Zinc finger protein LjRSDL regulates arbuscule degeneration of arbuscular mycorrhizal fungi in Lotus japonicus.

In arbuscular mycorrhizal (AM) symbiosis, appropriate regulation of the formation, maintenance, and degeneration of the arbuscule are essential for plants and fungi. In this study, we identified a Cysteine-2/Histidine-2 zinc finger protein (C2H2-ZFP)-encoding gene in Lotus japonicus named Regulator of Symbiosome Differentiation-Like (LjRSDL) that is required for arbuscule degeneration. Evolutionary analysis showed that homologs of LjRSDL exist in mycorrhizal flowering plants. We obtained ProLjRSDL::GUS transgenic hairy roots and showed that LjRSDL was strongly upregulated upon AM colonization, particularly at 18 days post AM fungi inoculation and specifically expressed in arbuscular-containing cells. The mycorrhization rate increased in the ljrsdl mutant but decreased in LjRSDL overexpressed L. japonicus. Interestingly, we observed higher proportions of large arbuscule in the ljrsdl mutant but lower proportions of larger arbuscule in LjRSDL overexpressing plants. Transcriptome analyses indicated that genes involved in arbuscule degeneration were significantly changed upon the dysregulation of LjRSDL and that LjRSDL-dependent regulation in AM symbiosis is mainly via the hormone signal transduction pathway. LjRSDL, therefore, represents a C2H2-ZFP that negatively regulates AM symbiosis. Our study provides insight into understanding plant-AM fungal communication and AM symbiosis development.

Genome-wide transcriptome and gene family analysis reveal candidate genes associated with potassium uptake of maize colonized by arbuscular mycorrhizal fungi.

BACKGROUND: Potassium (K) is an essential nutrient for plant growth and development. Maize (Zea mays) is a widely planted crops in the world and requires a huge amount of K fertilizer. Arbuscular mycorrhizal fungi (AMF) are closely related to the K uptake of maize. Genetic improvement of maize K utilization efficiency will require elucidating the molecular mechanisms of maize K uptake through the mycorrhizal pathway. Here, we employed transcriptome and gene family analysis to elucidate the mechanism influencing the K uptake and utilization efficiency of mycorrhizal maize. METHODS AND RESULTS: The transcriptomes of maize were studied with and without AMF inoculation and under different K conditions. AM symbiosis increased the K concentration and dry weight of maize plants. RNA sequencing revealed that genes associated with the activity of the apoplast and nutrient reservoir were significantly enriched in mycorrhizal roots under low-K conditions but not under high-K conditions. Weighted gene correlation network analysis revealed that three modules were strongly correlated with K content. Twenty-one hub genes enriched in pathways associated with glycerophospholipid metabolism, glycerolipid metabolism, starch and sucrose metabolism, and anthocyanin biosynthesis were further identified. In general, these hub genes were upregulated in AMF-colonized roots under low-K conditions. Additionally, the members of 14 gene families associated with K obtain were identified (ARF: 38, ILK: 4, RBOH: 12, RUPO: 20, MAPKK: 89, CBL: 14, CIPK: 44, CPK: 40, PIN: 10, MYB: 174, NPF: 79, KT: 19, HAK/HKT/KUP: 38, and CPA: 8) from maize. The transcript levels of these genes showed that 92 genes (ARF:6, CBL:5, CIPK:13, CPK:2, HAK/HKT/KUP:7, PIN:2, MYB:26, NPF:16, RBOH:1, MAPKK:12 and RUPO:2) were upregulated with AM symbiosis under low-K conditions. CONCLUSIONS: This study indicated that AMF increase the resistance of maize to low-K stress by regulating K uptake at the gene transcription level. Our findings provide a genome-level resource for the functional assignment of genes regulated by K treatment and AM symbiosis in K uptake-related gene families in maize. This may contribute to elucidate the molecular mechanisms of maize response to low K stress with AMF inoculation, and provided a theoretical basis for AMF application in the crop field.

