Sugarcane Species Research Articles

Bioinformatic insights into sugar signaling pathways in sugarcane growth

The SnRK1, hexokinase, and TORC1 (TOR, LST8, RAPTOR) are three pivotal kinases at the core of sugar level sensing, significantly impacting plant metabolism and development. We retrieved and analyzed protein sequences of these three kinase pathways from seven sugarcane transcriptome and genome datasets, identifying protein domains, phylogenetic relationships, sequence ancestry, and in silico expression levels. Additionally, we predicted HXK subcellular localization and assessed its enzymatic activity in sugarcane leaves and culms along development in the field. We retrieved 11 TOR, 23 RAPTOR, 55 LST8, 95 SnRK1α, 98 HXK, and 14 HXK-like putative full-length sequences containing all the conserved domains. Most of these transcripts seem to share a common origin with the three ancestral species of sugarcane: Saccharum officinarum, Saccharum spontaneum, and Saccharum barberi. We accessed the expression profile of sequences from one sugarcane transcriptome. We found the highest enzymatic activity of HXK in culms in the first month, which, at this stage, provides carbon (sucrose) and nitrogen (amino acids) for initial plant development. Our approach places novel sugar sensing sequences that work as a guideline for further research into the underlying signaling mechanisms and biotechnology applications in sugarcane.

Identification of male sterility-related genes in Saccharum officinarum and Saccharum spontaneum.

Candidate male sterility genes were identified in sugarcane, which interacts with kinase-related proteins, transcription factors, and plant hormone signaling pathways to regulate stamen and anther development. Saccharum officinarum is a cultivated sugarcane species that its predominant feature is high sucrose content in stems. Flowering is necessary for breeding new cultivars but will terminate plant growth and reduce sugar yield. The wild sugarcane species Saccharum spontaneum has robust and viable pollen, whereas most S. officinarum accessions are male sterile, which isa desirable trait of a maternal parent in sugarcane breeding. To study male sterility and related regulatory pathways in sugarcane, we carried out RNAseq using flowers in different developmental stages between male-sterile S. officinarum accession 'LA Purple' and fertile S. spontaneum accession 'SES208'. Gene expression profiles were used to detect how genes are differentially expressed between male sterile and fertile flowers and to identify candidate genes for male sterility. Weighted gene correlation networks analysis (WGCNA) was conducted to investigate the regulatory networks. Transcriptomic analyses showed that 988 genes and 2888 alleles were differentially expressed in S. officinarum compared to S. spontaneum. Ten differentially expressed genes and thirty alleles were identified as candidate genes and alleles for male sterility in sugarcane. The gene Sspon.03G0007630 and two alleles of the gene Sspon.08G0002270, Sspon.08G0002270-2B and Sspon.08G0014700-1A, were involved in the early stamen or carpel development stages, while the remaining genes were classified into the post-meiosis stage. Gibberellin, auxin, and jasmonic acid signaling pathways are involved in the stamen development in sugarcane. The results expanded our knowledge of male sterility-related genes in sugarcane and generated genomic resources to facilitate the selection of ideal maternal parents to improve breeding efficiency.

Identification and characterization of PAL genes involved in the regulation of stem development in Saccharum spontaneumL.

BackgroundSaccharum spontaneumL. is a closely related species of sugarcane and has become an important genetic component of modern sugarcane cultivars. Stem development is one of the important factors for affecting the yield, while the molecular mechanism of stem development remains poorly understanding in S. spontaneum. Phenylalanine ammonia-lyase (PAL) is a vital component of both primary and secondary metabolism, contributing significantly to plant growth, development and stress defense. However, the current knowledge about PAL genes in S. spontaneum is still limited. Thus, identification and characterization of the PAL genes by transcriptome analysis will provide a theoretical basis for further investigation of the function of PAL gene in sugarcane.ResultsIn this study, 42 of PAL genes were identified, including 26 SsPAL genes from S. spontaneum, 8 ShPAL genes from sugarcane cultivar R570, and 8 SbPAL genes from sorghum. Phylogenetic analysis showed that SsPAL genes were divided into three groups, potentially influenced by long-term natural selection. Notably, 20 SsPAL genes were existed on chromosomes 4 and 5, indicating that they are highly conserved in S. spontaneum. This conservation is likely a result of the prevalence of whole-genome replications within this gene family. The upstream sequence of PAL genes were found to contain conserved cis-acting elements such as G-box and SP1, GT1-motif and CAT-box, which collectively regulate the growth and development of S. spontaneum. Furthermore, quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis showed that SsPAL genes of stem had a significantly upregulated than that of leaves, suggesting that they may promote the stem growth and development, particularly in the + 6 stem (The sixth cane stalk from the top to down) during the growth stage.ConclusionsThe results of this study revealed the molecular characteristics of SsPAL genes and indicated that they may play a vital role in stem growth and development of S. spontaneum. Altogether, our findings will promote the understanding of the molecular mechanism of S. spontaneum stem development, and also contribute to the sugarcane genetic improving.

