Genus Saccharum Research Articles

Graph-based mitochondrial genomes of three foundation species in the Saccharum genus.

We reported the graph-based mitochondrial genomes of three foundation species (Saccharum spontaneum, S. robustum and S. officinarum) for the first time. The results revealed pan-structural variation and evolutionary processes in the mitochondrial genomes within Saccharum. Saccharum belongs to the Andropogoneae, and cultivars species in Saccharum contribute nearly 80% of sugar production in the world. To explore the genomic studies in Saccharum, we assembled 15 complete mitochondrial genomes (mitogenome) of three foundation species (Saccharum spontaneum, S. robustum and S. officinarum) using Illumina and Oxford Nanopore Technologies sequencing data. The mitogenomes of the three species were divided into a total of eight types based on contig numbers and linkages. All mitogenomes in the three species encoded 51 unique genes, including 32 protein-coding, 3 ribosomal RNA (rRNA) and 16 transfer RNA (tRNA) genes. The existence of long and short-repeat-mediated recombinations in the mitogenome of S. officinarum and S. robustum was revealed and confirmed through PCR validation. Furthermore, employing comparative genomics and phylogenetic analyses of the organelle genomes, we unveiled the evolutionary relationships and history of the major interspecific lineages in Saccharum genus. Phylogenetic analyses of homologous fragments between S. officinarum and S. robustum showed that S. officinarum and S. robustum are phylogenetically distinct and that they were likely parallel rather than domesticated. The variations between ancient (S. sinense and S. barberi) and modern cultivated species (S. hybrid) possibly resulted from hybridization involving different S. officinarum accessions. Lastly, this project reported the first graph-based mitogenomes of three Saccharum species, and a systematic comparison of the structural organization, evolutionary processes, and pan-structural variation of the Saccharum mitogenomes revealed the differential features of the Saccharum mitogenomes.

Assembly and comparative genome analysis of four mitochondrial genomes from Saccharum complex species.

Saccharum complex includes genera Saccharum, Miscanthus, Erianthus, Narenga, and Tripidium. Since the Saccharum complex/Saccharinae constitutes the gene pool used by sugarcane breeders to introduce useful traits into sugarcane, studying the genomic characterization of the Saccharum complex has become particularly important. Here, we assembled graph-based mitochondrial genomes (mitogenomes) of four Saccharinae species (T. arundinaceum, E. rockii, M. sinensis, and N. porphyrocoma) using Illumina and PacBio sequencing data. The total lengths of the mitogenomes of T. arundinaceum, M. sinensis, E. rockii and N. porphyrocoma were 549,593 bp, 514,248 bp, 481,576 bp and 513,095 bp, respectively. Then, we performed a comparative mitogenomes analysis of Saccharinae species, including characterization, organelles transfer sequence, collinear sequence, phylogenetics analysis, and gene duplicated/loss. Our results provided the mitogenomes of four species closely related to sugarcane breeding, enriching the mitochondrial genomic resources of the Saccharinae. Additionally, our study offered new insights into the evolution of mitogenomes at the family and genus levels and enhanced our understanding of organelle evolution in the highly polyploid Saccharum genus.

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.

Pharmacological Screening of Polyherbal Formulation for Diabetic Associated Hyperlipidemia


 Plant parts such as seeds, berries, roots, leaves, bark, or flowers can be used medicinally. This practise is known as herbal medicine, botanical medicine, or phytomedicine. Outside of mainstream treatment, herbalists have long used herbalism for conventional medicine. Grewia subinaequalisDC in the family Tiliaceae, only one genus, Grewia, yields edible fruit. Saccharum officinarum is a species of grass belonging to the genus Saccharum that is characterised by its robust growth and size. Neisosperma oppositifolium is a tree that typically exhibits a height range of 6 to 25 metres, although it has been observed to vary from as low as 2.5 metres to extraordinarily high heights of 45 metres, and in rare cases, even up to 60 metres. The presence of moisture in crude pharmaceuticals is an unavoidable factor that should be minimised to the greatest extent possible9,10. The process of drying significantly influences both the quality and purity of the material.

Flowering in sugarcane-insights from the grasses.

