In order to understand molecular basis of cross-fertilized kernel advantage and heterosis, improved differential display of mRNA was used in this study to analyze alterations in gene expression between cross-fertilized and self-fertilized kernels at 2, 4, 6, 8, 10 and 12 days after pollination (DAP) by using 3 wheat hybrids with different level of heterosis. Four patterns of differential expression were observed: (i) bands observed in cross-fertilized kernels but not in self-fertilized kernels (BCnS); (ii) bands occurring in only self-fertilized kernels but not in cross-fertilized kernels (BSnC); (iii) cDNA over-expressed in cross-fertilized kernels compared to self-fertilized kernels (OEC); (iv) cDNA under-expressed in cross-fertilized kernels compared to self-fertilized kernels (UEC). Further analysis showed that BCnS is positively correlated with heterosis, but BSnC is negatively correlated with heterosis. Four differentially expressed cDNA fragments were verified by reverse-northern blot and sequence homology search in GenBank showed that one of them was new sequences; the other exhibited higher similarity to NBS-LRR type resistance protein, 1,6-bisphosphatase and photosystem Ⅱ chlorophyll a-binding protein psbB, respectively, which indicated diverse pathways may be involved in heterosis formation.