Genome-wide identification of JAZ gene family in sugarcane and function analysis of ScJAZ1/2 in drought stress response and flowering regulation

The JASMONATE ZIM DOMAIN (JAZ) proteins are a key inhibitors of the jasmonic acid (JA) signaling pathway that play an important role in the regulation of plant growth and development and environmental stress responses. However, there is no systematic identification and functional analysis of JAZ gene family members in sugarcane. In this study, a total of 49 SsJAZ genes were identified from the wild sugarcane species Saccharum spontaneum genome that were unevenly distributed on 13 chromosomes. Phylogenetic analysis showed that all SsJAZ members can be divided into six groups, and most of the SsJAZ genes contained photoreactive and ABA-responsive elements. RNA-seq analysis revealed that SsJAZ1-1/2/3/4 and SsJAZ7-1 were significantly upregulated under drought stress. The transcript level of ScJAZ1 which is the homologous gene of SsJAZ1 in modern sugarcane cultivars was upregulated by JA, PEG, and abscisic acid (ABA). Moreover, ScJAZ1 can interact with three other JAZ proteins to form heterodimers. The spatial and temporal expression analysis showed that SsJAZ2-1/2/3/4 were highly expressed in different tissues and growth stages and during the day-night rhythm between 10:00 and 18:00. Overexpression of ScJAZ2 in Arabidopsis accelerated flowering through activating the expression of AtSOC1, AtFT, and AtLFY. Moreover, the transcription level of ScJAZ2 was about 30-fold in the early-flowering sugarcane variety than that of the non-flowering variety, indicating ScJAZ2 positively regulated flowering. This first systematic analysis of the JAZ gene family and function analysis of ScJAZ1/2 in sugarcane provide key candidate genes and lay the foundation for sugarcane breeding.

Identification of Stomata Types in Plants of The Genus Saccharum

Stomata are openings in the epidermis that are lined by two special epidermal cells known as guard cells. In sugarcane (Saccharum officinarum) leaves, stomata are found on both the upper (adaxial) and lower (abaxial) surfaces, making them amphistomatous. Stomata are classified into several types, including anomocytic, anisocytic, and parasitic. Studies on the structure of stomata in plants of the genus Saccharum are still limited. This study aims to provide scientific information about the types of stomata in Saccharum plants. This study was carried out from October to November 2023 at the biology laboratory. Qualitative descriptive method with the type of stomata considered under the microscope. Data collection techniques using observation. The results of this investigation show that the genus Saccharum has Graminae type stomata, guard cells shaped like a halter. Stomata size in sugarcane species is generally larger than in barnyard grass species.

Analysis of the 2OG‐Fe (II) oxygenase family reveals new insights associated with flowering regulation in Saccharum spontaneum

AbstractPlant flowering is a crucial phenomenon affecting crop yield and crossbreeding. Saccharum spontaneum is the closest wild relative sugarcane species widely used in sugarcane genetic improvement. However, the molecular interactions among the components of the flowering regulatory network of S. spontaneum are still unclear. In this study, we conducted a transcriptome sequencing approach to analyze the expressed genes on four developmental stages of panicle samples in S. spontaneum. The weighted gene co‐expression network analysis (WGCNA) results suggested that genes from the MEred (p = 3 × 10−4) module were significantly enriched in pathways of ABC transporters, photosynthesis, and circadian rhythm‐plant. Thus, 23 genes associated with flower development‐related genes were screened, including 7 AGL24, 1 FLC, 3 VIN3, 3 GA20ox, and 9 ELF3. In the gibberellic acid (GA) synthesis pathway, GA20ox, GA3ox, and GA2ox were the three enzymes that catalyzed later reactions of GA biosynthesis and belong to the 2‐oxoglutarate Fe (II) oxygenase (2OG) superfamily. Flavonoid synthesis was promoted by 2‐oxoglutarate Fe (II) oxygenase genes, a flowering hormone, which catalyzed and oxidized naringenin to dihydrokaempferol, an important precursor of florigen. Genome‐wide analysis revealed that 122 Ss2OG genes were identified in S. spontaneum, of which 99 genes were enriched in the panicle transcriptome. Furthermore, a quantitative reverse transcription polymerase chain reaction analysis showed that Ss2OG‐FeII_Oxy90, Ss2OG‐FeII_Oxy47, and Ss2OG‐FeII_Oxy89 could introduce the flower development, while Ss2OG‐FeII_Oxy34, Ss2OG‐FeII_Oxy37, and Ss2OG‐FeII_Oxy50 inhibited it. These findings provide new clues and resources for exploring the molecular mechanism of flowering regulation in S. spontaneum and promoting sugarcane breeding.