Genome-wide identification of nitrate transporter 1/peptide transporter family (NPF) induced by arbuscular mycorrhiza in the maize genome.

The Transporter 1/Peptide Transporter Family (NPF) is essential for the uptake and transport of nitrate nitrogen. Significant increases in nitrogen have been increasingly reported for many mycorrhizal plants, but there are few reports on maize. Here, we have identified the maize NPF family and screened for arbuscular mycorrhiza fungi (AMF) induced NPFs. In this study, a systematic analysis of the maize NPF gene family was performed. A total of 82 NPF genes were identified in maize. ZmNPF4.5 was strongly induced by AMF in both low and high nitrogen. Lotus japonicus hairy root-induced transformation experiments showed that ZmNPF4.5 promoter-driven GUS activity was restricted to cells containing tufts. Yeast backfill experiments indicate that ZmNPF4.5 functions in nitrate uptake. Therefore, we speculate that ZmNPF4.5 is a key gene for nitrate-nitrogen uptake in maize through the mycorrhizal pathway. This is a reference value for further exploring the acquisition of nitrate-nitrogen by maize through AMF pathway. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-024-01464-3.

The mechanism of promoting rhizosphere nutrient turnover for arbuscular mycorrhizal fungi attributes to recruited functional bacterial assembly.

Symbiosis with arbuscular mycorrhizal (AM) fungi improves plant nutrient capture from the soil, yet there is limited knowledge about the diversity, structure, functioning, and assembly processes of AM fungi-related microbial communities. Here, 16S rRNA gene sequencing and metagenomic sequencing were used to detect bacteria in the rhizosphere of Lotus japonicus inoculated with and without AM fungi, and the L. japonicus mutant ljcbx (defective in symbiosis) inoculated with AM fungi in southern grassland soil. Our results show that AM symbiosis significantly increased bacterial diversity and promoted deterministic processes of bacterial community construction, suggesting that mycorrhizal symbiosis resulted in the directional enrichment of bacterial communities. AM fungi promoted the enrichment of nine bacteria, including Ohtaekwangia, Niastella, Gemmatimonas, Devosia, Sphingomonas, Novosphingobium, Opitutus, Lysobacter, Brevundimonas, which are positively correlated with NPK-related parameters. Through a functional identification experiment, we found that six of these genera, including Brevundimonas, Lysobacter, Ohtaekwangia, Sphingomonas, Devosia, and Gemmatimonas, demonstrated the ability to mineralize organophosphate and dissolve inorganic phosphorus, nitrogen, and potassium. Our study revealed that AM fungi can regulate rhizosphere bacterial community assembly and attract specific rhizosphere bacteria to promote soil nutrient turnover in southern grasslands.

Construction of a virtual simulation laboratory for gene detection.

OBJECTIVE: The current paper aims to discuss the development of a virtual simulation experiment teaching system and review its effectiveness in improving the teaching of clinical skills to college medical students. METHODS: Collaborators used 3D Studio Max, Unity 3D and Visual Studio to develop four modules: laboratory thinking training, biosafety training, gene testing and experimental assessment. Teaching was conducted and a virtual software program was used for evaluation of the students. RESULTS: The laboratory safety training system, virtual gene experiment system and experimental assessment system were developed. The results of the questionnaire survey show that the software provides good interactivity and guidance. The interest of medical students in study is improved and they received training in clinical experimental thinking. Student evaluation assists their scientific research practice, and can improve the awareness of biosafety. CONCLUSION: The virtual simulation experiment teaching system, when applied in the teaching of undergraduate and postgraduate experiment courses, can bring about rapid improvements in the following areas: biosafety awareness, interest in learning about experiments and experimental skills, clinical experimental thinking, and comprehensive experimental ability.

Surface-enhanced Raman scattering from an electromagnetic induced transparency substrate for the determination of hepatocellular carcinoma.

