Silicon induces ROS scavengers, hormone signalling, antifungal metabolites, and silicon deposition against brown stripe disease in sugarcane.

Bipolaris setariae is known to cause brown stripe disease in sugarcane, resulting in significant yield losses. Silicon (Si) has the potential to enhance plant growth and biotic resistance. In this study, the impact of Si on brown stripe disease was investigated across susceptible and resistant sugarcane varieties, utilizing four Si concentrations (0, 15, 30, and 45 g per barrel of Na2SiO3·5H2O). Si significantly reduced the incidence of brown stripe disease (7.41-59.23%) and alleviated damage to sugarcane growth parameters, photosynthetic parameters, and photosynthetic pigments. Submicroscopic observations revealed that Si induced the accumulation of silicified cells in leaves, reduced spore accumulation, decreased stomatal size, and protected organelles from B. setariae damage. In addition, Si increased the activity of antioxidant enzymes (superoxide dismutase, peroxidase, and catalase), reduced reactive oxygen species production (malondialdehyde and hydrogen peroxide) and modulated the expression of genes associated with hormone signalling (PR1, TGA, AOS, AOC, LOX, PYL8, and SnRK2), leading to the accumulation of abscisic acid and jasmonic acid and inhibiting SA synthesis. Si also activated the activity of metabolism-related enzymes (polyphenol oxidase and phenylalanine ammonia lyase) and the gene expression of PAL-dependent genes (PAL, C4H, and 4CL), regulating the accumulation of metabolites, such as chlorogenic acid and lignin. The antifungal test showed that chlorogenic acid (15ug µL-1) had a significant inhibitory effect on the growth of B. setariae. This study is the first to demonstrate the inhibitory effect of Si on B. setariae in sugarcane, highlighting Si as a promising and environmentally friendly strategy for managing brown stripe disease.

Ultrasonic treatment alleviated cadmium stress in sugarcane via improving antioxidant activity and physiological and biochemical status.

Cadmium (Cd) is a toxic element that endangers crop growth and affects food safety and human health. Therefore, the study of Cd mitigation technology is important. Ultrasonic treatment can improve crop growth and enhance their ability to resist various abiotic stresses. In this study, the effect of ultrasonic treatment on alleviating sugarcane Cd stress was studied in a barrel experiment using sugarcane varieties 'ROC22' and 'LC05-136' as test materials. Sugarcane buds without ultrasonic treatment and with ultrasonic treatment (20-40 kHz mixed frequency ultrasound for 2 min, dry treatment) were planted in soil with Cd contents of 0, 50, 100, 250, and 500 mg·kg-1. Compared with non-ultrasonic treatment, Ultrasonic treatment significantly increased the activities of antioxidant enzymes in sugarcane, significantly increased the content of osmoregulation substances, significantly reduced the content of superoxide anion (the highest decreases reached 11.55%) and malondialdehyde (the highest decreases reached 20.59%), and significantly increased the expression level of metallothionein (MT)-related genes, with the expression of ScMT1 increased by 8.80-37.49% and the expression of ScMT2-1-5 increased by 1.55-69.33%. In addition, ultrasonic treatment significantly reduced the Cd contents in sugarcane roots, stems, leaves, bagasse, and juice (the highest reduction in Cd content was 49.18%). In general, ultrasonic treatment regulated the metabolism of reactive oxygen species and MT-related gene expression in sugarcane, increased the Cd tolerance of sugarcane, promoted photosynthesis in sugarcane leaves, improved root morphology, enhanced sugarcane growth, and increased cane and sugar yield.

Silicon enhancement for endorsement of Xanthomonas albilineans infection in sugarcane.