Pyrolysis of Energy Cane Bagasse: Investigating Kinetics, Thermodynamics, and Effect of Temperature on Volatile Products

Energy cane is a genotype derived from species of sugarcane (Saccharum officinarum and Saccharum spontaneum) with a lower sucrose content and higher fiber content for bioenergy purposes. It is a rustic plant that demands less fertile soils that do not compete with food crops. In this work, an analysis of energy cane bagasse pyrolysis products was performed, assessing the effect of reaction temperature and kinetic and thermodynamic parameters. Anhydrosugars, such as D-allose, were the primary compounds derived from the decomposition of energy cane at 500 °C. Methyl vinyl ketone and acetic acid were favored at 550 and 600 °C. At 650 °C, methyl glyoxal, acetaldehyde and hydrocarbons were favored. Among the hydrocarbons observed, butane, toluene and olefins such as 1-decene, 1-undecene, 1-tridecene and 1-tetradecene were the most produced. The Friedman isoconversional method was able to determine the average activation energies in the ranges 113.7−149.4, 119.9−168.0, 149.3−196.4 and 170.1−2913.9 kJ mol−1 for the decomposition of, respectively, pseudo-extractives, pseudo-hemicellulose, pseudo-cellulose and pseudo-lignin. The thermodynamic parameters of activation were determined within the ranges of 131.0 to 507.6 kJ mol−1 for ΔH, 153.7 to 215.2 kJ mol−1 for ΔG and −35.5 to 508.8 J mol−1 K−1 for ΔS. This study is very encouraging for the cultivation and use of high-fiber-content energy cane bagasse, after sucrose extraction, to produce biofuels as an alternative to the current method of conversion into electricity by low-efficiency burning.

Comparative physiological and transcriptome analysis in cultivated and wild sugarcane species in response to hydrogen peroxide-induced oxidative stress

BackgroundSugarcane is an important energy crop grown worldwide,supplementing various renewable energy sources. Cultivated and wild sugarcane species respond differently to biotic and abiotic stresses. Generally, wild species are tolerant to various abiotic stresses. In the present study, the physiological and molecular responses of cultivated and wild sugarcane species to oxidative stress at the transcriptional levels were compared. Transcriptional responses were determined using RNAseq. The representative RNA-seq transcript values were validated by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) and confirmed through physiological responses.ResultsOxidative stress causes leaf-rolling and -tip drying in cultivated sugarcane, but the wild species are tolerant. Higher chlorophyll fluorescence was observed in the wild species than that in the cultivated varieties under stress. Wild species can maintain a higher chlorophyll stability index than the cultivated species, which was confirmed by the lower transcripts of the chlorophyllase gene in the wild species than that in the cultivated variety. Transcription factor genes (NAC, MYB, and WRKY) were markedly expressed in response to oxidative stress, revealing their involvement in stress tolerance. The analysis revealed synchronized expression of acetyl-transferase, histone2A, cellulose synthase, and secondary cell wall biosynthetic genes in the wild species. The validation of selected genes and 15 NAC transcription factors using RT-qPCR revealed that their expression profiles were strongly correlated with RNA-seq. To the best of our knowledge, this is the first report on the oxidative stress response in cultivated and wild sugarcane species.ConclusionPhysiological and biochemical changes in response to oxidative stress markedly differ between cultivated and wild sugarcane species. The differentially expressed stress-responsive genes are grouped intothe response to oxidative stress, heme-binding, peroxidase activity, and metal ion binding categories. Chlorophyll maintenance is a stress tolerance response enhanced by the differential regulation of the chlorophyllase gene.There is a considerable difference in the chlorophyll stability index between wild and cultivated varieties. We observed a substantial regulation of secondary wall biosynthesis genes in the wild species compared with that in the cultivated variety, suggesting differences in stress tolerance mechanisms.

