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在转基因烟草和玉米中分离和鉴定玉米高亲和力磷酸盐转运基因 ZmPht1;5 的 295bp 强启动子。

Isolation and characterization of a 295-bp strong promoter of maize high-affinity phosphate transporter gene ZmPht1; 5 in transgenic Nicotiana benthamiana and Zea mays.

机构信息

The Key Laboratory of Plant Development and Environment Adaptation Biology, Ministry of Education, School of Life Science, Shandong University, Qingdao, 266237, China.

Biology Institute, Qilu University of Technology (Shandong Academy of Sciences), Jinan, Shandong, China.

出版信息

Planta. 2020 May 18;251(6):106. doi: 10.1007/s00425-020-03400-7.

DOI:10.1007/s00425-020-03400-7
PMID:32424449
Abstract

The small 295-bp ZmPht1; 5 promoter is sufficient to drive high-intensity expression of target genes, especially under phosphate deprivation conditions, and is therefore useful for crop improvement via transgenic techniques. Phosphate (Pi) deficiency has become a major challenge and limiting factor in world agricultural production. Manipulating the gene expression using appropriate promoters to improve the Pi absorption and utilization efficiency of crops could reduce the requirement for Pi fertilizers. In the study, a 295-bp strong promoter (M2P-7) of maize high-affinity phosphate transporter ZmPht1; 5 was isolated and functionally validated in transgenic Nicotiana benthamiana and maize by analyzing the ZmPht1; 5 promoter (M2P-1) and its 5' truncated variants (M2P-2 ~ M2P-8) in different sizes under normal and Pi-deprivation conditions. The M2P-7 displayed the highest promoter activities among 5' truncated fragments in all tested tissues of transgenic Nicotiana benthamiana at different development stages, which was 1.5 and 3 times higher than the well-used CaMV35S promoter under normal and Pi-deprivation conditions, respectively. In maize, the M2P-7 promoter activity was comparable to the maize ubiquitin1 promoter widely used in monocots under normal condition, which was about 1.3 times that of the ubiquitin1 promoter under Pi-deprivation environments. Moreover, the M2P-7 fragment is only 295 bp in length, thus reducing the construct size, and is therefore beneficial for genetic transformation. Thus, the small promoter M2P-7 of plant origin could be of great use for monocotyledonous and dicotyledonous crop improvement via transgenic techniques based on its promoter activities, expression patterns and small size.

摘要

该 295-bp 的ZmPht1;5 启动子足以驱动靶基因的高强度表达,尤其是在缺磷条件下,因此可通过转基因技术用于作物改良。磷(Pi)缺乏已成为全球农业生产的主要挑战和限制因素。通过使用适当的启动子来操纵基因表达,以提高作物对 Pi 的吸收和利用效率,可以减少对 Pi 肥料的需求。在这项研究中,从玉米高亲和力 Pi 转运蛋白 ZmPht1;5 中分离出一个 295-bp 的强启动子(M2P-7),并通过分析在正常和 Pi 剥夺条件下不同大小的ZmPht1;5 启动子(M2P-1)及其 5'截断变体(M2P-2~M2P-8),在转基因烟草原生质体和玉米中对其功能进行了验证。在不同发育阶段的转基因烟草原生质体的所有测试组织中,M2P-7 在所有 5'截断片段中显示出最高的启动子活性,在正常和 Pi 剥夺条件下分别比常用的 CaMV35S 启动子高 1.5 倍和 3 倍。在玉米中,M2P-7 启动子活性在正常条件下与广泛用于单子叶植物的玉米泛素 1 启动子相当,在 Pi 剥夺环境下约为泛素 1 启动子的 1.3 倍。此外,M2P-7 片段的长度仅为 295bp,因此减少了构建体的大小,有利于遗传转化。因此,基于其启动子活性、表达模式和小尺寸,这种来自植物的小启动子 M2P-7 对于通过转基因技术对单子叶植物和双子叶植物的改良可能非常有用。

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