Silicon (Si) is considered to be a plant growth and development regulator element as well as provide the regulatory response against various biotic stressors. However, the potential mechanism of Si enhancement to regulate plant disease resistance remains to be studied. Therefore, the current study evaluated the effects of Si application on the performance of sugarcane against Xanthomonas albilineans (Xa) infection. Si was applied exogenously (0, 3.85 and 7.70 g Si/kg soil) and the results show that plant height, stem circumference and leaf width of siliconized sugarcane have been improved, which effectively reduced the disease index (0.17-0.21) and incidence (58.2%-69.1%) after Xa infection. Lowest values of MDA (348.5 nmol g-1 FW) and H2O2 (3539.4 mmol/L) were observed in 7.70 g Si/kg soil followed by in 3.85 g Si/kg soil (MDA: 392.6 nmol g-1 FW and H2O2: 3134.6 mmol/L) than that of the control. Whereas, PAL enzyme activity (50.8 mmol/L), JA (230.2 mmol/L) and SA (2.7 ug mL-1) contents were significantly higher in 7.70 g Si/kg soil followed by in 3.85 g Si/kg soil (PAL: 46.3 mmol/L, JA: 182.7 mmol/L and SA: 2.4 ug mL-1) as compared to control. The lower MDA, H2O2 level and higher enzymatic activities were associated with the highest expression levels of their metabolic pathway associated genes i.e., ShMAPK1, ShLOX, ShPAL, ShAOS, ShAOC, ShC4H, ShCAT, Sh4CL and ShNPR1 (22.08, 15.56, 10.42, 3.35, 2.54, 2.14, 1.82, 1.67 and 1.22 folds, respectively) in 7.70 g Si/kg soil as compared to other experimental units and control. Overall, the results of current study indicates that siliconized sugarcane more actively regulates disease resistance through modulation of growth and MDA, H2O2, SA and JA associated metabolic pathways.

ScAOC1, an allene oxide cyclase gene, confers defense response to biotic and abiotic stresses in sugarcane.

KEY MESSAGE: An allene oxide cyclase gene which is involved in defense against biotic and abiotic stresses was cloned and characterized in sugarcane. Allene oxide cyclase (AOC), a key enzyme in jasmonate acid (JA) biosynthesis, affects the stereoisomerism and biological activity of JA molecules, and plays an important role in plant stress resistance. In this study, four SsAOC alleles (SsAOC1-SsAOC4), which shared similar gene structure and were located on Chr1A, Chr1B, Chr1C, and Chr1D, respectively, were mined from sugarcane wild species Saccharum spontaneum, and a homologous gene ScAOC1 (GenBank Accession Number: MK674849) was cloned from sugarcane hybrid variety Yacheng05-179 inoculated with Sporisorium scitamineum for 48 h. ScAOC1 and SsAOC1-SsAOC4 were alkaline, unstable, hydrophilic, and non-secretory proteins, which possess the same set of conserved motifs and were clustered into one group in the phylogenetic analysis. ScAOC1 was expressed in all sugarcane tissues, but with different levels. After infection by S. scitamineum, the transcripts of ScAOC1 were increased significantly both in the smut-susceptible (ROC22) and resistant (Yacheng05-179) varieties, but its transcripts were more accumulated and lasted for a longer period in the smut-resistant variety than in the smut-susceptible one. ScAOC1 was down-regulated under MeJA and NaCl treatments, but up-regulated under SA, ABA, PEG, and cold stresses. Transiently overexpressing ScAOC1 gene into Nicotiana benthamiana leaves regulated the responses of N. benthamiana to two pathogens Ralstonia solanacearum and Fusarium solani var. coeruleum. Furthermore, prokaryotic expression analysis showed overexpression of ScAOC1 in Escherichia coli BL21 could enhance its tolerance to NaCl, mannitol, and cold stimuli. These results indicated that ScAOC1 may play an active role in response to biotic and abiotic stresses in sugarcane.

Molecular Mechanisms Underlying Sugarcane Response to Aluminum Stress by RNA-Seq.