Global Responses of Autopolyploid Sugarcane Badila (Saccharum officinarum L.) to Drought Stress Based on Comparative Transcriptome and Metabolome Profiling.

Sugarcane (Saccharum spp. hybrid) is frequently affected by seasonal drought, which causes substantial declines in quality and yield. To understand the drought resistance mechanisms of S. officinarum, the main species of modern sugarcane, at a molecular level, we carried out a comparative analysis of transcriptome and metabolome profiling of the sugarcane variety Badila under drought stress (DS). Compared with control group (CG) plants, plants exposed to DS had 13,744 (6663 up-regulated and 7081 down-regulated) differentially expressed genes (DEGs). GO and KEGG analysis showed that the DEGs were enriched in photosynthesis-related pathways and most DEGs had down-regulated expression. Moreover, the chlorophyll content, photosynthesis (Photo), stomatal conductance (Cond), intercellular carbon dioxide concentration (Ci) and transpiration rate (Trmmol) were sharply decreased under DS. These results indicate that DS has a significant negative influence on photosynthesis in sugarcane. Metabolome analysis identified 166 (37 down-regulated and 129 up-regulated) significantly regulated metabolites (SRMs). Over 50% of SRMs were alkaloids, amino acids and their derivatives, and lipids. The five most significantly enriched KEGG pathways among SRMs were Aminoacyl-tRNA biosynthesis, 2-Oxocarboxylic acid metabolism, Biosynthesis of amino acids, Phenylalanine metabolism, and Arginine and proline metabolism (p < 0.05). Comparing CG with DS for transcriptome and metabolome profiling (T_CG/DS and M_CG/DS, respectively), we found three of the same KEGG-enriched pathways, namely Biosynthesis of amino acids, Phenylalanine metabolism and Arginine and proline metabolism. The potential importance of Phenylalanine metabolism and Arginine and proline metabolism was further analyzed for response to DS in sugarcane. Seven SRMs (five up-regulated and two down-regulated) and 60 DEGs (17 up-regulated and 43 down-regulated) were enriched in Phenylalanine metabolism under DS, of which novel.31261, Sspon.04G0008060-1A, Sspon.04G0008060-2B and Sspon.04G0008060-3C were significantly correlated with 7 SRMs. In Arginine and proline metabolism, eight SRMs (seven up-regulated and one down-regulated) and 63 DEGs (32 up-regulated and 31 down-regulated) were enriched, of which Sspon.01G0026110-1A (OAT) and Sspon.03G0002750-3D (P5CS) were strongly associated with proline (r > 0.99). These findings present the dynamic changes and possible molecular mechanisms of Phenylalanine metabolism as well as Arginine and proline metabolism under DS and provide a foundation for future research and sugarcane improvement.

Genome-Wide Identification of Auxin-Responsive GH3 Gene Family in Saccharum and the Expression of ScGH3-1 in Stress Response.

Gretchen Hagen3 (GH3), one of the three major auxin-responsive gene families, is involved in hormone homeostasis in vivo by amino acid splicing with the free forms of salicylic acid (SA), jasmonic acid (JA) or indole-3-acetic acid (IAA). Until now, the functions of sugarcane GH3 (SsGH3) family genes in response to biotic stresses have been largely unknown. In this study, we performed a systematic identification of the SsGH3 gene family at the genome level and identified 41 members on 19 chromosomes in the wild sugarcane species, Saccharum spontaneum. Many of these genes were segmentally duplicated and polyploidization was the main contributor to the increased number of SsGH3 members. SsGH3 proteins can be divided into three major categories (SsGH3-I, SsGH3-II, and SsGH3-III) and most SsGH3 genes have relatively conserved exon-intron arrangements and motif compositions. Diverse cis-elements in the promoters of SsGH3 genes were predicted to be essential players in regulating SsGH3 expression patterns. Multiple transcriptome datasets demonstrated that many SsGH3 genes were responsive to biotic and abiotic stresses and possibly had important functions in the stress response. RNA sequencing and RT-qPCR analysis revealed that SsGH3 genes were differentially expressed in sugarcane tissues and under Sporisorium scitamineum stress. In addition, the SsGH3 homolog ScGH3-1 gene (GenBank accession number: OP429459) was cloned from the sugarcane cultivar (Saccharum hybrid) ROC22 and verified to encode a nuclear- and membrane-localization protein. ScGH3-1 was constitutively expressed in all tissues of sugarcane and the highest amount was observed in the stem pith. Interestingly, it was down-regulated after smut pathogen infection but up-regulated after MeJA and SA treatments. Furthermore, transiently overexpressed Nicotiana benthamiana, transduced with the ScGH3-1 gene, showed negative regulation in response to the infection of Ralstonia solanacearum and Fusarium solani var. coeruleum. Finally, a potential model for ScGH3-1-mediated regulation of resistance to pathogen infection in transgenic N. benthamiana plants was proposed. This study lays the foundation for a comprehensive understanding of the sequence characteristics, structural properties, evolutionary relationships, and expression of the GH3 gene family and thus provides a potential genetic resource for sugarcane disease-resistance breeding.

