Drought stress at jointing–booting directly affects plant growth and productivity in rice. Limited by natural factors, the jointing and booting stages of short-growth-period rice varieties are highly overlapped in high-latitude areas, which are more sensitive to water deficit. However, little is known about the dry matter translocation in rice and the strategies of starch synthesis and filling of superior and inferior grains under different drought stress was unclear. In this study, the rice plants were subjected to three degrees of drought stress (−10 kPa, −25 kPa, −40 kPa) for 15 days during the jointing–booting stage; we investigated dry matter accumulation and translocation, grain filling and enzyme activities to starch synthesis of superior and inferior grains in rice with overlapping growth stages from 2016 to 2017. The results showed that drought stress significantly reduced dry matter accumulation in the stems and leaves. Mild and moderate drought increased dry matter translocation efficiency. However, severe drought stress largely limited the dry matter accumulation and translocation. A large amount of dry matter remains in vegetative organs under severe drought stress. The high content in NSC in stem and sheath plays a key role in resisting drought stress. The drought stress at jointing–booting directly caused a change in the grain filling strategy. Under moderate and severe drought, the grain-filling active period of the superior grains was shortened to complete the necessary reproductive growth. The grain-filling active period of the inferior grains was significantly prolonged to avoid a decrease in grain yield. The significant decrease in the grain-filling rate of the superior and inferior grains caused a reduction in the thousand-grain weight. In particular, the influence of the grain-filling rate of inferior grains on the thousand-grain weight was more significant. Drought stress changed the starch synthesis strategies of the superior and inferior grains. Soluble starch synthase and starch branching enzyme activities of inferior grains increased significantly under drought stress. GBSS activity was not sensitive to drought stress. Therefore, amylose content was decreased and amylopectin synthesis was enhanced under drought stress, especially in inferior grains.

Rice salt tolerance at the germination stage directly affects the germination rate and seedling establishment of rice directly seeded in saline soils, which in turn affects yield. In this study, we determined the relative germination potential (RGP) and relative germination index (RGI) under 200 mM salt stress and control conditions using 295 japonica rice accessions. Statistical analysis showed extensive phenotypic variations under salt stress conditions. Twenty-one varieties with an RGP ≥ 80% and an RGI ≥ 80% were screened. Based on genotypic data including, 788,396 single-nucleotide polymorphisms (SNPs), 40 quantitative trait loci (QTL) were localized on rice chromosomes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12, which were shown to be significantly associated with rice salt tolerance at the germination stage, including 20 for RGP and 20 for RGI, using genome-wide association analysis. Six QTL with ≥ 3 consecutive significant SNP loci and localized in the same LD interval were selected for further analysis. Four rice genes (LOC_Os01g04920, LOC_Os10g38350, LOC_Os10g38470, and LOC_Os10g38489) were selected as important candidates for salt tolerance based on haplotype analysis and functional annotation. The findings could facilitate the development of valuable rice varieties for direct seeding in salinized soil and improve japonica rice salt tolerance at the germination stage through molecular breeding.

Low-temperature chilling is a major abiotic stress leading to reduced rice yield and is a significant environmental threat to food security. Low-temperature chilling studies have focused on physiological changes or coding genes. However, the competitive endogenous RNA mechanism in rice at low temperatures has not been reported. Therefore, in this study, antioxidant physiological indices were combined with whole-transcriptome data through weighted correlation network analysis, which found that the gene modules had the highest correlation with the key antioxidant enzymes superoxide dismutase and peroxidase. The hub genes of the superoxide dismutase-related module included the UDP-glucosyltransferase family protein, sesquiterpene synthase and indole-3-glycerophosphatase gene. The hub genes of the peroxidase-related module included the WRKY transcription factor, abscisic acid signal transduction pathway-related gene plasma membrane hydrogen-ATPase and receptor-like kinase. Therefore, we selected the modular hub genes and significantly enriched the metabolic pathway genes to construct the key competitive endogenous RNA networks, resulting in three competitive endogenous RNA networks of seven long non-coding RNAs regulating three co-expressed messenger RNAs via four microRNAs. Finally, the negative regulatory function of the WRKY transcription factor OsWRKY61 was determined via subcellular localization and validation of the physiological indices in the mutant.

Cold stress impairs crop productivity through cascading inhibition of root growth, nitrogen (N) metabolism, and photosynthesis, yet the systematic linkages among these physiological disruptions remain poorly understood. It is crucial to elucidate the mechanisms by which cold-tolerant varieties maintain root growth and N-metabolizing enzyme homeostasis. This two-year field study investigated how cold duration at the tillering stage impacted root traits, N metabolism, photosynthesis, and their relationships with the yield of two japonica rice varieties differing in cold tolerance. A cold-tolerant (Dongnong 428) and a cold-sensitive variety (Songjing 10) were grown in a paddy field for two consecutive growing seasons in 2021 and 2022. Cold water (15 °C) was irrigated for 0 (denoted as D0), 5 (D5), 10 (D10), and 15 days (D15) during the tillering stage. Compared to D0, cold-water treatments significantly reduced root traits and total dry weight of both varieties. Cold stress significantly impaired N metabolism and photosynthesis, leading to significant reductions in N efficiency. The magnitude of these changes turned to greater with cold-water treatment duration. Dongnong 428 showed stronger cold tolerance, attributed to its maintenance of superior root traits and photosynthetic performance, as well as higher activities of enzymes in the roots, which sustained N assimilation and utilization. These factors primarily contributed to Dongnong 428 achieving 11.6–20.9% higher yields compared to Songjing 10. Cold stress during the tillering stage disrupts root growth and photosynthesis, impairs plant N acquisition ability, resulting in substantial yield loss. Cold-tolerant varieties maintain superior root morphology/functionality and photosynthetic performance.

Low-temperature stress profoundly impairs rice root physiology and reshapes rhizosphere microbial communities. This 2023–2024 study examined its effects on Oryza sativa var. japonica across key growth stages. All treatments significantly suppressed root morphology and function, with the greatest reductions under combined tillering–booting stress (T3), followed by tillering (T1) and booting (T2). Strain DN428 exhibited a stronger cold tolerance than SJ10, with milder declines in root traits. Low-temperature stress elevated soil organic matter and total nitrogen while decreasing available phosphorus and potassium, leading to notable shifts in the microbial community structure and metabolic pathways. Weighted Gene Co-expression Network Analysis identified lacZ, fucK, and rafA in the MEbrown module as potential regulators of varietal cold responses. Mechanistically, yield loss in DN428 was mainly linked to the suppression of microbial gene expression, while in SJ10 it was associated with broader declines in microbial diversity and functional potential. Both varieties experienced yield reductions, accompanied by decreased root activity and nitrogen uptake. These findings underscore the necessity of a “gene–microbe–function” strategy to enhance microbial metabolism and optimize root–soil interactions under cold stress.