Some metals are beneficial to plants and contribute to critical physiological processes. Some metals, however, are not. The presence of aluminum ions (Al3+) can be very toxic, especially in acidic soils. Considerable parts of the world's arable land are acidic in nature; mechanistically elucidating a plant's response to aluminum stress is critical to mitigating this stress and improving the quality of plants. To identify the genes involved in sugarcane response to aluminum stress, we generated 372 million paired-end RNA sequencing reads from the roots of CTC-2 and RB855453, which are two contrasting cultivars. Data normalization resulted in 162,161 contigs (contiguous sequences) and 97,335 genes from a de novo transcriptome assembly (trinity genes). A total of 4858 and 1307 differently expressed genes (DEGs) for treatment versus control were identified for the CTC-2 and RB855453 cultivars, respectively. The DEGs were annotated into 34 functional categories. The majority of the genes were upregulated in the CTC-2 (tolerant cultivar) and downregulated in RB855453 (sensitive cultivar). Here, we present the first root transcriptome of sugarcane under aluminum stress. The results and conclusions of this study are a crucial launch pad for future genetic and genomic studies of sugarcane. The transcriptome analysis shows that sugarcane tolerance to aluminum may be explained by an efficient detoxification mechanism combined with lateral root formation and activation of redox enzymes. We also present a hypothetical model for aluminum tolerance in the CTC-2 cultivar.

Comparative genomics revealed the gene evolution and functional divergence of magnesium transporter families in Saccharum.

BACKGROUND: Sugarcane served as the model plant for discovery of the C4 photosynthetic pathway. Magnesium is the central atom of chlorophyll, and thus is considered as a critical nutrient for plant development and photosynthesis. In plants, the magnesium transporter (MGT) family is composed of a number of membrane proteins, which play crucial roles in maintaining Mg homeostasis. However, to date there is no information available on the genomics of MGTs in sugarcane due to the complexity of the Saccharum genome. RESULTS: Here, we identified 10 MGTs from the Saccharum spontaneum genome. Phylogenetic analysis of MGTs suggested that the MGTs contained at least 5 last common ancestors before the origin of angiosperms. Gene structure analysis suggested that MGTs family of dicotyledon may be accompanied by intron loss and pseudoexon phenomena during evolution. The pairwise synonymous substitution rates corresponding to a divergence time ranged from 142.3 to 236.6 Mya, demonstrating that the MGTs are an ancient gene family in plants. Both the phylogeny and Ks analyses indicated that SsMGT1/SsMGT2 originated from the recent ρWGD, and SsMGT7/SsMGT8 originated from the recent σ WGD. These 4 recently duplicated genes were shown low expression levels and assumed to be functionally redundant. MGT6, MGT9 and MGT10 weredominant genes in the MGT family and werepredicted to be located inthe chloroplast. Of the 3 dominant MGTs, SsMGT6 expression levels were found to be induced in the light period, while SsMGT9 and SsMTG10 displayed high expression levels in the dark period. These results suggested that SsMGT6 may have a function complementary to SsMGT9 and SsMTG10 that follows thecircadian clock for MGT in the leaf tissues of S. spontaneum. MGT3, MGT7 and MGT10 had higher expression levels Insaccharum officinarum than in S. spontaneum, suggesting their functional divergence after the split of S. spontaneum and S. officinarum. CONCLUSIONS: This study of gene evolution and expression of MGTs in S. spontaneum provided basis for the comprehensive genomic study of the entire MGT genes family in Saccharum. The results are valuable for further functional analyses of MGT genes and utilization of the MGTs for Saccharum genetic improvement.

Encapsulation of S-nitrosoglutathione into chitosan nanoparticles improves drought tolerance of sugarcane plants.

