An all-in-one visual selection system for male-sterile line production in maize and rice

The utilization of heterosis has significantly improved crop yields, yet the hybrid seed production remains hindered by labor-intensive manual emasculation. Although current male-sterility systems, such as cytoplasmic and environment-sensitive genic male sterility, have improved the efficiency of hybrid seed production, their limited genetic adaptability and environmental dependence remain major challenges. Here we report an all-in-one seed production technology (ASPT) that integrates CRISPR/Cas9, RUBY and essential seed production technology (SPT) components into a single vector, enabling the efficient generation and production of male-sterile lines in both maize and rice. The engineered RUBY marker facilitates visual identification of male-sterile and maintainer lines, with an accuracy of 99.81% in automated seed sorting and 100% in secondary field screening. Notably, the ASPT was successfully introduced into 21 genetically diverse maize elite inbred lines, demonstrating its broad compatibility. ASPT facilitates scalable and precise propagation of male-sterile lines in both maize and rice, offering a broadly applicable solution for advancing hybrid seed production in crops.