Sugarcane, a cash crop, is easily affected by low temperature, which results in a decrease in yield and sugar production. Breeding a new variety with cold tolerance is an essential strategy to reduce loss from cold stress. The identification of germplasms and genes/proteins with cold tolerance is a vital step in breeding sugarcane varieties with cold tolerance via a conventional program and molecular technology. In this study, the physiological and biochemical indices of 22 genotypes of S. spontaneum were measured, and the membership function analysis method was used to comprehensively evaluate the cold tolerance ability of these genotypes. The physiological and biochemical indices of these S. spontaneum genotypes showed a sophisticated response to low temperature. On the basis of the physiological and chemical indices, the genotypes were classified into different cold tolerance groups. Then, the high-tolerance genotype 1027 and the low-tolerance genotype 3217 were selected for DIA-based proteomic analysis by subjecting them to low temperature. From the four comparison groups, 1123, 1341, 751, and 1693 differentially abundant proteins (DAPs) were identified, respectively. The DAPs based on genotypes or treatments participated in distinct metabolic pathways. Through detailed analysis of the DAPs, some proteins related to protein homeostasis, carbohydrate and energy metabolism, amino acid transport and metabolism, signal transduction, and the cytoskeleton may be involved in sugarcane tolerance to cold stress. Furthermore, five important proteins related to cold tolerance were discovered for the first time in this study. This work not only provides the germplasms and candidate target proteins for breeding sugarcane varieties with cold tolerance via a conventional program and molecular breeding, but also helps to accelerate the determination of the molecular mechanism underlying cold tolerance in sugarcane.

Sugarcan e is a major crop for sugar and biofuel production and is cultivated in tropical and subtropical areas worldwide. Sugarcane growth is constrained because of winter’s low-temperature stress, and cold resistance is an important limitation in sugarcane growth enhancement. Therefore, in this study, we identified a gene involved in the low-temperature stress response of sugarcane. Calcineurin B-like (CBL) protein is a calcium signal receptor involved in the cold stress response. Five sugarcane CBL genes were cloned, sequenced, and named SoCBL1, SoCBL3, SoCBL5, SoCBL6, and SoCBL9. The protein sequences of these genes were analyzed. The calculated molecular weight of these proteins was 24.5, 25.9, 25.2, 25.6, and 26.3 kD, respectively. Subcellular localization analysis revealed that SoCBL1, SoCBL3, SoCBL6, and SoCBL9 were situated in the cytoplasm, while SoCBL5 was present in mitochondria. Secondary structure analysis showed that these five CBL proteins had similar secondary structures. Conserved domain analysis displayed that each sugarcane CBL protein contained three conserved EF domains. According to the self-expanding values of the phylogenetic tree, the CBL gene family was divided into four groups. The CBL1 and CBL9 genes were classified into one group, illustrating that these two genes might possess a similar function. The expression analysis of the SoCBL gene under low temperatures showed that SoCBL3 and SoCBL5 were affected significantly, while SoCBL1 and SoCBL9 were less affected. These results demonstrate that the CBL genes in sugarcane have similar characteristics and present differences in genetic diversity and gene expression response to low temperatures. Therefore, these genes might be novel candidates for fighting cold stress in sugarcane.

Sugarcane (Saccharum officinarum L.) faces significant challenges in China, including labor-intensive cultivation, low yields, and environmental stresses. Enhancing root development and stress tolerance through phytohormones and molecular breeding is a promising approach to boosting productivity. Indole-3-butyric acid is a phytohormone known for promoting root development and stress resistance. However, its effects on sugarcane root development under low temperature remain poorly understood. This study demonstrated that IBA markedly promoted root initiation, elongation, and biomass under low temperature, and significantly increased the levels of phytohormones, including GA₃, ABA, JA, IAA, and ZT, suggesting the activation of multiple signaling pathways. Transcriptome analysis revealed numerous differentially expressed genes related to metabolic pathways such as glycolysis, the tricarboxylic acid cycle, and glutathione metabolism. Weighted gene co-expression network analysis identified core gene modules correlated with phytohormone activities, highlighting their role in the IBA-mediated stress response. Eleven core genes, including GSTU6, FAR1, and BCAT3, and nine hub genes, such as Ub-CEP52-1 and ACS1, were identified as critical components for IBA-induced root development and stress mitigation. These findings provide insights into the molecular mechanisms underlying IBA-induced root development and stress tolerance in sugarcane, offering candidate genes for breeding high-yield, stress-tolerant varieties and demonstrating IBA’s potential as a strategy to enhance productivity under challenging conditions.

