Ectopic Expression of Sugarcane ScAMT1.1 Has the Potential to Improve Ammonium Assimilation and Grain Yield in Transgenic Rice under Low Nitrogen Stress.

In China, nitrogen (N) fertilizer is excessively used in sugarcane planting areas, while the nitrogen use efficiency (NUE) of sugarcane is relatively low. Mining and identifying the key genes in response to low N stress in sugarcane can provide useful gene elements and a theoretical basis for developing sugarcane varieties with high NUE. In our study, RNA-Seq combined with qRT-PCR analysis revealed that the ScAMT1.1 gene responded positively to low N stress, resulting in the stronger low N tolerance and high NUE ability of sugarcane cultivar ROC22. Then, ScAMT1.1 was cloned from sugarcane. The full-length cDNA of the ScAMT1.1 gene is 1868 bp, containing a 1491 bp open reading frame (ORF), and encoding 496 amino acids. ScAMT1.1 belongs to the AMT superfamily and shares 91.57% homologies with AMT1.1 from Oryza sativa. Furthermore, it was stably overexpressed in rice (O. sativa). Under low N treatment, the plant height and the fresh weight of the ScAMT1.1-overexpressed transgenic rice were 36.48% and 51.55% higher than that of the wild-type, respectively. Both the activity of ammonium assimilation key enzymes GS and GDH, and the expression level of ammonium assimilation key genes, including GS1.1, GS1.2, GDH, Fd-GOGAT, and NADH-GOGAT2 in the transgenic plants, were significantly higher than that of the wild-type. The grain number and grain yield per plant in the transgenic rice were 6.44% and 9.52% higher than that of the wild-type in the pot experiments, respectively. Taken together, the sugarcane ScAMT1.1 gene has the potential to improve ammonium assimilation ability and the yield of transgenic rice under low N fertilizer conditions. This study provided an important functional gene for improving sugarcane varieties with high NUE.

Schottky barrier effect on plasmon-induced charge transfer.

Plasmon-induced charge transfer causes electron-hole spatial separation at the metal-semiconductor interface, which plays a key role in photocatalytic and photovoltaic applications. The Schottky barrier formed at the metal-semiconductor interface can modify the hot carrier dynamics. Taking the Ag-TiO2 system as an example, we have investigated plasmon-induced charge transfer at the Schottky junction using quantum mechanical simulations. We find that the Schottky barrier induced by n-type doping enhances the electron transfer and that induced by p-type doping enhances the hole transfer, which is attributed to the shift of the Fermi energy and the band bending of the Schottky junction at the interface. The Schottky barrier also modifies the layer distribution of hot carriers. In particular, for the system with a large band bending, there exists electron-hole spatial separation inside the TiO2 substrate. Our results reveal the mechanism and dynamics of charge transfer at the Schottky junction, and pave the way for manipulating plasmon-assisted photocatalytic and photovoltaic applications.

Identification of Low-Nitrogen-Related miRNAs and Their Target Genes in Sugarcane and the Role of miR156 in Nitrogen Assimilation.

Chemical nitrogen (N) fertilizer is widely used in sugarcane production, especially in China and India. Understanding the molecular mechanisms and mining miRNAs and their target genes associated with nitrogen use efficiency (NUE) in sugarcane can aid in developing the N-efficient varieties, and thus is beneficial to reduce N fertilizer application. In this study, the root miRNA database of N-efficient sugarcane variety ROC22 under low N stress (0.3 mM NH4NO3) for 3 h was constructed, along with their transcriptome-rearranged data. KEGG analysis indicated that those candidate target genes, corresponding to differentially expressed miRNAs, were mainly enriched in N metabolism, amino acid metabolism, carbohydrate metabolism, photosynthesis, and hormone signal transduction pathways. It was found that under low N stress for 0-24 h, there was a negative correlation between miR168 and SPX, along with miR396 and acnA. Furthermore, the expression of miR156 in the roots of ROC22 was significantly up-regulated under low N treatment. Compared with the wild-type, the Arabidopsis plants overexpressing sugarcane miR156 exhibited significantly improved length and surface area of roots, while the expression of one NO3- transporter gene NRT1.1, three N assimilation key genes (NR1, NIR1, and GS), and the activity of two N assimilation key enzymes (NR and GS) were up-regulated under low N treatment. It can be reasonably deduced that sugarcane miR156 can enhance the nitrogen assimilation ability of the overexpressed Arabidopsis plants under low N application, and thus has a potential ability for improving sugarcane NUE. The present study should be helpful for understanding the molecular regulatory network in the N-efficient sugarcane genotype responding to low N stress and could provide the candidate miRNAs with a potential function in improving sugarcane NUE.

