Summary: Heat stress (HS) has become a major factor limiting crop yields worldwide. HS inhibits plant growth by ROS accumulation, and NADPH oxidases (Rbohs) are major ROS producers in plants. Here, we show that CRISPR/Cas knockout of the OsRbohB (OsRbohB‐KO) significantly increased rice tolerance to HS imposed at various different growth stages. We produced OsRbohB‐KO and OsRbohB‐overexpression (OsRbohB‐OE) lines in a japonica cultivar, Nipponbare. Compared with nontransgenic wild‐type (WT) plants, the OsRbohB‐KO lines showed a significant increase in chlorophyll contents (5.2%–58.0%), plant growth (48.2%–65.6%) and grain yield (8.9%–20.5%), while reducing HS‐induced ROS accumulation in seeds (21.3%–33.0%), seedlings (13.0%–30.4%), anthers (13.1%–20.3%) and grains (9.7%–22.1%), under HS conditions. Analysis of yield components revealed that the increased yield of OsRbohB‐KO plants was due to increased starch synthetase activity, spikelets per panicle (2.0%–9.3%), filled spikelets (4.8%–15.5%), percentage of filled spikelets (2.4%–6.8%) and 1000‐grain weight (2.9%–7.4%) under HS conditions during the reproductive stage. Grain milling and appearance quality, and starch content were also significantly increased in OsRbohB‐KO plants under HS conditions during the mature stage. Furthermore, OsRbohB‐KO significantly upregulated the expression levels of heat shock‐related genes, OsHSP23.7, OsHSP17.7, OsHSF7 and OsHsfA2a, in rice seedlings and grains under long‐term HS conditions. Conversely, OsRbohB‐OE resulted in phenotypes that were opposite to OsRbohB‐KO in most cases. Our results suggest that suppression of OsRbohB provides an effective approach for alleviating heat damage and improving grain yield and quality of rice under long‐term HS conditions.

Background: Heat stress is a major restrictive factor that causes yield loss in rice. We previously reported the priming effect of abscisic acid (ABA) on rice for enhanced thermotolerance at the germination, seedling and heading stages. In the present study, we aimed to understand the priming effect and mechanism of ABA on grain filling capacity in rice under heat stress. Results: Rice plants were pretreated with distilled water, 50 μM ABA and 10 μM fluridone by leaf spraying at 8 d or 15 d after initial heading (AIH) stage and then were subjected to heat stress conditions of 38 °C day/30 °C night for 7 days, respectively. Exogenous ABA pretreatment significantly super-activated the ABA signaling pathway and improved the SOD, POD, CAT and APX enzyme activity levels, as well as upregulated the ROS-scavenging genes; and decreased the heat stress-induced ROS content (O₂^– and H₂O₂) by 15.0–25.5% in rice grain under heat stress. ABA pretreatment also increased starch synthetase activities in rice grain under heat stress. Furthermore, ABA pretreatment significantly improved yield component indices and grain yield by 14.4–16.5% under heat stress. ABA pretreatment improved the milling quality and the quality of appearance and decreased the incidence of chalky kernels and chalkiness in rice grain and improved the rice grain cooking quality by improving starch content and gel consistence and decreasing the amylose percentage under heat stress. The application of paraquat caused overaccumulation of ROS, decreased starch synthetase activities and ultimately decreased starch content and grain yield. Exogenous antioxidants decreased ROS overaccumulation and increased starch content and grain yield under heat stress. Conclusion: Taken together, these results suggest that exogenous ABA has a potential priming effect for enhancing rice grain filling capacity under heat stress at grain filling stage mainly by inhibiting ROS overaccumulation and improving starch synthetase activities in rice grain.

Nitrogen is a key element in promoting crop growth and development and improving photosynthesis. This study aimed to study the response of two rice genotypes to the restoration of N supply after varying periods of N deficiency. We used the low-nitrogen-tolerant rice Jijing 88 (JJ 88) and the nitrogen-sensitive rice variety Xinong 999 (XN 999) as test materials. The results of this study indicated that, compared to XN 999, JJ 88 has a higher content of the photosynthetic pigments. Photosynthesis in JJ 88 has strong adaptability under low-nitrogen conditions. Upon an increase in the nitrogen supply level, the maximum regeneration rate of ribulose biphosphate (RuBP, J_max) and the maximum carboxylation rate of RuBP (V_cmax) in JJ 88 showed a relatively large increase. The chlorophyll fluorescence parameters, including the effective quantum yield of photosystem II (Φ_PSII), the efficiency of excitation capture by open PSII centers (Fv′/Fm′), photochemical fluorescence quenching (qP), and the electron transfer rate (ETR) decreased slightly, while the non-photochemical fluorescence quenching (NPQ) increased slightly. Under low-nitrogen conditions, low-nitrogen-tolerant rice varieties maintain reasonable growth during the seedling stage. With an increase in the nitrogen supply level, the dry matter accumulation, photosynthetic pigment content, photosynthesis, and electron transfer ability of plants improve, but not to normal nitrogen supply levels. However, compared with XN 999, JJ 88 has a more proactive recovery ability. The research results provide valuable guidance for the breeding of nitrogen-efficient rice varieties and nitrogen fertilizer management.

Nitrogen (N) availability significantly influences plant metabolism and productivity. The aim of this study was to assess the effects of low N stress and subsequent N supplementation on key enzymes of nitrogen metabolism, nitrogen metabolism-related substances, and chlorophyll a fluorescence kinetic parameters in rice genotypes with different nitrogen utilization efficiencies. We used the Jijing 88 (low-N tolerant) and Xinong 999 (low-N sensitive) as test materials. During the seedling, tillering, and booting stages, the 1/2N and 1/4N treatments were restored to the 1N treatment level. Nine treatments were used in this experiment: CK (1N), A1 (1/2N), A2 (1/2N restored to 1N during the seedling stage), A3 (1/2N restored to 1N during the tillering stage), A4 (1/2N restored to 1N during the booting stage), B1 (1/4N), B2 (1/4N restored to 1N during the seedling stage), B3 (1/4N restored to 1N during the tillering stage), and B4 (1/4N restored to 1N during the booting stage). Key physiological responses, nitrogen compounds, enzymes activities, and chlorophyll a fluorescence kinetics were analyzed. Under low nitrogen conditions, the growth and nitrogen assimilation of rice were inhibited. Compared to XN 999, JJ 88 maintains higher levels of dry matter, nitrate reductase activity (NR), glutamine synthetase activity (GS), glutamate oxaloacetate transaminase activity (GOT), glutamate pyruvate transaminase activity (GPT), as well as nitrate (NO₃⁻) and ammonium (NH₄⁺) nitrogen contents. After N supplementation during the early growth stage, both JJ 88 and XN 999 exhibit recovery capabilities. However, in the late growth stage, JJ 88 demonstrates superior recovery capabilities. In addition to enhancing nitrogen metabolism levels, there is also an increase in the content of osmotic regulation substances such as soluble sugars, free amino acids, and proline, along with responses in chlorophyll fluorescence kinetic parameters. This was primarily manifested in the enhancement of performance index (PI_ABS, PI_total), and quantum yield (φ_EO, φ_RO, ψ_EO), which maintain photosynthetic performance and electron transport efficiency. The research findings indicated that reducing N supply during the early growth stage and restoring N levels in the later stage are beneficial for the recovery of low-nitrogen-tolerant rice varieties. Therefore, in the context of sustainable agricultural production, the breeding of low-nitrogen-tolerant rice varieties and the optimization of N fertilizer management are crucial.