The RNA-mediated silencing of one of the Pin genes in allohexaploid wheat simultaneously decreases the expression of the other, and increases grain hardness
The RNA-mediated silencing of one of the Pin genes in allohexaploid wheat simultaneously decreases the expression of the other, and increases grain hardness
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DOI：
10.1093/jxb/err103
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发表时间：
2011-07-01
影响因子：
6.9
通讯作者：
Nadolska-Orczyk, Anna
中科院分区：
文献类型：
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作者：
Gasparis, Sebastian;Orczyk, Waclaw;Nadolska-Orczyk, Anna
在两个小麦品种Kontesa和Torka中诱导并分析了Pina和Pinb（负责异源六倍体小麦的籽粒质地的两个基因）的RNAi介导的沉默。在非转基因植物中的两个基因的表征表明，Pinb携带点突变，指定Pinb-D1c在两个品种。这种突变不影响转录丰度或蛋白质含量。构建hpRNA类型的两个沉默盒，并用于通过农杆菌的稳定转化。总共获得43个代表两个栽培品种的转基因株系，用Pina或Pinb的沉默盒转化或用两种盒共转化。发现同一子代植物中两种基因的相对转录水平相似，与所使用的沉默盒无关。用沉默盒之一转化的Kontesa和Torka的分离T-1后代中Pina和Pinb转录物水平的降低超过80%。用两种基因的沉默盒共转化导致Kontesa的一些分离的T-1后代中超过91%的Pina和Pinb转录本减少。沉默被传递到T-3系的T-4籽粒世代。一个显着的减少或缺乏这两个嘌呤吲哚蛋白质的沉默线与一个重要的增加谷物硬度。讨论涵盖了Pin基因功能的一些新见解，包括与siRNA信号无关的同时沉默两者。
The RNAi-mediated silencing of Pina and Pinb, the two genes responsible for the grain texture of allohexaploid wheat, was induced and analysed in two wheat cultivars, Kontesa and Torka. A characterization of the two genes in non-transgenic plants revealed that Pinb carries a point mutation, designated Pinb-D1c in both cultivars. This mutation does not influence transcript abundance or protein content. Two silencing cassettes of the hpRNA type were constructed and used for stable transformation via Agrobacterium. In total, 43 transgenic lines representing the two cultivars were obtained, transformed with the silencing cassettes for Pina or for Pinb or co-transformed with both cassettes. The relative transcript levels of the two genes in the same progeny plant were found to be similar, independent of the silencing cassette used. The reduction in the Pina and Pinb transcript levels in the segregating T-1 progeny of Kontesa and Torka transformed with one of the silencing cassettes exceeded 80%. Co-transformation with the silencing cassettes for both genes resulted in a reduction of over 91% of Pina and Pinb transcripts in some segregating T-1 progeny of Kontesa. The silencing was transmitted to the T-4 kernel generation of the T-3 lines. A significant reduction or lack of both puroindoline proteins in the silenced lines correlated with an essential increase in grain hardness. The discussion covers some new insights into the function of the Pin genes, including the simultaneous silencing of both, independent of the siRNA signal.