Search bioRxiv⌕ Search

Biology subjects

Carneiro, M. S.

Publications and source records attributed to Carneiro, M. S..

7 recordsLinked to original sources

Mass Spectrometry-Based Characterization of Histone Post-Translational Modifications in Sugarcane

Histones, the core components of nucleosomes, play an important role in chromatin structure and function through post-translational modifications (PTMs) that regulate gene expression without altering the DNA sequence. Sugarcane (Saccharum spp.), a highly polyploid and economically significant crop, has a complex genome with a limited understanding of histone PTMs. In this study, we employed mass spectrometry to characterise the PTMs of histones in internodes of field-grown commercial sugarcane, harvested at different times of the day, with a focus on histones H2A and H2B, which had not been previously studied in this context. Using a sorghum histone catalog as a reference, we identified 26 H1, 91 H2A, 70 H2B, 5 CENH3, 36 H3.1, 23 H3.3, 18 H3.3L, and 57 H4 loci in the sugarcane genome. We detected extensive PTMs, including acetylation and methylation, across histones H2A, H2B, H3, and H4. Canonical and variant histones showed distinct post-translational modification (PTM) patterns, highlighting their functional specialisation. This study provides a comprehensive catalog of sugarcane histone PTMs, offering insights into their roles in chromatin dynamics and gene regulation, with potential applications in crop improvement and stress adaptation.

plant biology↗

Multiomic insights into sucrose accumulation in sugarcane

Sugarcane (Saccharum spp.) holds significant economic importance in sugar and biofuel production. Despite extensive research, understanding highly quantitative traits, such as sucrose content, remains challenging due to the complex genomic landscape of the crop. In this study, we conducted a multiomic investigation to elucidate the genetic architecture and molecular mechanisms governing sucrose accumulation in sugarcane. Using a biparental cross (IACSP95-3018 x IACSP93-3046) and a genetically diverse collection of sugarcane genotypes, we evaluated the soluble solids (Brix) and sucrose content (POL) across various years and environments. Both populations were genotyped using a genotyping-by-sequencing (GBS) approach, with single nucleotide polymorphisms (SNPs) identified via bioinformatics pipelines. Genotype-phenotype associations were established using a combination of traditional linear mixed-effect models and machine learning algorithms. Furthermore, we conducted an RNA sequencing (RNA-Seq) experiment on genotypes exhibiting distinct Brix and POL profiles across different developmental stages. Differentially expressed genes (DEGs) potentially associated with variations in sucrose accumulation were identified. All findings were integrated through a comprehensive gene coexpression network analysis. Strong correlations among the evaluated characteristics were observed, with estimates of modest to high heritabilities. By leveraging a broad set of SNPs identified for both populations, we identified several SNPs potentially linked to phenotypic variance. Our examination of genes close to these markers facilitated the association of such SNPs with DEGs in genotypes with contrasting sucrose levels. Through the integration of these results with a gene coexpression network, we delineated a set of genes potentially involved in the regulatory mechanisms of sucrose accumulation in sugarcane, collectively contributing to the definition of this critical phenotype. Our findings constitute a significant resource for biotechnology and plant breeding initiatives. Furthermore, our genotype-phenotype association models hold promise for application in genomic selection, offering valuable insights into the molecular underpinnings governing sucrose accumulation in sugarcane.

genetics↗

Temporal Gene Expression in Apical Culms Shows Early Changes in Cell Wall Biosynthesis Genes in Sugarcane

Multiple genes in sugarcane control sucrose accumulation and the biosynthesis of cell wall components; however, it is unclear how these genes are expressed in its apical culms. To better understand this process, we sequenced mRNA from +1 stem internodes collected from four genotypes with different concentrations of soluble solids. Culms were collected at four different time points, ranging from six to 12-month-old plants. Here we show differentially expressed genes related to sucrose metabolism and cell wall biosynthesis, including genes encoding invertases, sucrose synthase and cellulose synthase. Our results showed increased expression of invertases in IN84-58, the genotype with lower sugar and higher fiber content, as well as delayed expression of secondary cell wall-related cellulose synthase for the other genotypes. Interestingly, genes involved with hormone metabolism were differentially expressed across time points in the three genotypes with higher soluble solids content. A similar result was observed for genes controlling maturation and transition to reproductive stages, possibly a result of selection against flowering in sugarcane breeding programs. These results indicate that carbon partitioning in apical culms of contrasting genotypes is mainly associated with differential cell wall biosynthesis, and may include early modifications for subsequent sucrose accumulation. Co-expression network analysis identified transcription factors related to growth and development, showing a probable time shift for carbon partitioning occurred in 10-month-old plants.