Surface-enhanced Raman scattering (SERS) is a powerful analytical method that is especially suitable for the detection of protein molecules. Detection sensitivity of SERS is directly related to the enhancement factor of the substrate, which is dependent on the strength of a local surface electric field generated by surface plasmonic resonance from substrate. In this study, an electromagnetic induced transparency like (EIT-like) metamaterial was used as the SERS substrate. The corresponding plasmonic resonance structure not only produces stronger optical near field but also reduces the spectral line broadening due to radiation damping. This is very beneficial for SERS process, which is strongly dependent on electric field intensity, to improve the sensitivity of SERS detection. Compared with the single resonance mode substrate, the enhancement factor for SERS with the double-mode substrate was increased by an order of magnitude. The obtained EIT-like substrate was used as a SERS-active substrate to detect Lens culinaris agglutinin (LCA)-reactive fraction of AFP (AFP-L3), a hepatocellular carcinoma (HCC)-specific maker. Experimental results are in good agreement with the clinical diagnosis, which demonstrates the potential application of metamaterials in the SERS-based diagnosis and biosensing.

Determination of human COVID-19 total antibodies in serum using a time-resolved fluorescence immunoassay.

Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is spreading rapidly around the world. Antibody detection plays an important role in the diagnosis of COVID-19. Here, we established a new time-resolved fluorescence immunoassay (TRFIA) to determine COVID-19 total antibodies. A double-antigen sandwich TRFIA was optimized and established: recombinant nucleocapsid phosphoprotein (N protein) and spike protein (S protein) of COVID-19 immobilized on 96-well plates captured human COVID-19 antibodies and then banded together with the N/S proteins labeled with europium(III) (Eu3+ ) chelates, and finally, time-resolved fluorometry was used to measure the fluorescence values. We successfully established a TRFIA method for the detection of human COVID-19 total antibodies, and the cutoff value was 2.02. There was no cross-reactivity with the negative reference of the National Reference Panel for IgM and IgG antibodies to COVID-19. The CV of the precision assay was 3.19%, and the assay could be stored stably for 15 days at 37°C. Compared with that of the colloidal gold method and chemiluminescence method, the sensitivity of the TRFIA method was higher, and the false positive/negative rate was lower. This established TRFIA has high sensitivity, accuracy, and specificity, which indicates that this method provides a new detection method for the high-throughput routine diagnosis of COVID-19.

A novel SCARECROW-LIKE3 transcription factor LjGRAS36 in Lotus japonicus regulates the development of arbuscular mycorrhizal symbiosis.

The symbiosis with arbuscular mycorrhizal (AM) fungi improves plants' nutrient uptake. During this process, transcription factors have been highlighted to play crucial roles. Members of the GRAS transcription factor gene family have been reported involved in AM symbiosis, but little is known about SCARECROW-LIKE3 (SCL3) genes belonging to this family in Lotus japonicus. In this study, 67 LjGRAS genes were identified from the L. japonicus genome, seven of which were clustered in the SCL3 group. Three of the seven LjGRAS genes expression levels were upregulated by AM fungal inoculation, and our biochemical results showed that the expression of LjGRAS36 was specifically induced by AM colonization. Functional loss of LjGRAS36 in mutant ljgras36 plants exhibited a significantly reduced mycorrhizal colonization rate and arbuscular size. Transcriptome analysis showed a deficiency of LjGRAS36 led to the dysregulation of the gibberellic acid signal pathway associated with AM symbiosis. Together, this study provides important insights for understanding the important potential function of SCL3 genes in regulating AM symbiotic development. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01161-z.

Overexpression of a phosphate transporter gene ZmPt9 from maize influences growth of transgenic Arabidopsis thaliana.

