Sugarcane (Saccharum spp.) is one of the most important crops in the world. Drought stress is a major abiotic stress factor that significantly reduces sugarcane yields. However the gene network that mediates plant responses to water stress remains largely unknown in several crop species. Although several microRNAs that mediate post-transcriptional regulation during water stress have been described in other species, the role of the sugarcane microRNAs during drought stress has not been studied. The objective of this work was to identify sugarcane miRNAs that are differentially expressed under drought stress and to correlate this expression with the behavior of two sugarcane cultivars with different drought tolerances. The sugarcane cultivars RB867515 (higher drought tolerance) and RB855536 (lower drought tolerance) were cultivated in a greenhouse for three months and then subjected to drought for 2, 4, 6 or 8 days. By deep sequencing of small RNAs, we were able to identify 18 miRNA families. Among all of the miRNAs thus identified, seven were differentially expressed during drought. Six of these miRNAs were differentially expressed at two days of stress, and five miRNAs were differentially expressed at four days. The expression levels of five miRNAs (ssp-miR164, ssp-miR394, ssp-miR397, ssp-miR399-seq 1 and miR528) were validated by RT-qPCR (quantitative reverse transcriptase PCR). Six precursors and the targets of the differentially expressed miRNA were predicted using an in silico approach and validated by RT-qPCR; many of these targets may play important roles in drought tolerance. These findings constitute a significant increase in the number of identified miRNAs in sugarcane and contribute to the elucidation of the complex regulatory network that is activated by drought stress.

The ability to expand crop plantations without irrigation is a major goal to increase agriculture sustainability. To achieve this end, we need to understand the mechanisms that govern plant growth responses under drought conditions. In this study, we combined physiological, transcriptomic, and genomic data to provide a comprehensive picture of drought and recovery responses in the leaves and roots of sugarcane. Transcriptomic profiling using oligoarrays and RNA-seq identified 2898 (out of 21,902) and 46,062 (out of 373,869) transcripts as differentially expressed, respectively. Co-expression analysis revealed modules enriched in photosynthesis, small molecule metabolism, alpha-amino acid metabolism, trehalose biosynthesis, serine family amino acid metabolism, and carbohydrate transport. Together, our findings reveal that carbohydrate metabolism is coordinated with the degradation of amino acids to provide carbon skeletons to the tricarboxylic acid cycle. This coordination may help to maintain energetic balance during drought stress adaptation, facilitating recovery after the stress is alleviated. Our results shed light on candidate regulatory elements and pave the way to biotechnology strategies towards the development of drought-tolerant sugarcane plants.

Drought is one of the main factors affecting the productivity of agricultural crops, and plants respond to such stress by activating various physiological and biochemical mechanisms against dehydration. The present study investigated two varieties of sugarcane (Saccharum spp.) with contrasting responses to drought (RB867515, more tolerant; and RB855536, less tolerant) and subjected them to progressive drought conditions (2, 4, 6 and 8 days) followed by rehydration. Drought caused a decrease in water potential (ψw) and osmotic potential (ψos) in the leaves, which recovered to normal levels after rehydration only up to the fourth day of drought. Water stress changed the carbon metabolism of leaves by reducing starch and sucrose contents and increasing glucose and fructose contents in both varieties. Water deficit caused a significant reduction in the maximum quantum efficiency of photosystem II (Fv/Fm) and effective quantum yield (ΦPSII) in both varieties; however, RB867515 recovered faster after rehydration. Under water stress, the more tolerant variety RB867515 exhibited increased activity of the antioxidant enzymes catalase, ascorbate peroxidase and superoxide dismutase compared with the RB855536 variety. The results suggest that RB867515 is more tolerant to drought conditions because of a more efficient antioxidant system, which results in reduced photosynthesis photoinhibition during water stress, thus revealing itself as a potential physiological marker for drought tolerance studies.

The flowering of sugarcane (Saccharum spp. hybrids) is a developmental stage that is desirable for breeding programs but undesirable for commercial cultivation, since it causes losses in sugar content and cane yield. This study aimed to (1) characterise the flowering of four varieties of sugarcane with different flowering times and intensities, (2) determine the nutrient levels in their vegetative apexes and (3) to evaluate the effect of calcium on their flowering. The study was divided into two trials. In the first trial, we tested the effect of planting season on the flowering of four varieties of sugarcane that had contrasting flowering periods. Late or reluctant flowering varieties had higher concentrations of calcium in their vegetative apexes in relation to the early variety. At the same time, the early variety, with heavy flowering, had higher potassium in its vegetative apexes. In the second trial, the effect of calcium on their flowering was tested. The application of calcium on the leaves resulted in its accumulation in the vegetative apexes, causing a reduction in potassium levels in the same tissue. This changed the plant phenology, resulting in a month's delay in flowering and a 50 % reduction in the final flowering intensity. Calcium application by foliar sprays can be used to delay and reduce sugarcane flowering.