Unraveling Nitrogen Fixing Potential of Endophytic Diazotrophs of Different Saccharum Species for Sustainable Sugarcane Growth.

Sugarcane (Saccharum officinarum L.) is one of the world’s highly significant commercial crops. The amounts of synthetic nitrogen (N2) fertilizer required to grow the sugarcane plant at its initial growth stages are higher, which increases the production costs and adverse environmental consequences globally. To combat this issue, sustainable environmental and economic concerns among researchers are necessary. The endophytic diazotrophs can offer significant amounts of nitrogen to crops through the biological nitrogen fixation mediated nif gene. The nifH gene is the most extensively utilized molecular marker in nature for studying N2 fixing microbiomes. The present research intended to determine the existence of novel endophytic diazotrophs through culturable and unculturable bacterial communities (EDBCs). The EDBCs of different tissues (root, stem, and leaf) of five sugarcane cultivars (Saccharum officinarum L. cv. Badila, S. barberi Jesw.cv Pansahi, S. robustum, S. spontaneum, and S. sinense Roxb.cv Uba) were isolated and molecularly characterized to evaluate N2 fixation ability. The diversity of EDBCs was observed based on nifH gene Illumina MiSeq sequencing and a culturable approach. In this study, 319766 operational taxonomic units (OTUs) were identified from 15 samples. The minimum number of OTUs was recorded in leaf tissues of S. robustum and maximum reads in root tissues of S. spontaneum. These data were assessed to ascertain the structure, diversity, abundance, and relationship between the microbial community. A total of 40 bacterial families with 58 genera were detected in different sugarcane species. Bacterial communities exhibited substantially different alpha and beta diversity. In total, 16 out of 20 genera showed potent N2-fixation in sugarcane and other crops. According to principal component analysis (PCA) and hierarchical clustering (Bray–Curtis dis) evaluation of OTUs, bacterial microbiomes associated with root tissues differed significantly from stem and leaf tissues of sugarcane. Significant differences often were observed in EDBCs among the sugarcane tissues. We tracked and validated the plethora of individual phylum strains and assessed their nitrogenase activity with a culture-dependent technique. The current work illustrated the significant and novel results of many uncharted endophytic microbial communities in different tissues of sugarcane species, which provides an experimental system to evaluate the biological nitrogen fixation (BNF) mechanism in sugarcane. The novel endophytic microbial communities with N2-fixation ability play a remarkable and promising role in sustainable agriculture production.

Genomic Analyses of SUT and TST Sugar Transporter Families in Low and High Sugar Accumulating Sugarcane Species (Saccharum spontaneum and Saccharum officinarum)

Genomic Analyses of SUT and TST Sugar Transporter Families in Low and High Sugar Accumulating Sugarcane Species (Saccharum spontaneum and Saccharum officinarum)

High-Throughput Sequencing-Based Analysis of Rhizosphere and Diazotrophic Bacterial Diversity Among Wild Progenitor and Closely Related Species of Sugarcane (Saccharum spp. Inter-Specific Hybrids).

