Nitrate triggered phosphoproteome changes and a PIN2 phosphosite modulating root system architecture

Vega, Andrea; Fredes, Isabel; O’Brien, José Antonio; Shen, Zhouxin; Ötvös, Krisztina; Abualia, Rashed; Benková, Eva; Briggs, Steven P.; Gutiérrez, Rodrigo A.

doi:10.15252/embr.202051813

Cited by 28 publications

(20 citation statements)

References 134 publications

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“…Recently, it was shown that nitrate coordinately stimulates cell elongation and cell proliferation and lateral root development via effects on PIN2-mediated auxin transport ( Vega et al, 2021 ; Ötvös et al, 2021 ). This could be linked to nitrate regulated PIN2 phosphorylation changes that stimulated PIN2 secretion and recycling ( Ötvös et al, 2021 ).…”

Section: Discussionmentioning

confidence: 99%

Constitutive Active CPK30 Interferes With Root Growth and Endomembrane Trafficking in Arabidopsis thaliana

et al. 2022

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Calcium-dependent protein kinases (CPK) are key components of a wide array of signaling pathways, translating stress and nutrient signaling into the modulation of cellular processes such as ion transport and transcription. However, not much is known about CPKs in endomembrane trafficking. Here, we screened for CPKs that impact on root growth and gravitropism, by overexpressing constitutively active forms of CPKs under the control of an inducible promoter in Arabidopsis thaliana. We found that inducible overexpression of an constitutive active CPK30 (CA-CPK30) resulted in a loss of root gravitropism and ectopic auxin accumulation in the root tip. Immunolocalization revealed that CA-CPK30 roots have reduced PIN protein levels, PIN1 polarity defects and impaired Brefeldin A (BFA)-sensitive trafficking. Moreover, FM4-64 uptake was reduced, indicative of a defect in endocytosis. The effects on BFA-sensitive trafficking were not specific to PINs, as BFA could not induce aggregation of ARF1- and CHC-labeled endosomes in CA-CPK30. Interestingly, the interference with BFA-body formation, could be reverted by increasing the extracellular pH, indicating a pH-dependence of this CA-CPK30 effect. Altogether, our data reveal an important role for CPK30 in root growth regulation and endomembrane trafficking in Arabidopsis thaliana.

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Section: Discussionmentioning

confidence: 99%

Constitutive Active CPK30 Interferes With Root Growth and Endomembrane Trafficking in Arabidopsis thaliana

et al. 2022

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“…Local auxin distribution is achieved by the distinct localization of PIN proteins, which dynamically change in response to developmental cues, such as nutrient levels of the soil. PIN2, which localizes at the cortex and in epidermal cells, channels auxin flow from the root tip towards the elongation zone, creating the auxin gradient in roots together with other auxin transporting proteins [5,47,48]. Auxin efflux carriers PIN2 is important for modulation of root system architecture in response to different nutrient levels of the soil.…”

Section: Discussionmentioning

confidence: 99%

“…In Arabidopsis thaliana, phosphorus deficiency promotes root hair number and length via suppressing vacuolar degradation of PIN2, thus enhancing root absorption capacity and efficiency [29]. PIN2 also plays key role in regulating primary and lateral root growth in response to nitrate, as nitratetreated pin2 mutants showed longer primary roots and less lateral roots number than wild-type plants [48]. Furthermore, the phosphorylation status of PIN2, which regulates polar plasma membrane localization, is necessary for root system architecture modulation in response to nitrate.…”

Section: Discussionmentioning

confidence: 99%

Transporter NRT1.5/NPF7.3 suppresses primary root growth under low K+ stress by regulating the degradation of PIN-FORMED2

