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RNA干扰介导下调非香型水稻IR-64(L.)中2-乙酰-1-吡咯啉表达基因

RNAi-mediated down regulation of gene for expression of 2-acetyl-1-pyrroline in non-scented rice IR-64 ( L.).

作者信息

Khandagale Kiran S, Chavhan Rahul, Nadaf Altafhusain B

机构信息

1Department of Botany, Savitribai Phule Pune University, Pune, 411007 India.

Vilasrao Deshmukh Agricultural Biotechnology College, Latur, India.

出版信息

3 Biotech. 2020 Apr;10(4):145. doi: 10.1007/s13205-020-2131-8. Epub 2020 Mar 2.

Abstract

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.

摘要

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。

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