Phosphate transporters (PHTs) are well-known for their roles in phosphate uptake in plants. However, their actions in imparting plant growth in plants are still not so clear. In our previous study, we observed that maize PHT1 gene ZmPt9 plays a significant role in phosphate uptake. In this study, we further characterized ZmPt9 in response to low phosphate condition through ZmPt9 promoter inductive analysis by GUS staining and quantification. To elucidate the function of ZmPt9, we generated overexpression plant in Arabidopsis. ZmPt9 overexpressing Arabidopsis plants conferred small leaves and early flowering compared with the wild-type plants. In addition, ZmPt9 can complement the late flowering phenotype of Arabidopsis mutant pht1;2. The qRT-PCR analysis revealed that overexpression of ZmPt9 in Arabidopsis changed expression levels of some flowering-related genes. Further expressed detection of hormone related genes revealed that GA and auxin maybe the main determinant for growth influences of ZmPt9. In conclusion, these results suggest that apart from phosphate transport activity, ZmPt9 can be further exploited for improving crops growth.

Highly stable organic photothermal agent based on near-infrared-II fluorophores for tumor treatment.

BACKGROUND: The aim to develop a highly stable near-infrared (NIR) photoinduced tumor therapy agent stems from its considerable potential for biological application. Due to its long wavelength, biological imaging exhibits a high signal-to-background ratio, deep tissue penetration and maximum permissible light power, which can minimize damage to an organism during photoinduced tumor therapy. RESULTS: A class of stable NIR-II fluorophores (NIR998, NIR1028, NIR980, NIR1030, and NIR1028-S) based on aza-boron-dipyrromethene (aza-BODIPY) dyes with donor-acceptor-donor structures have been rationally designed and synthesized by harnessing the steric relaxation effect and intramolecular photoinduced electron transfer (IPET). These fluorophores exhibit an intense range of NIR-II emission, large Stokes shift (≥ 100 nm), excellent photothermal conversion performance, and superior stability against photobleaching. Among the NIR-II fluorophores, NIR998 possesses better NIR-II emission and photothermal conversion performance. NIR998 nanoparticles (NIR998 NPs) can be encapsulated by liposomes. NIR998 NPs show superior stability in the presence of light, heat, and reactive oxygen nitrogen species than that of indocyanine green NPs, as well as a higher photothermal conversion ability (η = 50.5%) compared to other photothermal agents. Finally, under the guidance of photothermal imaging, NIR998 NPs have been proven to effectively eliminate tumors via their excellent photothermal conversion performance while presenting negligible cytotoxicity. CONCLUSIONS: Utilizing IPET and the steric relaxation effect can effectively induce NIR-II emission of aza-BODIPY dyes. Stable NIR998 NPs have excellent photothermal conversion performance and negligible dark cytotoxicity, so they have the potential to act as photothermal agents in biological applications.

Rational Design of Near-Infrared-Absorbing Pt(II)-Chelated Azadipyrromethene Dyes as a New Generation of Photosensitizers for Synergistic Phototherapy.

Pt(II) photosensitizers are emerging as novel Pt anticancer agents for cancer photodynamic therapy (PDT) to avoid uncontrollable toxicity of cisplatin. However, the application of Pt(II) photosensitizers is limited by tumor hypoxia and the poor penetration depth of excitation light. To overcome these drawbacks, exploiting the next generation of Pt anticancer agents is of urgent need. According to theoretical calculations, novel near-infrared (NIR)-absorbing Pt(II)-chelated azadipyrromethene dyes (PtDP-X, where X = N, C, and S) were designed. Importantly, spin-orbit coupling of the Pt atom could promote the intersystem crossing of a singlet-to-triplet transition for converting oxygen to singlet oxygen (1O2), and the azadipyrromethene skeleton could provide a strong photothermal effect. As expected, PtDP-X exhibited intense NIR absorption and synergistic PDT and photothermal effects with low dark cytotoxicity. Furthermore, water-soluble and biocompatible PtDP-N nanoparticles (PtDP-N NPs) were prepared that achieved effective tumor cell elimination with low side effects under 730 nm light irradiation in vitro and in vivo. This pioneering work could push the exploitation of NIR-absorbing metal-chelated azadipyrromethene dyes, so as to promote the positive evolution of phototherapy agents.

APETALA 2 transcription factor CBX1 is a regulator of mycorrhizal symbiosis and growth of Lotus japonicus.

