Overexpression of the Maize ZmNLP6 and ZmNLP8 Can Complement the Arabidopsis Nitrate Regulatory Mutant nlp7 by Restoring Nitrate Signaling and Assimilation

Cao, Huairong; Qi, Shengdong; Sun, Ming; Li, Zehui; Yang, Yi; Crawford, Nigel M.; Wang, Yong

doi:10.3389/fpls.2017.01703

Cited by 63 publications

(75 citation statements)

References 47 publications

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“…Cooled samples were centrifuged at 13,000 rpm for 10 min, and 400 μL of each supernatant was transferred to a flow cell. NO 3 □ content in each cell was detected with the hydrazine reduction method using the AutoAnalyzer 3 continuous flow analytical system, as previously described (Cao et al, 2017).…”

Section: Methodsmentioning

confidence: 99%

The stability and ABA import activity of NRT1.2 inArabidopsisare regulated by CEPR2 via phosphorylation modification

Zhang

et al. 2020

Preprint

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ABA signaling and transport play important roles in plant systematic responses to environmental factors. Previously, we reported that C-terminally encoded peptide receptor 2 (CEPR2) balances plant growth and ABA responses by phosphorylating abscisic acid (ABA) receptors. Here, we showed that CEPR2 also directly phosphorylated the ABA transporter NRT1.2 by phosphaproteomics analysis using CEPR2 overexpression line and by physiological and biochemical experiments. Using transgenic seedlings, we demonstrated that NRT1.2 positively regulated the ABA response, and that CEPR2 acted on NRT1.2 epistatically. Specifically, CEPR2 phosphorylated NRT1.2 at serine 292 (Ser292) to promote the degradation of NRT1.2 via the 26S proteasome and vacuolar degradation pathways. However, ABA strongly inhibited the phosphorylation of NRT1.2 at Ser292, stabilizing NRT1.2. Transportation assays in yeast and Xenopus oocytes showed that the nonphosphorylation of NRT1.2 showed high ABA but very low nitrate import activity, while the phosphorylation of NRT1.2 by CEPR2 eliminated NRT1.2-mediated ABA import but slightly increased nitrate transport. These findings were consistent with our analyses of complement mutants in planta. Thus, our results provide detailed mechanisms of NRT1.2 in transporting ABA and nitrate in response to environmental conditions in plants. Besides, it is a better way for plants to overexpress the unphosphorylated NRT1.2 at Ser292 both under high and low nitrate conditions.

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Section: Methodsmentioning

confidence: 99%

The stability and ABA import activity of NRT1.2 inArabidopsisare regulated by CEPR2 via phosphorylation modification

Zhang

et al. 2020

Preprint

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“…Nitrate content was measured using the hydrazine reduction method as described by Cao et al . (). The assay of 15 NO 3 − uptake was performed as described previously (Wang & Tsay, ).…”

Section: Methodsmentioning

confidence: 99%

“…Nitrate content, 15 NO 3 À uptake, nitrate reductase activity and amino acid content assays Nitrate content was measured using the hydrazine reduction method as described by Cao et al (2017). The assay of 15 NO 3 À uptake was performed as described previously (Wang & Tsay, 2011).…”

Section: Qrt-pcr Assaymentioning

confidence: 99%

“…Interestingly, NLP7 is also involved in long-term effects; for example, its overexpression can promote root development under nitrate-rich conditions (Yu et al, 2016). Recently, two homologous genes (ZmNLP6 and ZmNLP8) in the maize (Zea mays) NLP family have been characterised as encoding proteins that play important roles in enhancing nitrate assimilation, root growth, and nitrogen use efficiency (NUE) when ectopically expressed in Arabidopsis (Cao et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

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The long noncoding RNA T5120 regulates nitrate response and assimilation in Arabidopsis