Flowering is a crucial phase for angiosperms to continue their species propagation and is highly regulated. In the current review, flowering in sugarcane and the associated mechanisms are elaborately presented. In sugarcane, flowering has two effects, wherein it is a beneficial factor from the breeder's perspective and crucial for crop improvement, but commercially, it depletes the sucrose reserves from the stalks; hence, less value is assigned. Different species of Saccharum genus are spread across geographical latitudes, thereby proving their ability to grow in multiple inductive daylengths of different locations according in the habituated zone. In general, sugarcane is termed an intermediate daylength plant with quantitative short-day behaviour as it requires reduction in daylength from 12h 55min to 12h or 12h 30min. The prime concern in sugarcane flowering is its erratic flowering nature. The transition to reproductive stage which reverts to vegetative stage if there is any deviation from ambient temperature and light is also an issue. Spatial and temporal gene expression patterns during vegetative to reproductive stage transition and after reverting to vegetative state could possibly reveal how the genetic circuits are being governed. This review will also shed a light on potential roles of genes and/or miRNAs in flowering in sugarcane. Knowledge of transcriptomic background of circadian, photoperiod, and gibberellin pathways in sugarcane will enable us to better understand of variable response in floral development.

A complete gap-free diploid genome in Saccharum complex and the genomic footprints of evolution in the highly polyploid Saccharum genus.

A diploid genome in the Saccharum complex facilitates our understanding of evolution in the highly polyploid Saccharum genus. Here we have generated a complete, gap-free genome assembly of Erianthus rufipilus, a diploid species within the Saccharum complex. The complete assembly revealed that centromere satellite homogenization was accompanied by the insertions of Gypsy retrotransposons, which drove centromere diversification. An overall low rate of gene transcription was observed in the palaeo-duplicated chromosome EruChr05 similar to other grasses, which might be regulated by methylation patterns mediated by homologous 24 nt small RNAs, and potentially mediating the functions of many nucleotide-binding site genes. Sequencing data for 211 accessions in the Saccharum complex indicated that Saccharum probably originated in the trans-Himalayan region from a diploid ancestor (x = 10) around 1.9-2.5 million years ago. Our study provides new insights into the origin and evolution of Saccharum and accelerates translational research in cereal genetics and genomics.

In Silico Dissection of Regulatory Regions of PHT Genes from Saccharum spp. Hybrid and Sorghum bicolor and Expression Analysis of PHT Promoters under Osmotic Stress Conditions in Tobacco

Phosphorus (P) is the second-most essential macronutrient required for the growth and development of plants. It is involved in a number of cellular processes that contribute to the plant’s growth and development. This study investigated Saccharum spp. hybrid and Sorghum bicolor promoter regions of Phosphate transporters (PHT), viz., PHT1, PHT2, PHT3, PHT4, and PHO1, through in silico analysis. The transcription start sites (TSS), conserved motifs, and CpG islands were studied using various computational techniques. The distribution of TSSs indicated the highest promoter prediction scores (1.0). MSh2 and MSb4 were recognized as the common promoter motifs for PHT promoters, found in with 85 to 100% percentage of distribution. The CpG analysis revealed that the promoter regions of most PHT genes had low CpG density, indicating a possible tissue-specific expression. The PHT promoters were investigated for the presence of biotic- and abiotic-stress-associated transcription factor binding sites (TFbs) that revealed the presence of binding motifs for major transcription factors (TFs), namely, AP2/ERF, bHLH, bZIP, MYB, NAC, and WRKY. Therefore, the in-silico analysis of the promoter regions helps us to understand the regulation mechanism of phosphate transporter promoters and gene expression under stress management. The 5′ regulatory region of the EaPHT gene was isolated from Erianthus, a wild relative of the genus Saccharum. The promoter construct was prepared and transformed in tobacco wherein the promoter drove the expression of GUS. Analysis of GUS expression in transgenic tobacco revealed enhanced expression of GUS under salt-stress conditions. This is the first report of the isolation and characterization of a phosphate transporter gene promoter from Erianthus and is expected to be useful for the development of salt-stress transgenic crop plants.

Reconstruction of karyotypic evolution in Saccharum spontaneum species by comparative oligo-FISH mapping

