Silencing of puroindoline a alters the kernel texture in transgenic bread wheat

Grain hardness is an important end-use quality parameter of bread wheat, and one of the most important characters for quality improvement. The objective of this study was to further understand the function of puroindolines and the underlying mechanism in the formation of kernel texture. The highly efficient expression vector pUBPa harboring puroindoline a (Pina) was introduced into the bread wheat cultivar Zhongyou 9507-60 via biolistic transformation and transgenic plants were obtained. The integration of the foreign Pina gene was confirmed by PCR and genomic DNA Southern blot analysis. The levels of friabilin on the surface of water-washed starch granules varied among the transgenic lines. SDS-PAGE analysis of Triton X-114 extracted protein showed that the PINA protein was absent in three transgenic lines, indicating that the endogenous Pina gene most likely had been co-suppressed by the over-expression of the Pina transgene. SKCS kernel hardness and scanning electron microscopy analysis further confirmed the changes of kernel texture in these lines.