genomics↗

Field microenvironments regulate crop diel transcript and metabolite rhythms

O_LIMost research in plant chronobiology was done in laboratory conditions. However, they usually fail to mimic natural conditions and their slight fluctuations, highlighting or obfuscating rhythmicity. High-density crops, such as sugarcane (Saccharum hybrid), generate field microenvironments with specific light and temperature due to mutual shading. C_LIO_LIWe measured the metabolic and transcriptional rhythms in the leaves of 4-month-old (4 mo) and 9 mo sugarcane grown in the field. Most of the assayed rhythms in 9 mo sugarcane peaked >1 h later than in 4 mo sugarcane, including rhythms of the circadian clock gene, LATE ELONGATED HYPOCOTYL (LHY). C_LIO_LIWe hypothesized that older sugarcane perceives dawn later than younger sugarcane due to self-shading. As a test, we measured LHY rhythms in plants on the east and the west side of a field. We also tested if a wooden wall built between lines of sugarcane changed their rhythms. The LHY peak was delayed in the plants in the west of the field or beyond the wall; both shaded at dawn. C_LIO_LIWe conclude that plants in the same field may have different phases due to field microenvironments, impacting important agronomical traits, such as flowering time, stalk weight and number. C_LI

plant biology↗

The sugarcane and sorghum kinomes: insights into evolutionary expansion and diversification

The protein kinase (PK) superfamily is one of the largest superfamilies in plants and is the core regulator of cellular signaling. Even considering this substantial importance, the kinomes of sugarcane and sorghum have not been profiled. Here we identified and profiled the complete kinomes of the polyploid Saccharum spontaneum (Ssp) and Sorghum bicolor (Sbi), a close diploid relative. The Sbi kinome was composed of 1,210 PKs; for Ssp, we identified 2,919 PKs when disregarding duplications and allelic copies, which were related to 1,345 representative gene models. The Ssp and Sbi PKs were grouped into 20 groups and 120 subfamilies and exhibited high compositional similarities and evolutionary divergences. By utilizing the collinearity between these species, this study offers insights about Sbi and Ssp speciation, PK differentiation and selection. We assessed the PK subfamily expression profiles via RNA-Seq, identifying significant similarities between Sbi and Ssp. Moreover, through coexpression networks, we inferred a core structure of kinase interactions with specific key elements. This study is the first to categorize the allele specificity of a kinome and provides a wide reservoir of molecular and genetic information, enhancing the understanding of the evolutionary history of Sbi and Ssp PKs. HighlightThis study describes the catalog of kinase gene family in Saccharum spontaneum and Sorghum bicolor, providing a reservoir of molecular features and expression patterns based on RNA-Seq and co-expression networks.

genomics↗

Alternative splicing of circadian clock genes is temperature regulated in field-grown sugarcane

Alternative Splicing (AS) is a mechanism that generates different mature transcripts from precursor mRNAs (pre-mRNAs) of the same gene. In plants, a wide range of physiological and metabolic events are related to AS, as well as fast responses to changes in temperature. AS is present in around 60% of intron-containing genes in Arabidopsis, 46% in rice and 38% in maize and it is widespread among the circadian clock genes. Little is known about how AS influences the circadian clock of C4 plants, like commercial sugarcane, a C4 crop with a complex hybrid genome. This work aims to test if the daily dynamics of AS forms of circadian clock genes are regulated by environmental factors, such as temperature, in the field. A systematic search for AS in five sugarcane clock genes, ScLHY, ScPRR37, ScPRR73, ScPRR95 and ScTOC1 using different organs of sugarcane sampled during winter, with 4 months old plants, and during summer, with 9 months old plants, revealed temperature- and organ-dependent expression of at least one alternatively spliced isoform in all genes. Expression of AS isoforms varied according to the season. Our results suggest that AS events in circadian clock genes are correlated with temperature.

plant biology↗

Rhythms of Transcription in Field-Grown Sugarcane Are Highly Organ Specific

We investigated whether different specialized organs in field-grown sugarcane follow the same temporal rhythms in transcription. We assayed the transcriptomes of three organs during the day: leaf, a source organ; internodes 1 and 2, sink organs focused on cell division and elongation; and internode 5, a sink organ focused on sucrose storage. The leaf had twice as many rhythmic transcripts (>68%) as internodes, and the rhythmic transcriptomes of the two internodes were more similar to each other than to those of the leaves. More transcripts were rhythmic under field conditions than under circadian conditions and most of their peaks were during the day. Among the transcripts that were considered expressed in all three organs, only 7.4% showed the same rhythmic time course pattern. The central oscillators of these three organs -- the networks that generate circadian rhythms -- had similar dynamics with different amplitudes. The differences between the rhythmic transcriptomes in circadian conditions and field conditions highlight the importance of field experiments to understand the plant circadian clock in natura. The highly specialized nature of the rhythmic transcriptomes in sugarcane organs probably arises from amplitude differences in tissue-specific circadian clocks and different sensitivities to environmental cues. One sentence summaryThe rhythmic transcriptome of field-grown sugarcane is highly organ-specific.

plant biology↗