BackgroundKaryotype dynamics driven by chromosomal rearrangements has long been considered as a fundamental question in the evolutionary genetics. Saccharum spontaneum, the most primitive and complex species in the genus Saccharum, has reportedly undergone at least two major chromosomal rearrangements, however, its karyotypic evolution remains unclear.ResultsIn this study, four representative accessions, i.e., hypothetical diploid sugarcane ancestor (sorghum, x = 10), Sa. spontaneum Np-X (x = 10, tetraploid), 2012–46 (x = 9, hexaploid) and AP85–441 (x = 8, tetraploid), were selected for karyotype evolution studies. A set of oligonucleotide (oligo)-based barcode probes was developed based on the sorghum genome, which allowed universal identification of all chromosomes from sorghum and Sa. spontaneum. By comparative FISH assays, we reconstructed the karyotype evolutionary history and discovered that although chromosomal rearrangements resulted in greater variation in relative lengths of some chromosomes, all chromosomes maintained a conserved metacentric structure. Additionally, we found that the barcode oligo probe was not applicable for chromosome identification in both Sa. robustum and Sa. officinarum species, suggesting that sorghum is more distantly related to Sa. robustum and Sa. officinarum compared with Sa. spontaneum species.ConclusionsOur study demonstrated that the barcode oligo-FISH is an efficient tool for chromosome identification and karyotyping research, and expanded our understanding of the karyotypic and chromosomal evolution in the genus Saccharum.

Identification of Sugarcane S. spontaneum (Poaceae) Germplasm: Evidence from rDNA-ITS and rDNA Locus Analyses

Sugarcane is a major crop for sugar production around the world. The complexity of the sugarcane genome creates challenges for the use of both conventional and molecular breeding methods to improve sugarcane at a genetic level. DNA sequencing is an important tool to assess how the genus Saccharum and the genera of the Saccharum complex are interrelated. Here, we identify the kinship of Nepal2013-6 (Saccharum spontaneum, x = 10) using a tetra-primer amplification refractory mutation system (ARMS) PCR. Based on rDNA-ITS sequence analysis, the accession Nepal2013-6 falls within a single cluster with S. spontaneum (Yunnan82-114 and SES208), which is consistent with the previous results. Moreover, fluorescence in situ hybridization (FISH) results indicate that the 5S rDNA spots are consistent with the chromosomal ploidy in the analytical Saccharum materials, whereas 35S rDNA has similar or fewer sites than the ploidy. Therefore, 5S rDNA FISH patterns would be more suitable than 35S rDNA for chromosomal ploidy analysis in S. spontaneum with varied basic chromosome number x = 8, 9, 10. Altogether, these results indicate that the rDNA sequences will be a useful marker for further rapidly identifying the relationship and ploidy of S. spontaneum in sugarcane breeding.

Optimal Deep Learning Driven Smart Sugarcane Crop Monitoring on Remote Sensing Images

Abstract Crop monitoring is a process that involves regular field visits that seem to be difficult since it needs a huge amount of time and manpower. Thus, in modern agriculture, with an extensive range of satellite data such as Landsat, Sentinel-2, Modis, and Palsar, data are readily available. Sugarcane is a tall perennial grass belonging to the genus Saccharum, utilized for producing sugar. These plants were generally 2–6 m tall with fibrous, stout, jointed stalks, rich in sucrose, that will be accumulated in the stalk internodes. Sugarcanes have a different growth pattern and phenology than many other crops; thus, the spectral and temporal features of satellite data are examined by utilizing statistical and machine learning (ML) techniques for optimal discrimination of sugarcane fields with other crops. In this study, we propose an Optimal Deep Learning Driven Smart Sugarcane Crop Monitoring (ODLD-SSCM) model on Remote Sensing Images. The presented ODLD-SSCM model mainly intends to estimate the crop yield of sugarcanes using RSIs. In the presented ODLD-SSCM technique, the sugarcane yield mapping can be derived by the use of the self-attentive deep learning (SADL) model. Besides, an oppositional spider colony optimization (OSCO) algorithm is used for the hyperparameter tuning of the ODLD-SSCM model. A detailed set of experimentations were performed to demonstrate the enhanced outcomes of the ODLDSSCM model. A comprehensive comparison study pointed out the enhancements of the ODLD-SSCM model over other recent approaches.

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.

Chromosome Painting Provides Insights Into the Genome Structure and Evolution of Sugarcane.

