Nitrogen availability remains a principal constraint to crop productivity. Plants cannot directly assimilate the abundant nitrogen available in our atmosphere; instead, they rely on the uptake of inorganic forms of nitrogen, such as ammonium and nitrate from the soil. Nitrogen is a limiting nutrient in wheat production, and wheat yields are very responsive to nitrogen fertilisation. Only diazotrophic bacteria can convert atmospheric nitrogen to ammonia via biological nitrogen fixation (BNF), and although improving BNF in wheat has been a longstanding objective, there have been no descriptions of successful modification of wheat crops showing increased BNF in the literature. Here we describe the use of polycistronic multiplexed CRISPR to modify the flavone biosynthetic pathway of hexaploid wheat (Triticum aestivum) plants, generating DNA-edited plants with increased apigenin content. The apigenin-enriched plants exude apigenin into the soil, inducing the colonisation of the roots and subsequent formation of biofilms in soil by diazotrophic bacteria. The low permeability of the biofilm to oxygen protected the bacterial nitrogenase and stimulated BN. Under nitrogen-limiting conditions, apigenin-enriched wheat lines exhibited increased nitrogen content, improved photosynthetic performance, and higher grain yield relative to wild-type controls. This work demonstrates the feasibility of engineering associative BNF in cereals via metabolic reprogramming of root exudation, offering a sustainable route to reduce dependence on synthetic nitrogen fertilisers.

Plants need to adapt to fluctuating atmospheric humidity and respond to both high and low humidity. Despite our substantial understanding of plant responses to low humidity, molecular mechanisms underlying the high humidity (HH) response are much less well understood. In this study, we investigated early responses to HH in Arabidopsis. Expression of CYP707A3, encoding an abscisic acid (ABA) 8 '- hydroxylase, is induced by HH within 10 min, which leads to a decrease in foliar ABA level. We identified that the combined action of CAMTA3 and CAMTA2 transcription factors regulate this response. This regulation requires a calmodulin (CaM)- binding domain of CAMTA3. Transcriptomes of HH- regulated genes are enriched in those related to calcium signaling, including cyclic nucleotide- gated ion channels (CNGCs). Moreover, HH induces CNGC2- and CNGC4- mediated increases in cytosolic Ca2+ concentrations in leaves within a few minutes. We also found that CNGC2, CNGC4, and CAMTAs participate in HH- induced hyponastic movement of petioles. Taken together, our results indicate that CNGC2/CNGC4- Ca2+-CaM- CAMTA3/CAMTA2 acts as a primary regulatory module to trigger downstream HH responses.

Gibberellins (GA) are phytohormones that regulate a number of key traits for crop performance, such as control of plant height, flowering time, and nitrogen use efficiency. We have curated gene family organization and expression of genes for GA biosynthesis, catabolism, and signaling in allopolyploid Brassica napus. Genes encoding early GA biosynthetic enzymes, including ent-copalyl diphosphate synthase (CPS), ent-kaurene synthase (KS), ent- kaurene oxidase (KO), and ent-kaurenoic acid oxidases (KAO), form small families that are composed of one or two homeologous gene pairs. Late GA biosynthesis, including GA 20-oxidase (GA20ox), and GA 3-oxidase (GA3ox) and GA catabolism GA 2-oxidase (GA2ox) enzymes, and signaling proteins, including GIBBERELLIN INSENSITIVE DWARF1 (GID1) receptors, GID2/SLEEPY(SLY) F-box proteins, and DELLA repressors are encoded by larger families of 5-15 pairs of homeologs (i. e., 10-30 genes). Overall, families of GA metabolism and signaling genes are well maintained among ten Brassica napus accessions regarding the number of family members. However, this is achieved by the equilibrium between active gene duplications and losses. The BnaGA20ox, BnaGA2ox, and BnaGID2/SLY2 families show remarkable diversification within accessions. The BnaGA20ox and BnaGA2ox families show gene duplications and losses more frequently than other families, and a number of truncated genes are annotated for these genes. Truncated GA2ox were frequently found in Brassica oleracea, suggesting that active diversification of BnaGA2ox is derived from the progenitor. Gene duplications and losses are frequently observed for the pairs of homeologs. Moreover, homeolog exchanges are enriched for family members showing gene duplications and/ or losses. These suggest that homeolog interaction is associated with gene family maintenance. Transcriptomes at various developmental stages in the ZS11 accession showed that most homeolog pairs are co-expressed, and gene expression divergence between homeologous genes is mostly tissue-specific. These analyses suggest that genomics-based breeding is suitable to effectively select GA-related traits from a limited source of genetic diversity in young polyploid B. napus.

