Sporisorium Scitamineum Research Articles

Histidine Kinase Sln1 and cAMP/PKA Signaling Pathways Antagonistically Regulate Sporisorium scitamineum Mating and Virulence via Transcription Factor Prf1.

Many prokaryotes and eukaryotes utilize two-component signaling pathways to counter environmental stress and regulate virulence genes associated with infection. In this study, we identified and characterized a conserved histidine kinase (SsSln1), which is the sensor of the two-component system of Sln1–Ypd1–Ssk1 in Sporisorium scitamineum. SsSln1 null mutant exhibited enhanced mating and virulence capabilities in S. scitamineum, which is opposite to what has been reported in Candida albicans. Further investigations revealed that the deletion of SsSLN1 enhanced SsHog1 phosphorylation and nuclear localization and thus promoted S. scitamineum mating. Interestingly, SsSln1 and cAMP/PKA signaling pathways antagonistically regulated the transcription of pheromone-responsive transcription factor SsPrf1, for regulating S. scitamineum mating and virulence. In short, the study depicts a novel mechanism in which the cross-talk between SsSln1 and cAMP/PKA pathways antagonistically regulates mating and virulence by balancing the transcription of the SsPRF1 gene in S. scitamineum.

The CaCA superfamily genes in Saccharum: comparative analysis and their functional implications in response to biotic and abiotic stress

BackgroundIn plants, Calcium (Ca2+) acts as a universal messenger in various signal transduction pathways, including responses to biotic and abiotic stresses and regulation of cellular and developmental processes. The Ca2+/cation antiporter (CaCA) superfamily proteins play vital roles in the transport of Ca2+ and/or other cations. However, the characteristics of these superfamily members in Saccharum and their evolutionary and functional implications have remained unclear.ResultsA total of 34 CaCA genes in Saccharum spontaneum, 5 CaCA genes in Saccharum spp. R570, and 14 CaCA genes in Sorghum bicolor were identified and characterized. These genes consisted of the H+/cation exchanger (CAX), cation/Ca2+ exchanger (CCX), EF-hand / CAX (EFCAX), and Mg2+/H+ exchanger (MHX) families, among which the CCX and EFCAX could be classified into three groups while the CAX could be divided into two groups. The exon/intron structures and motif compositions suggested that the members in the same group were highly conserved. Synteny analysis of CaCAs established their orthologous and paralogous relationships among the superfamily in S. spontaneum, R570, and S. bicolor. The results of protein-protein interactions indicated that these CaCA proteins had direct or indirect interactions. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis demonstrated that most members of Saccharum CaCA genes exhibited a similar expression pattern in response to hormonal (abscisic acid, ABA) treatment but played various roles in response to biotic (Sporisorium scitamineum) and abiotic (cold) stresses. Furthermore, ScCAX4, a gene encoding a cytoplasm, plasma membrane and nucleus positioning protein, was isolated from sugarcane. This gene was constitutively expressed in different sugarcane tissues and its expression was only induced at 3 and 6 h time points after ABA treatment, however was inhibited and indued in the whole process under cold and S. scitamineum stresses, respectively.ConclusionsThis study systematically conducted comparative analyses of CaCA superfamily genes among S. spontaneum, R570, and S. bicolor, delineating their sequence and structure characteristics, classification, evolutionary history, and putative functions. These results not only provided rich gene resources for exploring the molecular mechanism of the CaCA superfamily genes but also offered guidance and reference for research on other gene families in Saccharum.