The genus Saccharum is composed of species with high polyploidy and highly varied chromosome numbers, laying a challenge for uncovering its genomic structure and evolution. We developed a chromosome 2 painting (CP2) probe by designing oligonucleotides covering chromosome 2 of Saccharum spontaneum (2n = 8x = 64). Fluorescence in situ hybridization (FISH) using this CP2 probe revealed six types of ploidies from twenty S. spontaneum clones, including 6x, 8x, 10x, 11x, 12x, and 13x clones. The finding of S. spontaneum clones with uneven of ploid suggested that certain S. spontaneum clones come from hybridization. It renews our knowledge that S. spontaneum is derived from autopolyploidization. Combined with a S. spontaneum-specific probe, chromosome 2-derived chromosome or fragments from either S. spontaneum or Saccharum officinarum can be identified in sugarcane modern cultivars. We revealed unexpected high level of interspecific recombination from introgressive S. spontaneum chromosomes (>50.0%) in cultivars ROC22 and ZZ1, indicating frequent chromosome exchange in cultivars. Intriguingly, we observed interspecific recombination recurring among either homoeologous or non-homoeologous chromosomes in sugarcane cultivars. These results demonstrated that chromosome painting FISH is a powerful tool in the genome dissection of sugarcane and provide new insights into the genome structure and evolution of the complex genus Saccharum.

The reference genome of Miscanthus floridulus illuminates the evolution of Saccharinae

Miscanthus, a member of the Saccharinae subtribe that includes sorghum and sugarcane, has been widely studied as a feedstock for cellulosic biofuel production. Here, we report the sequencing and assembly of the Miscanthus floridulus genome by the integration of PacBio sequencing and Hi-C mapping, resulting in a chromosome-scale, high-quality reference genome of the genus Miscanthus. Comparisons among Saccharinae genomes suggest that Sorghum split first from the common ancestor of Saccharum and Miscanthus, which subsequently diverged from each other, with two successive whole-genome duplication events occurring independently in the Saccharum genus and one whole-genome duplication occurring in the Miscanthus genus. Fusion of two chromosomes occurred during rediploidization in M. floridulus and no significant subgenome dominance was observed. A survey of cellulose synthases (CesA) in M. floridulus revealed quite high expression of most CesA genes in growing stems, which is in agreement with the high cellulose content of this species. Resequencing and comparisons of 75 Miscanthus accessions suggest that M. lutarioriparius is genetically close to M. sacchariflorus and that M. floridulus is more distantly related to other species and is more genetically diverse. This study provides a valuable genomic resource for molecular breeding and improvement of Miscanthus and Saccharinae crops.

SSR Marker Based Molecular Genetic Diversity Analysis Among Saccharum spontaneum (L.) Collected from North Western Region of India

Saccharum spontaneum L. is the most diverse species of the genus Saccharum with the widest distribution from Africa-Mediterranean region to Far East including South Pacific islands. Its contribution in the development of present day sugarcane cultivars is well recognized. In the present study, 30 S. spontaneum L. accessions collected from North Western India including the states of Gujarat and Rajasthan were characterized with respect to their ecological distribution using 20 STMS (Sequence Tagged Microsatellite Site) primers. The study revealed the ability of STMS markers in discriminating the accessions with specific markers with 95.4% polymorphism. The pair wise genetic distance among 30 S. spontaneum L. accessions ranged from 0.17 to 0.48 with an average of 0.32. The maximum genetic distance was observed between the pairs IND 07-1462 and IND 08-1501 and IND 07-1464 and IND 08-1501. All the 30 S. spontaneum L. accessions were grouped into five major clusters. Among the five clusters, Cluster II had maximum of nine accessions viz., IND 07-1461, IND 07-1462, IND 07-1463, IND 07-1465, IND 07-1466, IND 07-1467, IND 07-1468, IND 07-1470 and IND 07-1474. All the accessions except IND 07-1474 were from Kachchh district of North West arid zone with low rainfall and high salinity. Majority of the accessions clustered as per the agro-climatic zones. Unique markers for the accession IND 07-1460, IND 07-1463 and IND 07-1482 were identified which could help in maintaining germplasm identity. Accessions from arid and semi-arid zones were grouped separately into different clusters. These accessions were collected from the regions where drought and salinity were prevalent hence could be the potential sources for developing climate resilient sugarcane varieties with drought and salinity tolerance.

<p><strong>A new combination in the genus <em>Tripidium </em>(Poaceae: Andropogoneae)</strong></p>

The genus Tripidium Scholz in Valdés & Scholz (2006: 664) (Poaceae: Andropogoneae) currently includes six species, distributed in Asia, southern Europe, and northern Africa, with the greatest diversity in India (Welker et al. 2019). The species of Tripidium have traditionally been treated as part of the “Saccharum complex”, formed by sugarcane [S. officinarum Linnaeus (1753a: 54)] and relatives, being included either in the genus Saccharum Linnaeus (1753a: 54) (e.g., Clayton & Renvoize 1986, Shouliang & Phillips 2006) or in Erianthus Michaux (1803: 54) (e.g., Mukherjee 1958, Besse et al. 1997). However, recent nuclear and plastome phylogenies (Welker et al. 2015, 2019, Lloyd Evans et al. 2019, Welker et al. in press) have unequivocally demonstrated that Tripidium belongs to a different lineage from sugarcane and relatives and thus should be accepted as a genus distinct from Saccharum or Erianthus. According to the plastome phylogeny of Welker et al. (in press), Tripidium is sister to the newly circumscribed subtribe Rottboelliinae, which comprises the genera Rottboellia Naezén (1779: 23) s.str. and Coix Linnaeus (1753b: 972).

