The small peptide CEP1 and the NIN-like protein NLP1 regulate NRT2.1 to mediate root nodule formation across nitrate concentrations.

Legumes acquire fixed nitrogen (N) from the soil and through endosymbiotic association with diazotrophic bacteria. However, establishing and maintaining N2-fixing nodules are expensive for the host plant, relative to taking up N from the soil. Therefore, plants suppress symbiosis when N is plentiful and enhance symbiosis when N is sparse. Here, we show that the nitrate transporter MtNRT2.1 is required for optimal nodule establishment in Medicago truncatula under low-nitrate conditions and the repression of nodulation under high-nitrate conditions. The NIN-like protein (NLP) MtNLP1 is required for MtNRT2.1 expression and regulation of nitrate uptake/transport under low- and high-nitrate conditions. Under low nitrate, the gene encoding the C-terminally encoded peptide (CEP) MtCEP1 was more highly expressed, and the exogenous application of MtCEP1 systemically promoted MtNRT2.1 expression in a compact root architecture 2 (MtCRA2)-dependent manner. The enhancement of nodulation by MtCEP1 and nitrate uptake were both impaired in the Mtnrt2.1 mutant under low nitrate. Our study demonstrates that nitrate uptake by MtNRT2.1 differentially affects nodulation at low- and high-nitrate conditions through the actions of MtCEP1 and MtNLP1.

Constitutive activation of a nuclear-localized calcium channel complex in Medicago truncatula.

Nuclear Ca2+ oscillations allow symbiosis signaling, facilitating plant recognition of beneficial microsymbionts, nitrogen-fixing rhizobia, and nutrient-capturing arbuscular mycorrhizal fungi. Two classes of channels, DMI1 and CNGC15, in a complex on the nuclear membrane, coordinate symbiotic Ca2+ oscillations. However, the mechanism of Ca2+ signature generation is unknown. Here, we demonstrate spontaneous activation of this channel complex, through gain-of-function mutations in DMI1, leading to spontaneous nuclear Ca2+ oscillations and spontaneous nodulation, in a CNGC15-dependent manner. The mutations destabilize a hydrogen-bond or salt-bridge network between two RCK domains, with the resultant structural changes, alongside DMI1 cation permeability, activating the channel complex. This channel complex was reconstituted in human HEK293T cell lines, with the resultant calcium influx enhanced by autoactivated DMI1 and CNGC15s. Our results demonstrate the mode of activation of this nuclear channel complex, show that DMI1 and CNGC15 are sufficient to create oscillatory Ca2+ signals, and provide insights into its native mode of induction.

Axial resolution analysis in compressive digital holographic microscopy.

Digital holographic microscopy with compressive sensing (CDHM) has successfully achieved tomography and has been applied in many fields. However, the enhancement of axial resolution in CDHM remains to be elucidated. By deducing accurate formulas for the lateral and axial resolutions without paraxial approximation, we quantized the elongation effect of a digital holography (DH) system in this study. Thus, we revealed that the elongation effect, which is affected only by the system's numerical aperture (NA), is an inherent property of DH systems. We present a detailed analysis herein on the physical significance of the coherence parameter, which is the ratio of a system's limit axial resolution to the interlayer spacing more thoroughly than in previous research. Further, we achieved the tomography of a fiber by using a DH system with a 10 × microscope, with CS to eliminate the elongation effect, and experimentally validated our theoretical results. By applying these theoretical guidelines, we distinguished crossed fibers at distances of 36.4 µm and 48.5 µm, respectively, using the same experimental setup. There would be potential applications of this theory in tomography and observation of microscale objects in the areas of biological and fluid.

Influence of sparse constraint functions on compressive holographic tomography.

In this paper, we quantified and analyzed the impact of the l1 norm and total variation (TV) norm sparse constraints on the reconstruction quality under different interlayer spacings, sampling rates, and signal-to-noise ratios. For high-quality holograms, the results of compressive-sensing reconstruction using l1 norm achieved higher quality than those by the TV norm. In contrast, for low-quality holograms, the quality of TV-norm-based reconstruction results was relatively stable and better than that of l1 norm. In addition, we explained why interlayer spacing cannot be smaller and recommend the use of axial resolution of the digital holography system as the interlayer spacing. The conclusions are valuable in the choice of sparse constraints in compressive holographic tomography.

Digital holographic phase imaging based on phase iteratively enhanced compressive sensing.