Liu

Chang

et al. 2019

New Phytologist

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Summary Long noncoding RNAs (lncRNAs) are crucial regulators in many plant biological processes. However, it remains unknown whether lncRNAs can respond to nitrate or function in nitrate regulation. We detected 695 lncRNAs, 480 known and 215 novel, in Arabidopsis seedling roots; six showed altered expression in response to nitrate treatment, among which T5120 showed the highest induction. Overexpression of T5120 in Arabidopsis promoted the response to nitrate, enhanced nitrate assimilation and improved biomass and root development. Biochemical and molecular analyses revealed that NLP7, a master nitrate regulatory transcription factor, directly bound to the nitrate‐responsive cis‐element (NRE)‐like motif of the T5120 promoter and activated T5120 transcription. In addition, T5120 partially restored the nitrate signalling and assimilation phenotypes of nlp7 mutant, suggesting that T5120 is involved in NLP7‐mediated nitrate regulation. Interestingly, the expression of T5120 was regulated by the nitrate sensor NRT1.1. Therefore, T5120 is modulated by NLP7 and NRT1.1 to regulate nitrate signalling. Our work reveals a new regulatory mechanism in which lncRNA T5120 functions in nitrate regulation, providing new insights into the nitrate signalling network. Importantly, lncRNA T5120 can promote nitrate assimilation and plant growth to improve nitrogen use efficiency.

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“…Disruption of AtNLP7 or suppression of AtNLP6 results in a N-starved phenotype and impaired nitrate signaling (Castaings et al, 2009; Konishi and Yanagisawa, 2013). Constitutive expression of the maize ZmNLP6 or ZmNLP8 can complement the Arabidopsis nlp7 mutant by restoring nitrate signaling and assimilation (Cao et al, 2017). AtNLP7 is regulated by nitrate via a nuclear retention mechanism, which is controlled by the phosphorylation of the conserved serine205 by Ca 2+ -sensor protein kinases10/30/32 (AtCPK10/30/32) in the N-terminal region of AtNLP7 (Liu et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Rice NIN-LIKE PROTEIN 4 is a master regulator of nitrogen use efficiency

Zhang

Xia

et al. 2020

Preprint

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1Nitrogen (N) is one of the key essential macronutrients that affects rice growth 3 2 and yield. Inorganic N fertilizers are excessively used to boost yield and generate 3 3 serious collateral environmental pollution. Therefore, improving crop N use 3 4 efficiency (NUE) is highly desirable and has been a major endeavor in crop 3 5 improvement. However, only a few regulators have been identified that can be used to 3 6 improve NUE in rice to date. Here we show that the NIN-like protein OsNLP4 3 7 significantly improves the rice NUE and yield. Field trials consistently showed that 3 8 loss-of-OsNLP4 dramatically reduced yield and NUE compared with wild type under 3 9 different N regimes. In contrast, the OsNLP4 overexpression lines remarkably 4 0 increased yield by 30% and NUE by 47% under moderate N level compared with wild 4 1 type. Transcriptomic analyses revealed that OsNLP4 orchestrates the expression of a 4 2 majority of known N uptake, assimilation and signaling genes by directly binding to 4 3 the nitrate-responsive cis-element in their promoters to regulate their expression. 4 4 Moreover, overexpression of OsNLP4 can recover the phenotype of Arabidopsis nlp7 4 5 mutant and enhance its biomass. Our results demonstrate that OsNLP4 is a master 4 6 regulator of NUE in rice and sheds light on crop NUE improvement. 4 7 4 8

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Overexpression of the Maize ZmNLP6 and ZmNLP8 Can Complement the Arabidopsis Nitrate Regulatory Mutant nlp7 by Restoring Nitrate Signaling and Assimilation

Cited by 63 publications

References 47 publications

The stability and ABA import activity of NRT1.2 inArabidopsisare regulated by CEPR2 via phosphorylation modification

The stability and ABA import activity of NRT1.2 inArabidopsisare regulated by CEPR2 via phosphorylation modification

The long noncoding RNA T5120 regulates nitrate response and assimilation in Arabidopsis

Rice NIN-LIKE PROTEIN 4 is a master regulator of nitrogen use efficiency

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