Sugarcane is the most important sugar crop and one of the leading energy-producing crops in the world. Ratoon stunting disease (RSD), caused by the bacterium Leifsonia xyli subsp. xyli, poses a huge threat to ratoon crops, causing a significant yield loss in sugarcane. Breeding resistant varieties is considered the most effective and fundamental approach to control RSD in sugarcane. The exploration of resistance genes forms the foundation for breeding resistant varieties through molecular technology. The pglA gene is a pathogenicity gene in L. xyli subsp. xyli, encoding an endopolygalacturonase. In this study, the pglA gene from L. xyli subsp. xyli and related microorganisms was analyzed. Then, a non-toxic, non-autoactivating pglA bait was successfully expressed in yeast cells. Simultaneously the yeast two-hybrid library was generated using RNA from the L. xyli subsp. xyli-infected sugarcane. Screening the library with the pglA bait uncovered proteins that interacted with pglA, primarily associated with ABA pathways and the plant immune system, suggesting that sugarcane employs these pathways to respond to L. xyli subsp. xyli, triggering pathogenicity or resistance. The expression of genes encoding these proteins was also investigated in L. xyli subsp. xyli-infected sugarcane, suggesting multiple layers of regulatory mechanisms in the interaction between sugarcane and L. xyli subsp. xyli. This work promotes the understanding of plant–pathogen interaction and provides target proteins/genes for molecular breeding to improve sugarcane resistance to L. xyli subsp. xyli.

Although thiamethoxam is frequently used in sugarcane production in Guangxi, China as a systemic insecticide and bioactivator, little is known about its field control effectiveness and residue dissipation across commercially significant genotypes grown in the region. In this study, we have addressed this gap by assessing the residue dynamics and borer control effectiveness of thiamethoxam in three significant sugarcane types (GT 40, Haizhe 22, and ROC22). Three application doses (Low: 750 mL/hm²; Medium: 1500 mL/hm²; High: 2250 mL/hm²) were sprayed as a soil drench along with urea for field and greenhouse studies. Over the course of 75 days, residues in juice and leaf tissues were measured using LC-MS/MS, and stem borer damage and control effectiveness were recorded. Thiamethoxam residues in juice peaked at 45 days after treatment (DAT), with ROC22 exhibiting the highest concentration (11.07 mg/kg). All treated plots continued to have thiamethoxam residues above the Chinese maximum residue limit (MRL) of 0.3 mg/kg at 75 DAT, suggesting an inadequate pre-harvest interval. Leaf residues remained a long-term reservoir for more than 56 days and were an order of magnitude greater. While the low dose demonstrated increased efficacy over time, the medium and high doses produced statistically equal borer control (50–75% efficacy), exhibiting a saturation effect. There were notable varietal differences: Haizhe 22 achieved steady control with fewer residues, while ROC22 was most vulnerable and needed greater doses. These results demonstrate that a 75-day pre-harvest period is insufficient for consumer safety, requiring additional experiments for longer intervals for susceptible cultivars. The ideal economic threshold is determined to be the middle dose (1500 mL/hm²), which balances environmental load, chemical input, and efficacy. To enable precision application techniques suited to certain sugarcane genotypes, future research should concentrate on clarifying the genetic basis of varietal uptake discrepancies.