Plasmonic Photocatalysis with Nonthermalized Hot Carriers.

Hot carriers generated by plasmonic damping have been suggested to promote photocatalysis, yet it remains unclear how the nonthermalized hot carriers dynamically activate and promote the energy transfer processes. Here, we present an Anderson-Newns model to describe the vibrational excitation and bond dissociation induced by plasmonic hot carriers. The nonthermal distribution of the hot carriers generated by plasmon damping is accounted for on equal footing with thermal carriers at a given temperature in the electron-molecule scattering. We found that the nonthermal electrons in the high energy region can, albeit in much smaller populations, provide an efficient and dominant channel for photodissociation especially in the low-temperature and quantum plasmon regime. Our model captures the wavelength dependence and reproduces the enhancement factors observed by experiments for oxygen dissociation on silver nanoparticles. It also paves a way to harvesting nonthermal plasmonic energy for photocatalysis in the quantum regime.

Characterization and Manipulation of Intervalley Scattering Induced by an Individual Monovacancy in Graphene.

Intervalley scattering involves microscopic processes that electrons are scattered by atomic-scale defects on the nanoscale. Although central to our understanding of electronic properties of materials, direct characterization and manipulation of range and strength of the intervalley scattering induced by an individual atomic defect have so far been elusive. Using scanning tunneling microscope, we visualize and control intervalley scattering from an individual monovacancy in graphene. By directly imaging the affected range of monovacancy-induced intervalley scattering, we demonstrate that it is inversely proportional to the energy; i.e., it is proportional to the wavelength of massless Dirac fermions. A giant electron-hole asymmetry of the intervalley scattering is observed because the monovacancy is charged. By further charging the monovacancy, the bended electronic potential around the monovacancy softens the scattering potential, which, consequently, suppresses the intervalley scattering of the monovacancy.

Effect of light polarization on plasmon-induced charge transfer.

Plasmonic nanoclusters can strongly absorb light energy and generate hot carriers, which have great potentials in photovoltaic and photocatalytic applications. A vital step for those plasmonic applications is the charge transfer at the metal-semiconductor interface. The effect of the light polarization on the charge transfer has not been theoretically investigated so far. Here, we take the Ag-TiO2 system as a model system to study the polarization effect using time-dependent density functional theory simulations. We find that the charge transfer is sensitive to the light polarization, which has its origin in the polarization-dependent hot carrier distributions. For the linearly polarized light, it shows a sine-square dependence on the polar angle, indicating that the charge transfer response to the linear polarization can be decomposed into components perpendicular and parallel to the interface. We also find that there exists directional charge transfer with a circular light polarization. Our results demonstrate that the light polarization can significantly affect the charge transfer behavior and, thus, offer a new degree of freedom to manipulate the plasmonic applications.

Characterization of silicon transporter gene family in Saccharum and functional analysis of the ShLsi6 gene in biotic stress.