Resistance evaluation of sugarcane mutants to Sporisorium scitamineum, the causal agent of sugarcane smut disease

Sugarcane is one of the important crops in the world. It can produce a broad range of valuable products in the area of food, health, agriculture, and bioplastic. However, sugarcane is vulnerable to Sporisorium scitamineum infection. The use of resistant varieties is believed to be a proper control method for the disease. The induced mutation is one of breeding program methods that can be considered to acquire resistant varieties of sugarcane to smut disease. In 2018, we did mutation for some sugarcane varieties then in 2019 the mutants were screened for their resistance to smut disease. This research aimed to identify the resistance character of sugarcane mutants to smut disease. The inoculation method was conducted by dipping the bud into S. scitamineum teliospore suspension for 10 minutes at 30 °C. The buds were then planted and maintained in the polybags. The results showed that out of 41 evaluated mutants, 11 mutants were highly resistant, 9 mutants were resistant, 8 mutants were susceptible and 13 mutants were highly susceptible to smut infection. It seems that we could obtain resistant sugarcane varieties through induced mutation, however the resistance of the cane is still being evaluated during the ratoon stage.

Identification of Sugarcane Germplasm in Sri Lanka for Breeding of Varieties Resistant to Smut Disease (c.a. Sporisorium scitamineum)

Identification of Sugarcane Germplasm in Sri Lanka for Breeding of Varieties Resistant to Smut Disease (c.a. Sporisorium scitamineum)

Comparative expression analysis of potential pathogenicity-associated genes of high- and low-virulent Sporisorium scitamineum isolates during interaction with sugarcane.

The online version contains supplementary material available at 10.1007/s13205-021-02893-7.

Survey of sugarcane smut (Sporisorium scitamineum) and association of factors influencing disease epidemics in sugarcane plantations of Ethiopia

Survey of sugarcane smut (Sporisorium scitamineum) and association of factors influencing disease epidemics in sugarcane plantations of Ethiopia

English

Sugarcane smut caused by Sporisorium scitanmineumis the most severe sugarcane disease that causes major economic losses in sugarcane production in China, and disease resistance breeding is an important way of preventing and controlling this disease. In this study, BC3F1lines derived from the cross between YC 73-226 and YCE 06-111 were used to generate sugarcane smut-resistant and -susceptible gene pools using bulked segregant analysis (BSA). Eighty-nine random primers of start codon targeted (SCoT) polymorphisms were screened, whereas only primer SCoT44 could stably amplify the specific fragment (HE-Ss44) in the resistant pool. Then, several primer pairs of sequence characterized amplified regions (SCARs) were designed based on the sequence alignment of HE-Ss44 (920 bp), which was recovered after purification, and only one pair of SCAR primers (Ss44-F2/R2, forward: 5'-GGCGGGCACCGTCGAGTCCACAT-3'; reverse: 5'-CCGTCCGTCGG TCTCGTCCTTACG-3') could stably amplify a 400-bp specific band in resistant gene pool and its individuals. A validation test of SCAR marker Ss44-F2/R2 was performed using 34 sugarcane cultivars with known smut resistance, which revealed a selection accuracy of 82.35% between marker detection and known smut resistance. Moreover, Pearson’s correlation analysis also showed that the SCAR marker Ss44-F2/R2 was significantly correlated (r= 0.583, P= 0.0003 < 0.01) with the smut resistance trait in sugarcane. In addition, the nucleotide sequence of HE-Ss44 linked with smut-resistancewas not aligned to the homologous sequence in GenBank (NCBI), and the accession number was MG740763. The SCAR marker Ss44-F2/R2 developed in this study can be used for the rapid detection of smut resistance in sugarcane and may be utilized as reference for the improvement of sugarcane smut resistance based on molecular marker-assisted selection.© 2021 Friends Science Publishers

MAP kinase Hog1 mediates a cytochrome P450 oxidoreductase to promote the Sporisorium scitamineum cell survival under oxidative stress