Considering the significant role of genetic background in plant-microbe interactions and that most crop rhizospheric microbial research was focused on cultivars, understanding the diversity of root-associated microbiomes in wild progenitors and closely related crossable species may help to breed better cultivars. This study is aimed to fill a critical knowledge gap on rhizosphere and diazotroph bacterial diversity in the wild progenitors of sugarcane, the essential sugar and the second largest bioenergy crop globally. Using a high-throughput sequencing (HTS) platform, we studied the rhizosphere and diazotroph bacterial community of Saccharum officinarum L. cv. Badila (BRS), Saccharum barberi (S. barberi) Jesw. cv Pansahi (PRS), Saccharum robustum [S. robustum; (RRS), Saccharum spontaneum (S. spontaneum); SRS], and Saccharum sinense (S. sinense) Roxb. cv Uba (URS) by sequencing their 16S rRNA and nifH genes. HTS results revealed that a total of 6,202 bacteria-specific operational taxonomic units (OTUs) were identified, that were distributed as 107 bacterial groups. Out of that, 31 rhizobacterial families are commonly spread in all five species. With respect to nifH gene, S. barberi and S. spontaneum recorded the highest and lowest number of OTUs, respectively. These results were validated by quantitative PCR analysis of both genes. A total of 1,099 OTUs were identified for diazotrophs with a core microbiome of 9 families distributed among all the sugarcane species. The core microbiomes were spread across 20 genera. The increased microbial diversity in the rhizosphere was mainly due to soil physiochemical properties. Most of the genera of rhizobacteria and diazotrophs showed a positive correlation, and few genera negatively correlated with the soil properties. The results showed that sizeable rhizospheric diversity exists across progenitors and close relatives. Still, incidentally, the rhizosphere microbial abundance of progenitors of modern sugarcane was at the lower end of the spectrum, indicating the prospect of Saccharum species introgression breeding may further improve nutrient use and disease and stress tolerance of commercial sugarcane. The considerable variation for rhizosphere microbiome seen in Saccharum species also provides a knowledge base and an experimental system for studying the evolution of rhizobacteria-host plant association during crop domestication.

Population Structure and Genetic Diversity Analysis in Sugarcane (Saccharum spp. hybrids) and Six Related Saccharum Species

Sugarcane (Saccharum spp. hybrids) is one of the most important commercial crops for sugar, ethanol, and other byproducts production; therefore, it is of great significance to carry out genetic research. Assessing the genetic population structure and diversity plays a vital role in managing genetic resources and gene mapping. In this study, we assessed the genetic diversity and population structure among 196 Saccharum accessions, including 34 S. officinarum, 69 S. spontaneum, 17 S. robustum, 25 S. barberi, 13 S. sinense, 2 S. edule, and 36 Saccharum spp. hybrids. A total of 624 polymorphic SSR alleles were amplified by PCR with 22 pairs of fluorescence-labeled highly polymorphic SSR primers and identified on a capillary electrophoresis (CE) detection system including 109 new alleles. Three approaches (model-based clustering, principal component analysis, and phylogenetic analysis) were conducted for population structure and genetic diversity analyses. The results showed that the 196 accessions could be grouped into either three (Q) or eight (q) sub-populations. Phylogenetic analysis indicated that most accessions from each species merged. The species S. barberi and S. sinense formed one group. The species S. robustum, S. barberi, S. spontaneum, S. edule, and sugarcane hybrids merged into the second group. The S. officinarum accessions formed the third group located between the other two groups. Two-way chi-square tests derived a total of 24 species-specific or species-associated SSR alleles, including four alleles each for S. officinarum, S. spontaneum, S. barberi, and S. sinense, five alleles for S. robustum. and three alleles for Saccharum spp. hybrids. These species-specific or species-associated SSR alleles will have a wide application value in sugarcane breeding and species identification. The overall results provide useful information for future genetic study of the Saccharum genus and efficient utilization of sugarcane germplasm resources in sugarcane breeding.

Variação genotípica de cana-de-açúcar para tolerância a salinidade: Respostas morfológicas e fisiológicas