KEY MESSAGE: An AP2 family gene CBX1 is involved in mycorrhizal symbiosis and growth of Lotus japonicus. APETALA 2 (AP2) transcriptional regulator is highly conserved in plants. CBX1 from Lotus japonicus is a member of AP2 family. AMF (Arbuscular mycorrhizal fungi) inoculation experiment demonstrated that expression of CBX1 was significantly induced by AMF. Further promoter analysis showed that the - 764 to - 498 bp region of the CBX1 promoter containing CTTC motif is the AMF responsive region. Functional analysis of cbx1 mutant suggested CBX1 is critical for mycorrhizal symbiosis, especially for arbuscule formation. Moreover, under noncolonized condition, overexpression of CBX1 reduced the root length of L. japonicus but increased the size of root system and shoot length, whereas cbx1 mutant reduced the root size and shoot length, but not effect on root length. In addition, cbx1 altered activity of monolignol biosynthetic gene and increased lignin levels. Collectively, these data indicated that CBX1 is a positive regulator of symbiotic activity and plays roles in the growth of L. japonicus.

DNA Repair Gene ZmRAD51A Improves Rice and Arabidopsis Resistance to Disease.

RAD51 (DNA repair gene) family genes play ubiquitous roles in immune response among species from plants to mammals. In this study, we cloned the ZmRAD51A gene (a member of RAD51) in maize and generated ZmRAD51A overexpression (ZmRAD51A-OE) in rice, tobacco, and Arabidopsis. The expression level of ZmRAD51A was remarkably induced by salicylic acid (SA) application in maize, and the transient overexpression of ZmRAD51A in tobacco induced a hypersensitive response. The disease resistance was significantly enhanced in ZmRAD51A- OE (overexpressing) plants, triggering an increased expression of defense-related genes. High-performance liquid chromatography (HPLC) analysis showed that, compared to control lines, ZmRAD51A-OE in rice plants resulted in higher SA levels, and conferred rice plants resistance to Magnaporthe oryzae. Moreover, the ZmRAD51A-OE Arabidopsis plants displayed increased resistance to Pseudomonas syringae pv. tomato DC3000 when compared to wild types. Together, our results provide the evidence that, for the first time, the maize DNA repair gene ZmRAD51A plays an important role in in disease resistance.

Identification of Long Non-Coding RNAs and the Regulatory Network Responsive to Arbuscular Mycorrhizal Fungi Colonization in Maize Roots.

Recently, long noncoding RNAs (lncRNAs) have emerged as vital regulators of many biological processes in animals and plants. However, to our knowledge no investigations on plant lncRNAs which respond to arbuscular mycorrhizal (AM) fungi have been reported thus far. In this study, maize roots colonized with AM fungus were analyzed by strand-specific RNA-Seq to identify AM fungi-responsive lncRNAs and construct an associated regulatory network. A total of 1837 differentially expressed protein coding genes (DEGs) were identified from maize roots with Rhizophagus irregularis inoculation. Many AM fungi-responsive genes were homologs to MtPt4, STR, STR2, MtFatM, and enriched pathways such as fatty acid biosynthesis, response to phosphate starvation, and nitrogen metabolism are consistent with previous studies. In total, 5941 lncRNAs were identified, of which more than 3000 were new. Of those, 63 lncRNAs were differentially expressed. The putative target genes of differentially expressed lncRNAs (DELs) were mainly related to phosphate ion transmembrane transport, cellular response to potassium ion starvation, and lipid catabolic processes. Regulatory network analysis showed that DELs might be involved in the regulation of bidirectional nutrient exchange between plant and AM fungi as mimicry of microRNA targets. The results of this study can broaden our knowledge on the interaction between plant and AM fungi.

Identification and Functional Characterization of a Maize Phosphate Transporter Induced by Mycorrhiza Formation.