Abscisic acid (ABA) is a plant hormone that regulates numerous plant processes, including plant growth, development, and stress physiology. ABA plays an important role in enhancing plant stress tolerance. This involves the ABA-mediated control of gene expression to increase antioxidant activities for scavenging reactive oxygen species (ROS). ABA is a fragile molecule that is rapidly isomerized by ultraviolet (UV) light and catabolized in plants. This makes it challenging to apply as a plant growth substance. ABA analogs are synthetic derivatives of ABA that alter ABA's functions to modulate plant growth and stress physiology. Modifying functional group(s) in ABA analogs alters the potency, selectivity to receptors, and mode of action (i.e., either agonists or antagonists). Despite current advances in developing ABA analogs with high affinity to ABA receptors, it remains under investigation for its persistence in plants. The persistence of ABA analogs depends on their tolerance to catabolic and xenobiotic enzymes and light. Accumulated studies have demonstrated that the persistence of ABA analogs impacts the potency of its effect in plants. Thus, evaluating the persistence of these chemicals is a possible scheme for a better prediction of their functionality and potency in plants. Moreover, optimizing chemical administration protocols and biochemical characterization is also critical in validating the function of chemicals. Lastly, the development of chemical and genetic controls is required to acquire the stress tolerance of plants for multiple different uses.

The plant hormone (S)-abscisic acid (ABA) is a signalling molecule found in all plants that triggers plants' responses to environmental stressors such as heat, drought, and salinity. Metabolism-resistant ABA analogs that confer longer lasting effects require multi-step syntheses and high costs that prevent their application in crop protection. To solve this issue, we have developed a two-step, efficient and scalable synthesis of (+)-tetralone ABA from (S)-ABA methyl ester. A challenging three-carbon insertion and a bicyclic ring formation on (S)-ABA methyl ester was achieved through a highly regioselective Knoevenagel condensation, cyclization, and oxidation in one-pot. Further we have studied the biological activity and metabolism of (+)-tetralone ABA in planta and found the analog is hydroxylated similarly to ABA. The biologically active hydroxylated tetralone ABA has greater persistence than 8 '-hydroxy ABA as cyclization to the equivalent of phaseic acid is prevented by the aromatic ring. (+)-tetralone ABA complemented the growth retardation of an Arabidopsis ABA-deficient mutant more effectively than (+)-ABA. Taken together, this new synthesis allows the production of the potent ABA agonist efficiently on an industrial scale.

Nitrogen is the most abundant element in the atmosphere and serves as the foundation block of life, including plants on earth. Unlike carbon fixation through photosynthesis, plants rely heavily on external supports to acquire nitrogen. To this end, plants have adapted various strategies such as forming mutualistic relationships with nitrogen-fixing bacteria and evolving a large regulatory network that includes multiple transporters, sensors, and transcription factors for fine-tuning nitrate sensing and signaling. Nodule Inception (NIN) and NIN-like protein (NLP) are central in this network by executing multiple functions such as initiating and regulating the nodule symbiosis for nitrogen fixation, acting as the intracellular sensor to monitor the nitrate fluctuations in the environment, and activating the transcription of nitrate-responsive genes for optimal nitrogen uptake, assimilation, and usage. The involvement of NLPs in intracellular nitrate binding and early nitrate responses highlight their pivotal role in the primary nitrate response (PNR). Genome-wide reprogramming in response to nitrate by NLP is highly transient and rapid, requiring regulation in a precise and dynamic manner. This review aims to summarize recent progress in the study of NIN/NLP for a better understanding of the molecular basis of their roles and regulations in nitrate sensing and signaling, with the hope of shedding light on increasing biological nitrogen fixation and improving nitrogen use efficiency (NUE) to minimize fertilizer input in agriculture.

