A chromosomal-scale genome assembly of modern cultivated hybrid sugarcane provides insights into origination and evolution.

Sugarcane is a vital crop with significant economic and industrial value. However, the cultivated sugarcane's ultra-complex genome still needs to be resolved due to its high ploidy and extensive recombination between the two subgenomes. Here, we generate a chromosomal-scale, haplotype-resolved genome assembly for a hybrid sugarcane cultivar ZZ1. This assembly contains 10.4 Gb genomic sequences and 68,509 annotated genes with defined alleles in two sub-genomes distributed in 99 original and 15 recombined chromosomes. RNA-seq data analysis shows that sugar accumulation-associated gene families have been primarily expanded from the ZZSO subgenome. However, genes responding to pokkah boeng disease susceptibility have been derived dominantly from the ZZSS subgenome. The region harboring the possible smut resistance genes has expanded significantly. Among them, the expansion of WAK and FLS2 families is proposed to have occurred during the breeding of ZZ1. Our findings provide insights into the complex genome of hybrid sugarcane cultivars and pave the way for future genomics and molecular breeding studies in sugarcane.

Identification of gene modules and hub genes associated with Colletotrichum siamense infection in mango using weighted gene co-expression network analysis.

Colletotrichum siamense is a hemibiotrophic ascomycetous fungus responsible for mango anthracnose. The key genes involved in C. siamense infection remained largely unknown. In this study, we conducted weighted gene co-expression network analysis (WGCNA) of RNA-seq data to mine key genes involved in Colletotrichum siamense-mango interactions. Gene modules of Turquoise and Salmon, containing 1039 and 139 respectively, were associated with C. siamense infection, which were conducted for further analysis. GO enrichment analysis revealed that protein synthesis, organonitrogen compound biosynthetic and metabolic process, and endoplasmic reticulum-related genes were associated with C. siamense infection. A total of 568 proteins had homologs in the PHI database, 370 of which were related to virulence. The hub genes in each module were identified, which were annotated as O-methyltransferase (Salmon) and Clock-controlled protein 6 (Turquoise). A total of 24 proteins exhibited characteristics of SCRPs. By using transient expression in Nicotiana benthamiana, the SCRPs of XM_036637681.1 could inhibit programmed cell death (PCD) that induced by BAX (BCL-2-associated X protein), suggesting that it may play important roles in C. siamense infection. A mango-C. siamense co-expression network was constructed, and the mango gene of XM_044632979.1 (auxin-induced protein 15A-like) was positively associated with 5 SCRPs. These findings help to deepen the current understanding of necrotrophic stage in C. siamense infection.

Identification of Gene Modules and Hub Genes Associated with Sporisorium scitamineum Infection Using Weighted Gene Co-Expression Network Analysis.

Sporisorium scitamineum is a biotrophic fungus responsible for sugarcane smut disease. To investigate the key genes involved in S. scitamineum infection, we conducted RNA sequencing of sugarcane sprouts inoculated with S. scitamineum teliospores. A weighted gene co-expression network analysis (WGCNA) showed that two co-expressed gene modules, MEdarkturquoise and MEpurple-containing 66 and 208 genes, respectively-were associated with S. scitamineum infection. The genes in these two modules were further studied using Gene Ontology (GO) enrichment analysis, pathogen-host interaction (PHI) database BLASTp, and small secreted cysteine-rich proteins (SCRPs) prediction. The top ten hub genes in each module were identified using the Cytohubba plugin. The GO enrichment analysis found that endoplasmic reticulum-related and catabolism-related genes were expressed during S. scitamineum infection. A total of 83 genes had homologs in the PHI database, 62 of which correlated with pathogen virulence. A total of 21 proteins had the characteristics of small secreted cysteine-rich proteins (SCRPs), a common source of fungal effectors. The top ten hub genes in each module were identified, and seven were annotated as Mig1-Mig1 protein, glycosyl hydrolase, beta-N-acetylglucosaminidase, secreted chorismate mutase, collagen, mRNA export factor, and pleckstrin homology domain protein, while the remaining three were unknown. Two SCRPs-SPSC_06609 and SPSC_04676-and three proteins-SPSC_01958, SPSC_02155, and SPSC_00940-identified in the PHI database were also among the top ten hub genes in the MEdarkturquoise and MEpurple modules, suggesting that they may play important roles in S. scitamineum infection. A S. scitamineum infection model was postulated based on current findings. These findings help to deepen the current understanding of early events in S. scitamineum infection.

