以RNA干扰γ-氨基丁酸转氨酶1基因(OsGABA-T1)表达提高稻米γ-氨基丁酸(GABA)含量

doi:10.3724/SP.J.1006.2015.01305

摘要/Abstract

摘要： γ-氨基丁酸(GABA)是一种四碳非蛋白质氨基酸, 具有降血压等功能。为提高稻米中GABA含量, 利用RNA干扰技术, 构建水稻中GABA代谢关键酶GABA转氨酶1基因(OsGABA-T1)的干扰载体, 通过农杆菌介导法, 将其转化至粳稻品种宁粳1号中。实时荧光定量PCR检测结果表明导入的RNA干扰结构成功地降低了目的基因OsGABA-T1的表达, 且干扰家系中OsGABA-T2基因表达也随之下降。对转基因T₃代稻米GABA含量测定发现, 糙米中GABA含量相对于对照增加了13倍以上, 精米中GABA含量也显著增加, 而其他主要氨基酸含量则没有明显变化。测定储藏4个月的转基因稻米发现, GABA含量仍具有较高水平。所以, 利用RNA干扰技术可有效提高稻米γ-氨基丁酸(GABA)含量, 为培育富含GABA的降血压功能性水稻品种提供基础。

关键词: γ, -氨基丁酸, RNA干扰, γ, -氨基丁酸转氨酶1, 转基因水稻

Abstract: γ-Aminobutyric acid (GABA) is a four-carbon non-protein amino acid and has been identified to have a function in reducing blood pressure. In order to increase GABA contents in rice grains, we constructed a RNA interference (RNAi) vector suppressing the expression of GABA transaminase 1 gene (OsGABA-T1). GABA transaminase is a key enzyme that converts GABA to succinate semialdehyde (SSA). The RNA interference vector was transformed into the callus of japonica cv. Ningjing 1. qRT-PCR analysis showed that OsGABA-T1 transcript abundance in the RNAi transgenic lines was effectively reduced, so did the homolog gene OsGABA-T2. The GABA contents of brown rice in T₃ generation of RNAi lines were 13 times higher than these in Ningjing 1, so did in polished rice as well as brown rice and polished rice stored for four months of post-harvest in transgenic rice, while other main amino acid contents did not change much. In summary, this research provides a basis for breeding a rice variety which could contribute to reduce blood pressure due to the enrichment of GABA contents in rice seeds though effectively suppressing the expression of OsGABA-T1 gene by the RNA interference technique.

Key words: GABA, RNA interference, OsGABA-T1, Transgenic rice

周露, 沈贝贝, 白苏阳, 刘喜, 江玲, 翟虎渠, 万建民. 以RNA干扰γ-氨基丁酸转氨酶1基因(OsGABA-T1)表达提高稻米γ-氨基丁酸(GABA)含量[J]. 作物学报, 2015, 41(09): 1305-1312.

ZHOU Lu, SHEN Bei-Bei, BAI Su-Yang, LIU Xi, JIANG Ling, ZHAI Hu-Qu, WAN Jian-Min. RNA Interference of OsGABA-T1 Gene Expression Induced GABA Accumulation in Rice Grain[J]. Acta Agron Sin, 2015, 41(09): 1305-1312.

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