency of the oscillating signal rather than its amplitude. This could be attributed to the inhibition on meristematic activity by Cd, which resulted in decreased LRC cell number and LRC-PCD frequency. Genetic evidence further showed that LRC cell number is positively correlated with root resistance to Cd. Our study reveals root cap dynamics as a novel mechanism mediating root responses to Cd, providing insight into the signalling pathways of the root clock responding to environmental cues.

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Section: Methodsmentioning

confidence: 99%

“…Kymographs (http://www.embl.de/eamnet/html/body_kymograph.html) were generated by Fiji (Xuan et al, ) to visualize the spatiotemporal changes of DR5:Luciferase signals in the root tips during PR growth.…”

Section: Methodsmentioning

confidence: 99%

Cadmium stress suppresses lateral root formation by interfering with the root clock

Xie

Wang

Zheng

et al. 2019

Plant Cell & Environment

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“…pDR5:Luciferase expression along the primary root was analysed by using Lumazon (Xuan, Opdenacker, Vanneste, & Beeckman, 2018).…”

Section: Luciferase Assaymentioning

confidence: 99%

Volatile compounds from beneficial rhizobacteria Bacillus spp. promote periodic lateral root development in Arabidopsis

Shao

Xie

et al. 2021

Plant Cell & Environment

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Lateral root formation is coordinated by both endogenous and external factors. As biotic factors, plant growth-promoting rhizobacteria can affect lateral root formation, while the regulation mechanism is unclear. In this study, by applying various marker lines, we found that volatile compounds (VCs) from Bacillus amyloliquefaciens SQR9 induced higher frequency of DR5 oscillation and prebranch site formation, accelerated the development and emergence of the lateral root primordia and thus promoted lateral root development in Arabidopsis. We demonstrated a critical role of auxin on B. amyloliquefaciens VCs-induced lateral root formation via respective mutants and pharmacological experiments. Our results showed that auxin biosynthesis, polar transport and signalling pathway are involved in B. amyloliquefaciens VCs-induced lateral roots formation. We further showed that acetoin, a major component of B. amyloliquefaciens VCs, is less active in promoting root development compared to VC blends from B. amyloliquefaciens, indicating the presence of yet uncharacterized/unknown VCs might contribute to B. amyloliquefaciens effect on lateral root formation. In summary, our study revealed an auxin-dependent mechanism of B. amyloliquefaciens VCs in regulating lateral root branching in a non-contact manner, and further efforts will explore useful VCs to promote plant root development.

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“…The Luciferase imaging of whole seedlings and oscillation expression analysis was performed as described 44…”

Section: Oscillation and Prebranch Sitementioning

confidence: 99%

ABCB-mediated auxin transport in outer root tissues regulates lateral root spacing inArabidopsis

Chen

Hao

et al. 2020

Preprint

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Root branching is an important strategy to explore efficiently large volumes of soil. To economize this process, lateral roots (LR) are formed along the growing root at discrete positions that are instructed by oscillating auxin signals derived from the lateral root cap (LRC). This assumes that auxin moves from the LRC across multiple layers to accumulate in the pericycle. Here, we identified, using gene silencing and CRISPR based approaches, a group of five genetically linked, closely related ABCBs that control LR spacing by modulating the amplitude of the auxin oscillation. The transporters localize to the plasma membrane and reveal significant auxin export activity. These ABCBs are mainly expressed in the LRC and epidermis where they contribute to auxin transport towards the root oscillation zone. Our findings highlight the importance of auxin transport in the outer tissues of the root meristem to regulate LR spacing.

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Long-Term In Vivo Imaging of Luciferase-Based Reporter Gene Expression in Arabidopsis Roots

Cited by 18 publications

References 15 publications

Cadmium stress suppresses lateral root formation by interfering with the root clock

Cadmium stress suppresses lateral root formation by interfering with the root clock

Volatile compounds from beneficial rhizobacteria Bacillus spp. promote periodic lateral root development in Arabidopsis

ABCB-mediated auxin transport in outer root tissues regulates lateral root spacing inArabidopsis

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