低磷条件下过表达OsPHF1基因对粳稻农艺性状的影响
收稿日期: 2018-03-23
修回日期: 2018-06-04
网络出版日期: 2018-11-10
基金资助
国家重点研发计划资助项目(2016YFD0100700);农业部转基因专项(2016ZX08009003-004, 2014ZX08001-005);国家自然科学基金资助项目(31570269, 31570279)
Agronomic Traits of Marker-free Transgenic japonica Rice with Overexpression of OsPHF1 Under Low Phosphorus Environment
Received date: 2018-03-23
Revised date: 2018-06-04
Online published: 2018-11-10
【目的】 磷酸盐转运体运输协助因子(PHF1)通过转录后调节特定磷转运蛋白,影响磷酸盐的利用效率。本研究通过培育过表达OsPHF1的无选择标记转基因粳稻空育131,研究在不同磷浓度环境中OsPHF1的过表达对粳稻空育131产量的影响,为培育可商品化的磷高效转基因水稻品种提供依据。方法 利用双T-DNA方法构建OsPHF1的过表达载体,通过农杆菌侵染法和后续筛选获得了无选择标记的转基因空育131纯合株系,通过对T3和T4代转基因植株的田间试验,研究转基因品系在低磷浓度(75或112.5 kg/hm2过磷酸钙)、中低磷浓度(225或300 kg/hm2过磷酸钙)和正常磷浓度(450 kg/hm2过磷酸钙)下的农艺性状。结果 获得了3个无筛选标记的纯合OsPHF1过表达转基因空育131株系F18-18、F22-32和F25-6。其中,F22-32和F25-6的OsPHF1的表达量远高于野生型。大田试验显示,F22-32和F25-6株系的T3代在中低磷(300 kg/hm2过磷酸钙)环境中,分蘖数比对照分别增加了55%和25%,增产幅度分别为38%和34%;F22-32和F25-6株系T4代在低磷条件下(112.5 kg/hm2过磷酸钙)产量的增幅最大,增产了30%~35%;在中低磷条件下(225 kg/hm2过磷酸钙)分蘖数和产量也有明显增加。结论 双T-DNA法能用于培育过表达OsPHF1的无筛选标记转基因水稻。田间试验显示,高表达OsPHF1的转基因株系在中低磷条件下(112.5、225或300 kg/hm2过磷酸钙)分蘖数和产量稳定增加。
关键词： OsPHF1; 过表达; 无选择标记的转基因水稻; 低磷浓度; 产量
胡张华, 刘秀艳, 王玉锋, 彭瑜, 史晓亮, 滕胜 . 低磷条件下过表达OsPHF1基因对粳稻农艺性状的影响[J]. 中国水稻科学, 2018 , 32(6) : 557 -564 . DOI: 10.16819/j.1001-7216.2018.8034
【Objective】PHF1 (phosphate transporter assistance factor) regulates specific phosphorous transporters after transcriptional regulation and affects the utilization efficiency of phosphate. The main purpose is to develop selective-marker-free OsPHF1-overexpressed japonica rice (Kongyu 131), and study the agronomic traits of transgenic rice at different low phosphorus concentrations. It contributes to the cultivation of transgenic rice varieties with high phosphorus utilization efficiency.【Method】Marker-free OsPHF1-overexpressed transgenic lines were obtained by using double T-DNA methods, Agrobacterium-mediated infection and subsequent screening. The field experiments for T3 and T4 generations of transgenic plants were carried out under low phosphorus concentration (75 or 112.5 kg /hm2 calcium superphosphate) medium-to-low phosphorus concentrations (225 or 300 kg /hm2 calcium superphosphate) and normal phosphorus concentration (450 kg /hm2 calcium superphosphate).【Result】Three marker-free homozygous transgenic lines F18-18, F22-32 and F25-6 were obtained. The expression levels of OsPHF1 in lines F22-32 and F25-6 were much higher than that of wild type. Field experiments showed that compared with the wild type, the tiller number of F22-32 and F25-6 (T3 generation) were increased by 55% and 25%, respectively and the yield per plant of F22-32 and F25-6 were increased by 38%–34%, respectively under medium-to-low phosphorus environment (300 kg/hm2 calcium superphosphate). The yield of F22-32 and F25-6 transgenic lines (T4) increased by 30% to 35%, the biggest improvement under low phosphorus (112.5 kg /hm2 calcium superphosphate). Meantime, the tiller number and yield of F22-32 and F25-6 (T4) also showed obvious boost under medium-to-low phosphorus (225 kg /hm2 calcium superphosphate) levels. 【Conclusion】Marker-free OsPHF1 transgenic lines were cultivated using double T-DNA method. Two generation of homozygous transgenic lines showed significantly increased tiller number and yield at medium-to-low phosphorus level (112.5, 225 or 300 kg /hm2 calcium superphosphate).
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