ABSTRACT Sugarcane (Saccharum spp.) is an important crop due to sugar, ethanol and bioenergy production. Its cultivation may occur in tropical regions exposed to high salinity. The aim was to identify cultivars tolerant to salinity to allow the cultivation of sugarcane (Saccharum spp.) in saline soils. To test the hypothesis that sugarcane show natural genotypic variation to salinity tolerance, we tested ten cultivars (SP80-3280, RB855453 RB966928, RB855156, SP80-1842, SP80-1816, RB928064, RB867515, RB92579, RB855536) and two sugarcane species: IM76-228 (S. robustum) and IN84-82 (S. spontaneum) under two concentrations of sodium chloride (NaCl): control (concentration found naturally in the soil used: electrical conductivity of 0.083 dS m-1) and soil enriched with NaCl: EC of 7.2 dS m-1. Biometry and photosynthesis traits were evaluated. The plants were collected and leaf (LDM), stem (SDM), root and total dry matter were used to estimate the tolerance index (TI). A cluster analysis was done to identify phenotypic dissimilarity. Three distinct groups regarding salinity tolerance on biomass-basis were formed. The cultivars RB855156, SP80-1842, SP80-1816 and species IM76-228 showed no reduction in LDM and SDM. Nonetheless, the cultivars SP80-3280, RB928064, RB92579 and species IN84-82 were impaired by salinity. The cultivar SP80-1816 showed the highest biomass accumulation and the highest TI. Therefore, we found a great genotypic variation regarding salinity tolerance in sugarcane, which can be explored by growers to cultivate in saline soils. Also can be used by the Sugarcane Breeding Programs to improve the salinity tolerance.

Partial gene sequencing of phosphoenol pyruvate carboxylase (PEPc) gene from Saccharum spp. hybrid CoLk 94184

Phosphoenol pyruvate carboxylase (PEPc), a key enzyme in photosynthesis, an important carboxylase for plant cell metabolism, fixes CO2 in mesophyll cells to form dicarboxylic acids, and being more specific for inorganic carbon, improves the photosynthetic rate of C4 plants. In the present study, partial gene sequencing of PEPc was done using Saccharum spp. hybrid, CoLk 94184 of sugarcane, one of the most popular variety released by ICAR-IISR, Lucknow. Utilizing PEPc of different cultivars of sugarcane species, the primers were designed for PEPc gene and the first ever nucleotide sequence was determined, especially for Saccharum spp. hybrid CoLk 94184 (accession no: MW766370.1). Sequence based phylogenetic tree showed close relatedness between the PEPc sequences for various Saccharum cultivars, thus pointing to little genetic change, while that for sorghum and maize were relatively distantly related. Results may help in understanding the photosynthetic process using sugarcane hybrid specific genes

Root-Derived Endophytic Diazotrophic Bacteria Pantoea cypripedii AF1 and Kosakonia arachidis EF1 Promote Nitrogen Assimilation and Growth in Sugarcane.

Excessive, long-term application of chemical fertilizers in sugarcane crops disrupts soil microbial flora and causes environmental pollution and yield decline. The role of endophytic bacteria in improving crop production is now well-documented. In this study, we have isolated and identified several endophytic bacterial strains from the root tissues of five sugarcane species. Among them, eleven Gram-negative isolates were selected and screened for plant growth-promoting characteristics, i.e., production of siderophores, indole-3-acetic acid (IAA), ammonia, hydrogen cyanide (HCN), and hydrolytic enzymes, phosphorus solubilization, antifungal activity against plant pathogens, nitrogen-fixation, 1-aminocyclopropane-1-carboxylic acid deaminase activity, and improving tolerance to different abiotic stresses. These isolates had nifH (11 isolates), acdS (8 isolates), and HCN (11 isolates) genes involved in N-fixation, stress tolerance, and pathogen biocontrol, respectively. Two isolates Pantoea cypripedii AF1and Kosakonia arachidis EF1 were the most potent strains and they colonized and grew in sugarcane plants. Both strains readily colonized the leading Chinese sugarcane variety GT42 and significantly increased the activity of nitrogen assimilation enzymes (glutamine synthetase, NADH glutamate dehydrogenase, and nitrate reductase), chitinase, and endo-glucanase and the content of phytohormones gibberellic acid, indole-3-acetic acid, and abscisic acid. The gene expression analysis of GT42 inoculated with isolates of P. cypripedii AF1 or K. arachidis EF1 showed increased activity of nifH and nitrogen assimilation genes. Also, the inoculated diazotrophs significantly increased plant nitrogen content, which was corroborated by the 15N isotope dilution analysis. Collectively, these findings suggest that P. cypripedii and K. arachidis are beneficial endophytes that could be used as a biofertilizer to improve plant nitrogen nutrition and growth of sugarcane. To the best of our knowledge, this is the first report of sugarcane growth enhancement and nitrogen fixation by Gram-negative sugarcane root-associated endophytic bacteria P. cypripedii and K. arachidis. These strains have the potential to be utilized as sugarcane biofertilizers, thus reducing nitrogen fertilizer use and improving disease management.