The entrapment of NO donors in nanomaterials has emerged as a strategy to protect these molecules from rapid degradation, allowing a more controlled release of NO and prolonging its effect. On the other hand, we have found beneficial effects of S-nitrosoglutathione (GSNO) - a NO donor - supplying to sugarcane plants under water deficit. Here, we hypothesized that GSNO encapsulated into nanoparticles would be more effective in attenuating the effects of water deficit on sugarcane plants as compared to the supplying of GSNO in its free form. The synthesis and characterization of chitosan nanoparticles containing GSNO were also reported. Sugarcane plants were grown in nutrient solution, and then subjected to the following treatments: control (well-hydrated); water deficit (WD); WD + GSNO sprayed in its free form (WDG) or encapsulated (WDG-NP). In general, both GSNO forms attenuated the effects of water deficit on sugarcane plants. However, the encapsulation of this donor into chitosan nanoparticles caused higher photosynthetic rates under water deficit, as compared to plants supplied with free GSNO. The root/shoot ratio was also increased when encapsulated GSNO was supplied, indicating that delayed release of NO improves drought tolerance of sugarcane plants. Our results provide experimental evidence that nanotechnology can be used for enhancing NO-induced benefits for plants under stressful conditions, alleviating the negative impact of water deficit on plant metabolism and increasing biomass allocation to root system.

Excess Copper-Induced Changes in Antioxidative Enzyme Activity, Mineral Nutrient Uptake and Translocation in Sugarcane Seedlings.

Sugarcane is a potential species for use in heavy metal remediation. To analyze the effect of excess copper on sugarcane, the biomass, mineral nutrient content and activities of antioxidative enzymes were measured under copper stress. The results revealed that the biomass of roots and shoots significantly decreased with increasing copper concentration in solution. Most copper accumulated in the roots, and the translocation factor of copper decreased with an increase in copper stress. The MDA content in sugarcane roots notably increased under copper stress. The POD activity in sugarcane roots increased, and CAT activity decreased under copper stress. The Zn, Fe and Mn contents in shoots increased significantly under 200 µmol L-1 Cu2+ treatments. The Zn and Mg contents in roots notably decreased under copper stress, while the Zn and Mg translocation factors increased. These results indicated that the increase in POD activity and the modification of mineral nutrient uptake and transfer might play an important role in reducing the detrimental effects of excess copper.

CfPDIP1, a novel secreted protein of Colletotrichum falcatum, elicits defense responses in sugarcane and triggers hypersensitive response in tobacco.

Colletotrichum falcatum, a hemibiotrophic fungal pathogen, causes one of the major devastating diseases of sugarcane-red rot. C. falcatum secretes a plethora of molecular signatures that might play a crucial role during its interaction with sugarcane. Here, we report the purification and characterization of a novel secreted protein of C. falcatum that elicits defense responses in sugarcane and triggers hypersensitive response (HR) in tobacco. The novel protein purified from the culture filtrate of C. falcatum was identified by MALDI TOF/TOF MS and designated as C. falcatum plant defense-inducing protein 1 (CfPDIP1). Temporal transcriptional profiling showed that the level of CfPDIP1 expression was greater in incompatible interaction than the compatible interaction until 120 h post-inoculation (hpi). EffectorP, an in silico tool, has predicted CfPDIP1 as a potential effector. Functional characterization of full length and two other domain deletional variants (CfPDIP1ΔN1-21 and CfPDIP1ΔN1-45) of recombinant CfPDIP1 proteins has indicated that CfPDIP1ΔN1-21 variant elicited rapid alkalinization and induced a relatively higher production of hydrogen peroxide (H2O2) in sugarcane suspension culture. However, in Nicotiana tabacum, all the three forms of recombinant CfPDIP1 proteins triggered HR along with the induction of H2O2 production and callose deposition. Further characterization using detached leaf bioassay in sugarcane revealed that foliar priming with CfPDIP1∆1-21 has suppressed the extent of lesion development, even though the co-infiltration of CfPDIP1∆1-21 with C. falcatum on unprimed leaves increased the extent of lesion development than control. Besides, the foliar priming has induced systemic expression of major defense-related genes with the concomitant reduction of pathogen biomass and thereby suppression of red rot severity in sugarcane. Comprehensively, the results have suggested that the novel protein, CfPDIP1, has the potential to trigger a multitude of defense responses in sugarcane and tobacco upon priming and might play a potential role during plant-pathogen interactions.