Improving biological nitrogen fixation (BNF) in cereal crops is a long-sought objective; however, no successful modification of cereal crops showing increased BNF has been reported. Here, we described a novel approach in which rice plants were modified to increase the production of compounds that stimulated biofilm formation in soil diazotrophic bacteria, promoted bacterial colonization of plant tissues and improved BNF with increased grain yield at limiting soil nitrogen contents. We first used a chemical screening to identify plant-produced compounds that induced biofilm formation in nitrogen-fixing bacteria and demonstrated that apigenin and other flavones induced BNF. We then used CRISPR-based gene editing targeting apigenin breakdown in rice, increasing plant apigenin contents and apigenin root exudation. When grown at limiting soil nitrogen conditions, modified rice plants displayed increased grain yield. Biofilm production also modified the root microbiome structure, favouring the enrichment of diazotrophic bacteria recruitment. Our results support the manipulation of the flavone biosynthetic pathway as a feasible strategy for the induction of biological nitrogen fixation in cereals and a reduction in the use of inorganic nitrogen fertilizers.

Jasmonic acid (JA) signaling plays a pivotal role in plant development and defense. MYC2 is a master transcription factor in JA signaling, and was found to be phosphorylated and negatively regulated by MAP kinase and receptor-like kinase. However, the kinases that positively regulate MYC2 through phosphorylation and promote MYC2-mediated activation of JA response have not been identified. Here, we identified CK2 as a kinase that phosphorylates MYC2 and thus regulates the JA signaling. CK2 holoenzyme can interact with MYC2 using its regulatory subunits and phosphorylate MYC2 at multiple sites with its catalytic subunits. Inhibition of CK2 activity in a dominant-negative plant line, CK2mut, repressed JA response. On the other hand, increasing CK2 activity by overexpression of CKB4, a regulatory subunit gene of CK2, enhanced JA response in a MYC2-dependent manner. Substitution of the Ser and Thr residues at phosphorylation sites of MYC2 by CK2 with Ala impaired MYC2 function in activating JA response. Further investigations evidenced that CK2 facilitated the JA-induced increase of MYC2 binding to the promoters of JA-responsive genes in vivo. Our study demonstrated that CK2 plays a positive role in JA signaling, and reveals a previously undiscovered mechanism that regulates MYC2 function.

We herein explored a smart Fe-HPPy@Au/DOX theranostic agent for CT diagnosis and PTT/chemotherapy/CDT synergistic treatment of cancer. When the Fe-HPPy@Au/DOX theranostic agent entered the tumor, the tumor environment accelerated the trapped Fe ions release to catalyze the production of ˙OH for CDT. NIR irradiation drove the PTT, and at the same time improved the CDT by increasing the production of ˙OH and triggered DOX release for chemotherapy. In addition, the Au nanoparticles on the surface of Fe-HPPy@Au nanocomposites could be used as a CT imaging agent and catalyzer to produce H2O2 for enhanced CDT.

通过野外试验采集稻田田面水和稻田排水径流的水样,研究了胶体磷流失贡献和流失规律。根据实际天气情况,选择1次典型暴雨事件（48 mm/h）和8次不同强度（8,9,19,23,23,32,36,49 mm/h）的降雨事件采集样品。结果表明:（1）田面水和稻田排水中磷素流失形式以颗粒磷为主。在过1μm滤膜的总磷中,胶体磷的流失贡献为21%～73%,超过真溶解磷的流失贡献。（2）降雨后田面水中胶体磷的贡献范围为9%～44%,稻田排水中胶体磷流失贡献范围为10%～16%,金属氧化物胶体、有机质胶体以及金属氧化物—有机质胶体浓度与胶体磷流失贡献呈正向相关性,田面水中皮尔森相关系数分别为0.544,0.635,0.781（p＜0.05）,稻田排水中分别为0.734,0.350,0.747（p＜0.05）,三者结合磷为径流中胶体磷的重要赋存形态。（3）施肥通过影响电导率和离子强度来削减胶体磷的流失贡献,而降雨强度则通过影响pH来增加胶体磷的流失贡献,降雨强度与稻田排水胶体磷流失浓度的线性回归系数R~2=0.75,但两者对田面水和稻田排水中胶体磷流失贡献的影响程度不同。