Digital holography has been widely applied in quantitative phase imaging (QPI) for monolayer objects within a limited depth. For multilayer imaging, compressive sensing is employed to eliminate defocused images but with missing phase information. A phase iteratively enhanced compressive sensing (PIE-CS) algorithm is proposed to achieve phase imaging and eliminate defocused images simultaneously. Linear filtering is first applied to the off-axis hologram in Fourier domain, and an intermediate reconstructed complex image is obtained. A periodic phase mask is then superimposed on the intermediate reconstructed image to iteratively eliminate the defocused images and recover the object with phase information. The experimental recovery of amplitude and phase of a two-layer sample with as little as 7% random measurement is demonstrated. The average phase error of the PIE-CS algorithm is analyzed, and the results show the feasibility for QPI.

Cellular and molecular basis of symbiotic nodule development.

Root nodule development plays a vital role in establishing the mutualistic relationship between legumes and nitrogen-fixing rhizobia. Two primary processes are involved in nodule development: formative cell divisions in the root cortex and the subsequent differentiation of nodule cells. The first process involves the mitotic reactivation of differentiated root cortex cells to form nodule primordium after perceiving symbiotic signals. The second process enables the nascent nodule primordium cells to develop into various cell types, leading to the creation of a functional nodule capable of supporting nitrogen fixation. Thus, both division and differentiation of nodule cells are crucial for root nodule development. This review provides an overview of the most recent advancements in comprehending the cellular and molecular mechanisms underlying symbiotic nodule development in legumes.

NLP1 reciprocally regulates nitrate inhibition of nodulation through SUNN-CRA2 signaling in Medicago truncatula.

Most legume plants can associate with diazotrophic soil bacteria called rhizobia, resulting in new root organs called nodules that enable N2 fixation. Nodulation is an energy-consuming process, and nodule number is tightly regulated by independent systemic signaling pathways controlled by CLE/SUNN and CEP/CRA2. Moreover, nitrate inhibits legume nodulation via local and systemic regulatory pathways. In Medicago truncatula, NLP1 plays important roles in nitrate-induced inhibition of nodulation, but the relationship between systemic and local pathways in mediating nodulation inhibition by nitrate is poorly understood. In this study, we found that nitrate induces CLE35 expression in an NLP1-dependent manner and that NLP1 binds directly to the CLE35 promoter to activate its expression. Grafting experiments revealed that the systemic control of nodule number involves negative regulation by SUNN and positive regulation by CRA2 in the shoot, and that NLP1's control of the inhibition of rhizobial infection, nodule development, and nitrogenase activity in response to nitrate is determined by the root. Unexpectedly, grafting experiments showed that loss of CRA2 in the root increases nodule number at inhibitory nitrate levels, probably because of CEP1/2 upregulation in the cra2 mutants, suggesting that CRA2 exerts active negative feedback regulation in the root.

NIN interacts with NLPs to mediate nitrate inhibition of nodulation in Medicago truncatula.

Legume plants can assimilate inorganic nitrogen and have access to fixed nitrogen through symbiotic interaction with diazotrophic bacteria called rhizobia. Symbiotic nitrogen fixation is an energy-consuming process and is strongly inhibited when sufficient levels of fixed nitrogen are available, but the molecular mechanisms governing this regulation are largely unknown. The transcription factor nodule inception (NIN) is strictly required for nodulation and belongs to a family of NIN-like proteins (NLPs), which have been implicated in the regulation of nitrogen homeostasis in Arabidopsis. Here, we show that mutation or downregulation of NLP genes prevents nitrate inhibition of infection, nodule formation and nitrogen fixation. We find that NIN and NLPs physically interact through their carboxy-terminal PB1 domains. Furthermore, we find that NLP1 is required for the expression of nitrate-responsive genes and that nitrate triggers NLP1 re-localization from the cytosol to the nucleus. Finally, we show that NLP1 can suppress NIN activation of CRE1 expression in Nicotiana benthamiana and Medicago truncatula. Our findings highlight a central role for NLPs in the suppression of nodulation by nitrate.

Author Correction: NIN interacts with NLPs to mediate nitrate inhibition of nodulation in Medicago truncatula.

In the version of this Article originally published, the name of author Zhenpeng Luo was incorrectly listed as Zhenpeng Luo Luo. Additionally, nitrate in Fig. 6b was incorrectly shown as 'NO3+' rather than 'NO3-'. This has now been amended.