The MAP kinase high osmolarity glycerol 1 (Hog1) plays a central role in responding to external oxidative stress in budding yeast Saccchromyces cerevisiae. However, the downstream responsive elements regulated by Hog1 remain poorly understood. In this study, we report that a Sporisorium scitamineum orthologue of Hog1, named as SsHog1, induced transcriptional expression of a putative cytochrome P450 oxidoreductase encoding gene SsCPR1, to antagonize oxidative stress. We found that upon exposure to hydrogen peroxide (H2 O2 ), SsHog1 underwent strikingly phosphorylation, which was proved to be critical for transcriptional induction of SsCPR1. Loss of SsCPR1 led to hypersensitive to oxidative stress similar as the sshog1Δ mutant did, but was resistant to osmotic stress, which is different from the sshog1Δ mutant. On the other hand, overexpression of SsCPR1 in the sshog1Δ mutant could partially restore its ability of oxidative stress tolerance, which indicated that the Hog1 MAP kinase regulates the oxidative stress response specifically through cytochrome P450 (SsCpr1) pathway. Overall, our findings highlight a novel MAPK signalling pathway mediated by Hog1 in regulation of the oxidative stress response via the cytochrome P450 system, which plays an important role in host-fungus interaction.

New 24-Membered Macrolactins Isolated from Marine Bacteria Bacillus siamensis as Potent Fungal Inhibitors against Sugarcane Smut.

Sugarcane smut, caused by Sporisorium scitamineum, is one of the most devastating fungal diseases affecting sugarcane worldwide. To develop a potent sugarcane smut fungicide, secondary metabolites of marine-derived Bacillus siamensis were isolated and screened for inhibitory activities, which led to the discovery of five new 24-membered macrolactins, bamemacrolactins A-E (1-5), with 3 being the most potent inhibitor. The antifungal mechanism of 3 was studied by assessing its effects on mycelial morphology and the cell wall. Differential proteomics were used to analyze proteins in S. scitamineum upon treatment with bamemacrolactin C and to elucidate its antifungal mechanism. A total of 533 differentially expressed proteins were found. After the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses, eight target proteins were selected, and their functions were discussed. Six of the eight proteins were reported as antifungal targets. The target proteins are involved in the oxidative phosphorylation pathway. Therefore, the potent inhibition of S. scitamineum by compound 3 is most likely through oxidative phosphorylation and targeting a series of enzymes.

EMS-Based In Vitro Mutagenesis and Mutant Screening for Smut Resistance with Agronomic Traits in Sugarcane

Induced mutagenesis offers a useful method to improve desirable characters in sugarcane (Saccharum spp. hybrids). In this study, eleven Ethyl Methyl Sulfonate (EMS)-induced mutants of the Indian sugarcane cultivar CoC 671 were evaluated for smut disease resistance along with agronomic and quality traits viz. early maturity, high sucrose, high cane yield and for quality. PCR-based early detection assay for smut resistance was successfully adopted using the Internal Transcribed Spacer sequence of Sporisorium scitamineum and the same was correlated with field-grown mutants exhibiting resistance upon artificial smut pathogen inoculation. The smut resistant mutants were also found superior for juice and sugar quality parameters such as Brix %, Sucrose %, Purity % as well as Commercial Cane Sugar content at 10th and 12th month maturity. Mutants TC 2819 and TC 2826 exhibited superior for sucrose content (20–24%) than parent CoC 671 (18–21%), respectively, at 10th and 12th month of maturity. The study highlights potential of EMS-induced genetic variability for the mutants with smut resistance and agronomic traits in sugarcane.

Genome-wide identification, characterization and expression analysis of the carotenoid cleavage oxygenase (CCO) gene family in Saccharum