The reuse of water in agriculture has become more common in water management worldwide. However, there is very limited information about nutrient retention in water reclamation management. In this study, an improved low impact development (LID) practice was constructed to investigate the synergistic effects of three substrates amendment on nitrogen (N) and phosphorus (P) retention under two irrigation modules: spray and drip irrigation. The orthogonal combination of the three substrates was controlled during four leaching events, with polyacrylamide (PAM), peat soil, and straw biochar application rates of 1, 2, and 4 g kg(-1); 5, 10, and 20 g kg(-1); and 10, 20, and 40 g kg(-1), respectively. Results showed that the optimum treatments for N and P were 2 g kg(-1) of PAM; 2 g kg(-1 )of PAM, 10 g kg(-1) of peat soil, and 40 g kg(-1) of straw biochar, respectively. The highest amounts of N and P retention under spray and drip irrigation were 83.12 mg N kg(-1) and 50.09 mg N.kg(-1), and 11.88 mg P.kg(-1) and 7.47 mg P.kg(-1), respectively. The analysis of variance indicated that PAM, biochar, and peat soil affected the retention of leachate, N, and P differently. PAM application could not only improve the water, N, P retention capacity of soil, but also significantly increase the content of > 2 mm water-stable soil aggregate (WSA) (p < 0.05), and there is an advisable linear relation between N, P retention and the content of > 2 mm WSA (R-2 = 0.79, 0.67, respectively). Overall, this study concludes that a combined application of PAM and biochar could reduce P loss and increase the > 2 mm WSA under leaching condition.

Multifunctional nanocomposites for image-guided cancer therapy are highly desired in clinical application. Herein, a novel theranostic agent based on gold and ferroferric oxide nanoparticles coating polypyrrole particles (PPy@Fe3O4/Au nanocomposites) for computed tomography (CT) and magnetic resonance (MR) imaging guided photothermal therapy was successfully assembled by a very facile electrostatic adsorption method. PPy@Fe3O4/Au nanocomposites exhibit good biocompatibility in vitro and in vivo. Because of high r2 relaxivity of Fe3O4 and high X-ray attenuation ability of Au, the PPy@Fe3O4/Au nanocomposites exhibited desirable CT and MR imaging performance, which provide more comprehensive and accurate diagnostic information. Moreover, PPy@Fe3O4/Au nanocomposites can efficiently kill cancer cells by hyperthermia with the guiding of CT and MR imaging, even completely ablate tumours. Hence, the electrostatic adsorption assembled PPy@Fe3O4/Au nanocomposites have great potential in clinical application for diagnosing and treating tumour in the future.

Plant growth and development are dependent on chloroplast development and function. Constitutive high level accumulation of a chloroplast stress signal, 3'-phosphoadenosine-5'-phosphate (PAP), confers drought tolerance to plants, but slow downs and alters plant growth and development. PAP, a by-product of sulfur metabolism, is maintained at very low levels by the SAL1 phosphatase during vegetative growth of Arabidopsis and accumulates in rosettes during drought and excess light. Eight independent forward genetic screens in Arabidopsis identified SAL1 as the regulator of multiple phenotypes related to stress responses, hormonal signaling and/or perception. In this perspective article, we collate all the sal1 phenotypes published in the past two decades, and distill the different pathways affected. Our meta-analysis of publicly available sal1 microarray data coupled to preliminary hormonal treatment and profiling results on sal1 indicate that homeostasis and responses to multiple hormones in sal1 are altered during rosette growth, suggesting a potential connection between SAL1-PAP stress retrograde pathway and hormonal signaling. We propose the SAL1-PAP pathway as a case study for integrating chloroplast retrograde signaling, light signaling and hormonal signaling in plant growth and morphogenesis.

Here we report a draft genome sequence of Azoarcus communis SWub3, a nitrogen-fixing bacterium isolated from root tissues of Kallar grass in Pakistan.

Nitrate promotes seed germination at low concentrations in many plant species, and functions as both a nutrient and a signal. As a nutrient, it is assimilated via nitrite to ammonium, which is then incorporated into amino acids. Nitrate reductase (NR) catalyses the reduction of nitrate to nitrite, the committed step in the assimilation. Seed sensitivity to nitrate is affected by other environmental factors, such as light and after-ripening, and by genotypes. Mode of nitrate action in seed germination has been well documented in Arabidopsis thaliana and the hedge mustard Sisymbrium officinale. In these species nitrate promotes seed germination independent of its assimilation by NR, suggesting that it acts as a signal to stimulate germination. In Arabidopsis, maternally applied nitrate affects the degree of primary dormancy in both wild-type and mutants defective in NR. This indicates that nitrate acts not only during germination, but also during seed development to negatively regulate primary dormancy. Functional genomics studies in Arabidopsis have revealed that nitrate elicits downstream events similar to other germination stimulators, such as after-ripening, light and stratification, suggesting that these distinct environmental signals share the same target(s). In Arabidopsis, the NIN-like protein 8 (NLP8) transcription factor, which acts downstream of nitrate signalling, induces nitrate-dependent gene expression. In particular, a gene encoding the abscisic acid (ABA) catabolic enzyme CYP707A2 is directly regulated by NLP8. This regulation triggers a nitrate-induced ABA decrease that permits seed germination. This review article summarizes an update of our current understanding of the regulation of seed dormancy and germination by nitrate.

