Huabing Deng

Heat stress has emerged as one of the major environmental factors constraining rice growth. In our previous studies, OsPUP1, a member of the purine transporter protein family in rice, has been verified to be involved in the transport of cytokinin (CK). Unfortunately, the role of the OsPUP1 gene in the heat stress response has not been fully elucidated to date. In this study, we show that overexpression of OsPUP1, encoding a purine permease in rice, leads to severe growth inhibition and oxidative damage under heat stress. OsPUP1 overexpression lines exhibited increased accumulation of peroxides and malondialdehyde (MDA), along with significantly reduced activities of superoxide dismutase (SOD) and peroxidase (POD). Together with previous findings, these results suggest that OsPUP1 modulates CK transport, thereby influencing the spatial distribution of CK within the plant, which in turn regulates antioxidant defense mechanisms and heat stress response. This study uncovers a key role for OsPUP1 in controlling thermotolerance in rice and highlights the importance of CK distribution in plant adaptation to heat stress.

Rice (Oryza sativa) is a staple crop that is highly susceptible to heat stress (HS), which severely impairs growth and yield. In this study, we identified the rice Ovate Family Protein OsOFP3 as a novel negative regulator in response to heat. Our results demonstrate that the expression of OsOFP3 is suppressed at both the transcriptional and protein levels under HS. Overexpression of OsOFP3 significantly reduces the survival rate of rice seedlings under HS and exacerbates chlorophyll degradation, membrane damage, and the accumulation of reactive oxygen species (H₂O₂ and O₂⁻). In contrast, OsOFP3 mutants exhibit enhanced heat tolerance. Moreover, OsOFP3-overexpressing plants display increased stomatal opening and decreased stomatal closure under HS. Molecular interaction analysis further reveals that OsOFP3 interacts with the C-terminal domain of OsHTAS, a known positive regulator of heat tolerance encoding an E3 ubiquitin ligase, and this interaction depends on the RING domain of OsHTAS. Taken together, our findings indicate that OsOFP3 negatively regulates rice heat tolerance by disrupting ROS homeostasis, inhibiting stomatal closure, and potentially antagonizing the OsHTAS-mediated signaling pathway. This research provides new insights into the molecular mechanisms underlying HS tolerance in rice.

Screening and breeding more resistant heat stress restorer lines represent an effective approach to addressing the decline in hybrid rice seed production caused by heat stress (HS). However, the molecular mechanisms affecting the differences in the heat resistance of anthers under HS remain unclear. This study compared the gene expression patterns of two hybrid rice restorer lines with differing heat resistances under HS and discusses the mechanisms of the heat response in rice. Under heat stress, 247 DEGs were co-expressed across varieties and were involved in biological processes such as protein processing and carbon metabolism, with heat shock proteins being the most ubiquitous. Interestingly, a substantial enrichment of genes related to non-structural carbohydrates and ATP was observed among the unique DEGs in R996 and R4628. Simultaneously, the contents of non-structural carbohydrates and ATP levels in the young spikes of R996 were significantly higher than those in R4628. This suggests that starch, soluble sugars and ATP play significant roles in heat tolerance during the flowering stage of rice. Overall, this study provides novel insights into the molecular mechanisms underlying heat stress resistance in indica rice restorer lines and informs future strategies for the genetic improvement of heat tolerance in these varieties.

There are many factors affecting rice yield and quality during cultivation, including temperature, light, water, and fertilization, among which high temperature (HT) is one of the main factors affecting rice yield and quality. However, less is known about the effects and potential mechanisms of different durations of HT stress during the grain filling stage on grain quality. In this study, the differences in rice quality and starch rapid viscosity analyzer (RVA) characteristics of eight indica rice varieties under different high-temperature treatment times were studied by simulating high temperature in an artificial climate chamber. The prolonged duration of HT leads to an overall deterioration in the milling quality, appearance quality, and cooking quality of rice. The impact of HT duration on the starch RVA characteristics of rice is more complex and is mainly related to the varieties. Among them, the starch RVA characteristics of R313 were more stable. It is worth noting that there is a significant difference in the sensitivity of the appearance quality of 8XR274 and 5W0076 to HT duration, with 8XR272 being more sensitive and 5W0076 being the opposite. We selected these two varieties for transcriptome analysis after 14 days of HT treatment and found that the number of differentially expressed genes (DEGs) in 8XR274 was significantly less than that in 5W0076. The DEGs of 8XR274 were mainly enriched in pathways related to carbohydrates, while 5W0076 was mainly enriched in pathways related to photosynthesis. Our study provides a new perspective on the molecular response and related genes of different rice varieties under high temperature, as well as the high-quality rice breeding under high temperature.