Silicon, one of the most prevalent elements in the soil, is beneficial for plant growth and defense against different stresses. The silicon transporter gene (Lsi) plays an important role in the uptake and transport of silicon in higher plants. In this study, a total of 32 Lsi genes, including 20 SsLsi in sugarcane wild species Saccharum spontaneum, 5 ShLsi in Saccharum hybrid cultivar R570 and 7 SbLsi in sugarcane related species Sorghum bicolor, were identified and classified into three groups. Bioinformatics analysis showed that instability, hydrophobicity, localization of cell membranes and vacuoles were the main features of the Lsi proteins. Whole genome and segmental duplication contributed to the main expansion of Lsi gene family. Collinearity analysis of the Lsi genes showed that S. spontanum and R570 had a collinear relationship with monocotyledonous plants S. bicolor and Oryza sativa, but not with dicotyledonous plants Arabidopsis thaliana and Vitis vinifera. The replicated Lsi genes were mainly subjected to strong selection pressure for purification. The diverse cis-regulatory elements in the promoter of SsLsi, ShLsi and SbLsi genes suggested that they were widely involved in the response of plants to various stresses and the regulation of the growth and development. Transcriptome data and real time quantitative PCR analysis showed that the Lsi genes exhibited different expression profiles in sugarcane tissues and under Sporisorium scitamineum, drought and cold stresses. In addition, the cDNA and genomic DNA sequences of ShLsi6 that was homologous to SsLsi1b gene was cloned from Saccharum hybrid cultivar ROC22. Transient expression analysis showed that, compared with the control, Nicotiana benthamiana leaves which overexpressed the ShLsi6 gene showed a high sensitivity after inoculation with tobacco pathogens Ralstonia solanacearum and Fusarium solani var. coeruleum. This study provides important information for further functional analysis of Lsi genes and resistant breeding in sugarcane.

Tunable electron and hole injection channels at plasmonic Al-TiO2 interfaces.

Metallic nanostructures can strongly absorb light through their plasmon excitations, whose nonradiative decay generates hot electron-hole pairs. When the metallic nanostructure is interfaced with a semiconductor, the spatial separation of hot carriers plays the central and decisive roles in photovoltaic and photocatalytic applications. In recent years, free-electron metals like Al have attracted tremendous attentions due to the much higher plasmon frequencies that could extend to the ultraviolet regime. Here, the plasmon excitations and charge separations at the Al-TiO2 interfaces have been investigated using quantum-mechanical calculations, where the atomic structures and electronic dynamics are all treated from first-principles. It is found that the high-frequency plasmon of Al produces abundant and broad-band hot-carrier distributions, where the electron-hole symmetry is broken by the presence of the semiconductor band gap. Such an asymmetric hot-carrier distribution provides two competing channels, which can be controlled either by tuning the laser frequency, or by harnessing the plasmon frequency through the geometry and shape of the metallic nanostructure. Our study suggests that the Al plasmon offers a versatile and tunable pathway for the charge transfer and separation, and has general implications in plasmon-assisted photovoltaics and photocatalysis.

Integrated Plasmonics: Broadband Dirac Plasmons in Borophene.

The past decade has witnessed numerous discoveries of two-dimensional (2D) semimetals and insulators, whereas 2D metals were rarely identified. Borophene, a monolayer boron sheet, has recently emerged as a perfect 2D metal with unique electronic properties. Here we study collective excitations in borophene, which exhibit two major plasmon modes with low damping rates extending from the infrared to ultraviolet regime. The anisotropic 1D plasmon originates from electronic transitions of tilted Dirac cones in borophene, analogous to that in extreme doped graphene. These features enable borophene as an integrated platform of 1D, 2D, and Dirac plasmons, promising for directional polariton transport and broadband optical communication in next-generation optoelectronic devices.

Tunable magnetism of a single-carbon vacancy in graphene.

Creating a single-carbon vacancy introduces (quasi-)localized states for both σ and π electrons in graphene. Theoretically, interactions between the localized σ electrons and quasilocalized π electrons of a single-carbon vacancy in graphene are predicted to control its magnetism. However, experimentally confirming this prediction through manipulating the interactions remains an outstanding challenge. Here we report the manipulation of magnetism in the vicinity of an individual single-carbon vacancy in graphene by using a scanning tunnelling microscopy (STM) tip. Our spin-polarized STM measurements, complemented by density functional theory calculations, indicate that the interactions between the localized σ and quasilocalized π electrons could split the π electrons into two states with opposite spins even when they are well above the Fermi level. Via the STM tip, we successfully manipulate both the magnitude and direction of magnetic moment of the π electrons with respect to that of the σ electrons. Three different magnetic states of the single-carbon vacancy, exhibiting magnetic moments of about 1.6 µB, 0.5 µB, and 0 µB respectively, are realized in our experiment.

