Dattgonde Nagesh, Tiwari Sharad, Sapre Swapnil, Gontia-Mishra Iti
Biotechnology Centre, Jawaharlal Nehru Agriculture University, Jabalpur, India.
Iran J Biotechnol. 2019 Jan 11;17(1):e1563. doi: 10.21859/ijb.1563. eCollection 2019 Jan.
Oat ( ) with high nutritive value and fiber content is used as the main food grain in many countries for human diet as well as animal feed. Recently, it became difficult to transfer new genes through the conventional breeding due to the lack of desirable traits.
The current study aimed at achieving a standardized protocol for Agrobacterium-mediated transformation in oat.
For oat transformation, mature seeds were sterilized, germinated, and used for explants generation. GV3101 with the binary vector pCAMBIA 1305.1, which carries as reporter gene, was utilized in the transformation. The co-cultivation treatment assisted with sonication, and vacuum infiltration, and their combination was employed for transformation with different incubation periods of 48, 72, and 96 hours under the dark conditions.
Among the different transformation treatments, the vacuum treatment with 72 hours dark incubation had the best results. Vacuum infiltration of the cultures from leaf base produced a maximum of 25% hygromycin-resistant explants. These explants upon GUS assay and PCR analysis revealed 21.85% and 19.04% transformation efficiency, respectively.
It could be concluded that vacuum infiltration assisted Agrobacterium-mediated transformation is the most efficient method to conduct the genetic improvement of the oat using transformation protocol.
燕麦营养价值高且纤维含量丰富，在许多国家被用作人类饮食和动物饲料的主要粮食作物。近来，由于缺乏理想性状，通过传统育种转移新基因变得困难。
本研究旨在实现燕麦农杆菌介导转化的标准化方案。
用于燕麦转化的成熟种子经消毒、萌发后用于外植体的制备。携带作为报告基因的二元载体pCAMBIA 1305.1的GV3101用于转化。采用超声处理、真空渗透及其组合辅助共培养处理，在黑暗条件下进行48、72和96小时不同培养时间的转化。
在不同转化处理中，黑暗培养72小时的真空处理效果最佳。从叶基部对培养物进行真空渗透产生了最多25%的潮霉素抗性外植体。这些外植体经GUS检测和PCR分析，转化效率分别为21.85%和19.04%。
可以得出结论，真空渗透辅助农杆菌介导的转化是利用转化方案对燕麦进行遗传改良的最有效方法。