Cytokinins (CKs) play important functions in plant growth and development and in response to adversity stress. However, little is known about the role CK plays in rice grain quality. We hypothesized that exogenous cytokinins could improve rice grain quality by regulating physiological traits and genes related to starch synthesis. Therefore, we exogenously applied different concentrations of kinetin (KT), an exogenous CK, during the grain-filling period. Our results show that all the different concentrations of exogenous KT treatments resulted in a significant increase in thousand-grain weight. In particular, chalkiness and chalky grain rate were significantly reduced, and gel consistency (GC) content and alkali spreading value (ASV) were significantly increased in 10⁻⁸ M KT treatment. Meanwhile, the exogenous application of 10⁻⁸ M KT positively affected the transcription of some starch synthesis-related genes, which was in contrast to the 10⁻⁵ M KT treatment. In conclusion, the exogenous application of appropriate concentrations of KT during the grain-filling period can ultimately affect rice grain quality by regulating the changes in the relevant indicators, such as appearance quality (AQ) and eating and cooking qualities (ECQ).

Hybrid rice needs to prevent lodging due to excessive grain weight during maturity, which can lead to yield loss, grain quality deterioration and difficulty in harvesting. Breeding excellent parents (restorer and male-sterile lines) is the foundation for cultivating lodging-resistant hybrid rice. However, there is still limited information about restorers with improved lodging resistance. To identify a new restorer with significantly enhanced resistance to lodging, the restorer line Huazhan, which has been used for many years in hybrid rice breeding in China, was used as the control, to evaluate the lodging resistance of the new restorer R382. A two-year field experiment was executed to investigate the difference on lodging-related traits of the basal two internodes above-ground, such as out diameters of stems with or without leaf sheath (DWS, DWOS), culm thickness (CT), the section modulus (SM), bending stress (BS), and bending moment at break (M). The results indicated that DWS, CT, BS, M of R382 was at least 19.9%, 13.3%, 36.5% and 52.6% higher than that of Huazhan. However, the negative indicator lodging index of R382 was over 17.5% lower. The number and area of vascular bundles, and the contents of the chemical components of the cell wall including cellulose, lignin, and hemicellulose, together with the accumulation of starch granules, were significantly greater in R382. Furthermore, in the basal stems during the jointing stage, a series of genes that promote cell wall formation exhibited significantly higher expression levels in R382. Meanwhile, there was no significant difference in the yield of R382 and Huazhan. Thus, this new restorer could be a valuable donor in the future hybrid rice breeding programs for improving lodging resistance.

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.

High temperature is a significant environmental stress that limits plant growth and agricultural productivity. GDSL lipase is a hydrolytic enzyme with a conserved GDSL sequence at the N-terminus, which has various biological functions, such as participating in plant growth, development, lipid metabolism, and stress resistance. However, little is known about the function of the GDSL lipase gene in the heat tolerance of rice. Here, we characterized a lipase family protein coding gene HTA1, which was significantly induced by high temperature in rice. Rice seedlings in which the mutant hta1 was knocked out showed enhanced heat tolerance, whereas the overexpressing HTA1 showed more sensitivity to heat stress. Under heat stress, hta1 could reduce plant membrane damage and reactive oxygen species (ROS) levels and elevate the activity of antioxidant enzymes. Moreover, real-time quantitative PCR (RT-qPCR) analysis showed that mutant hta1 significantly activated gene expression in antioxidant enzymes, heat response, and defense. In conclusion, our results suggest that HTA1 negatively regulates heat stress tolerance by modulating the ROS accumulation and the expression of heat-responsive and defense-related genes in rice seedlings. This research will provide a valuable resource for utilizing HTA1 to improve crop heat tolerance.

Heat stress (HS) has become one of the major abiotic stresses that severely constrain rice growth. Abscisic acid (ABA) plays an important role in plant development and stress response. However, the effect of different concentrations of exogenous ABA on HS tolerance in rice still needs to be further elucidated. Here, we found that high concentrations of exogenous ABA increased HS damage in seedlings, whereas 10⁻¹² M ABA treatment increased fresh and dry weight under HS relative to mock seedlings. Our further data showed that, in response to HS, 10⁻⁵ M, ABA-treated seedlings exhibited a lower chlorophyll content, as well as transcript levels of chlorophyll biosynthesis and antioxidant genes, and increased the accumulation of reactive oxygen species (ROS). In addition, the transcript abundance of some heat-, defense-, and ABA-related genes was downregulated on 10⁻⁵ M ABA-treated seedlings under HS. In conclusion, high concentrations of exogenous ABA reduced the HS tolerance of rice seedlings, and this negative effect could be achieved by regulating the accumulation of ROS, chlorophyll biosynthesis, and the transcription levels of key genes in seedlings under HS.

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.