Abstract
To address the critical challenges of low embryogenic induction efficiency and limited system stability in winter wheat microspore culture, this study systematically optimized key technical parameters involved in microspore isolation, induction, and differentiation, while also assessing genotypic responses. Using three winter wheat cultivars as experimental materials, a series of culture trials was conducted under varying pretreatment regimes, isolation solutions, inoculation densities, and differentiation media. The results indicated that an extended period of low-temperature pretreatment notably improved microspore viability and embryogenic competence. Among the tested isolation solutions, one specific formulation yielded the highest proportion of viable microspores. An intermediate inoculation density was found to be optimal for callus induction, significantly enhancing tissue proliferation. Furthermore, a specially formulated differentiation medium substantially promoted organogenesis from callus tissues. Significant to highly significant differences among genotypes were observed during microspore culture, particularly at the stages of callus formation and differentiation. This study established a simplified, efficient, and reproducible microspore culture system with wide genotypic applicability, clarified the major influencing factors and their interactions in microspore embryogenesis, and provided a robust cellular engineering platform for accelerating the breeding process of winter wheat.