Phosphorus (P) is an essential macronutrient for plant life, although it is frequently not readily available to crops. Arbuscular mycorrhiza fungi (AMF) can improve plant P levels by inducing the expression of some phosphate (Pi) transporters. Symbiotic Pi uptake by Pi transporters is crucial for AMF colonization and arbuscule dynamics. However, the functions of mycorrhiza-inducible maize Pi transporters are largely unclear. We focused on the interaction between the Pi concentration and AMF colonization in maize, and detecting the induction of a Pi transporter. We investigated AMF colonization and arbuscular development in maize under high and low Pi environments. Low Pi increased AMF colonization and promoted arbuscular development. Further measurement of P concentration showed that AMF significantly improved the maize P status under low Pi conditions. Here, we identified the Pi transporter gene, ZmPt9, which was induced by mycorrhiza formation. In addition, ZmPt9-overexpressing roots were difficult to colonize by AMF. Pi response analysis showed that ZmPt9 complements a yeast mutant defective in Pi transporter activity and improves the P concentration in rice. Together, these data indicated that ZmPt9 is a mycorrhiza-inducible Pi transporter gene involved in Pi uptake.

Maize similar to RCD1 gene induced by salt enhances Arabidopsis thaliana abiotic stress resistance.

Plant SRO (SIMILAR TO RCD-ONE) proteins play important roles in regulating oxidation and metal ion metabolism. Numbers of SRO proteins have been functional identified in Arabidopsis and rice, but little is known in maize. In this study, we identified a salt induced SRO gene, ZmSRO1b, from maize and analyzed its characteristics. ZmSRO1b expressed mainly in leaf tissues. The ZmSRO1b is encoded by 595 amino acid residues and shared conserved protein models with AtRCD1 and AtSRO1 from Arabidopsis. Promoter-elements analysis showed ZmSRO1b promoter harbored salt and metal stress responsive elements, DRE, GT-like and MRE. Further promoter inductive analysis by GUS staining and quantification confirmed that ZmSRO1b promoter was induced by salt and cadmium (Cd). Methylviologen (MV) simulated oxidative stress showed ZmSRO1b promoter was also induced by MV. Overexpression of ZmSRO1b in Arabidopsis plants showed increased resistance to salt, Cd and oxidative stress. Our results for the first time experimentally validate the function of ZmSRO1b and contribute to the better understanding of SRO genes across different plant species.

Mutations within the major hydrophilic region (MHR) of Hepatitis B virus from individuals with simultaneous HBsAg and anti-HBs in Guangzhou, Southern China.

The mechanism of the coexistence of HBsAg and anti-HBs is still unclear. This study investigated the variations located in the major hydrophilic region (MHR) of HBV from individuals with simultaneous HBsAg and anti-HBs in Guangzhou, southern China. Among 4455 samples analyzed, 179 (4.02%) patients were discovered with both HBsAg and anti-HBs. Finally, 44 individuals with concurrent HBsAg and anti-HBs (defined as group I), and 88 patients with positive HBsAg and negative anti-HBs (defined as group II, served as control) were enrolled in the study. The number of residue changes per 100 residues within the MHR in group I was 7.1 times more frequent than group II (P < 0.001) and was discovered mostly in the MHR1 (aa99-119) (P < 0.001). Two or more residue changes in the MHR were discovered in 15 patients (34.1%) of group I, but were found in only one (1.1%) patient of group II (P < 0.001). The most common variants in group I were at positions s101Q, s133M, s126T/I, s131T, s145G, s120P, and s129Q. In addition, sQ101 K, sT131N, and sM133L were more frequently discovered in group I with significant difference (P < 0.05). In chronic hepatitis B (CHB) patients, the simultaneous of HBsAg and anti-HBs were accompanied with an increase of MHR variants, and suggested that the HBsAg mutants were selected by naturally acquired anti-HBs during chronic carriage.

Expression of a maize NBS gene ZmNBS42 enhances disease resistance in Arabidopsis.

