Rice (Oryza sativa L.) is a staple food that is consumed worldwide, and hybrid rice has been widely employed in many countries to greatly increase yield. However, the frequency of extreme temperature events is increasing, presenting a serious challenge to rice grain quality. Improving hybrid rice grain quality has become crucial for ensuring consumer acceptance. This study compared the differences in milling quality, appearance quality, and physical and chemical starch properties of rice grains of five restorer lines (the male parent of hybrid rice) when they encountered naturally unfavorable temperatures during the filling period under field conditions. High temperatures (HTs) and low temperatures (LTs) had opposite effects on grain quality, and the effect was correlated with rice variety. Notably, R751, R313, and Yuewangsimiao (YWSM) were shown to be superior restorer lines with good resistance to both HT and LT according to traits such as head rice rate, chalkiness degree, chalky rice rate, amylose content, alkali spreading value, and pasting properties. However, Huazhan and 8XR274 were susceptible to sub-optimal temperatures at the grain-filling stage. Breeding hybrid rice with adverse-temperature-tolerant restorer lines can not only ensure high yield via heterosis but also produce superior grain quality. This could ensure the quantity and taste of rice as a staple food in the future, when extreme temperatures will occur increasingly frequently.

Understanding the signaling pathways activated in response to these combined stresses and their crosstalk is crucial to breeding crop varieties with dual or multiple tolerances. However, most studies to date have predominantly focused on individual stress factors, leaving a significant gap in understanding plant responses to combined biotic and abiotic stresses. The bHLH family plays a multifaceted regulatory role in plant response to both abiotic and biotic stresses. In order to comprehensively identify and analyze the bHLH gene family in rice, we identified putative OsbHLHs by multi-step homolog search, and phylogenic analysis, molecular weights, isoelectric points, conserved domain screening were processed using MEGAX version 10.2.6. Following, integrative transcriptome analysis using 6 RNA-seq data including Xoo infection, heat, and cold stress was processed. The results showed that 106 OsbHLHs were identified and clustered into 17 clades. Os04g0301500 and Os04g0489600 are potential negative regulators of Xoo resistance in rice. In addition, Os04g0301500 was involved in non-freezing temperatures (around 4°C) but not to 10°C cold stresses, suggesting a complex interplay with temperature signaling pathways. The study concludes that Os04g0301500 may play a crucial role in integrating biotic and abiotic stress responses in rice, potentially serving as a key regulator of plant resilience under changing environmental conditions, which could be important for further multiple stresses enhancement and molecular breeding through genetic engineering in rice.

Chilling stress is one of the main abiotic factors affecting rice growth and yield. In rice, chlorophyllide a oxygenase encoded by OsCAO1 is responsible for converting chlorophyllide a to chlorophyllide b, playing a crucial role in photosynthesis and thus rice growth. However, little is known about the function of OsCAO1 in chilling stress responses. The presence of the cis-acting element involved in low-temperature responsiveness (LTR) in the OsCAO1 promoter implied that OsCAO1 probably is a cold-responsive gene. The gene expression level of OsCAO1 was usually inhibited by low temperatures during the day and promoted by low temperatures at night. The OsCAO1 knockout mutants generated by the CRISPR-Cas9 technology in rice (Oryza sativa L.) exhibited significantly weakened chilling tolerance at the seedling stage. OsCAO1 dysfunction led to the accumulation of reactive oxygen species and malondialdehyde, an increase in relative electrolyte leakage, and a reduction in antioxidant gene expression under chilling stress. In addition, the functional deficiency of OsCAO1 resulted in more severe damage to chloroplast morphology, such as abnormal grana thylakoid stacking, caused by low temperatures. Moreover, the rice yield was reduced in OsCAO1 knockout mutants. Therefore, the elevated expression of OsCAO1 probably has the potential to increase both rice yield and chilling tolerance simultaneously, providing a strategy to cultivate chilling-tolerant rice varieties with high yields.

Stem rot caused by Sclerotinia sclerotiorum poses a significant threat to global agriculture, leading to substantial economic losses. To explore innovative integrated pest management strategies and elucidate the underlying mechanisms, this study examined the impact of nano-silicon on enhancing resistance to Sclerotinia sclerotiorum in Brassica napus. Bacteriostatic assays revealed that nano-silicon effectively inhibited the mycelial growth of Sclerotinia sclerotiorum in a dose-dependent manner. Field trials corroborated the utility of nano-silicon as a fertilizer, substantially bolstering resistance in the Brassica napus cultivar Xiangyou 420. Specifically, the disease index was reduced by 39–52% across three distinct geographical locations when compared to untreated controls. This heightened resistance was attributed to nano-silicon’s role in promoting the accumulation of essential elements such as silicon (Si), potassium (K), and calcium (Ca), while concurrently reducing sodium (Na) absorption. Furthermore, nano-silicon was found to elevate the levels of soluble sugars and lignin, while reducing cellulose content in both leaves and stems. It also enhanced the activity levels of antioxidant enzymes. Transcriptomic analysis revealed 22,546 differentially expressed genes in Si-treated Brassica napus post-Sclerotinia inoculation, with the most pronounced transcriptional changes observed one day post-inoculation. Weighted gene co-expression network analysis identified a module comprising 45 hub genes that are implicated in signaling, transcriptional regulation, metabolism, and defense mechanisms. In summary, nano-silicon confers resistance to Brassica napus against Sclerotinia sclerotiorum by modulating biochemical defenses, enhancing antioxidative activities, and rapidly reprogramming key resistance-associated genes. These findings contribute to our mechanistic understanding of Si-mediated resistance against necrotrophic fungi and offer valuable insights for the development of stem-rot-resistant Brassica napus cultivars.