Carotenoid cleavage oxygenases (CCOs) play crucial roles in plant growth and development, as well as in the response to phytohormonal, biotic and abiotic stresses. However, comprehensive and systematic research on the CCO gene family has not yet been conducted in Saccharum. In this study, 47 SsCCO and 14 ShCCO genes were identified and characterized in Saccharum spontaneum and Saccharum spp. R570 cultivar, respectively. The SsCCOs consisted of 38 SsCCDs and 9 SsNCEDs, while ShCCOs contained 11 ShCCDs and 3 ShNCEDs. The SsCCO family could be divided into 7 groups, while ShCCO family into 5 groups. The genes/proteins contained similar compositions within the same group, and the evolutionary mechanisms differed between S. spontaneum and R570. Gene Ontology annotation implied that CCOs were involved in many physiological and biochemical processes. Additionally, 41 SsCCOs were regulated by 19 miRNA families, and 8 ShCCOs by 9 miRNA families. Cis-regulatory elements analysis suggested that CCO genes functioned in the process of growth and development or under the phytohormonal, biotic and abiotic stresses. qRT-PCR analysis indicated that nine CCO genes from different groups exhibited similar expression patterns under abscisic acid treatment, while more divergent profiles were observed in response to Sporisorium scitamineum and cold stresses. Herein, comparative genomics analysis of the CCO gene family between S. spontaneum and R570 was conducted to investigate its evolution and functions. This is the first report on the CCO gene family in S. spontaneum and R570, thus providing valuable information and facilitating further investigation into its function in the future.

Influence of whip smut (Sporisorium scitamineum) infection on quantitative parameters of sugarcane cultivars

Whip smut is one of the most prevalent diseases in almost all sugarcane growing regions of the world. To find out the quantitative losses to sugarcane caused by Sporisorium scitamineum, trials were conducted during the crop season 2014/15 and 2015/16 at sugarcane section, Agriculture Research Institute Tandojam, Pakistan. A total of 104 cultivars of different origins were grown in split plot design with 6 replications. A significant reduction in cane height by 34.33%, cane girth by 18.26%, millable canes by 28.87% and yield per hectare by 41.61% was observed. The most susceptible cultivar was CP-29-120, whereas, the most immune and productive variety was Hoth-318 with the mean yield of 153.33 ± 0.75 t/ha. The statistical analysis results showed highly significant (P = 0.000) impact of disease incidence on different cultivars. In the light of results obtained from the study, it can be concluded that smut disease can cause high reduction in all the quantitative parameters especially in millable canes and yield.

Whole Genome Analysis of Sugarcane Root-Associated Endophyte Pseudomonas aeruginosa B18-A Plant Growth-Promoting Bacterium With Antagonistic Potential Against Sporisorium scitamineum.

Sugarcane smut is a significant fungal disease that causes a major loss in sugar yield and quality. In this study, we isolated an endophytic strain B18 from a sugarcane root, which showed plant growth-promotion, hydrolytic enzyme production, antifungal activity against sugarcane pathogens (Sporisorium scitamineum, Ceratocystis paradoxa, Fusarium verticillioides), and the presence of nifH, acdS, and antibiotic genes (hcn, prn, and phCA) under in vitro conditions. BIOLOG(R) phenotypic profiling of B18 established its ability to use various carbon and nitrogen sources and tolerate a range of pH and osmotic and temperature stresses. Whole-genome analysis of B18, identified as Pseudomonas aeruginosa, showed that it consists of a single circular chromosome of 6,490,014 bp with 66.33% GC content. Genome annotation has identified 5,919 protein-coding genes, and 65 tRNA, and 12 rRNA genes. The P. aeruginosa B18 genome encodes genes related to ethylene, nitrogen (nifU, norBCDERQ, gltBDPS, and aatJMPQ), and phosphate (pstABCS and phoBDHRU) metabolism and produce indole-3-acetic acid and siderophores. This also includes genes encoding hydrolases and oxidoreductases, those associated with biocontrol mechanisms (hcnABC, phzA_B, phzDEFGMS, and pchA), colonization (minCDE and lysC), and biofilm formation (efp, hfq, flgBCDEFGHI, and motAB), and those associated with metabolism of secondary metabolites. Collectively, these results suggest a role for P. aeruginosa B18 in plant growth enhancement and biocontrol mechanisms. The P. aeruginosa B18 strain was found to be an efficient colonizer in sugarcane; it can improve growth through modulation of plant hormone production and enhanced host-plant resistance to smut pathogen S. scitamineum in a smut-susceptible sugarcane variety (Yacheng71-374). These biocontrol and plant growth promotion properties of P. aeruginosa B18 area are discussed in this report.