KEY MESSAGE: Expression of the ZmNBS42 in Arabidopsis plants conferred resistance to bacterial pathogens, providing potential resistance enhancement of maize in further genetic breeding. Nucleotide-binding site (NBS) domain proteins play critical roles in disease resistance. In this study, we isolate a novel NBS gene ZmNBS42 from maize and systematically investigate its function on disease resistance. We find that the expression levels of ZmNBS42 in maize leaf were strikingly increased in response to Bipolaris maydis inoculation and SA treatment. The spatial expression pattern analysis reveals that, during development, ZmNBS42 is ubiquitously highly expressed in maize root, leaf, stem, internode and seed, but lowly expressed in pericarp and embryo. To better understand the roles of ZmNBS42, we overexpressed ZmNBS42 in heterologous systems. Transient overexpression of ZmNBS42 in the leaves of Nicotiana benthamiana induces a hypersensitive response. ZmNBS42 overexpression (ZmNBS42-OE) Arabidopsis plants produced more SA content than Col-0 plants, and increased the expression levels of some defense-responsive genes compared to Col-0 plants. Moreover, the ZmNBS42-OE Arabidopsis plants displayed enhanced resistance against Pseudomonas syringae pathovar tomato DC3000 (Pst DC3000). These results together suggest that ZmNBS42 can serve as an important regulator in disease resistance, thus better understanding of ZmNBS42 would benefit the resistance enhancement in maize breeding programs.

Genome-wide identification and comparative analysis of phosphate starvation-responsive transcription factors in maize and three other gramineous plants.

KEY MESSAGE: The present study identified several important candidate Pi regulation genes of maize and provides a better understanding on the generation of PHR genes in gramineous plants. Plants have evolved adaptive responses to cope with low phosphate (Pi) soils. The previous studies have indicated that phosphate starvation response (PHR) genes play central roles in regulating plant Pi starvation responses. However, the investigation of PHR family in gramineous plants is limited. In this study, we identified 64 PHR genes in four gramineous plants, including maize, rice, sorghum, and brachypodium, and conducted systematical analyses on phylogenetic, structure, collinearity, and expression pattern of these PHR genes. Genome synteny analysis revealed that a number of PHR genes were present in the corresponding syntenic blocks of maize, rice, sorghum, and brachypodium, indicating that large-scale duplication events contributed significantly to the expansion and evolution of PHR genes in these gramineous plants. Gene expression analysis showed that many PHR genes were expressed in various tissues, suggesting that these genes are involved in Pi redistribution and allocation. In addition, the expression levels of PHR genes from maize and rice under low Pi stress conditions revealed that some PHRs may play an important role in Pi starvation response. Our results provided a better understanding on the generation of PHR genes in gramineous plants and identified several important candidate Pi regulation genes of maize.

Identification of Arbuscular Mycorrhiza Fungi Responsive microRNAs and Their Regulatory Network in Maize.

Maize can form symbiotic relationships with arbuscular mycorrhiza (AM) fungus to increase productivity and resistance, but the miRNAs in maize responsible for this process have not been discovered. In this study, 155 known and 28 novel miRNAs were identified by performing high-throughput sequencing of sRNA in maize roots colonized by AM fungi. Similar to the profiles in other AM-capable plants, a large proportion of identified maize miRNAs were 24 nt in length. Fourteen and two miRNAs were significantly down- and up-regulated in response to AM fungus Glomus intraradices inoculation, respectively, suggesting potential roles of these miRNAs in AM symbiosis. Interestingly, 12 of 14 significantly down-regulated known maize miRNAs belong to the miR399 family, which was previously reported to be involved in the interaction between Medicago truncatula and AM fungi. This result indicated that the miR399 family should regulate AM symbiosis conservatively across different plant lineages. Pathway and network analyses showed that the differentially expressed miRNAs might regulate lipid metabolism and phosphate starvation response in maize during the symbiosis process via their target genes. Several members of the miR399 family and the miR397 family should be involved in controlling the fatty acid metabolism and promoting lipid delivering from plants to AM fungi. To the best of our knowledge, this is the first report on miRNAs mediating fatty acids from plant to AM fungi. This study provides insight into the regulatory roles of miRNAs in the symbiosis between plants and AM fungi.