Anomalous Dirac Plasmons in 1D Topological Electrides.

The plasmon opens up the possibility to efficiently couple light and matter at subwavelength scales. In general, the plasmon frequency, intensity, and damping are dependent on the carrier density. These dependencies, however, are disadvantageous for stable functionalities of plasmons and render fundamentally a weak intensity at low frequency, especially for the Dirac plasmon (DP) widely studied in graphene. Here we demonstrate a new type of DP, emerging from a Dirac nodal-surface state, which can simultaneously exhibit a density-independent frequency, intensity, and damping. Remarkably, we predict the realization of anomalous DP (ADP) in 1D topological electrides, such as Ba_{3}CrN_{3} and Sr_{3}CrN_{3}, by first-principles calculations. The ADPs in both systems have a density-independent frequency and high intensity, and their frequency can be tuned from terahertz to midinfrared by changing the excitation direction. Furthermore, the intrinsic weak electron-phonon coupling of anionic electrons in electrides affords an added advantage of low-phonon-assisted damping and hence a long lifetime of the ADPs. Our Letter paves the way to developing novel plasmonic and optoelectronic devices by combining topological physics with electride materials.

Plasmon-Induced Electron-Hole Separation at the Ag/TiO2(110) Interface.

Plasmon-induced electron-hole separation at metal-semiconductor interfaces is an essential step in photovoltaics, photochemistry, and optoelectronics. Despite its importance in fundamental understandings and technological applications, the mechanism and dynamics of the charge separation under plasmon excitations have not been well understood. Here, the plasmon-induced charge separation between a Ag20 nanocluster and a TiO2(110) surface is investigated using time-dependent density functional theory simulations. It is found that the charge separation dynamics consists of two processes: during the first 10 fs an initial charge separation resulting from the plasmon-electron coupling at the interface and a subsequent charge redistribution governed by the sloshing motion of the charge-transfer plasmon. The interplay between the two processes determines the charge separation and leads to the inhomogeneous layer-dependent distribution of hot carriers. The hot electrons are more efficient than the hot holes in the charge injection, resulting in the charge separation. Over 40% of the hot electron-hole pairs are separated spatially from the interface. Finally, the second TiO2 layer receives the most net charges from the Ag nanocluster rather than the interfacial layer. These results reveal the mechanism and dynamics of the charge separation driven by the surface plasmon excitation and have broad implications in plasmonic applications.

Comparative Analysis of two Sugarcane Ancestors Saccharum officinarum and S. spontaneum based on Complete Chloroplast Genome Sequences and Photosynthetic Ability in Cold Stress.

Polyploid Saccharum with complex genomes hindered the progress of sugarcane improvement, while their chloroplast genomes are much smaller and simpler. Chloroplast (cp), the vital organelle, is the site of plant photosynthesis, which also evolves other functions, such as tolerance to environmental stresses. In this study, the cp genome of two sugarcane ancestors Saccharum officinarum and S. spontaneum were sequenced, and genome comparative analysis between these two species was carried out, together with the photosynthetic ability. The length is 141,187 bp for S. officinarum and that is 7 bp longer than S. spontaneum, with the same GC content (38.44%) and annotated gene number (134), 13 with introns among them. There is a typical tetrad structure, including LSC, SSC, IRb and IRa. Of them, LSC and IRa/IRb are 18 bp longer and 6 bp shorter than those in S. spontaneum (83,047 bp and 22,795 bp), respectively, while the size of SSC is same (12,544 bp). Five genes exhibit contraction and expansion at the IR junctions, but only one gene ndhF with 29 bp expansion at the border of IRb/SSC. Nucleotide diversity (Pi) based on sliding window analysis showed that the single copy and noncoding regions were more divergent than IR- and coding regions, and the variant hotspots trnG-trnM, psbM-petN, trnR-rps14, ndhC-trnV and petA-psbJ in the LSC and trnL-ccsA in the SSC regions were detected, and petA-psbJ with the highest divergent value of 0.01500. Genetic distances of 65 protein genes vary from 0.00000 to 0.00288 between two species, and the selective pressure on them indicated that only petB was subjected to positive selection, while more genes including rpoC2, rps3, ccsA, ndhA, ndhA, psbI, atpH and psaC were subjected to purifying or very strong purifying selection. There are larger number of codons in S. spontaneum than that in S. officinarum, while both species have obvious codon preference and the codons with highest-(AUG) and lowest frequency (AUA) are same. Whilst, the most abundant amino acid is leucine in both S. officinarum and S. spontaneum, with number of 2175 (10.88% of total) and 2228 (10.90% of total) codons, respectively, and the lowest number is cysteine, with only 221 (1.105%) and 224 (1.096%), respectively. Protein collinearity analysis showed the high collinearity though several divergences were present in cp genomes, and identification of simple sequence repeats (SSRs) were included in this study. In addition, in order to compare cold tolerance and explore the expanding function of this environmental stress, the chlorophyll relative content (SPAD) and chlorophyll fluorescence Fv/Fm were measured. The significantly higher SPAD were observed in S. spontaneum than those in S. officinarum, no matter what the control conditions, exposure to low temperature or during recovery, and so was for Fv/Fm under exposure to low temperature, together with higher level of SPAD in S. spontaneum in each measurement. Aforementioned results suggest much stronger photosynthetic ability and cold tolerance in S. spontaneum. Our findings build a foundation to investigate the biological mechanism of two sugarcane ancestor chloroplasts and retrieve reliable molecular resources for phylogenetic and evolutionary studies, and will be conducive to genetic improvement of photosynthetic ability and cold resistance in modern sugarcane.

