Liu R, Zhang H H, Chen Z X, Shahid M Q, Fu X L, Liu X D
College of Agriculture, South China Agricultural University, Guangzhou, Guangdong, China.
College of Agriculture, South China Agricultural University, Guangzhou, Guangdong, China
Genet Mol Res. 2015 Oct 29;14(4):13667-78. doi: 10.4238/2015.October.28.29.
Oryza officinalis has proven to be a natural gene reservoir for the improvement of domesticated rice as it carries many desirable traits; however, the transfer of elite genes to cultivated rice by conventional hybridization has been a challenge for rice breeders. In this study, the conserved sequence of plant stress-related NAC transcription factors was selected as a probe to screen the O. officinalis genomic transformation-competent artificial chromosome library by Southern blot; 11 positive transformation-competent artificial chromosome clones were subsequently detected. By Agrobacterium-mediated transformation, an indica rice variety, Huajingxian 74 (HJX74), was transformed with a TAC clone harboring a NAC gene-positive genomic fragment from O. officinalis. Molecular analysis revealed that the O. officinalis genomic fragment was integrated into the genome of HJX74. The transgenic lines exhibited high tolerance to drought stress. Our results demonstrate that the introduction of stress-related transformation-competent artificial chromosome clones, coupled with a transgenic validation approach, is an effective method of transferring agronomically important genes from O. officinalis to cultivated rice.
药用野生稻已被证明是改良栽培稻的天然基因库，因为它具有许多优良性状；然而，通过传统杂交将优良基因转移到栽培稻中对水稻育种者来说一直是一项挑战。在本研究中，选择植物胁迫相关NAC转录因子的保守序列作为探针，通过Southern杂交筛选药用野生稻基因组转化人工染色体文库；随后检测到11个阳性转化人工染色体克隆。通过农杆菌介导的转化，用携带来自药用野生稻的NAC基因阳性基因组片段的TAC克隆转化籼稻品种华粳籼74（HJX74）。分子分析表明，药用野生稻基因组片段已整合到HJX74的基因组中。转基因株系对干旱胁迫表现出高耐受性。我们的结果表明，引入胁迫相关的转化人工染色体克隆，结合转基因验证方法，是将药用野生稻中具有重要农艺性状的基因转移到栽培稻中的有效方法。
