Khandagale Kiran S, Chavhan Rahul, Nadaf Altafhusain B
1Department of Botany, Savitribai Phule Pune University, Pune, 411007 India.
Vilasrao Deshmukh Agricultural Biotechnology College, Latur, India.
3 Biotech. 2020 Apr;10(4):145. doi: 10.1007/s13205-020-2131-8. Epub 2020 Mar 2.
2-acetyl-1-pyrroline (2AP) is a principal aroma compound in scented rice and a mutation in 2 () is responsible aroma in scented rice. The present study was aimed at inducing 2AP production in non-scented rice cultivar IR-64 by silencing via technique. A vector pBSK was used for the construction of RNAi cassette and pRI101ON as a binary vector. (GV3101)-mediated transformation was done using embryogenic calli of IR-64. The resultant transgenic lines showed up to 14-fold reduction in expression of gene and 50% inhibition in enzyme activity. Gas chromatography (GC-MS) analyses showed a significant amount of 2AP production in RNAi callus, leaves, and seeds of IR-64. A total 39 volatile compounds were identified from the control and RNAi seeds of IR-64. Among them, octanal and 2-pentylfuron were found to be increased (30-40%) in RNAi seeds of IR-64. The content of precursors, proline, and methylglyoxal increased substantially, whereas GABA content reduced up to 25% in transgenic IR-64 lines. The study demonstrated that RNAi approach could be successfully used for imparting pleasant aroma character in non-scented rice cultivars.
2-乙酰基-1-吡咯啉（2AP）是香稻中的一种主要香气化合物，2（）中的一个突变导致了香稻的香气。本研究旨在通过RNA干扰技术沉默基因，从而在非香稻品种IR-64中诱导2AP的产生。使用载体pBSK构建RNA干扰盒，并使用pRI101ON作为二元载体。利用IR-64的胚性愈伤组织进行了农杆菌（GV3101）介导的转化。所得转基因株系的基因表达降低了14倍，酶活性受到50%的抑制。气相色谱（GC-MS）分析表明，IR-64的RNA干扰愈伤组织、叶片和种子中产生了大量的2AP。从IR-64的对照种子和RNA干扰种子中总共鉴定出39种挥发性化合物。其中，IR-64的RNA干扰种子中壬醛和2-戊基呋喃增加（30-40%）。转基因IR-64株系中前体、脯氨酸和甲基乙二醛的含量大幅增加，而γ-氨基丁酸含量降低了25%。该研究表明，RNA干扰方法可成功用于赋予非香稻品种宜人的香气特性。