Sequence Evolution, Abundance, and Chromosomal Distribution of Ty1-copia Retrotransposons in the Saccharum spontaneum Genome

Saccharum spontaneum is a wild germplasm resource of the genus Saccharum that has many valuable traits. Ty1-copia retrotransposons constitute a large proportion of plant genomes and affect genome sequence organization and evolution. This study aims to analyze the sequence heterogeneity, phylogenetic diversity, copy number, and chromosomal dispersion patterns of Ty1-copia retrotransposons in S. spontaneum. A total of 44 Ty1-copia reverse transcriptase subclones isolated from S. spontaneum showed a range of heterogeneity, and all sequences were A-T rich, averaging approximately 54.59%. Phylogenetic analysis divided the 44 reverse transcriptase sequences into 5 distinct lineages (Retrofit/Ale, Sire/Maximus, Bianca, Tork/TAR, and Ty1-copia like). Dot-blot hybridization revealed that Ty1-copia retrotransposons consisted of a significant component of approximately 38,900 copies and 16,300 copies per genome in the accessions YN82-114 (2n = 10x = 80) and AP85-441 (2n = 4x = 32), respectively. The results of a local blast analysis showed that there are 15,069 Ty1-copia retrotransposon copies in the genome of AP85-441, of which the Retrofit/Ale lineage had the highest copy number, followed by the Tork/TAR, Sire/Maximus, and Bianca lineages. Furthermore, both FISH and the local blast analysis with AP85-441 genomic data demonstrated that the Ty1-copia retrotransposons were unevenly distributed throughout the chromosomes. Taken together, this study provides insights into the role of Ty1-copia retrotransposons in the evolution and organization of the S. spontaneum genome.

Assessing genetic diversity and population structure of sugarcane cultivars, progenitor species and genera using microsatellite (SSR) markers.

Sugarcane is one among the most important commercial crops used to produce sugar, ethanol, and other byproducts, which significantly contributes in the GDP of India and many other countries around the world. Genetic diversity is a platform for any breeding program of a plant species. Estimation of the genetic variability and population structure play a vital role for conservation planning and management of plant genetic resources. Genetic variability serves as a source of noble alleles responsible for key agronomic and quality traits, which ultimately form basis for identification and selection of promising parents for breeding programs. In the present study genetic diversity and population structure of 139 accessions of the genus Saccharum, allied genera of family Poaceae and cultivars were assessed using informative microsatellite (SSR) markers. A sum of 427 alleles was produced using 61 polymorphic primers and number of alleles generated was ranged from 2 to 13 with an average of 7 alleles per locus. PIC values were ranged from 0.35 to 0.90, with a mean value of 0.66 for all the markers evaluated. Cluster analysis based on UPGMA method revealed three major clusters which were further subdivided into nine subclusters. Population structure analysis also established three subpopulations of used accession set, however there were no correlation of sub-groupings with that of place of origin. AMOVA analysis also confirmed that 83% and 17% of total variations were attributed to the within- and between-populations, correspondingly, demonstrating greater exchange of gene pool across places of origin. The principal component analysis (PCA) demonstrated the distribution of accessions in the scatter-plot was substantially dispersed, revealing rich genetic diversity among accessions of different species. The findings from this study will be useful in breeding programs for introgression of noble alleles into modern cultivars by exploiting natural genetic variation existing in sugarcane genetic resources.

A flow cytometry-based analysis to establish a cell cycle synchronization protocol for Saccharum spp.

