Seedling-derived leaf and root tip as alternative explants for callus induction and plant regeneration in maize

While being one of the world's most important crops, maize (Zea mays L.) is still relatively difficult to regenerate in tissue culture, which severely limits its improvement by genetic engineering. Currently, immature zygotic embryos provide the predominant material for transformation and regeneration. However, the procedures involved are often laborious and season-dependent. Therefore, new explants to replace or complement immature embryos are desirable. Here, we exploited root tips and young leaves isolated from 3-day-old dark-grown seedlings as alternative explant sources for establishing plant regeneration. As novel explants, the root tips could generate embryogenic calli similar to that from the young leaves. The rate of primary callus induction from root tips reached 97.2% and almost as high as 98.8% from immature embryos. The difference in callus induction rates among these explants may be closely related to the differences in expression level of stem cell-related genes in callus tissue. Moreover, the alternative explants are easy to obtain in large quantities. These combined results indicate that explants from seedling-derived root tips and leaf tissue have the potential to replace immature embryos for plant regeneration and transformation.