High-lysine corn generated by endosperm-specific suppression of lysine catabolism using RNAi

被引:90
作者
Hournard, Nancy M. [1 ]
Mainville, Jonnelle L. [1 ]
Bonin, Christopher P. [1 ]
Huang, Shihshieh [1 ]
Luethy, Michael H. [1 ]
Malvar, Thomas M. [1 ]
机构
[1] Monsanto Co, Mystic Res, Mystic, CT 06355 USA
关键词
lysine; high-lysine corn; RNAi; lysine catabolism; lysine-ketoglutarate recluctase/saccharopine dehydrogenase; (LKR/SDH);
D O I
10.1111/j.1467-7652.2007.00265.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Because of the limited lysine content in corn grain, synthetic lysine supplements are added 1 to corn meal-based rations for animal feed. The development of biotechnology, combined with the understanding of plant lysine metabolism, provides an alternative solution for increasing corn lysine content through genetic engineering. Here, we report that by suppressing lysine catabolism, transgenic maize kernels accumulated a significant amount of lysine. This was achieved by RNA interference (RNAi) through the endosperm-specific expression of an inverted-repeat (IR) sequence targeting the maize bifunctional lysine degradation enzyme, lysine-ketoglutarate reductase/saccharopine dehydrogenase (ZLKR/SDH). Although plant-short interfering RNA (siRNA) were reported to lack tissue specificity due to systemic spreading, we confirmed that the suppression of ZLKR/SDH in developing transgenic kernels was restricted to endosperm tissue. Furthermore, results from our cloning and sequencing of siRNA suggested the absence of transitive RNAi. These results support the practical use of RNAi for plant genetic engineering to specifically target gene suppression in desired tissues without eliciting systemic spreading and the transitive nature of plant RNAi silencing.
引用
收藏
页码:605 / 614
页数:10
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