SCREENING OF SUGARCANE CLONES UNDER ZONAL YIELD TRIALS III, II, I AND ADVANCED YIELD TRIALS AGAINST SMUT DISEASE (USTALIGO SCITAMINEA) IN PLANT CANE

A field trial was conducted on sugarcane genotype /clones against sugarcane smut disease (Ustilago Scitaminea) at Bangladesh sugarcane Research Institute (BSRI) farm, Ishurdi, Pubna during the cropping season 2010 -11 for plant cane. Forty-seven clones were inoculated by I g smut teliospore/liter of water equivalent to 105 -106 spore /ml, for 30 minutes mixed with tween 20 for the inoculation of smut spore by dipping method. Among them 32 clones were (R) resistant ,6 Clones were (MR) moderately Resistant, 6 were (MS) Moderately susceptible,2 were (S) Susceptible and only one clone 15-08 highly susceptible found to sugarcane smut disease.

Physiological Basis of Smut Infectivity in the Early Stages of Sugar Cane Colonization.

Sugar cane smut (Sporisorium scitamineum) interactions have been traditionally considered from the plant’s point of view: How can resistant sugar cane plants defend themselves against smut disease? Resistant plants induce several defensive mechanisms that oppose fungal attacks. Herein, an overall view of Sporisorium scitamineum’s mechanisms of infection and the defense mechanisms of plants are presented. Quorum sensing effects and a continuous reorganization of cytoskeletal components, where actin, myosin, and microtubules are required to work together, seem to be some of the keys to a successful attack.

Ustilago maydis effector Jsi1 interacts with Topless corepressor, hijacking plant jasmonate/ethylene signaling.

Summary Ustilago maydis is the causal agent of maize smut disease. During the colonization process, the fungus secretes effector proteins that suppress immune responses and redirect the host metabolism in favor of the pathogen. As effectors play a critical role during plant colonization, their identification and functional characterization are essential to understanding biotrophy and disease.Using biochemical, molecular, and transcriptomic techniques, we performed a functional characterization of the U. maydis effector Jasmonate/Ethylene signaling inducer 1 (Jsi1).Jsi1 interacts with several members of the plant corepressor family Topless/Topless related (TPL/TPR). Jsi1 expression in Zea mays and Arabidopsis thaliana leads to transcriptional induction of the ethylene response factor (ERF) branch of the jasmonate/ethylene (JA/ET) signaling pathway. In A. thaliana, activation of the ERF branch leads to biotrophic susceptibility. Jsi1 likely activates the ERF branch via an EAR (ET‐responsive element binding‐factor‐associated amphiphilic repression) motif, which resembles EAR motifs from plant ERF transcription factors, that interacts with TPL/TPR proteins.EAR‐motif‐containing effector candidates were identified from different fungal species, including Magnaporthe oryzae, Sporisorium scitamineum, and Sporisorium reilianum. Interaction between plant TPL proteins and these effector candidates from biotrophic and hemibiotrophic fungi indicates the convergent evolution of effectors modulating the TPL/TPR corepressor hub.

Sustainable Control of the Sugarcane Smut Disease Caused by <i>Sporisorium scitamineum</i> Piep. Using an Essential Oil of <i>Cymbopogon citratus</i>