Analytical analysis of spectral sensitivity of plasmon resonances in a nanocavity.

Metallic nanocavities exhibit extremely high spectral sensitivity to geometrical variations and are promising for sensing applications. Here, the sensitivity of a cubic dimer cavity, to picometer gap variation, is analysed in a model, which takes into account the phase shift of scattering at the boundaries and the quantum tunnelling effect in the small gap limit. The resonance wavelengths are expressed in terms of the plasmon frequency, the medium dielectric function, and the geometry of the gap. The sensitivity of the resonance wavelength to the gap width variation is found to be as high as 1 nm pm-1. While the resonance wavelengths depend on the materials' dielectric functions, the sensitivity is found to scale universally as a function of gap distance. In the sub-nanometer regime, electron tunnelling across the gap starts to suppress the plasmonic field, setting the limit of sensitivity of such a dimer cavity. The results given by the analytical model are complemented by numerical simulations using Comsol. Our model reveals the origin and universal behaviours of the sensitivity of the cavity plasmon and provides guidance for the design of new sensitive rulers at the picometer scale.

The Physiological and Agronomic Responses to Nitrogen Dosage in Different Sugarcane Varieties.

Nitrogen (N) is very important for sugarcane yield improvement, but the excessive application of N fertilizer brings about N pollution and a cost increase. Through distinguishing the difference of nitrogen use efficiency (NUE), we can reasonably apply N fertilizer according to the NUE characteristics of sugarcane varieties, and thus reduce N loss and maintain high yield. The present study showed the pot experiment results of identifying NUE types of nine main sugarcane varieties in the main sugarcane producing areas of China under controlled conditions, and identified the key physiological and agronomic indictors which can help to determine the NUE types of sugarcane. The test clones were exposed to varying levels of N fertilizer and 15 parameters that are likely to impact NUE were measured. The key results are (1) Sugarcane variety ROC22 has the high plant dry weight (PDW) and NUE among nine varieties under different N rates, it can take advantages under low N supply (225 kg/hm2 urea), and less N fertilizer can be applied properly in production. (2) Varieties of GT32 was good performing genotype for PDW and NUE under low N supply (225 kg/hm2 urea), GT42 was more suitable for moderate N environment (450 kg/hm2 urea), while YT94-128 was at middle N and high N supply (450-675 kg/hm2 urea). (3) Late stage of shoot elongation is suitable for differentiating sugarcane clones for NUE. (4) Leaf glutamine synthetase activity is the most reliable predictor of NUE in sugarcane. The result of pot experiment is sufficient to differentiate clonal variation for NUE in sugarcane as it reflects field experimental results. This study can set up a basis for identification the NUE types of sugarcane varieties and the development of reasonable N fertilizer application.