Wang

Zhao

et al. 2022

BMC Plant Biol

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Background The availability of potassium is one of the main environmental factors for modifying the plasticity of root architecture. Many potassium channels and transporters are involved in regulating primary root growth in response to low potassium stress. NRT1.5/NPF7.3 transporter is a NO3−/H+ and K+/H+ cotransporter, and participates in NO3− and K+ translocation from the roots to the shoots. However, the underlying mechanism of NRT1.5-regulated primary root growth under low potassium stress is unclear. Results We show that NRT1.5/NPF7.3 inhibited primary root growth under low potassium conditions by regulating the accumulation of PIN2 protein and auxin levels. Under low potassium conditions, the mutants nrt1.5 and lks2 exhibited longer primary roots, longer meristem regions and elongation zones of primary roots, and more cell activity in the meristem region compared to WT plants, revealing the involvement of NRT1.5 in LK (low potassium)-inhibition primary root growth. In addition, exogenous auxin (IAA), auxin analogue (NAA, 2.4-D) or auxin precursor (IBA) promoted the primary root growth of WT and the complementation line NRT1.5 COM plants. In addition, the application of NPA inhibited the primary root growth of the nrt1.5 and lks2 mutants. Auxin accumulation was higher in the root tip of nrt1.5 plants than in WT plants, indicating that NRT1.5 regulates root growth inhibition by regulating auxin distribution. Furthermore, PIN2 was degraded more quickly in nrt1.5 plants under LK stress. Conclusions Our findings reveal that NRT1.5 inhibits primary root growth by modulating the auxin level in the root tip via the degradation of PIN2.

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“…Nitrate mediates these functions largely through its impacts on endogenous auxin pathways. Numerous aspects of nitrate transport and signaling interact with auxin biosynthesis, signaling and transport (Vega et al, 2019; 2021; Asim et al, 2020; Otvos et al 2021; Liu and von Wirén, 2022). Multiple studies have identified auxin-related genes as differentially expressed in response to nitrate (Gifford et al, 2008; Vidal et al, 2010; 2013).…”

Section: Discussionmentioning

confidence: 99%

“…Nitrate assimilation and signaling have significant crosstalk with auxin transport and signaling pathways in the control of root development (Vega et al, 2019; Asim et al, 2020; Liu and von Wirén, 2022). For example, nitrate regulates root growth in part by modulating the phosphorylation status of the PIN2 auxin transporter (Otvos et al 2021; Vega et al 2021). Nitrate also induces the expression of the auxin receptor AUXIN SIGNALING F-BOX3 (AFB3) (Vidal et al, 2010; Vidal et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

The NIN-LIKE PROTEIN 7 (NLP7) transcription factor modulates auxin pathways to regulate root cap development

Kumar

Caldwell

Iyer‐Pascuzzi

2022

Preprint

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The root cap surrounds the root tip and promotes root growth by protecting the root apical meristem, influencing root branching, and sensing environmental signals like nitrate. The root cap maintains a constant size through the coordination of cell production in the root meristem with cell release at the tip of the root, a process that requires an auxin minima in the last layer of the root cap. To perform its functions, the root cap must maintain a constant size and synchronize external cues with development, but mechanisms underlying such coordination are not well understood. Mutations in the NIN LIKE PROTEIN 7 (NLP7) transcription factor, a master regulator of nitrate signaling, lead to defects in root cap cell release and cell production. Nitrate impacts root development through crosstalk with auxin. Therefore, we hypothesized that NLP7 regulates root cap cell release and cell production by modulating auxin pathways. Here we show that mutations in NLP7 abolish the auxin minima required for root cap cell release and alter root cap expression levels of the auxin carriers PIN-LIKES 3 (PILS3) and PIN-FORMED 7 (PIN7). We find that NLP7 is required for proper root cap cell production and differentiation and for expression of transcription factors that regulate these processes. Nitrate deficiency impacts auxin pathways in the last layer of the root cap, and this is mediated in part by NLP7. Together, our data suggest that NLP7 integrates nitrate signaling with auxin pathways to optimize root cap development in response to external nitrate cues.

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Nitrate triggered phosphoproteome changes and a PIN2 phosphosite modulating root system architecture

Cited by 28 publications

References 134 publications

Constitutive Active CPK30 Interferes With Root Growth and Endomembrane Trafficking in Arabidopsis thaliana

Constitutive Active CPK30 Interferes With Root Growth and Endomembrane Trafficking in Arabidopsis thaliana

Transporter NRT1.5/NPF7.3 suppresses primary root growth under low K+ stress by regulating the degradation of PIN-FORMED2

The NIN-LIKE PROTEIN 7 (NLP7) transcription factor modulates auxin pathways to regulate root cap development

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