Eissa Hala F, Hassanien Sameh E, Ramadan Ahmed M, El-Shamy Moustafa M, Saleh Osama M, Shokry Ahmed M, Abdelsattar Mohamed, Morsy Yasser B, El-Maghraby Maher A, Alameldin Hussien F, Hassan Sabah M, Osman Gamal H, Mahfouz Hesham T, Gad El-Karim Gharib A, Madkour Magdy A, Bahieldin Ahmed
Agricultural Genetic Engineering Research Institute (AGERI), Agriculture Research Center (ARC), Giza, 12619 Egypt.
Faculty of Biotechnology, Misr University for Science and Technology (MUST), Post Box 77, 6th October City, Egypt.
Plant Methods. 2017 May 22;13:41. doi: 10.1186/s13007-017-0191-5. eCollection 2017.
The main aim of this study was to improve fungal resistance in bread wheat via transgenesis. Transgenic wheat plants harboring barley () gene, driven by maize promoter, were obtained using biolistic bombardment, whereas the herbicide resistance gene, driven by the promoter was used as a selectable marker.
Molecular analysis confirmed the integration, copy number, and the level of expression of the gene in four independent transgenic events. Chitinase enzyme activity was detected using a standard enzymatic assay. The expression levels of gene in the different transgenic lines, compared to their respective controls, were determined using qRT-PCR. The transgene was silenced in some transgenic families across generations. Gene silencing in the present study seemed to be random and irreversible. The homozygous transgenic plants of T4, T5, T6, T8, and T9 generations were tested in the field for five growing seasons to evaluate their resistance against rusts and powdery mildew. The results indicated high chitinase activity at T0 and high transgene expression levels in few transgenic families. This resulted in high resistance against wheat rusts and powdery mildew under field conditions. It was indicated by proximate and chemical analyses that one of the transgenic families and the non-transgenic line were substantially equivalent.
Transgenic wheat with barley was found to be resistant even after five generations under artificial fungal infection conditions. One transgenic line was proved to be substantially equivalent as compared to the non-transgenic control.
本研究的主要目的是通过转基因提高面包小麦的真菌抗性。利用生物弹道轰击法获得了携带由玉米启动子驱动的大麦()基因的转基因小麦植株,而由启动子驱动的除草剂抗性基因用作选择标记。
分子分析证实了该基因在四个独立转基因事件中的整合、拷贝数和表达水平。使用标准酶促测定法检测几丁质酶活性。使用qRT-PCR测定不同转基因株系中该基因相对于各自对照的表达水平。该转基因在一些转基因家系的几代中发生沉默。本研究中的基因沉默似乎是随机且不可逆的。对T4、T5、T6、T8和T9代的纯合转基因植株进行了五个生长季节的田间试验,以评估它们对锈病和白粉病的抗性。结果表明,在T0代几丁质酶活性高,少数转基因家系中转基因表达水平高。这导致在田间条件下对小麦锈病和白粉病具有高抗性。近似分析和化学分析表明,其中一个转基因家系与非转基因品系基本等同。
发现携带大麦的转基因小麦即使在人工真菌感染条件下经过五代后仍具有抗性。与非转基因对照相比,一个转基因株系被证明基本等同。