Thermodynamic stability of magnetic states of monovacancy in graphene revealed by ab initio molecular dynamics simulations.

The stability of magnetic states is essential for potential spintronic applications. Here we report on the thermal stability of magnetic states of monovacancy graphene using ab initio molecular dynamics simulations. At room temperature, thermal fluctuations of the graphene lattice induce a rapid magnetic switching between two states with a high and low magnetic moment, indicating that due to the instability of the atomic structure of the vacancy, the associated magnetic moment is thermodynamically unstable. Lowering the temperature can significantly reduce the rate of the switching process and enhance the resident time on the high magnetic state. It stabilizes in the high magnetic state at as low as 30 K. Analyzing the atomic trajectories and the instant electronic structures confirms that these two magnetic states in MD simulations correspond to the magnetic and nonmagnetic states reported in the literatures. Such fluctuations of local magnetic moments are associated with the vertical displacement of the carbon atoms with the unsaturated dangling bond. This study reveals the dynamical correlation between atomic movement and the magnetic switching, and a comprehensive picture of vacancy magnetism in graphene. It has implications in graphene based spintronic devices.

Foreign cry1Ac gene integration and endogenous borer stress-related genes synergistically improve insect resistance in sugarcane.

BACKGROUND: Sugarcane (Saccharum spp. hybrids) is considered the most globally important sugar-producing crop and raw material for biofuel. Insect attack is a major issue in sugarcane cultivation, resulting in yield losses and sucrose content reductions. Stem borer (Diatraea saccharalis F.) causes serious yield losses in sugarcane worldwide. However, insect-resistant germplasms for sugarcane are not available in any collections all over the world, and the molecular mechanism of insect resistance has not been elucidated. In this study, cry1Ac transgenic sugarcane lines were obtained and the biological characteristics and transgene dosage effect were investigated and a global exploration of gene expression by transcriptome analysis was performed. RESULTS: The transgene copies of foreign cry1Ac were variable and random. The correlation between the cry1Ac protein and cry1Ac gene copies differed between the transgenic lines from FN15 and ROC22. The medium copy lines from FN15 showed a significant linear relationship, while ROC22 showed no definite dosage effect. The transgenic lines with medium copies of cry1Ac showed an elite phenotype. Transcriptome analysis by RNA sequencing indicated that up/down regulated differentially expressed genes were abundant among the cry1Ac sugarcane lines and the receptor variety. Foreign cry1Ac gene and endogenous borer stress-related genes may have a synergistic effect. Three lines, namely, A1, A5, and A6, were selected for their excellent stem borer resistance and phenotypic traits and are expected to be used directly as cultivars or crossing parents for sugarcane borer resistance breeding. CONCLUSIONS: Cry1Ac gene integration dramatically improved sugarcane insect resistance. The elite transgenic offspring contained medium transgene copies. Foreign cry1Ac gene integration and endogenous borer stress-related genes may have a synergistic effect on sugarcane insect resistance improvement.

Heterologous expression of a Glyoxalase I gene from sugarcane confers tolerance to several environmental stresses in bacteria.

Glyoxalase I belongs to the glyoxalase system that detoxifies methylglyoxal (MG), a cytotoxic by-product produced mainly from triose phosphates. The concentration of MG increases rapidly under stress conditions. In this study, a novel glyoxalase I gene, designated as SoGloI was identified from sugarcane. SoGloI had a size of 1,091 bp with one open reading frame (ORF) of 885 bp encoding a protein of 294 amino acids. SoGloI was predicted as a Ni2+-dependent GLOI protein with two typical glyoxalase domains at positions 28-149 and 159-283, respectively. SoGloI was cloned into an expression plasmid vector, and the Trx-His-S-tag SoGloI protein produced in Escherichia coli was about 51 kDa. The recombinant E. coli cells expressing SoGloI compared to the control grew faster and tolerated higher concentrations of NaCl, CuCl2, CdCl2, or ZnSO4. SoGloI ubiquitously expressed in various sugarcane tissues. The expression was up-regulated under the treatments of NaCl, CuCl2, CdCl2, ZnSO4 and abscisic acid (ABA), or under simulated biotic stress conditions upon exposure to salicylic acid (SA) and methyl jasmonate (MeJA). SoGloI activity steadily increased when sugarcane was subjected to NaCl, CuCl2, CdCl2, or ZnSO4 treatments. Sub-cellular observations indicated that the SoGloI protein was located in both cytosol and nucleus. These results suggest that the SoGloI gene may play an important role in sugarcane's response to various biotic and abiotic stresses.