Overexpression of Sugarcane ScDIR Genes Enhances Drought Tolerance in Nicotiana benthamiana.

Dirigent proteins (DIRs) are known to function in lignin biogenesis and to be involved in stress resistance in plants. However, the sugarcane DIRs have not been functionally characterized. In this study, we investigated the DIR-protein-encoding genes in Saccharum spp. (ScDIR) by screening collections of sugarcane databases, monitoring the responses of these genes to drought stress by real-time quantitative PCR, and identifying their heterologous expression in tobacco. Of the 64 ScDIRs identified, four belonging to the DIR-b/d (ScDIR5 and ScDIR11) and DIR-c (ScDIR7 and ScDIR40) subfamilies showed a significant transcriptional response when subjected to drought stress. ScDIR5, ScDIR7, and ScDIR11 are localized in the cell membrane, whereas ScDIR40 is found in the cell wall. The overexpression of these ScDIR genes in tobacco generally increased the drought tolerance of the transgenic lines, with ScDIR7 conferring the highest degree of drought tolerance. The characterization of the physiological and biochemical indicators (superoxide dismutase, catalase, malondialdehyde, and H2O2) confirmed that the ScDIR-overexpressing lines outperformed the wild type. These results demonstrated that specific ScDIRs in sugarcane respond and contribute to tolerance of drought stress, shedding light on potential means of improving drought tolerance in this crop.

A Plant-Derived Alkanol Induces Teliospore Germination in Sporisorium scitamineum.

Sugarcane smut caused by the basidiomycetes fungus Sporisorium scitamineum is a devastating disease for the sugarcane industry worldwide. As the initial step, the smut teliospores germinate on sugarcane buds, and subsequently, the mycelium infects the bud tissues. However, chemical signals that induce spore germination are still unknown. By comparison of the behavior of the teliospores on the buds of both resistant and susceptible varieties, we found that spore germination rates were significantly lower on the buds of resistant cultivars ZZ1, ZZ6, and ZZ9 than on the susceptible varieties GT42 and ROC22. It was found that the levels of hexacosanol and octacosanol were higher on the buds of smut-susceptible varieties than on the smut-resistant varieties. These observations were extended to the smut-resistant and smut-susceptible sub-genetic populations derived from the cross of ROC25 and YZ89-7. In artificial surface assays, we found that hexacosanol and octacosanol promoted smut teliospore germination. Transcriptome analysis of smut teliospores under the induction by octacosanol revealed that genes in the MAPK signaling pathway and fatty acid metabolism were significantly differentially expressed. Overall, our results provide evidence that alkanol plays important roles in smut teliospore germination and thus could be used as a potential marker for smut resistance in sugarcane breeding programs.

Biocontrol ability of killer yeasts (Saccharomyces cerevisiae) isolated from wine against Colletotrichum gloeosporioides on grape.

A total of 216 killer yeasts Saccharomyces cerevisiae, isolated from wine, were evaluated in controlling Colletotrichum gloeosporioides, a pre-harvest anthracnose agent of grape. Three of these yeast isolates were tested positive for antagonizing C. gloeosporioides and were further evaluated for their mechanisms as biological control agents (BCAs): production of antifungal compounds, production of hydrolytic enzymes, inhibition of C. gloeosporioides conidia germination, colonization on grape berry, and efficiency in controlling anthracnose of grape. The results showed that all three S. cerevisiae isolates produced antifungal compounds, inhibited C. gloeosporioides conidia germination and produced ß-1,3-glucanase and chitinase. All isolates colonized grape berry in large quantities and controlled C. gloeosporioides when artificially inoculated on grape berry. Among the three isolates, application of isolate GA8 resulted in 69.7% of disease reductions for C. gloeosporioides on grape berry. The antagonistic isolates of S. cerevisiae could represent important BCAs of anthracnose of grape caused by C. gloeosporioides that are responsible for economic losses in viticulture.