Hu Tingzhang, Zeng Hua, Hu Zongli, Qv Xiaoxiao, Chen Guoping
Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University , Chongqing 400044, People's Republic of China.
J Agric Food Chem. 2014 Dec 10;62(49):11988-93. doi: 10.1021/jf503801u. Epub 2014 Dec 1.
α-Linolenic and linoleic acids are essential fatty acids (EFAs) for humans and required for maintenance of optimal health, but they cannot be synthesized by the human body and must be obtained from dietary sources. Using TomloxC fragment, TomloxD fragment, and partial TomloxA sequence that is highly identical with TomloxB and TomloxE, a RNAi expression vector was constructed. The construct was used to transform tomato cotyledon explants with the Agrobacterium-mediated co-cultivation method. The real-time reverse transcription polymerase chain reaction analysis showed that the expression of TomloxA, TomloxB, TomloxC, TomloxD, and TomloxE in transgenic tomato plants was drastically repressed, which led to a marked decrease in the levels of lipoxygenase activity. Finally, higher accumulations of the endogenous α-linolenic and linoleic acids were detected in the transgenic tomato fruits, which were 1.65-3.99 and 2.91-4.98 times that of the non-transformed tomato fruits, respectively.
α-亚麻酸和亚油酸是人体必需脂肪酸(EFAs),对维持最佳健康状态至关重要,但人体无法合成,必须从饮食中获取。利用TomloxC片段、TomloxD片段以及与TomloxB和TomloxE高度同源的部分TomloxA序列,构建了一个RNAi表达载体。采用农杆菌介导的共培养方法,将该构建体用于转化番茄子叶外植体。实时逆转录聚合酶链反应分析表明,转基因番茄植株中TomloxA、TomloxB、TomloxC、TomloxD和TomloxE的表达受到显著抑制,这导致脂氧合酶活性水平显著降低。最后,在转基因番茄果实中检测到内源性α-亚麻酸和亚油酸的积累量更高,分别是非转基因番茄果实的1.65 - 3.99倍和2.91 - 4.98倍。