Modern sugarcane is an unusually complex heteroploid crop, and its genome comprises two or three subgenomes. To reduce the complexity of sugarcane genome research, the ploidy level and number of chromosomes can be reduced using flow chromosome sorting. However, a cell cycle synchronization (CCS) protocol for Saccharum spp. is needed that maximizes the accumulation of metaphase chromosomes. For flow cytometry analysis in this study, we optimized the lysis buffer, hydroxyurea(HU) concentration, HU treatment time and recovery time for sugarcane. We determined the mitotic index by microscopic observation and calculation. We found that WPB buffer was superior to other buffers for preparation of sugarcane nuclei suspensions. The optimal HU treatment was 2 mM for 18 h at 25 °C, 28 °C and 30 °C. Higher recovery treatment temperatures were associated with shorter recovery times (3.5 h, 2.5 h and 1.5 h at 25 °C, 28 °C and 30 °C, respectively). The optimal conditions for treatment with the inhibitor of microtubule polymerization, amiprophos-methyl (APM), were 2.5 μM for 3 h at 25 °C, 28 °C and 30 °C. Meanwhile, preliminary screening of CCS protocols for Badila were used for some main species of genus Saccharum at 25 °C, 28 °C and 30 °C, which showed that the average mitotic index decreased from 25 °C to 30 °C. The optimal sugarcane CCS protocol that yielded a mitotic index of >50% in sugarcane root tips was: 2 mM HU for 18 h, 0.1 X Hoagland’s Solution without HU for 3.5 h, and 2.5 μM APM for 3.0 h at 25 °C. The CCS protocol defined in this study should accelerate the development of genomic research and cytobiology research in sugarcane.

Characterization of a Saccharum spontaneum with a basic chromosome number of x = 10 provides new insights on genome evolution in genus Saccharum.

A novel tetraploid S. spontaneum with basic chromosome x = 10 was discovered, providing us insights in the origin and evolution in Saccharum species. Sugarcane (Saccharum spp., Poaceae) is a leading crop for sugar production providing 80% of the world's sugar. However, the genetic and genomic complexities of this crop such as its high polyploidy level and highly variable chromosome numbers have significantly hindered the studies in deciphering the genomic structure and evolution of sugarcane. Here, we developed the first set of oligonucleotide (oligo)-based probes based on the S. spontaneum genome (x = 8), which can be used to simultaneously distinguish each of the 64 chromosomes of octaploid S. spontaneum SES208 (2n = 8x = 64) through fluorescence in situ hybridization (FISH). By comparative FISH assay, we confirmed the chromosomal rearrangements of S. spontaneum (x = 8) and S. officinarum (2n = 8x = 80), the main contributors of modern sugarcane cultivars. In addition, we examined a S. spontaneum accession, Np-X, with 2n = 40 chromosomes, and we found that it was a tetraploid with the unusual basic chromosome number of x = 10. Assays at the cytological and DNA levels demonstrated its close relationship with S. spontaneum with basic chromosome number x = 8 (the most common accessions in S. spontaneum), confirming its S. spontaneum identity. Population genetic structure and phylogenetic relationship analyses between Np-X and 64 S. spontaneum accessions revealed that Np-X belongs to the ancient Pan-Malaysia group, indicating a close relationship to S. spontaneum with basic chromosome number of x = 8. This finding of a tetraploid S. spontaneum with basic chromosome number of x = 10 suggested a parallel evolution path of genomes and polyploid series in S. spontaneum with different basic chromosome numbers.

Transcriptome analysis of the effect of GA3 in sugarcane culm.

Our earlier studies have indicated that GA3, being a growth hormone, increases internodal length, in turn increasing sink strength and improving sucrose accumulation in sugarcane. In this study, transcriptomic level analysis was carried out on internodal samples of a high sugar accumulating variety (CoLk 94184) of sugarcane, to determine the effect of exogenous application of GA3 vis a vis functional analysis of differentially expressing transcripts. Overall, a total of 201,184 transcripts were identified, with median contig length of 450bp and N50 length of 1029bp. Analyzing the data from control and GA3-treated canes, at 0.01 significance, a total of 1516 differentially expressing transcripts were identified in bottom internodes and 1589 in top internodes. A KEGG (enrichment) analysis grouped the transcripts into 153 plant-related functional categories. From among these, the transcripts which were functionally relevant to sugar metabolism and photosynthesis were sieved out. Starch and sucrose metabolizing genes showed maximum fold change of 5.0 and 3.0 among top and bottom internodal samples. A homology match using Blastx analysis tool yielded 65 transcripts/differentially expressed genes (DEGs) which were found to share homology with C4 plants like Saccharum, Sorghum and Zea mays. Differentially expressing transcripts from both top and bottom internodes were validated by qRT-PCR, indicating their importance in such study. Results also enriched sugarcane transcriptome resources useful for omics study in genus Saccharum and family Poaceae.