Pollen–pistil interactions in divergent wide crosses lead to spatial and temporal pre-fertilization reproductive barrier in flax (Linum usitatissimum L.)

Linseed, has been a source of natural fiber for textile industries since its domestication. However, despite being the potential source of trait reservoir, the use of Linum wild genetic resources for the improvement of economic traits are not exploited widely. This is mainly due to the degree of genetic divergence that exists among the interspecific ecotypes causing crossability issues. Self-incompatibility due to the occurrence of heterostyly is very well reported in distantly related crop wild relatives of Linum and, the mechanism of self-incompatibility between different floral morphs is also studied. However, pollen germination and tube growth responses in the interspecific crosses are rarely studied. Thus, the present study was exclusively carried out to assess the major pre-zygotic barriers and their effect on pollen germination on foreign stigma using fluorescent microscopy of aniline blue stain-aided technology, to understand how the species barriers operate on pollen germination and pollen tube growth. The study revealed that the pollen-pistil interaction in the wide crosses among L. usitatissimum X L. grandiflorum was regulated by both temporal and spatial pre-fertilization barriers. Callose deposition within 2 h after pollination (HAP) at the stigma surface, was the major cause inhibiting pollen germination. Various kinds of aberrations started appearing during the 2-4 HAP. The complexity of interspecific hybridization was observed in terms of arrest of pollen tube (PT) growth in the ovary, ruptured, twisted and swollen pollen tube tip, tube growth in reverse direction, convoluted and terminated growth patterns. Inconsistent growth rates of pollen tubes to reach various stylar regions emphasizes the importance of studying these wild relatives for potential agricultural advancements. The results show that while distant hybridization with L. grandiflorum is less efficient, pollen tubes can still navigate the ovular tissues, albeit with some delay. This finding opens avenues for investigating factors that hinder viable seed formation, enhancing our understanding of reproductive success in distant hybridization with this species.