Karla Cordero-Lara

Rice is a staple food grown worldwide. While white rice varieties have been extensively studied, there is limited information on the performance of pigmented rice genotypes and their tolerance to water deficit. This study evaluated nineteen black rice genotypes and one white cultivar over three years under contrasting water regimes: traditional flooding and non-flood irrigation (NFI). Genotype–environment interactions and their impact on agronomic, yield, and grain quality traits were assessed. Black genotypes under NFI showed reduced flowering and grain quality. The average yield was 31% lower than the white cultivar. Significant genetic correlations were found between grain yield and days to anthesis (DSA), grain weight (TGW), chalkiness (CHA), and translucency (TRAN), with high broad-sense heritability (H² > 0.9). Most traits exhibited high heritability (H² > 0.7), indicating strong genetic stability. Grain yield (GR) was highly and negatively correlated with percent sterility (PS) (r = −0.84) and had a heritability of 0.76. Environmental conditions significantly influenced yield, confirming the potential for selecting water stress–tolerant genotypes. These findings provide valuable insights into black rice breeding and optimizing water management practices to support sustainable production. To our knowledge, this study is the first to evaluate the performance of a diverse set of black rice genotypes across multiple seasons under contrasting water regimes in a Mediterranean environment.

Rice (Oryza sativa L.), a staple food for over half of the global population, plays a pivotal role in food security. Among its two primary groups, japonica and indica, temperate japonica varieties are particularly valued for their high-quality grain and culinary uses. Although some of these varieties are adapted to cooler climates, they often suffer from reduced productivity or increased disease susceptibility when cultivated in warmer productive environments. These limitations underscore the need for breeding programs to incorporate biotechnological tools that can enhance the adaptability and resilience of the plants. However, New Genomic Techniques (NGTs), including CRISPR-Cas9, require robust in vitro systems, which are still underdeveloped for temperate japonica genotypes. In this study, we developed a reproducible and adaptable protocol for protoplast isolation and regeneration from the temperate japonica cultivars ‘Ónix’ and ‘Platino’ using somatic embryos as the starting tissue. Protoplasts were isolated via enzymatic digestion (1.5% Cellulase Onozuka R-10 and 0.75% Macerozyme R-10) in 0.6 M AA medium over 18–20 h at 28 °C. Regeneration was achieved through encapsulation in alginate beads and coculture with feeder extracts in 2N6 medium, leading to embryogenic callus formation within 35 days. Seedlings were regenerated in N6R and N6F media and acclimatized under greenhouse conditions within three months. The isolated protoplast quality displayed viability rates of 70–99% within 48 h and supported transient PEG-mediated transfection with GFP. Additionally, the transient expression of a gene editing CRISPR-Cas9 construct targeting the DROUGHT AND SALT TOLERANCE (OsDST) gene confirmed genome editing capability. This protocol offers a scalable and genotype-adaptable system for protoplast-based regeneration and gene editing in temperate japonica rice, supporting the application of NGTs in the breeding of cold-adapted cultivars.

Rice (Oryza sativa) varieties are generated through breeding programs focused on local requirements. In Chile, the southernmost rice producer, rice productivity relies on the use and generation of temperate japonica germplasms, which need to be adapted to the intensifying effects of climate change. Advanced biotechnological tools can contribute to these breeding programs; new technologies associated with precision breeding, including gene editing, rely on procedures such as regeneration and gene transfer. In this study, the local rice varieties Platino, Cuarzo, Esmeralda, and Zafiro were evaluated for somatic embryogenesis potential using a process that involved the combined use of auxins and cytokinins. An auxin-based (2,4-D) general medium (2N6) allowed for the induction of embryogenic masses in all the genotypes. After induction, masses required culturing either in N6R (kinetin; Platino) or N6RN (BAP, kinetin, IBA, and 2,4-D; Cuarzo, Esmeralda, and Zafiro) to yield whole plants using regeneration medium (N6F, no hormone). The sprouting rates indicated Platino as the most responsive genotype; for this reason, this variety was evaluated for gene transfer. Fifteen-day-old embryo masses were assayed for Agrobacterium-mediated transformation using the bacterial strain EHA105 harboring pFLC-Myb/HPT/GFP, a modified T-DNA vector harboring a geminivirus-derived replicon. The vector included the green fluorescent protein reporter gene, allowing for continuous traceability. Reporter mRNA was produced as early as 3 d after agroinfiltration, and stable expression of the protein was observed along the complete process. These achievements enable further biotechnological steps in these and other genotypes from our breeding program.

Rice is an essential diet component for a significant portion of the population worldwide. Due to the high water demand associated with rice production, improving water use efficiency and grain quality is critical to increasing the sustainability of the crop. This species includes rice varieties with diverse pigmentation patterns. Grain quality, including industrial, nutritional, and functional quality traits, of two black rice genotypes and a commercial white rice cultivar were evaluated in different locations and under different water regimes. Flooding produced higher grain weight compared to alternate wetting and drying irrigation. A high correlation was found between grain color, total phenolic content (TPC), and antioxidant activity. The black rice genotypes showed higher TPC levels and antioxidant capacity, mainly due to higher levels of cyanidin 3-O-glucoside. The phenolic profile varied between whole and polished grains, while mineral composition was influenced by location and irrigation regime. In turn, the environment influenced grain quality in terms of industrial and nutritional characteristics, with significant differences in quality between whole and polished grains. This study provides valuable information on the genotype–environment relationship in rice and its effect on grain quality, which could contribute to selecting genotypes for an appropriate environment.

Rice is a staple food mainly cultivated in the tropics, however ~20% of global production originates from temperate regions. The japonica rice (tropical and temperate), is the unique rice type adapted to the temperate conditions through evolution. Nevertheless, at present, there is limited information regarding to breeding of temperate japonica rice because most of the global research and production has been focused on indica rice, which is the most cultivated type (~80% of global production). However, there are many countries where rice is grown under temperate conditions, such as Japan, Russia, Australia, USA, Chile, South Korea, Uzbekistan, Hungary, among others. More information and collaboration between these countries are required to develop better japonica rice cultivars to meet the current agricultural challenges. Here, the japonica rice breeding history, current status and future challenges are compiled to be available to japonica rice breeders, researchers and growers.