The use of endosperm balance number for predicting gamete selection in complex polyploid interspecific Trifolium repens × T. nigrescens crosses

Endosperm balance number (EBN) was used to predict the outcome of crosses among a series of complex polyploid interspecific hybrids between Trifolium repens and T. nigrescens. Normal endosperm development, and hence seed‐set, requires a 2 : 1 maternal : paternal EBN ratio in endosperm. Based on their crossability, T. repens (2n = 4x = 32) and T. nigrescens (2n = 2x = 16) were assigned the same EBN of 4. This number was used to predict the relative success of different backcrosses and intercrosses. The method was further developed to successfully predict which gametes would effect fertilization when complex polyploid hybrids with uneven genomic constitutions (5x, 7x) produced mixtures of euploid and aneuploid gametes. This was achieved by allocating EBNs to all potential gametes of complex polyploids. Allocation of gamete EBNs enabled not only prediction of the success or failure of a cross, but also which gametes would be filtered to effect fertilization in complex crosses. This enhanced ability to understand the potential outcomes of complex crosses will increase the efficiency of plant breeding programmes that integrate secondary gene‐pools into economic species by interspecific hybridization.