Bread Wheat Microspore Culture.

Microspore culture is one of the doubled-haploid-producing techniques that can speed up the breeding process and shorten the time for releasing new cultivars. Growing world population and changing climate demand to expedite the development of new resilient and high-yielding cultivars for staple crops such as bread wheat (Triticum aestivum L.). Here, we describe a comprehensive protocol for obtaining bread wheat doubled haploids through microspore culture. Key steps including donor plant growth and spike pretreatment, microspore isolation and density-gradient-based purification, microspore-ovary coculture and developmental stage tracking, embryo transferring and plantlet regeneration, as well as ploidy level determination and chromosome doubling of microspore-developed plantlets are covered in detail.