Transcriptional analysis identifies major pathways as response components to Sporisorium scitamineum stress in sugarcane.

BACKGROUND: Sugarcane smut, which is caused by Sporisorium scitamineum, is a severe fungal disease affecting sugarcane. However, the major pathways involved in the interaction between sugarcane and S. scitamineum remains unclear. RESULTS: In the present study, suppression subtractive hybridization (SSH) library construction, together with reverse northern blotting, was conducted on the most prevalent sugarcane genotype ROC22 challenged with S. scitamineum. After alignment and homologous expressed sequence tag (EST) assembly, a total of 155 differentially expressed unigenes were identified from SSH libraries. Totally, 26 of 155 differentially expressed unigenes were analyzed by qRT-PCR in sugarcane smut-resistant genotype YC05-179 and susceptible genotype ROC22. Genes encoded two unknown protein (Q1 and Q11), serine/threonine kinase (Q2), fiber protein (Q3), eukaryotic translation initiation factor 5A (Q23), and Sc14-3-3-like protein (Q24) were induced in sugarcane smut-resistant genotype YC05-179 but inhibited in susceptible genotype ROC22. Based on the differential expression data achieved from SSH libraries and qRT-PCR, we found that, serine/threonine kinases, Ca2+ sensors, mitogen-activated protein genes and some NBS-LRR genes may involve in the signal recognition and transduction of smut fungus infection in sugarcane. While in the plant hormone signaling pathways, the genes related to auxin, abscisic acid, salicylic acid and ethylene were more apparently in response to smut fungus invasion. The hypersensitive response, protein metabolism, polyamine synthesis, and cell wall formation may play an important role in sugarcane defense against smut fungus colonization. Additionally, the Sc14-3-3 might serve as a molecular modulator in sugarcane being immune to smut disease by interacting with proteins like ScGAPN (Q10), which have been further verified by BiFC assay. CONCLUSIONS: The findings of the present study could provide a general view about gene pathways involving in sugarcane defense against smut disease and facilitate a better understanding of the molecular mechanism underlying sugarcane-S. scitamineum interaction.

Particle Bombardment of the cry2A Gene Cassette Induces Stem Borer Resistance in Sugarcane.

Sugarcane borer is the most common and harmful pest in Chinese sugarcane fields, and can cause damage to the whole plant during the entire growing season. To improve borer resistance in sugarcane, we constructed a plant expression vector pGcry2A0229 with the bar gene as the marker and the cry2A gene as the target, and introduced it into embryogenic calli of most widely cultivated sugarcane cultivar ROC22 by particle bombardment. After screening with phosphinothricin in vitro and Basta spray, 21 resistance-regenerated plants were obtained, and 10 positive transgenic lines harboring the cry2A gene were further confirmed by conventional PCR detection. Real-time quantitative PCR (RT-qPCR) analysis showed that the copy number of the cry2A gene varied among different transgenic lines but did not exceed four copies. Quantitative ELISA analysis showed that there was no linear relationship with copy number but negatively correlated with the percentage of borer-infested plants. The analysis of industrial and agronomic traits showed that the theoretical sugar yields of transgenic lines TR-4 and TR-10 were slightly lower than that of the control in both plant cane and ratoon cane; nevertheless, TR-4 and TR-10 lines exhibited markedly lower in frequency of borer-infested plants in plant cane and in the ratoon cane compared to the control. Our results indicate that the introduction of the cry2A gene via bombardment produces transgenic lines with obviously increased stem borer resistance and comparable sugar yield, providing a practical value in direct commercial cultivation and crossbreeding for ROC22 has been used as the most popular elite genitor in various breeding programs in China.