Agronomic evaluation and molecular characterisation of the acetolactate synthase gene in imazapyr tolerant sugarcane (Saccharum hybrid) genotypes.

KEY MESSAGE: Mutagenesis had no effect on number of stalks/plot, stalk height, fibre and sucrose content of mutants. Imazapyr tolerance is likely due to a S622N mutation in the acetolactate synthase gene. The herbicidal compound imazapyr is effective against weeds such as Cynodon and Rottboellia species that constrain sugarcane production. This study aimed to compare agronomic characteristics of three imazapyr tolerant mutants (Mut 1, Mut 6 and Mut 7) with the non-mutated N12 control after 18 months of growth, and to sequence the acetolactate synthase (ALS) gene to identify any point mutations conferring imazapyr tolerance. There were no significant differences in the number of stalks/plot, stalk height, fibre and sucrose contents of the mutants compared with the N12 control. However, Mut 1 genotype was more susceptible to the Lepidopteran stalk borer, Eldana saccharina when compared with the non-mutated N12 (11.14 ± 1.37 and 3.89 ± 0.52% internodes bored, respectively), making Mut 1 less desirable for commercial cultivation. Molecular characterisation of the ALS gene revealed non-synonymous mutations in Mut 6. An A to G change at nucleotide position 1857 resulted in a N513D mutation, while a G to A change at nucleotide position 2184 imposed a S622N mutation. Molecular dynamics simulations revealed that the S622N mutation renders an asparagine side chain clash with imazapyr, hence this mutation is effective in conferring imazapyr tolerance.

Euclidean distance can identify the mannitol level that produces the most remarkable integral effect on sugarcane micropropagation in temporary immersion bioreactors.

Plant scientists usually record several indicators in their abiotic factor experiments. The common statistical management involves univariate analyses. Such analyses generally create a split picture of the effects of experimental treatments since each indicator is addressed independently. The Euclidean distance combined with the information of the control treatment could have potential as an integrating indicator. The Euclidean distance has demonstrated its usefulness in many scientific fields but, as far as we know, it has not yet been employed for plant experimental analyses. To exemplify the use of the Euclidean distance in this field, we performed an experiment focused on the effects of mannitol on sugarcane micropropagation in temporary immersion bioreactors. Five mannitol concentrations were compared: 0, 50, 100, 150 and 200 mM. As dependent variables we recorded shoot multiplication rate, fresh weight, and levels of aldehydes, chlorophylls, carotenoids and phenolics. The statistical protocol which we then carried out integrated all dependent variables to easily identify the mannitol concentration that produced the most remarkable integral effect. Results provided by the Euclidean distance demonstrate a gradually increasing distance from the control in function of increasing mannitol concentrations. 200 mM mannitol caused the most significant alteration of sugarcane biochemistry and physiology under the experimental conditions described here. This treatment showed the longest statistically significant Euclidean distance to the control treatment (2.38). In contrast, 50 and 100 mM mannitol showed the lowest Euclidean distances (0.61 and 0.84, respectively) and thus poor integrated effects of mannitol. The analysis shown here indicates that the use of the Euclidean distance can contribute to establishing a more integrated evaluation of the contrasting mannitol treatments.

Effect of cadmium on morphometric traits, antioxidant enzyme activity and phytochelatin synthase gene expression (SoPCS) of Saccharum officinarum var. cp48-103 in vitro.

