Department of Genetics and Genetic Engineering Research Center (GERC); Faculty of Agriculture Cairo University; Giza, Egypt.
Agriculture Genetic Engineering Research Institute (AGERI); Agriculture Research Center (ARC); Giza, Egypt.
GM Crops Food. 2014 Apr-Jun;5(2):132-8. doi: 10.4161/gmcr.28941. Epub 2014 Jul 9.
We present an efficient method for the production of transgenic salt tolerant hexaploid wheat plants expressing the Arabidopsis AtNHX1 gene. Wheat mature zygotic embryos were isolated from two hexaploid bread wheat (Triticum aestivum) cultivars (namely: Gemmeiza 9 and Gemmeiza 10) and were transformed with the A. tumefaciens LBA4404 harboring the pBI-121 vector containing the AtNHX1 gene. Transgenic wheat lines that express the gus intron was obtained and used as control. The results confirmed that npt-II gene could be transmitted and expressed in the T2 following 3:1 Mendelian segregation while the control plant couldn't. The data indicate that, the AtNHX1 gene was integrated in a stable manner into the wheat genome and the corresponding transcripts were expressed. The transformation efficiency was 5.7 and 7.5% for cultivars Gemmeiza 10 and Gemmeiza 9, respectively. A greenhouse experiment was conducted to investigate the effect of AtNHX1 gene in wheat salt tolerance. The transgenic wheat lines could maintain high growth rate under salt stress condition (350 mM NaCl) while the control plant couldn't. The results confirmed that Na(+)/H(+) antiporter gene AtNHX1 increased salt tolerance by increasing Na(+) accumulation and keeping K+/Na(+) balance. Thus, transgenic plants showed high tolerance to salt stress and can be considered as a new genetic resource in breeding programs.
我们提出了一种高效的生产方法,用于生产表达拟南芥 AtNHX1 基因的耐盐性六倍体小麦转基因植株。从小麦六倍体面包小麦(Triticum aestivum)两个品种(即:Gemmeiza 9 和 Gemmeiza 10)中分离成熟的合子胚,并将含有 AtNHX1 基因的 pBI-121 载体的农杆菌 LBA4404 转化。获得了表达 gus 内含子的转基因小麦株系作为对照。结果证实,npt-II 基因可以在 T2 代中以 3:1 的孟德尔分离方式进行传递和表达,而对照植株则不能。数据表明,AtNHX1 基因以稳定的方式整合到小麦基因组中,并表达相应的转录物。Gemmeiza 10 和 Gemmeiza 9 两个品种的转化效率分别为 5.7%和 7.5%。进行了温室实验以研究 AtNHX1 基因在小麦耐盐性中的作用。转基因小麦株系在盐胁迫条件(350mM NaCl)下仍能保持较高的生长速度,而对照植株则不能。结果证实,Na+/H+逆向转运蛋白基因 AtNHX1 通过增加 Na+积累和维持 K+/Na+平衡来提高耐盐性。因此,转基因植物对盐胁迫具有较高的耐受性,可以作为育种计划中的一种新的遗传资源。