Chromosomal Characterization of Tripidium arundinaceum Revealed by Oligo-FISH.

Sugarcane is of important economic value for producing sugar and bioethanol. Tripidium arundinaceum (old name: Erianthus arundinaceum) is an intergeneric wild species of sugarcane that has desirable resistance traits for improving sugarcane varieties. However, the scarcity of chromosome markers has hindered the cytogenetic study of T. arundinaceum. Here we applied maize chromosome painting probes (MCPs) to identify chromosomes in sorghum and T. arundinaceum using a repeated fluorescence in situ hybridization (FISH) system. Sequential FISH revealed that these MCPs can be used as reliable chromosome markers for T. arundinaceum, even though T. arundinaceum has diverged from maize over 18 MYs (million years). Using these MCPs, we identified T. arundinaceum chromosomes based on their sequence similarity compared to sorghum and labeled them 1 through 10. Then, the karyotype of T. arundinaceum was established by multiple oligo-FISH. Furthermore, FISH results revealed that 5S rDNA and 35S rDNA are localized on chromosomes 5 and 6, respectively, in T. arundinaceum. Altogether, these results represent an essential step for further cytogenetic research of T. arundinaceum in sugarcane breeding.

Analytical Quantification of mycotoxin by LC-ESI-MS/MS, and MALDI-TOF MS in phoma sorghina species of sugarcane

Mycotoxin s are secondary metabolites produced by fungi, which are harmful to humans and animals. Phoma is a common fungal plant pathogen that can produce mycotoxins in sugarcane. Phoma sorgina belonging to phoma species that cause devastating fungal disease twisted leaf on sugarcane, The present study helps to carry out research on potential metabolites of fungal strain belongs to the phoma sorghina species such as P. sorghina (BS2-1), P. sorghina (BS11-1), P. sorghina (BSQP) isolated from sugarcane. These metabolites may influence the functioning of the fungus and are thus important indirectly for its growth and survival. Mass spectrometry (MS) is commonly used for the detection, characterization and quantitation of mycotoxins in agricultural crops. Phoma species were assessed to provide a risk assessment of their secondary metabolites in sugarcane by describing their global metabolome data obtained from the LC-MS instrument. Another study examined through using matrix-assisted laser desorption ionisation – time of flight mass spectrometry (MALDI-TOF MS) to identify phoma species rapidly and accurately. Different protein mass spectra obtained with MALDI-TOF MS were able to separate the phoma species reported in sugarcane. Therefore, the discovery of new conjugated mycotoxins and secondary metabolites produced phoma species was made possible by combining high mass accuracy data with analytic approaches.

Silicon attenuates the effects of water deficit in sugarcane by modifying physiological aspects and C:N:P stoichiometry and its use efficiency

Climate change has increased the occurrence of droughts and the use of deficient irrigation associated with silicon (Si) can be a strategy to mitigate water deficit in crops; however, the mechanisms involved have to be understood. The objective of this study was to evaluate whether the application of Si via fertigation associated with leaf spray mitigates the effects of water deficit by improving the physiological aspects and modifying C: N: P stoichiometry, inducing an increase in nutrient use efficiency in the ratoon of two sugarcane species (Saccharum officinarum- conventional sugarcane and S. spontaneum- energy cane). Two experiments were carried out in a greenhouse, using conventional sugar cane (variety-RB 966928) and energy cane (variety-VX2). In both experiments, the treatments were arranged in a 2 × 2 factorial scheme, without water deficit and with severe water deficit (30% of soil water retention capacity) without and with application of Si via fertigation associated with leaf spray, both in a concentration 2.5 mmol L−1, arranged in randomized blocks. At 80 days after ratoon regrowth, the following parameters were evaluated: Si content and accumulation, efficiency of using C, N and P, C: Si, C: N and C: P ratios, quantum efficiency of PS II, relative content of water, leaf water potential, leaf area, number of leaves and tillers and dry matter production. In the absence of Si, water deficit affected the physiological variables and the growth of the two sugarcane species. The supply of Si via fertigation associated with foliar spraying of soluble Si increased the absorption of this element in the ratoon of conventional sugarcane and energy cane, regardless of water condition. Si attenuated the effects caused by severe water deficit because of the increase in the relative water content and water potential, thus decreasing oxidative stress, impacting photosynthesis efficiency and also modifying the C: N: P stoichiometry, increasing use efficiency for these nutrients and the growth of the two study species.