Identification of allelic variations of puroindoline genes controlling grain hardness in wheat using a modified denaturing PAGE
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文摘
Puroindoline genes (Pina and Pinb) form the molecular basis of wheat grain hardness or texture. Variations in either gene are reported to be associated with grain hardness in wheat. Here, a modified denaturing PAGE was used to detect Pina and Pinb allelic variations in 102 common wheat cultivars and other species related to wheat. Two variations of Pina (Pina-D1b and Pina-D1p) and five variations of Pinb (null, Pinb-D1b, Pinb-D1u, Pinb-D1v and Pinb-D1w) were identified in common (T. aestivum) and spelt wheats (T. aestivum ssp. spelta). No allelic variation was found in Tibet semi-wild wheat (T. aestivum ssp. tibetanum), club wheat (T. compactum), or Aegilops tauschii. Puroindoline genes were absent in wild␣emmer (T. turgidum var. dicoccoides). The sequencing results of the PCR fragments from Pina and Pinb (except for null type) indicated allelic variants carrying single base mutation, such as Pina-D1p, Pinb-D1u, Pinb-D1v and Pinb-D1w were novel types. Our results showed that the modified PAGE used in this study provided a satisfactory resolving power for identifying single nucleotide mutations; therefore, it is a practical and simple tool to study allelic variation of Pina and Pinb in wheat and related species.

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