Sugarcane smut disease, caused by <i>Sporisorium scitamineum</i>, is one of the most damaging fungal diseases in the world. This study aims to evaluate the effect of treatment with a formulation based on <i>Cymbopogon citratus</i> essential oil on the incidence of <i>S. scitamineum</i> in sugarcane cultivation. The study was conducted under controlled conditions at the research site of the Bingerville scientific pole in Côte d'Ivoire. Some sugarcane cuttings of NCo376 variety were inoculated by dipping in a teliospores solution of 5.106 teliospores/ml. Before planting, the cuttings were cold treated in a <i>C. citratus</i> essential oil. Propiconazole-treated cuttings served as a reference control and untreated cuttings served as a negative control. Agronomic parameters of the cane and descriptors of smut disease were monitored for eight months to assess the health status of treated and untreated plants. Analysis of variance and comparison of means were performed using the Newman-Keuls test at 5% significance level using Statistica 7.1 software. The results showed that, the pre-treatment of cuttings with <i>C. citratus</i> oil had a benefic effect on the cutting’s germination, height, tillering, stem diameter, internode length, number of internodes and biomass of sugarcane plants with the doses of 500-ppm and 1000-ppm. Moreover, the incidence of smut disease was greatly reduced from the 500-ppm of <i>C. citratus </i>essential oil. In addition, the essential oil-based treatment had similar effects to propiconazole and much better than the untreated controls. Therefore, this pre-treatment of cuttings with <i>C. citratus</i> essential oil could be an alternative to the use of chemicals in the fight against sugarcane smut disease.

Comparative analysis of protein and differential responses of defense-related gene and enzyme activity reveals the long-term molecular responses of sugarcane inoculated with Sporisorium scitamineum

ABSTRACT Smut, the most important sugarcane fungal disease in China, is caused by Sporisorium scitamineum. Smut infection causes severe cane and sugar yield loss. Whole-plant resistance response is characterized by the expression of a large number of pathogenesis-related proteins and genes. Here, proteomic analysis by isobaric tag for relative and absolute quantification was conducted to identify differentially expressed proteins of two sugarcane varieties with contrasting resistance to smut, i.e. Yacheng71-374 (smut-susceptible) and GT29 (smut-resistant) following S. scitamineum infection. A total of 125 and 290 proteins were found differentially expressed after whip appearance in Yacheng71-374 and GT29 sugarcane varieties. Increased up-regulation of peroxidase and catalase and proteins involved in photosynthesis, calcium signaling, abscisic acid biosynthesis, and protein metabolism, were observed in GT29 as compared to Yacheng71-374, suggesting their potential involvement in smut defense mechanism(s) in sugarcane. The expression level of five important defense-related enzymes i.e. chitinase, glucanase, catalase, superoxide dismutase, and phenylalanine ammonia-lyase was analyzed by RT-qPCR. The expression of these enzymes was found enhanced considerably in GT29 as compared to Yacheng71-374 for up to six months following pathogen infection. These results provide new molecular insights into long-term responses of sugarcane to smut infection, a much over-looked aspect of sugarcane smut disease.

Resistance mechanisms and expression of disease resistance-related genes in sugarcane (Sacchrum officinarum) to Sporisorium scitamineum infection.

Resistance of sugarcane (Saccharum officinarum L.) to smut disease (caused by Sporisorium scitamineum) is driven by two separate mechanisms, external and internal resistance. Two progenies generated from an introgression cross, with contrasting responses to smut infection were used to investigate this interaction. Histopathological screening at different stages of the plant growth was used to determine the extent of mycelium growth within sugarcane tissues. Ten disease resistance-related genes were selected, and the relative expression determined using quantitative real-time reverse transcription PCR (real-time RT-qPCR). The results revealed that PR10, HCT1 and ScChi were down-regulated in the susceptible progeny and up-regulated in the resistant progeny early infection process. This may reflect an early attempt to halt pathogen development by increasing the lignin deposition at the infection site. At 8 weeks post-inoculation, they were highly up-regulated in the susceptible progeny coincided with whip development. This reveals a major role for these genes to whip development in the susceptible progeny and indicates that while PR10 is involved in the resistance mechanism of resistant progeny early infection process it also has a role in susceptibility. These results on genetically related progeny with different responses to smut infection reveal a complex interaction of genes and gene networks being induced in response to fungal invasion.

Evaluation of native Trichoderma isolates for the management of sugarcane smut (Ustilago scitaminea) in sugar plantations of Ethiopia

Biological control of sugarcane smut (Ustilago scitaminea Syd.) using Trichoderma isolates is one of the untapped potentials in Ethiopian sugarcane plantations. To this end, this study was carried ...