Rice–fish co-cultures have been practiced for over 2000 years, and they have tremendous potential in terms of increasing food security and economic benefits. However, little research has been conducted into achieving stable yields and high lodging resistance with regard to rice while simultaneously promoting the harmonious and healthy growth of fish in rice–fish co-culture paddy fields. We conducted a field study aimed at selecting suitable rice varieties for rice–fish co-culture systems (encompassing both ratoon and main crop). This selection process was grounded in an evaluation of lodging resistance and grain yield among 33 rice varieties used throughout the studied region. The results revealed a range of lodging indices of the main crop for the second internode, spanning from 62.43 to 138.75, and the annual grain yield (main crop and ratoon crop) ranged from 7.17 to 13.10 t ha⁻¹ within rice–fish co-culture systems. We found that the use of rice–fish co-culture farming could improve the milling quality, nutrient quality, and appearance quality of rice, though the improvement gained through co-culturing varied across rice varieties. Moreover, the lodging index of the three basal internodes of rice plants was significantly and positively correlated with the plant height and the culm fresh weight, but it was negatively correlated with the bending strength of the rice basal internodes. Additionally, the 33 tested rice varieties were clustered in accordance with their lodging resistance (i.e., high resistance with lodging indices 62.43–75.42; medium resistance with lodging indices 80.57–104.62; and low resistance with lodging indices 113.02–138.75) according to the hierarchical cluster analysis. The 33 rice varieties were also clustered in accordance with the annual (main crop and ratoon crop) grain yield (i.e., high yield with 11.17–13.10 t ha⁻¹; medium yield with 10.15–10.83 t ha⁻¹; and low yield with 7.16–9.88 t ha⁻¹). In all, 11 rice varieties were identified by a comprehensive evaluation as suitable varieties for grain production in the rice–fish co-culture system. These varieties displayed favorable traits, including a high annual rice yield, strong lodging resistance, and good grain quality. This is the first study to systematically evaluate rice varieties based on grain yield, lodging resistance, and grain quality in rice–fish co-culture systems.

Seed development plays a critical role in determining both crop yield and grain quality in rice. As a key nutrient storage organ, the rice endosperm development not only contributes to grain filling but also plays an essential role during the early stages of seed germination. Amino acid metabolism is active during the process of seed development and seed germination. Asparagine is a primary amino acid responsible for long-distance organic nitrogen transport in plants. Asparagine synthetase catalyzes the synthesis of asparagine from aspartate and glutamine. In this study, CRISPR/Cas9-mediated knockout mutants of the OsASN2 gene of rice were generated. Homozygous mutants exhibited complete failure of seed germination, and heterozygotes could not produce homozygous offspring. Endosperm development of homozygous mutant seeds showed severe defects. Additionally, interacting protein screening combined with pull-down and co-immunoprecipitation (Co-IP) assays confirmed that OsASN2 physically interacted with pyruvate phosphate dikinase OsPPDKB, the mutants of which showed impaired endosperm development. These findings collectively indicate that OsASN2 plays a critical role in seed development and germination in rice.