Cadmium is an important environmental pollutant which genetically, physiologically and biochemically affects the cell. Phytochelatins (PC) are involved in one of the phytoremediation mechanisms, in which they are able to bind heavy metals, such as cadmium. The objective of this study was to evaluate morphometric, antioxidant enzyme activity, and SoPCS gene expression in sugarcane growing under cadmium stress. After propagation, samples were cultured in triplicate for 14 days in modified MS medium containing CdCl2 (100, 250, 500 µmol). The morphometric traits, pigments, quantity and antioxidant enzyme activity were studied in treated plantlets. SoPCS gene expression was analyzed by qRT-PCR. Growth traits decreased following cadmium treatment. The amount of Chla, Chlb and ChlT decreased in treated samples, whereas carotenoids increased significantly. A rapid increase in antioxidant enzyme activity was detected. Enhanced SoPCS gene expression was observed in treated roots, whereas gene expression pattern in leaves was irregular. In conclusion, cadmium decreases the photosynthetic mechanism and growth rate in sugarcane. Antioxidative enzymes and SoPCS gene expression were significantly unregulated in sugarcane roots compared to the leaves. Cadmium concentration in shoots and roots of sugarcane significantly increased. The management of cadmium bioaccumulation in non-edible tissues of sugarcane such as leaves and roots that are sometimes burned after harvest can be applied for environmental protection.

In silico prediction, phylogenetic and bioinformatic analysis of SoPCS gene, survey of its protein characterization and gene expression in response to cadmium in Saccharum officinarum.

Phytochelatin synthase isolated from microorganisms, yeasts, algae and plant, serve a fundamental role in reducing heavy metals. In this research the in silico PCS gene structure (SoPCS) of sugarcane, its secondary and 3D protein structure, physicochemical properties, cell localization and phylogenetic tree were predicted utilizing bioinformatics tools. SoPCS expression in the leaves and roots of sugarcane in tissue culture treated with cadmium was also studied utilizing real time PCR. The predicted SoPCS gene contains 1524 nucleotides, a protein encoded with 508 amino acids of which the molecular weight is 55953.3 Da, 6 exons and 5 introns. The subcellular position of the enzyme is mitochondrion or cytoplasmic. Two domains belonging to the phytochelatin synthase family with similar features was found in Pfam having more than 97% similarity with the predicted SoPCS protein. Phylogeny analyses of plant species were well isolated from other organisms. Ten disulfide-bonded cysteines were excluded from the structure of SoPCS. The predicted 3D structure of SoPCS showed that it is able to bind to L-gamma-glutamylcysteine as substrate. The binding site sequence of PCS included amino acids 52(Q),55(P),56(A),57(F), 58(C),103(G),104(I),151(S),163(G),165(F),206(D), 213(R). The common amino acid with conserved sequence in the binding site of the plant was 103Gly. Gene expression indicated that SoPCS has an important role in the response of sugarcane to cadmium with potential use in genetic engineering to remove metal contaminants in the environment. This is the first characterization of a PCS from sugarcane.

Impact of percentage and particle size of sugarcane biochar on the sorption behavior of clomazone in Red Latosol.

Biochar is a carbonaceous material that has excellent potential as a fertilizer and soil conditioner. However, there is a lack of information concerning the effects of the amount and particle size of this pyrogenic material on the soil sorption capacity. In this work, evaluation was made of changes in clomazone (CMZ) sorption in a Red Latosol following soil conditioning using different percentages (0.25, 0.5, and 1% w/w) of sugarcane biochar in three particle sizes (<106, 106-508, and 508-610 µm). The conditioned soils presented apparent sorption coefficients (Kd) up to 1300 times higher than that of pure soil, besides changes in the behavior of CMZ sorption. The biochar particle size and percentage influenced sorption of the herbicide as well as its retention in the amended soil during desorption processes. Both sorption and desorption Freundlich constants were linearly correlated with the external surface area of the biochar present in the soil.

Absorption and translocation of sulfometuron-methyl in sugarcane (Saccharum officinarum L.) at different growth stages.