Plants perceive and integrate information from the environment to time critical transitions in their life cycle. Some mechanisms underlying this quantitative signal processing have been described, whereas others await discovery. Seeds have evolved a mechanism to integrate environmental information by regulating the abundance of the antagonistically acting hormones abscisic acid (ABA) and gibberellin (GA). Here, we show that hormone metabolic interactions and their feedbacks are sufficient to create a bistable developmental fate switch in Arabidopsis seeds. A digital single-cell atlas mapping the distribution of hormone metabolic and response components revealed their enrichment within the embryonic radicle, identifying the presence of a decision-making center within dormant seeds. The responses to both GA and ABA were found to occur within distinct cell types, suggesting cross-talk occurs at the level of hormone transport between these signaling centers. We describe theoretically, and demonstrate experimentally, that this spatial separation within the decision-making center is required to process variable temperature inputs from the environment to promote the breaking of dormancy. In contrast to other noise-filtering systems, including human neurons, the functional role of this spatial embedding is to leverage variability in temperature to transduce a fate-switching signal within this biological system. Fluctuating inputs therefore act as an instructive signal for seeds, enhancing the accuracy with which plants are established in ecosystems, and distributed computation within the radicle underlies this signal integration mechanism.

Seeds respond to multiple different environmental stimuli that regulate germination. Nitrate stimulates germination in many plants but how it does so remains unclear. Here we show that the Arabidopsis NIN-like protein 8 (NLP8) is essential for nitrate-promoted seed germination. Seed germination in nlp8 loss-of-function mutants does not respond to nitrate. NLP8 functions even in a nitrate reductase-deficient mutant background, and the requirement for NLP8 is conserved among Arabidopsis accessions. NLP8 reduces abscisic acid levels in a nitrate-dependent manner and directly binds to the promoter of CYP707A2, encoding an abscisic acid catabolic enzyme. Genetic analysis shows that NLP8-mediated promotion of seed germination by nitrate requires CYP707A2. Finally, we show that NLP8 localizes to nuclei and unlike NLP7, does not appear to be activated by nitrate-dependent nuclear retention of NLP7, suggesting that seeds have a unique mechanism for nitrate signalling.

In angiosperms, a double fertilization event initiates the development of two distinct structures, the embryo and endosperm. The endosperm plays an important role in supporting embryonic growth by supplying nutrients, protecting the embryo and controlling embryo growth by acting as a mechanical barrier during seed development and germination. Its structure and function in the mature dry seed is divergent and specialized among different plant species. A subset of endospermic tissues are composed of living cells even after seed maturation, and play an active role in the regulation of seed germination. Transcriptome analysis has provided new insights into the regulatory functions of the endosperm during seed germination. It is well known that the embryo secretes signals to the endosperm to induce the degradation of the seed reserve and to promote endosperm weakening during germination. Recent advances in seed biology have shown that the endosperm is capable of sensing environmental signals, and can produce and secrete signals to regulate the growth of the embryo. Thus, germination is a systemic response that involves bidirectional interactions between the embryo and endosperm.

In angiosperms, a double fertilization event initiates the development of two distinct structures, the embryo and endosperm. The endosperm plays an important role in supporting embryonic growth by supplying nutrients, protecting the embryo and controlling embryo growth by acting as a mechanical barrier during seed development and germination. Its structure and function in the mature dry seed is divergent and specialized among different plant species. A subset of endospermic tissues are composed of living cells even after seed maturation, and play an active role in the regulation of seed germination. Transcriptome analysis has provided new insights into the regulatory functions of the endosperm during seed germination. It is well known that the embryo secretes signals to the endosperm to induce the degradation of the seed reserve and to promote endosperm weakening during germination. Recent advances in seed biology have shown that the endosperm is capable of sensing environmental signals, and can produce and secrete signals to regulate the growth of the embryo. Thus, germination is a systemic response that involves bidirectional interactions between the embryo and endosperm.