《植物生理学报》 2018, 54(11): 1703-1710

铵硝营养对枳橙幼苗谷氨酰胺合成酶活性及相关基因表达的影响

孙敏红^1,2,3,4,*, 吴炼^1,2,3, 谢深喜⁴

¹中南林业科技大学经济林培育与保护教育部重点实验室, 长沙410004; ²中南林业科技大学经济林培育与利用湖南省2011协同创新中心, 长沙410004; ³中南林业科技大学经济林育种与栽培国家林业局重点实验室, 长沙410004; ⁴湖南农业大学园艺园林学院, 长沙410128

通信作者：孙敏红；E-mail: sunminhongcaddie@126.com

摘　要：

本文采用水培方法, 研究不同铵硝配比营养液对枳橙(Citrus sinensis×Poncirus trifoliate)幼苗体内谷氨酰胺合成酶(GS)活性变化及相关基因的表达量变化的影响。结果表明: 不同处理的枳橙幼苗叶片GS酶活性均随着培养时间的延长而有一定增加, 其中NO₃^-、NO₃^-:NH₄⁺=7:3、NO₃^-:NH₄⁺=1:1和NO₃^-:NH₄⁺=3:7处理的GS酶活性大小无显著差异, NH₄⁺处理最小。各处理幼苗根系的GS活性显著高于叶片, 其中NO₃^-:NH₄⁺=7:3和NO₃^-:NH₄⁺=1:1最高。3个GS相关基因CitGln 1.3、CitGln HZ、CitGln 2在幼苗各部位中均有表达。其中根系中Cis-Gln HZ的表达量最高, 叶片中CisGln HZ和CitGln 2表达量显著高于CisGln 1.3。由此推测CisGln HZ是存在于幼苗根系和叶片中GS酶的主要组成基因, 并且受到氮素形态及配比的调控, 对该基因进行系统进化树分析结果表明该基因与GS1家族功能最为相近。

关键词：枳橙; 硝态氮; 铵态氮; 谷氨酰胺合成酶

收稿：2018-05-18   修定：2018-11-11

资助：经济林培育与利用湖南省高校‘2011’协同创新中心开放基金资助项目(2014B-02)、湖南省教育厅科学研究项目一般项目(13c1140)、中南林业科技大学引进高层次人才科研启动基金(2017YJ031)和中南林业科技大学校级青年基金一般项目。

Effect of nitrate and ammonium nutrition on glutamine synthetase activities and relative genes for citrange

SUN Min-Hong^1,2,3,4,*, WU Lian^1,2,3, XIE Shen-Xi⁴

¹The Key Laboratory of Cultivation and Protection for Non-wood Forest Trees of Education Ministry, Central South University of Forestry and Technology, Changsha 410004, China; ²Collaborative Innovation Center of Cultivation and Utilization for Non-wood Forest Tree, Central South University of Forestry and Technology, Changsha 410004, China; ³The Key Laboratory of Non-wood Forest Products of State Forestry Administration, Central South University of Forestry and Technology, Changsha 410004, China; ⁴Horticulture and Landscape College, Hunan Agricultural University, Changsha 410128, China

Corresponding author: SUN Min-Hong; E-mail: sunminhongcaddie@126.com

Abstract:

Hydroponic experiments were carried out to study the influences of the different ratio of NO₃^-:NH₄⁺ on the citrange (Citrus sinensis×Poncirus trifoliate) seedling glutamine synthetase (GS) activity changes and relative genes expression. The results showed that the GS activities of citrange leaves in all treatments were increasing as the culture time continuing. The GS activities in treatments of NO₃^-, NO₃^-:NH₄⁺=7:3, NO₃^-:NH₄⁺=1:1 and NO₃^-:NH₄⁺=3:7 had no significant difference in each other, and that in treatment of NH₄⁺ was lowest. Under all treatments the GS activities in roots were significantly higher than those in leaves, and the GS activities of NO₃^-:NH₄⁺=7:3 and NO₃^-:NH₄⁺=1:1 were highest. Three GS relative genes (CitGln 1.3, CitGln HZ, CitGln 2) were all detected in citrange seedling. The expression of CitGln HZ was highest in roots, while the expression of CisGln HZ and CitGln 2 in leaves were higher than CitGln 1.3. And it was induced that CisGln HZ was the corn genes of GS in the roots and leaves. Phylogenetic trees analysed indicated that CitGln HZ function was similar to GS1 family.

Key words: citrange (Citurs sinensis×Poncirus trifoliate); nitrate; ammonium; glutamine synthetase