In Brazil, weed management in sugarcane fields is mainly done with the use of selective herbicide formulations. For many years, diuron+hexazinone was one of the main herbicide mixture formulations used in sugarcane. Later, sulfometuron-methyl was included in the same mixture, which was marketed as a new herbicide formulation for residual in-season weed control in sugarcane. The mixture diuron+hexazinone+sulfometuron-methyl has been widely used in commercial sugarcane fields in Brazil. However, recent field observations have shown that sugarcane plants at different growth stages varied in their phytotoxicity levels after treatment with diuron+hexazinone+sulfometuron-methyl. Greenhouse and laboratory studies were conducted to determine 14Csulfometuron-methyl absorption and translocation, as well as 14C distribution in sugarcane at two growth stages, 2 to 3 leaves and 5 to 6 leaves. 14Csulfometuron-methyl absorption by sugarcane did not differ between the two growth stages. Different patterns of 14C accumulation were observed, which may explain variations in sulfometuron-methyl phytotoxic responses observed in the field.

S-nitrosoglutathione spraying improves stomatal conductance, Rubisco activity and antioxidant defense in both leaves and roots of sugarcane plants under water deficit.

Water deficit is a major environmental constraint on crop productivity and performance and nitric oxide (NO) is an important signaling molecule associated with many biochemical and physiological processes in plants under stressful conditions. This study aims to test the hypothesis that leaf spraying of S-nitrosoglutathione (GSNO), an NO donor, improves the antioxidant defense in both roots and leaves of sugarcane plants under water deficit, with positive consequences for photosynthesis. In addition, the roles of key photosynthetic enzymes ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and phosphoenolpyruvate carboxylase (PEPC) in maintaining CO2 assimilation of GSNO-sprayed plants under water deficit were evaluated. Sugarcane plants were sprayed with water or GSNO 100 µM and subjected to water deficit, by adding polyethylene glycol (PEG-8000) to the nutrient solution. Sugarcane plants supplied with GSNO presented increases in the activity of antioxidant enzymes such as superoxide dismutase in leaves and catalase in roots, indicating higher antioxidant capacity under water deficit. Such adjustments induced by GSNO were sufficient to prevent oxidative damage in both organs and were associated with better leaf water status. As a consequence, GSNO spraying alleviated the negative impact of water deficit on stomatal conductance and photosynthetic rates, with plants also showing increases in Rubisco activity under water deficit.

Exogenous nitric oxide improves sugarcane growth and photosynthesis under water deficit.

MAIN CONCLUSION: Nitric oxide (NO)-mediated redox signaling plays a role in alleviating the negative impact of water stress in sugarcane plants by improving root growth and photosynthesis. Drought is an environmental limitation affecting sugarcane growth and yield. The redox-active molecule nitric oxide (NO) is known to modulate plant responses to stressful conditions. NO may react with glutathione (GSH) to form S-nitrosoglutathione (GSNO), which is considered the main reservoir of NO in cells. Here, we investigate the role of NO in alleviating the effects of water deficit on growth and photosynthesis of sugarcane plants. Well-hydrated plants were compared to plants under drought and sprayed with mock (water) or GSNO at concentrations ranging from 10 to 1000 µM. Leaf GSNO sprayed plants showed significant improvement of relative water content and leaf and root dry matter under drought compared to mock-sprayed plants. Additionally, plants sprayed with GSNO (≥ 100 µM) showed higher leaf gas exchange and photochemical activity as compared to mock-sprayed plants under water deficit and after rehydration. Surprisingly, a raise in the total S-nitrosothiols content was observed in leaves sprayed with GSH or GSNO, suggesting a long-term role of NO-mediated responses to water deficit. Experiments with leaf discs fumigated with NO gas also suggested a role of NO in drought tolerance of sugarcane plants. Overall, our data indicate that the NO-mediated redox signaling plays a role in alleviating the negative effects of water stress in sugarcane plants by protecting the photosynthetic apparatus and improving shoot and root growth.

Elucidating modes of activation and herbicide resistance by sequence assembly and molecular modelling of the Acetolactate synthase complex in sugarcane.

Acetolactate synthase (ALS) catalyzes the first portion of the biosynthetic pathway leading to the generation of branched-chain amino acids. As such it is essential for plant health and is a major target for herbicides. ALS is a very poorly characterized molecule in sugarcane. The enzyme is activated and inhibited by a regulatory subunit (known as VAT1 in plants) whose mode of action is entirely unknown. Using Saccharum halepense as a template we have assembled the ALS gene of sugarcane (Saccharum hybrid) and have modelled the structure of ALS based on an Arabidopsis template (the first ALS model for a monocot). We have also assembled the ALS regulatory proteins (VAT1 and VAT2) from sugarcane and show that VAT2 is specific to true grasses. Employing a bacterial model, we have generated a structural model for VAT1, which explains why the separate domains of the proteins bind to either leucine or valine but not both. Using co-evolution studies we have determined molecular contacts by which we modelled the docking of VAT1 to ALS. In conclusion, we demonstrate how the binding of VAT1 to ALS activates ALS and show how VAT1 can also confer feedback inhibition to ALS. We validate our ALS model against biochemical data and employ this model to explain the function of a novel herbicide binding mutant in sugarcane.

Steam pretreatment of Saccharum officinarum L. bagasse by adding of impregnating agents for advanced bioethanol production.

The main byproduct of the sugarcane industry, Saccharum officinarum L. bagasse (sugarcane bagasse, SCB), is widely used as lignocellulose biomass for bio-ethanol (EtOH) production. In this research study, SCB was pretreated by steam explosion (SE) method using two different impregnating agents: sulfur dioxide (SD) and hydrogen peroxide (HP). As matter of fact, the use of impregnating agents improves the performance of SE method, increasing the concentrations of fermentable sugars after enzymatic saccharification, and decreasing the inhibitor compounds produced during the steam pretreatment step. The aim of this study was to investigate and compare the use of the two impregnating agents in various SE-conditions in order to optimize pretreatment parameters. For every pretreatment condition, it has been evaluated: concentration of fermentable sugars, glucose and xylose yields, and the effects of the inhibitor compounds on enzymatic hydrolysis step. The obtained results allow to improve the efficiency of the whole process of bio-EtOH synthesis enhancing the amount of fermentable sugars produced and the eco-sustainability of the whole process. Indeed, the optimization of steam pretreatment leads to a reduction of energy requirements and to a lower environmental impact.

Overexpression of EaDREB2 and pyramiding of EaDREB2 with the pea DNA helicase gene (PDH45) enhance drought and salinity tolerance in sugarcane (Saccharum spp. hybrid).

KEY MESSAGE: EaDREB2 overexpressed in sugarcane enhanced tolerance to drought and salinity. When co-transformed with plant DNA helicase gene, DREB2 showed greater level of salinity tolerance than in single-gene transgenics. Drought is one of the most challenging agricultural issues limiting sustainable sugarcane production and can potentially cause up to 50 % yield loss. DREB proteins play a vital regulatory role in abiotic stress tolerance in plants. We previously reported that expression of EaDREB2 is enhanced by drought stress in Erianthus arundinaceus. In this study, we have isolated the DREB2 gene from E. arundinaceus, transformed one of the most popular sugarcane variety Co 86032 in tropical India with EaDREB2 through Agrobacterium-mediated transformation, pyramided the EaDREB2 gene with the gene coding for PDH45 driven by Port Ubi 2.3 promoter through particle bombardment and evaluated the V1 transgenics for soil deficit moisture and salinity stresses. Soil moisture stress was imposed at the tillering phase by withholding irrigation. Physiological, molecular and morphological parameters were used to assess drought tolerance. Salinity tolerance was assessed through leaf disc senescence and bud sprout assays under salinity stress. Our results indicate that overexpression of EaDREB2 in sugarcane enhances drought and salinity tolerance to a greater extent than the untransformed control plants. This is the first report of the co-transformation of EaDREB2 and PDH45 which shows higher salinity tolerance but lower drought tolerance than EaDREB2 alone. The present study seems to suggest that, for combining drought and salinity tolerance together, co-transformation is a better approach.